Abstract

Liposomes are intensively used as nanocarriers for biology, biochemistry, medicine, and in the cosmetics industry and their non-toxic and biocompatible nature makes these vesicles attractive systems for biomedical applications. Moreover, the conjugation of specific ligands to liposomes increases their cellular uptake and therapeutic efficiency. Considering these aspects, the aim of the present study was to obtain new formulations of cationic liposomes coated with dual-peptide functionalized carboxymethyl chitosan (CMCS) for the treatment of inner ear diseases. In order to achieve efficient active targeting and ensuring a high efficacy of the treatment, CMCS was functionalized with Tet1 peptide, to target specific ear cells, and TAT peptide, to ensure cellular penetration. Furthermore, dexamethasone phosphate was loaded as a model drug for the treatment of ear inflammation. The infrared spectroscopy confirmed the functionalization of CMCS with the two specific peptides. The mean diameter of the uncovered liposomes varied between 167 and 198 nm whereas the CMCS-coated liposomes ranged from 179 to 202 nm. TEM analysis showed the spherical shape and unilamellar structure of liposomes. The release efficiency of dexamethasone phosphate after 24 h from the uncoated liposomes was between 37 and 40% and it appeared that the coated liposomes modulated this release. The obtained results demonstrated that the liposomes are hemocompatible since, for a tested concentration of 100 mu g/mL, the liposome suspension had a lysis of erythrocytes lower than 2.5% after 180 min of incubation. In addition, the peptide-functionalized CMCS-coated liposomes induced a non-significant effect on the viability of normal V79-4 cells after 48 h, at the highest doses. Values of 71.31% were recorded (CLCP-1), 77.28% (CLCP-2) and 74.36% (CLCP-3), correlated with cytotoxic effects of 28.69%, 22.72%, and 25.64%.

Abstract

Skin defects caused by injuries such as burns that exceed a certain diameter no longer heal spontaneously and require more complex interventions. Hydrogels have received special attention due to their unique ability to block the penetration of bacteria into wounds as they can absorb contaminated exudates and exhibit a high degree of flexibility. This study is focused on the preparation and characterization of novel antibacterial biocomposite hydrogels based on two carbohydrates, alginate and hyaluronic acid (HA), with immobilized ZnO NPs and curcumin-loaded electrospun nanofibers to improve the efficacy of the wound healing process. The hydrogel matrix was obtained after crosslinking HA and alginate in the presence of the DMT-MM activator. Hydrogels were physicochemically characterized by FT-IR, scanning electron microscopy (SEM), UV-Vis spectroscopy, and the swelling degree was also investigated. It appeared that the swelling behavior of the biocomposite hydrogels was influenced by the amount of ZnO NPs and by the presence of nanofibers. Biodegradability, hemolysis, cell viability, antimicrobial, and in vitro irritability tests were carried out to assess their biological properties. The obtained results highlighted that these hydrogels are biodegradable, hemocompatible, non-cytotoxic, and non-irritating and have good antimicrobial activity. Based on the obtained results, these materials might be interesting candidates for wound healing treatment.

Abstract

Curcumin (CRC) is a natural active principle with important anti-inflammatory, antioxidant, antibacterial, and antitumor properties but has some limitations, such as poor bioavailability, low water solubility, and rapid metabolism. To preserve CRC's benefits and eliminate its limitations, novel CRC-loaded core-shell electrospun nanofibers were designed. The nanofibers were prepared by co-axial electrospinning method using poly(vinyl alcohol)/CRC as core and poly(vinyl alcohol)/sodium alginate as shell. Polymer coating protects the CRC, increasing its stability. The swelling degree of CRC- loaded nanofibers at pH 5.4 was around 326% higher than at pH 7.4 (297%) due to the repulsions of the anion-anion COO- groups. The release efficiency of CRC at pH 7.4 was 81%, while at pH 5.4 was about 96%. In the first 6 h, there was a slower release of CRC from the nanofibers in both acidic and slightly alkaline environments. Nanofibers showed good hemocompatibility, the values being between 2.36-3.22% after the first 90 min of contact, and after 180 min of treatment, the degree of erythrocyte lysis was between 3.78 and 4.93%. Cell viability assay on V79-4 Chinese hamster fibroblasts demonstrated that treatment with free CRC led to a value of 39% whereas for CRC-loaded nanofibers, the cell viability value increased to 59.66%. The results of the present study indicated that CRC-loaded electrospun nanofibers can have great potential for biomedical applications.

Abstract

Conventional administration of drugs to the inner ear involves therapeutic instability and non-specificity of release, and to overcome these limitations, various drug delivery systems have been developed. The aim of this study was to prepare and characterize peptides-functionalized oligochitosan microcapsules loaded with Dexamethasone and magnetic nanoparticles, which can be used for dual active targeted treatment of inner ear inflammation. The diameter of spherical microcapsules in aqueous solutions was found in the micrometer range. In vitro dexamethasone release kinetics, capsules biodegradation, haemolysis and cellular viability on V79-4 normal cells were also investigated. The release efficiency of dexamethasone from the microcapsules was between 74 % and 99.8 % after 24 h. Obtained results indicated that all analyzed microcapsules showed hemolysis degrees lower than 3 %, which demonstrated their non-hemolytic character. The viability and morphology tests on V79-4 cells depended on the administered dose and after a 48-h treatment, all analyzed capsules showed a non-toxic, weak or moderately cytotoxic effect.

Abstract

Abstract

Lung infections, such as: pneumonia, chronic obstructive cystic fibrosis, tuberculosis are generally caused by viruses, bacteria and fungi. As these infections are very difficult to treat, new therapeutic approaches are investigated in order to maximize the efficiency of the treatment and to reduce the major complications that can occur. The main objective of this study was focused on the preparation of drug-loaded peptides-functionalized microcapsules, obtained by a double emulsion, based on carboxylated chitosan (CMCS), poly(vinyl alcohol) (PVA) and an activator [4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride] (DMT-MM), for the dual active targeting and treatment of pulmonary infections. The microcapsules were functionalized on the surface with both CGSPGWVRC and indolicidin (IN) peptides, as effective ligands for the active targeting of both alveolar capillary endothelial cells and bacterial cells. FTIR spectroscopy confirmed the formation of ester and amide bonds into the structure of prepared microcapsules. Microcapsules diameter varied between 893 and 965 nm. The swelling degree in PBS, at pH 7.4, ranged between 1760 %- 2100 %. All the analyzed samples showed hemolysis degrees lower than 2 %, which demonstrated their non-hemolytic character. Evaluation of the impact of microcapsules on WI-38 normal human lung cells and RAW 264.7 mouse macrophages revealed a non-toxic or slightly cytotoxic effect. Internalization assay proved that microcapsules were localized at intracellular level.

Abstract

Chitosan is a natural polymer with numerous biomedical applications. The cellular activity of chitosan has been studied in various types of cancer, including melanoma, and indicates that these molecules can open new perspectives on antiproliferative action and anticancer therapy. This study analyzes how different chitosan conformations, such as alpha-chitosan (CH) or beta-oligochitosan (CO), with various degrees of deacetylation (DDA) and molar mass (MM), both in different concentrations and in CH-CO mixtures, influence the cellular processes of SK-MEL-28 melanocytes, to estimate the reactivity of these cells to the applied treatments. The in vitro evaluation was carried out, aiming at the cellular metabolism (MTT assay), cellular morphology, and chitinase-like glycoprotein YKL-40 expression. The in vitro effect of the CH-CO mixture application on melanocytes is obvious at low concentrations of alpha-chitosan/beta-oligochitosan (1:2 ratio), with the cell's response supporting the hypothesis that beta-oligo-chitosan amplifies the effect. This oligochitosan mixture, favored by the beta conformation and its small size, penetrates faster into the cells, being more reactive when interacting with some cellular components. Morphological effects expressed by the loss of cell adhesion and the depletion of YKL-40 synthesis are significant responses of melanocytes. beta-oligochitosan (1.5 kDa) induces an extension of cytophysiological effects and limits the cell viability compared to alpha-chitosan (400-900 kDa). Statistical analysis using multivariate techniques showed differences between the CH samples and CH-CO mixtures.

Abstract

The morphological ontogeny of Oribatula pannonica Willmann, 1949 is described and illustrated. Most prodorsal setae of juveniles are of medium size, and bothridial seta is clavate. The larva has 11 pairs of gastronotal setae, including h2, most are of medium size and barbed, nymphs have 13 pairs (p3 absent), of which most are short and smooth. Basal excentrosclerites are present at nine pairs of gastronotal setae (c1, c2, d-, l-series, h1) in the larva, and at 12 pairs (c1, c2, d-, l-, h-series, p1) in the nymphs. The morphology of adult O. pannonica is compared with congeners with 13 pairs or an unknown number of notogastral setae.

Abstract

This study describes the comparison between the interaction of a series of peptide-functionalized chitosan-based nanocapsules and liposomes with two cell lines, i.e., mouse macrophages RAW 264.7 and human endothelial cells EA.hy926. Both types of nanocarriers are loaded with magnetic nanoparticles and designed for anti-inflammatory therapy. The choice of these magnetic nanostructures is argued based on their advantages in terms of size, morphology, chemical composition, and the multiple possibilities of modifying their surface. Moreover, active targeting might be ensured by using an external magnetic field. To explore the impact of chitosan-based nanocapsules and liposomes on cell cytophysiology, the cell viability, using the MTT assay, and cell morphology were investigated. The results revealed low to moderate cytotoxicity of free nanocapsules and significant cytotoxicity induced by chitosan-coated liposomes loaded with dexamethasone, confirming its release from the delivery system. Thus, after 48 h of treatment with nanocapsules, the viability of RAW 264.7 cells varied between 88.18% (OCNPM-1I, 3.125 mu g/mL) and 76.37% (OCNPM-1, 25 mu g/mL). In the same conditions, EA.hy926 cell viability was between 99.91% (OCNPM-3, 3.125 mu g/mL) and 75.15% (OCNPM-3, 25 mu g/mL) at the highest dose (25 mu g/mL), the values being comparable for both cell lines. Referring to the cell reactivity after dexamethasone-loaded liposome application, the lowest viability of RAW 264.7 cells was 41.25% (CLDM5CP-1, 25 mu g/mL) and 58.20% (CLDMM2CP-1 1.25 mu g/mL) in the endothelial cell line, proving a selective character of action of nanocarriers. The cell morphology test, performed to support and confirm the results obtained by the MTT test, revealed a differentiated response for the two types of nano-carriers. As expected, an intense cytotoxic effect in the case of dexamethasone-loaded liposomes and a lack of cytotoxicity for drug-free nanocapsules were noticed. Therefore, our study demonstrated the biocompatible feature of the studied nanocarriers, which highlights them for future research as potential drug delivery systems for pharmacological applications, including anti-inflammatory therapy.

Abstract

The recruitment life stage, including germination and early seedling establishment, is the most vulnerable life stage of plants and has cascading effects on plant performance at later life stages. However, surprisingly little is known on the eco-evolutionary processes that determine the success of biological invasions at this life stage. We performed germination experiments with and without simulated drought stress and monitored early seedling growth in diploid and tetraploid Centaurea stoebe. . While diploids are the major cytotype in the native European range, only tetraploids became invasive in North America. Thus, C. stoebe is an excellent model species to simultaneously study both, pre-adaptive differences in the native range (diploids vs. tetraploids) and post-introduction evolution in the non-native range (native tetraploids vs. non-native tetraploids). To account for broad spatial-environmental variation within cytotypes and ranges, we germinated 23,928 seeds from 208 widely distributed populations. Tetraploids germinated better than diploids. Within tetraploids, invasive populations outperformed native populations in germination. However, these differences were not evident under simulated drought stress. Seedlings of invasive tetraploids had a higher biomass and developed the first true leaf earlier than those from the native range, while the native cytotypes did not differ in these early seedling traits. Our results suggest that a combination of pre-adaptation related to superior performance of polyploids (greater and faster germination) and post-introduction evolution towards higher performance in the invasive range (greater and faster germination, greater and faster accumulation of seedling biomass) may have contributed to the invasion success of tetraploid C. stoebe in North America.

Abstract

Biological nitrogen fixation by rhizobia-nodulated legumes reduces the dependence on synthetic nitrogen fertilizers. Identification of locally adapted rhizobia may uncover economically valuable strains for sustainable agriculture. This study investigated the diversity and symbiotic potential of rhizobia associated with three Medicago species from Eastern Romania's ecosystems. Phenotypic screening ensured that only rhizobial species were retained for molecular characterization. 16S rDNA sequencing clustered the isolates into four distinct groups: Sinorhizobium meliloti, Sinorhizobium medicae, Rhizobium leguminosarum, and Mesorhizobium spp. The chromosomal genes (atpD, glnII, recA) and nifH phylogenies were congruent, while the nodA phylogeny grouped the Mesorhizobium spp. isolates with R. leguminosarum. Medicago sativa was the most sampled plant species, and only S. meliloti and R. leguminosarum were found in its nodules, while Medicago falcata nodules hosted S. meliloti and Mesorhizobium spp. Medicago lupulina was the only species that hosted all four identified rhizobial groups, including S. medicae. This study provides the first report on the Mesorhizobium spp. associated with M. falcata nodules. Additionally, R. leguminosarum and two Mesorhizobium genospecies were identified as novel symbionts for Medicago spp. Comparative analysis of Medicago-associated rhizobia from other studies revealed that differences in 16S rDNA sequence type composition were influenced by Medicago species identity rather than geographic region.

Abstract

Wild Iris plants are usually found in spontaneous flora, but due to their ornamental characteristics, they can also be used for ornamental purposes, which means that it is very important to find the perfect conditions for plant growth. This research aimed to evaluate the ornamental value and adaptive behavior of wild Iris aphylla L. in ex situ conditions. Plants from wild flora were cultivated experimentally in the Floriculture field at the Faculty of Horticulture, IULS, Iasi, Romania. The biometric determinations revealed the significantly higher ornamental value of conserved plants grown in ex situ conditions compared to wild plants. In ex situ conditions, the plants displayed more vigorous growth (similar to 100%) and had a higher number of flowers per stem (5-9 flowers), whereas, in wild conditions, this species has from two to a maximum of five flowers. Given the absence of anatomical studies in the literature, detailed anatomical investigations of the leaf structure were performed, complemented by analyses of the photosynthetic pigment content to assess the plant's physiological performance. Additionally, the molecular phylogenetic analyses conducted using two plastid markers (rbcL and trnL-F) confirmed the taxonomic classification of the native I. aphylla L. species. To the best of our knowledge, this is the first report on the molecular phylogeny of the wild Iridaceae species in Romania. These findings provide insights into the taxonomy, morphology, cultivation potential, and ornamental value of the species, supporting future conservation and horticulture development programs.

Abstract

Wild Iris plants are usually found in spontaneous flora, but due to their ornamental characteristics, they can also be used for ornamental purposes, which means that it is very important to find the perfect conditions for plant growth. This research aimed to evaluate the ornamental value and adaptive behavior of wild Iris aphylla L. in “ex situ” conditions. Plants from wild flora were cultivated experimentally in the Floriculture field at the Faculty of Horticulture, IULS, Iași, Romania. The biometric determinations revealed the significantly higher ornamental value of conserved plants grown in “ex situ” conditions compared to wild plants. In “ex situ” conditions, the plants displayed more vigorous growth (~100%) and had a higher number of flowers per stem (5–9 flowers), whereas, in wild conditions, this species has from two to a maximum of five flowers. Given the absence of anatomical studies in the literature, detailed anatomical investigations of the leaf structure were performed, complemented by analyses of the photosynthetic pigment content to assess the plant’s physiological performance. Additionally, the molecular phylogenetic analyses conducted using two plastid markers (rbcL and trnL-F) confirmed the taxonomic classification of the native I. aphylla L. species. To the best of our knowledge, this is the first report on the molecular phylogeny of the wild Iridaceae species in Romania. These findings provide insights into the taxonomy, morphology, cultivation potential, and ornamental value of the species, supporting future conservation and horticulture development programs. © 2024 by the authors.

Abstract

5% of the human population uses levothyroxine as a treatment for hypothyroidism. This research is targeted to observe the impact of a commonly used drug on the seeds of the most frequently consumed plants. The chia seeds and wheat seeds were treated with a concentrated solution of levothyroxine over the course of 4 weeks. After the seedlings grew, the plants were preserved in ethanol. Following a month of preservation, the plants were calcinated then their ashes were examined for their mineral composition. This study has shown a big influence of levothyroxine on the growth and sprouting of the Salvia hispanica and Triticum aestivum seeds.

Abstract

This research is focused on cellular response to sodium metabisulphite (E223), sodium benzoate (E211), and sodium nitrite (E250) application on two normal mammalian cells, namely MCF-12A (ATCC CRL-3598) and Vero (ATCC CCL-81). The monitored parameters were cell viability (MTT assay), cell morphology (optical microscopy) and cell survival (clonogenic assay). The treatment was applied in doses of 12.5; 25; 50 and 100 µg/mL. According to all tests assessed, our results proved a dose-response relationship, the most sensibility presented the MCF-12A cell line.

Abstract

This paper presents a part of the results of a study which we conducted in the Bârnova forest. Two ichneumon wasps species: Allophatnus aetnensis (Rudow, 1883) and Hoplocryptus besseianus (Seyrig, 1926) (belonging to Cryptinae subfamily) are report for the first time in the Romanian fauna

Abstract

In recent years, silicon dioxide (SiO2) particles have drawn increasing attention due to their potential to enhance plant tolerance to various stresses by modulating physiological and biochemical processes. Salinity is an important abiotic stress with a harmful impact on crop growth and productivity. In this context, the aim of this study was to investigate the influence of silicon dioxide (SiO2) addition on milk thistle (Silybum marianum (L.) Gaertn.) under salinity stress. Thus, individual and combined effects of the application of sodium chloride (NaCl, 50 and 100 mM) and SiO2 particles (0.1 mM) on some physiological and oxidative stress parameters, such as growth variables, photosynthetic pigments and soluble protein contents, the levels of an oxidative stress biomarker (malondialdehyde), and the specific activity of several antioxidant enzymes (superoxide dismutase, catalase and peroxidase) were evaluated. The results revealed that salt stress (100 mM) inhibited plant growth and decreased chlorophyll and carotenoid levels, whereas the added SiO2 partially mitigated these negative effects. In addition, the 100 mM NaCl treatment significantly increased peroxidise (POD) activity and decreased soluble protein content after 22 days, while no significant differences were found for group subjected to salt stress in the presence of SiO2. Results suggest that application of SiO2 on salt-stressed milk thistle may modify the seedling metabolism and increase the plant salt tolerance capacity.

Abstract

In recent years, phytofunctionalized AgNPs have attracted great interest due to their remarkable biological activities. In the present study, AgNPs were synthesized using Abies alba and Pinus sylvestris bark extracts. The chemical profile of these bark extracts was analyzed by LC-HRMS/MS. As a first step, the synthesis parameters (pH, AgNO3 concentration, ratio of bark extract and AgNO3, temperature, and reaction time) were optimized. The synthesized AgNPs were characterized by ATR-FTIR spectroscopy, DLS, SEM, EDX, and TEM. Their antioxidant, cytotoxic, and antibacterial properties were evaluated by the DPPH, ABTS, MTT, and broth microdilution assays, respectively. Abies alba and Pinus sylvestris bark extract-derived AgNPs were well-dispersed, spherical, small (average particle size of 9.92 and 24.49 nm, respectively), stable (zeta potential values of -10.9 and -10.8 mV, respectively), and cytotoxic to A-375 human malignant melanoma cells (IC50 = 2.40 +/- 0.21 and 6.02 +/- 0.61 mu g/mL, respectively). The phytosynthesized AgNPs also showed antioxidant and antibacterial effects.

Abstract

The morphological ontogeny of Eubelba danubedeltaica sp. nov. is described and illustrated. The adult of E. danubedeltaica is most similar to E. sculpta (Mihelcic, 1957) which differs from E. danubedeltaica by the presence of tubercle E2p, slightly longer seta c1 and leg IV, and thicker and clearly barbed epimeral, genital, aggenital and anal setae whereas in E. danubedeltaica tubercle E2p is absent and these setae are thinner and most are smooth, except for finely barbed genital and anal setae that are clearly visible at a high SEM magnification. In the juveniles of E. danubedeltaica, most prodorsal setae are of medium size, except for short seta in in the nymphs and very long setiform bothridial seta which are strongly curved in the distal part. Most gastronotal setae of juveniles are long, or very long, except for medium-sized c3, la, lm and h2, and short h3 in the larva, and medium-sized c3 and p2 in nymphs. The nymphs are quadrideficient and eupheredermous, i.e. they have lost the d-series setae and carry the exuvial scalps of previous instars using a cornicle located between setal pair h3. In all juveniles, the famulus e on tarsus I is sunken but is emergent in the adult. Seta d accompanies solenidion & sigma; on genua I-III and & phi; on tibiae II-IV.

Abstract

The present study aimed to evaluate the morphological, cytogenetic and biochemical changes in wheat seedlings as affected by seed exposure to a proton beam at the Bragg peak. The average energy of the proton beam was of 171 MeV at the entrance into the irradiator room while at the point of sample irradiation the beam energy was of 150 MeV, with the average value of the Linear Energy Transfer of 0.539 keV/mu m and the dose rate of 0.55 Gy/min, the radiation doses being of the order of tens of Gy. Cytogenetic investigation has revealed the remarkable diminution of the mitotic index as linear dose-response curve as well as the spectacular linear increase of the aberration index. Analyzing some biometric parameters, it was found that neither dry matter nor water content of wheat seedlings was influenced by proton beam exposure. Studying the biochemical parameters related to the antioxidant defense system, we found that the irradiation caused the slight increasing tendency of peroxidase activity as well as the decreasing trend in the activity of superoxidedismutase in the seedlings grown from the irradiated seeds. The level of malonedialdehyde (MDA) and total polyphenols showed an increasing tendency in all seedling variants corresponding to irradiated seeds, compared to the control. We conclude that the irradiation clearly induced dose-response curves at the level of cytogenetic parameters together with relatively slight variation tendency of some biochemical parameters related to the antioxidant defense system while imperceptible changes could be noticed in the biometric parameters.

Abstract

The morphological ontogeny of Amerobelba decedens Berlese, 1908 is described and illustrated. In the adult of this species, the bothridial seta is setiform, unilaterally ciliate, notogastral setae are short and barbed, and sexual dimorphism is present on tarsus IV. Most prodorsal setae of juveniles are short and smooth, except for mediumsized ro, barbed in the nymphs. The bothridial seta is long, setiform and barbed in distal part. The gastronotal setae are of medium size and barbed, except for smooth c1 and c2 in all juveniles, and smooth of d-series and short h2 in the larva. Setae c1 and c2 are inserted on large macrosclerite, and other gastronotal setae are inserted on individual microsclerites, except for h2 in the larva. The anal and anogenital regions are reticulate, and adanal setae are inserted on individual microsclerites. The nymphs are quadrideficient and eupheredermous, i.e. they lose setae of d-series and carry the exuvial scalps of previous instars. In this species, seta d accompanies solenidion & sigma; on genu III and & phi; on tibia III and IV, both in the juveniles and adults. It is a xero-thermophilous species, with one generation per year.

Abstract

Selected seeds were exposed to radiation dose array of tens of Grey the seedlings being investigated during their early ontogenetic stages. The changes in the activity of some antioxidant biomolecules, either enzyme - like peroxidase, catalase, superoxidedismutase or non-enzymes like the flavonoids and polyphenols were emphasized. The levels of photosynthesis pigments were also estimated to search for the influence of antioxidant enzyme impairing on the plant growth.

Abstract

Silver nanoparticles synthesized using plant extracts as reducing and capping agents showed various biological activities. In the present study, colloidal silver nanoparticle solutions were produced from the aqueous extracts of Picea abies and Pinus nigra bark. The phenolic profile of bark extracts was analyzed by liquid chromatography coupled to mass spectrometry. The synthesis of silver nanoparticles was monitored using UV-Vis spectroscopy by measuring the Surface Plasmon Resonance band. Silver nanoparticles were characterized by attenuated total reflection Fourier transform infrared spectroscopy, Raman spectroscopy, dynamic light scattering, scanning electron microscopy, energy dispersive X-ray and transmission electron microscopy analyses. The antimicrobial and cytogenotoxic effects of silver nanoparticles were evaluated by disk diffusion and Allium cepa assays, respectively. Picea abies and Pinus nigra bark extract derived silver nanoparticles were spherical (mean hydrodynamic diameters of 78.48 and 77.66 nm, respectively) and well dispersed, having a narrow particle size distribution (polydispersity index values of 0.334 and 0.224, respectively) and good stability (zeta potential values of -10.8 and -14.6 mV, respectively). Silver nanoparticles showed stronger antibacterial, antifungal, and antimitotic effects than the bark extracts used for their synthesis. Silver nanoparticles obtained in the present study are promising candidates for the development of novel formulations with various therapeutic applications.

Abstract

Currently there is a growing interest both in obtaining large agricultural yields in order to provide food for a growing population as well as in increasing the sustainability of production systems by improving the health of the soil. The overall objective of this study was to evaluate the effect of organic versus chemical fertilization versus irrigation on runner bean crop and soil health. A bi-factorial field experiment was organized in a split plot design with three replicates during two years (2019 and 2020). The first factor, irrigation, had two graduations: irrigated with 3000 m(3).ha(-1) (Ir) and non-irrigated (Non-Ir). The second factor, fertilization, had four graduations: biosolids (1200 kg.ha(-1)) (B), chemical (360 kg.ha(-1)) (Ch), microorganism (80 kg.ha(-1)) (M) and unfertilized (Unfert). The results showed that, regarding the physiological state of the plants, both the photosynthesis and the chlorophyll content were higher at Ir x M (5.31 mu mol CO2 m(-)(2) s(-2) and 45.23 SPAD units, respectively). The yield of dried beans ranged between 1476-4811 kg.ha(-1) in 2019 and 1498-4727 kg.ha(-1) in 2020 and was highest for Ir x Ch. Soil health status estimated by determining the dehydrogenase activity in the soil, has been improved for irrigated (Ir) and fertilized (B, M, Ch) variants.

Abstract

Featured Application Potential use in flap surgery, towards growing flap viability, diminishing marginal, or partial flap necrosis, as well as shortening the waiting period for angiogenesis in pedicled or tubulised flaps before second stage reconstruction. The evolution of reconstructive methods for defects of the human body cannot yet replace the use of flap surgery. Research is still preoccupied with the ideal techniques for offering the best chances of survival of the flaps. In our study, we investigated the effects of cold atmospheric plasma (CAP), N-nitro-L-arginine methyl ester (L-NAME), and platelet-rich plasma (PRP) injectable solutions on flap survival using an in vivo model. Twenty-four Wistar rats (four groups) had the McFarlane flap raised and CAP, L-NAME, and PRP substances tested through a single dose subcutaneous injection. The control group had only a saline solution injected. To the best of our knowledge, this is the first study that evaluated a CAP activated solution through injection on flaps. The flap survival rate was determined by clinical examination (photography documented), hematology, thermography, and anatomopathological tests. The image digital analysis performed on the flaps showed that the necrosis area (control-49.64%) was significantly lower for the groups with the three investigated solutions: CAP (14.47%), L-NAME (18.2%), and PRP (23.85%). Thermography exploration revealed less ischemia than the control group on the CAP, L-NAME, and PRP groups as well. Anatomopathological data noted the best degree of angiogenesis on the CAP group, with similar findings on the L-NAME and PRP treated flaps. The blood work did not indicate infection or a strong inflammatory process in any of the subjects. Overall, the study shows that the CAP activated solution has a similar (better) impact on the necrosis rate (compared with other solutions with known effects) when injected on the modified dorsal rat skin flap, and on top of that it can be obtained fast, in unlimited quantities, non-invasively, and through a standardized process.

Abstract

This study aims to complete our research on Usnea barbata (L.) Weber ex F.H. Wigg (U. barbata) from the Calimani Mountains, Romania, with an elemental analysis and to explore its antibacterial and antifungal potential. Thus, we analyzed twenty-three metals (Ca, Fe, Mg, Mn, Zn, Al, Ag, Ba, Co, Cr, Cu, Li, Ni, Tl, V, Mo, Pd, Pt, Sb, As, Pb, Cd, and Hg) in dried U. barbata lichen (dUB) by inductively coupled plasma mass spectrometry (ICP-MS). For the second study, we performed dried lichen extraction with five different solvents (ethyl acetate, acetone, ethanol, methanol, and water), obtaining five U. barbata dry extracts (UBDE). Then, using an adapted disc diffusion method (DDM), we examined their antimicrobial activity against seven bacterial species-four Gram-positive (Staphylococcus aureus, Enterococcus casseliflavus, Streptococcus pyogenes, and Streptococcus pneumoniae) and three Gram-negative (Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa)-and two fungi species (Candida albicans and Candida parapsilosis). Usnic acid (UA) was used as a positive control. The ICP-MS data showed a considerable Ca content (979.766 mu g/g), followed by, in decreasing order, Mg, Mn, Al, Fe, and Zn. Other elements had low levels: Ba, Cu, Pb, and Cr (3.782-1.002 mu g/g); insignificant amounts (<1 mu g/g) of Hg and V were also found in dUB. The trace elements Ag, As, Cd, Co, Li, Tl, Mo, Pd, Pt, and Sb were below detection limits (<0.1 mu g/g). The DDM results-expressed as the size (mm) of the inhibition zone diameter (IZs)-proved that the water extract did not have any inhibitory activity on any pathogens (IZs = 0 mm). Gram-positive bacteria displayed the most significant susceptibility to all other UBDE, with Enterococcus casseliflavus showing the highest level (IZs = 20-22 mm). The most susceptible Gram-negative bacterium was Pseudomonas aeruginosa (IZs = 16-20 mm); the others were insensitive to all U. barbata dry extracts (IZs = 0 mm). The inhibitory activity of UBDE and UA on Candida albicans was slightly higher than on Candida parapsilosis.

Abstract

The study aimed to evaluate the effects of non-thermal plasma activated water (PAW) in combination with low temperature (15 degrees C) on the biosynthetic potential, antioxidant activity and enzymes activity in wheat (Triticum aestivum L. cv. Glosa) sprouts. The exposure to three different types of PAW (PAW1, PAW2 or PAW3) and low temperature (15 degrees C) had positive influence on wheat sprouts. Content in proteins and photosynthetic pigments and activity of superoxide dismutase increased in wheat sprouts at day 3 and shoots at day 8. Peroxidase was stimulated in wheat sprouts (day 3) while catalase activation was detected in shoots (day 8). Polyphenol oxidase, responsible for enzymatic browning, was inhibited in shoots at day 8. PAW1 boosted free phenolic content in shoots at day 8. An increment in the antioxidant activity of bound phenolic fraction of shoots (day 8) under treatment with PAW1 was also detected. Overall, non-thermal PAW in combination with low temperature (15 degrees C) is an efficient approach to enhance the quality of wheat (Triticum aestivum L. cv. Glosa) sprouts.

Abstract

Low thermal microwave bioeffects are subject of multidisciplinary research with biomedical and environmental issues. In this study some of such effects, with focus enzyme activity changes in Phanerochaete chrysosporium fungus were assayed, with addressing to microwave exposure risks. Very low thermal microwaves (Specific Absorption Rate = 2.32 W/kg) have induced non-linear variation in the antioxidant biochemical indicators with significantly increased for 1 hour exposure time while low thermal microwaves (Specific Absorption Rate = 8.61 W/kg) elicited linear increasing with exposure times. Lignin peroxidase activity was found to decrease remarkably with more than 30%, indicating the impairing of specific metabolic functions of the environmental fungus. The results sustain the hypothesis of reactive oxygen species generation and lipid peroxidation triggered by microwave absorption with consequences on the fungus ability to degrade lignin from wood waste.

Abstract

The morphological ontogeny of Punctoribates ghilarovi Shaldybina, 1969 is described and illustrated. This species is most similar to P. mundus Shaldybina, 1973, but is larger than the latter species. The juveniles of P. ghilarovi are light brown, with most prodorsal setae of medium size, except for long seta ro in the larva, and short and smooth seta ex in all instars. The larva has 12 pairs of gastronotal setae, including h(3), the nymphs have 15 pairs. Most of them are medium sized and barbed, except for slightly longer dm, dp, lm, lp and h(1), and short and smooth h(3) in the larva, and short and smooth da and dm, slightly longer dp, and clearly longer h(1), h(3) and lp in the nymphs. In all juveniles, a humeral organ is present.

Abstract

The present study aimed to investigate the effects of non-thermal plasma-activated water (PAW) on Triticum aestivum L. cv. Glosa sprouts. Two types of PAW were generated by exposing distilled water to a high-voltage electric discharge. Wheat caryopses were treated either with PAW1 (25 mg/L NO3-, 4 mg/L NO2-, and 6 mg/L H2O2) or PAW2 (35 mg/L NO3-, 5 mg/L NO2-, and 7.5 mg/L H2O2) for 8 days, with samples being collected at days 1, 2, 3, and 8. The germination rate, growth parameters, protein, photosynthetic pigments, total phenolic contents, antioxidant activity of free and bound phenolic fractions, and activity of antioxidant enzymes were evaluated. Both PAW1 and PAW2 had positive effects on Triticum aestivum L. cv. Glosa sprouts. PAW2 had a better impact on sprouts with respect to growth parameters, free phenolic content, and antioxidant activity of the free phenolic fraction (day 3), the accumulation of proteins, and photosynthetic pigments (day 8). In conclusion, exposure to PAW increases the quality of Triticum aestivum L. cv. Glosa sprouts. Further optimization of PAW treatment is required for the most favorable impact on the accumulation of biomass and health-promoting compounds in sprouts.

Abstract

The paper presents a biostimulant (codified Bios) containing algae extract (Ascophyllum nodosum), hydrolysed proteins, as well as humic substances with effects on plant growth and mineral nutrition, thus reducing the negative effect of climate and technological stress. The application of the biostimulant indirectly stimulated the plants for the additional absorption of nutrients as well as the increase of the photosynthesis process. The foliar application of the Bios product (as a 0.5% solution) led to the increase of the yield of wheat and tomato crops compared to the unfertilized control. The production increases were 36.46% for wheat and 50.12% for tomatoes. The stimulation of the photosynthesis process for both crops tested is evidenced by the increases of the concentrations of chlorophyll a and b measured in the treated leaves as compared to the control. The same trend was found for the carotene content. The nitrogen, phosphorus and potassium content of the leaves were also determined, and there were found significant increases compared to the unfertilized control.

Abstract

The paper presents some aspects about the attack and biology of Apethymus filiformis (Klug.), a non-indigenous defoliator pest of the sessile oak forest stands, Quercus petraea (Matt.) Liebl., 1784. It was recorded for the first time in Romania in 1999, but misidentified as Apethymus abdominalis Lep., a synonim of A. cereus Klug., an accepted species, but not present in the Romanian fauna. We established five new host – parasitoid relationships. In the laboratory we obtained from Apethymus filiformis larvae, by rearings, five new ichneumonoid parasitoid species: Charmon extensor (Linnaeus) (Hymenoptera: Braconidae: Charmontinae), Temelucha ophtalmica (Holmgren) (Hymenoptera: Ichneumonidae: Cremastinae), Gelis cinctus (Linnaeus) (Hymenoptera: Ichneumonidae: Cryptinae), Diplazon laetatorius (Fabricius) (Hymenoptera: Ichneumonidae: Diplazontinae), and Phytodietus ornatus (Desvignes) (Hymenoptera: Ichneumonidae: Tryphoninae).

Abstract

New topical gel formulations based on sodium alginate and hyaluronic acid containing AS1411 aptamer-functionalized polymeric nanocapsules loaded with an antitumoral drug (5-Fluorouracil) were designed as an innovative approach for the skin cancer treatment. Several important analyses were used to characterize these obtained topical gel formulations, namely: rheological tests, permeation assays across Strat-M (R) artificial membrane, ex-vivo permeation assays across chicken skin membrane, haemolysis tests, skin irritation tests, in vitro cytotoxicity assay on human basal carcinoma cells and in vivo tests. Rheological tests revealed that apparent viscosity decreases with the increase of the shear rate, for analyzed samples, which demonstrates a shear thinning behavior. Low levels of hemolysis values which ranged between 0.03 and 0.55% suggested that the tested formulations did not induce red blood cell lysis.. The gel formulations containing nanocapsules loaded with 5-FU proved to be non-irritant. Furthermore, by study the ex-vivo diffusion properties across the chicken skin membrane, it was proved that nanoencapsulation enhance the permeability properties of 5-FU. In vitro cytotoxicity assay on TE 354.T (ATCC (R) CRL-7762 (TM)) human basal carcinoma cell line showed that the obtained formulations loaded with 5-Fluorouracil manifest an important cytotoxic effect. Finally, the presence of Langerhans CD68 cells-positive in the epidermis and epithelial sheath of dermal hair follicles suggests a specific activation, migration and retrieval of nanoparticles by these cells. Following the results obtained in this study we can appreciate that the obtained topical gel formulations have a favourable biosafety and good antitumor effects which makes them attractive for skin cancer treatment.

Abstract

Topical liposomal drug formulations containing AS1411-aptamer conjugated liposomes were designed to deliver in a sustained way the 5-fluorouracil to the tumor site but also to increase the compliance of patients with basal cell carcinoma. The 5-fluorouracil penetrability efficiency through the Strat-M membrane and the skin irritation potential of the obtained topical liposomal formulations were evaluated in vitro and the Korsmeyer Peppas equation was considered as the most appropriate to model the drug release. Additionally, the efficiency of cytostatic activity for targeted antitumor therapy and the hemolytic capacity were performed in vitro. The obtained results showed that the optimal liposomal formulation is a crosslinked gel based on sodium alginate and hyaluronic acid containing AS1411-aptamer conjugated liposomes loaded with 5-fluorouracil, which appeared to have favorable biosafety effects and may be used as a new therapeutic approach for the topical treatment of basal cell carcinoma.

Abstract

The silver nanoparticles (AgNPs) were synthesized by both bio- and chemical-based approaches in which celandine (Chelidonium majus) aqueous extract and trisodium citrate were used as reducing agents and silver nitrate (AgNO3) as metal precursor, respectively. The formation of AgNPs was confirmed by color change and absorption spectroscopy. The antioxidant response of Phanerochaete chrysosporium, following exposure to various concentrations of biologically and chemically synthesized AgNPs and depending on the mycelium age (7 and 14 days), was further studied. Results revealed that the effect of AgNPs synthesized by biological reduction determined an increase of antioxidant enzymes activities (catalase and superoxide dismutase) with the increase of nanoparticles concentrations. Both, biological and chemical reduction revealed a very similar CAT activity at both intervals studied and at almost the same scale value in all P. chrysosporium variants.

Abstract

The effect of Tanacetum extracts on cell viability was assessed by MTT method on HeLa (human cervical epithelioid carcinoma), A375 (human malignant melanoma) and V79 (Chinese hamster pulmonary fibroblasts). Apoptosis, cell cycle analysis, and genotoxicity tests were performed to identify the possible mechanism of action. Also, the antimicrobial activity was investigated, and LC-MS analysis of extracts was carried out. Tanacetum extracts substantially reduced the viability of all tested cells, normal cells being more sensitive than cancer cells. T. vulgare and T. macrophyllum extracts induced apoptosis in normal cells V79, while none of the extracts induced apoptosis on HeLa or A375 neoplastic cells. T. vulgare extract arrested cell cycle progression of V79 and A375 cells at the G(2)/M phase and exhibited mild genotoxicity on melanoma cells A375. All extracts showed moderate antimicrobial activity against tested Gram-positive bacteria and fungi.

Abstract

Lichens represent a significant source of antioxidants due to numerous metabolites that can reduce free radicals. Usnea barbata (L.) F.H. Wigg. has been recognized and used since ancient times for its therapeutic effects, some of which are based on its antioxidant properties. The present study aims to analyze the phytochemical profile and to evaluate the antioxidant and cytotoxic potential of this lichen species. Five dry extracts of U. barbata (UBDE) in different solvents (acetone, ethyl acetate, ethanol, methanol, water) were prepared by refluxing at Soxhlet to achieve these proposed objectives and to identify which solvent is the most effective for the extraction. The usnic acid content (UAC) was quantified by ultra-high performance liquid chromatography (UHPLC). The total polyphenols content (TPC) and tannins content (TC) were evaluated by spectrophotometry, and the total polysaccharides (PSC) were extracted by a gravimetric method. The 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) free radical method was used to assess the antioxidant activity (AA) and the Brine Shrimp Lethality (BSL) assay was the biotest for cytotoxic activity evaluation. The ethyl acetate extract had the highest usnic acid content, and acetone extract had the highest content of total polyphenols and tannins. The most significant antioxidant effect was reported to methanol extract, and all the extracts proved high cytotoxicity. The water extract has the lowest cytotoxicity because usnic acid is slightly soluble in this solvent, and it was not found at UHPLC analysis. All extracts recorded a moderate correlation between the content of usnic acid, polyphenols, tannins, and AA; furthermore, it has been observed that the cytotoxicity varies inversely with the antioxidant effect.

Abstract

Lichens represent an important resource for common traditional medicines due to their numerous metabolites that can exert diverse pharmacological activities including anticancer effects. To find new anticancer compounds with fewer side effects and low tumor resistance, a bioprospective study of Usnea barbata (L.) F.H. Wigg. (U. barbata), a lichen from the Calimani Mountains (Suceava county, Romania) was performed. The aim of this research was to investigate the anticancer potential, morphologic changes, wound healing property, clonogenesis, and oxidative stress biomarker status of four extracts of U. barbata in different solvents (methanol, ethanol, acetone, and ethyl acetate), and also of usnic acid (UA) as a positive control on the CAL-27 (ATCC(R) CRL-2095 (TM)) oral squamous carcinoma (OSCC) cell line and V79 (ATCC(R) CCL-93 (TM)) lung fibroblasts as normal cells. Using the MTT assay and according to IC50 values, it was found that the most potent anticancer property was displayed by acetone and ethyl acetate extracts. All U. barbata extracts determined morphological modifications (losing adhesion capacity, membrane shrinkage, formation of abnormal cellular wrinkles, and vacuolization) with higher intensity in tumor cells than in normal ones. The most intense anti-migration effect was established in the acetone extract treatment. The clonogenic assay showed that some U. barbata extracts decreased the ability of cancer cells to form colonies compared to untreated cells, suggesting a potential anti-tumorigenic property of the tested extracts. Therefore, all the U. barbata extracts manifest anticancer activity of different intensity, based, at least partially, on an imbalance in antioxidant defense mechanisms, causing oxidative stress.

Abstract

Morphological ontogeny of Oribatella similesuperbula Weigmann, 2001 based on specimens from Romania, is described and illustrated. The nymphs of this species are apopheredermous, i.e. they carry the exuvial scalps of previous instars away from the dorsal integument, using modified setal pair da. The prodorsal and most gastronotal setae of juveniles are long and barbed, except for medium sized h(2) and minute h(3) in the larva, and medium sized c(1), p(2) and p(3) in the nymphs. The larva has 12 pairs of gastronotal setae, the nymphs have 15 pairs. In all juveniles, a humeral organ is present.

Abstract

The morphological ontogeny of Perlohmannia nasuta Schuster, 1958 is described and illustrated. The adult of this species is the most similar to that of P. dissimilis (Hewitt, 1908), but differs from it by having alveolar seta exp (versus setiform in P. dissimilis), which is unique in Perlohmannia Berlese, 1916. It also has more spines on the bothridial seta and shorter seta c(1) than has P. dissimilis. The juveniles of P. nasuta have the prodorsal seta exp alveolar, 7-8 anterior spines on the bothridial seta, and seta e(2) longer than f(2). Claparede's organ of larva is long and cudgel-shaped. This species differs clearly from P. dissimilis in the ontogeny of leg setae. In both species, hypertrichy occurs on tarsi, especially on tarsus I, but P. nasuta has more setae on tarsus I of deutonymph, tritonymph and adult than has P. dissimilis.

Abstract

Nowadays, numerous biomedical studies performed on natural compounds and plant extracts aim to obtain highly selective pharmacological activities without unwanted toxic effects. In the big world of medicinal plants, Usnea barbata (L) F.H. Wigg (U. barbata) and usnic acid (UA) are well-known for their therapeutical properties. One of the most studied properties is their cytotoxicity on various tumor cells. This work aims to evaluate their cytotoxic potential on normal blood cells. Three dry U. barbata extracts in various solvents: ethyl acetate (UBEA), acetone (UBA), and ethanol (UBE) were prepared. From UBEA we isolated usnic acid with high purity by semipreparative chromatography. Then, UA, UBA, and UBE dissolved in 1% dimethyl sulfoxide (DMSO) and diluted in four concentrations were tested for their toxicity on human blood cells. The blood samples were collected from a healthy non-smoker donor; the obtained blood cell cultures were treated with the tested samples. After 24 h, the cytotoxic effect was analyzed through the mechanisms that can cause cell death: early and late apoptosis, caspase 3/7 activity, nuclear apoptosis, autophagy, reactive oxygen species (ROS) level and DNA damage. Generally, the cytotoxic effect was directly proportional to the increase of concentrations, usnic acid inducing the most significant response. At high concentrations, usnic acid and U. barbata extracts induced apoptosis and DNA damage in human blood cells, increasing ROS levels. Our study reveals the importance of prior natural products toxicity evaluation on normal cells to anticipate their limits and benefits as potential anticancer drugs.

Abstract

The morphological ontogeny of Eremobelba geographica Berlese, 1908 is described and illustrated. Thi s species was investigated mainly in ecological and biological aspects. It inhabits forest soils and cultivated areas , but is not abundant, and adults dominate in extracted samples. In the juveniles, the prodorsal seta in is short, an d the bothridial seta is setiform. The nymphs are quadrideficient and eupheredermous, i.e. they carry exuvial scalps of the previous instars, using a cornicle. Paraproctal setae occur in all juvenile instars, which is rare in Brachypylina, and hypertrichy occurs in the aggenital region of the deutonymph, tritonymph and adult, an d adanal region of the adult. In all instars, seta don all genua and tibiae is present, except for tibia I of adult.

Abstract

The morphological ontogeny of Ctenobelba pilosella Jeleva, 1962 is described and illustrated. This species was investigated from its description mainly in ecological aspect. It was recorded from the forest and meadow soils, with rather low density, and small percent of juveniles.The adult is of medium size and has setiform bothridial seta, with 6-10 anterior spines. Notogaster is covered with asteriform granules and has 10 pairs of medium size, barbed setae. Seta d is absent from genua I-III and all tibia. Bothridial seta of juveniles is setiform and barbed, prodorsal and gastronotal setae are short, except of medium size ro and long, curved inwards lp. Nymphs are quadrideficient and eupheredermous i.e. they carry the exuvial scalps of previous instars. Anal valves of protonymph and deutonymph have two pairs of alveolar setae, which is rare in Brachypylina. The juveniles have seta d on all genua and tibiae.

Abstract

The morphological ontogeny of Oribatella reticulata Berlese, 1916 is described and illustrated. The nymphs of this species are apopheredermous, i.e. they carry the exuvial scalps of previous instars away from the dorsal integument, using modified setal pair da. The larva has 12 pairs of gastronotal setae, including short h(3), the nymphs have 15 pairs, and most are long and barbed. In all juveniles, a humeral organ is absent, which is unique in Oribatella. The tritonymph of this species has more thickened setae on leg segments (d on all femora and genu IV, ev' on femora III and IV, l on femora I-III and genua and tibiae I-IV) than the adult (l on genua and tibiae I and II, l' on genu and tibia IV).

Abstract

World Health Organization estimates that 30-50% of cancers are preventable by healthy lifestyle choices, early detection and adequate therapy. When the conventional therapeutic strategies are still regulated by the lack of selectivity, multidrug resistance and severe toxic side effects, nanotechnology grants a new frontier for cancer management since it targets cancer cells and spares healthy tissues. This review highlights recent studies using biotin molecule combined with functional nanomaterials used in biomedical applications, with a particular attention on biotinylated chitosan-based nanosystems. Succinctly, this review focuses on five areas of recent advances in biotin engineering: (a) biotin features, (b) biotinylation approaches, (c) biotin functionalized chitosan based nanosystems for drug and gene delivery functions, (d) diagnostic and theranostic perspectives, and (e) author's inputs to the biotin-chitosan based tumour-targeting drug delivery structures. Precisely engineered biotinylated-chitosan macromolecules shaped into nanosystems are anticipated to emerge as next-generation platforms for treatment and molecular imaging modalities applications.

Abstract

The Artemisia genus includes a large number of species with worldwide distribution and diverse chemical composition. The secondary metabolites of Artemisia species have numerous applications in the health, cosmetics, and food sectors. Moreover, many compounds of this genus are known for their antimicrobial, insecticidal, parasiticidal, and phytotoxic properties, which recommend them as possible biological control agents against plant pests. This paper aims to evaluate the latest available information related to the pesticidal properties of Artemisia compounds and extracts and their potential use in crop protection. Another aspect discussed in this review is the use of nanotechnology as a valuable trend for obtaining pesticides. Nanoparticles, nanoemulsions, and nanocapsules represent a more efficient method of biopesticide delivery with increased stability and potency, reduced toxicity, and extended duration of action. Given the negative impact of synthetic pesticides on human health and on the environment, Artemisia-derived biopesticides and their nanoformulations emerge as promising ecofriendly alternatives to pest management.

Abstract

Eight wild rose hip genotypes from different altitudes varying from 3m to 902m were analyzed in order to evaluate the total polyphenols, flavonoids content and the antioxidant activity. The Rosa spp.rosehips collected from the Northeastern and the Southeasternof Romanianwere, as follows: Rosa canina, R. caesia, R. corymbifera, R. micrantha, R. nitidula, R. rubiginosa, R. subcanina, and R. vosagiaca. In some genotypes, the level of flavonoids and polyphenol content increased with the increasing altitude while in other it was observed a decrease. Polyphenol content reached a maximum of 144.36 mg GAE/g DW in R. rubiginosa whereas the lowest content of 61.72 mg GAE/g DW was recorded in R. caesia. The highest polyphenol content was reached at altitude of 860 m. The amount of flavonoids content ranged between 7.32 mg CE/g DW and 19.45 CE/g DW in R. caesia and in R. nitidula, respectively. The radical scavenging capacities of Rosa genotypes extracts were not positively correlated with altitude, except the R. corymbifera extractswhere the antioxidant activity increased with the increase of altitude.

Abstract

The aim of this study was to investigate the cytotoxic effects of a hydromethanolic extract obtained from cembran pine needles in HeLa cell line. In this respect, the effects of needle extract on protein synthesis, viability, proliferation and cell cycle in HeLa cells were evaluated after 48 h treatment. Cembran pine needle extract dose-dependently decreased protein synthesis in HeLa cells causing 44.26% reduction in protein synthesis at 100µg/ml. At 25, 50 and 100µg/ml, it increased cell death in comparison with the control (20.99%, 21.49% and 23.63%, respectively vs. 9.83%). In addition, at 100µg/ ml, cembran pine needle extract showed a remarkable antiproliferative effect whereas at 25 and 50µg/ml, it induced sub-G1 phase cells accumulation (11.68 ± 0.81% and 14.69 ± 0.56%, respectively in comparison with control, 6.03 ± 0.55%), an indicator of proapoptotic effects. Taken together, these results indicate that cembran pine needles are a source of compounds with antitumor potential which needs to be further investigated and exploited

Abstract

This study aimed to describe the histo-anatomy of Tussilago farfara L. species from the Asteraceae family, with medicinal importance in Romania for the alternative treatment of respiratory diseases (asthma, laryngitis, cough, emphysema) and other disorders. The chemical composition of Coltsfoot includes more than 150 chemical substances (triterpenoids, sesquiterpenoids, alkaloids) with different medicinal proprieties (expectorant, antimicrobial, antitussive) and contraindications (pregnancy, lactation, hepatic disorders). The vegetal material used in this study was collected from the waterside of river Sirețel in the village Sirețel from Sirețel commune in Iași County. The cross-sections were performed manually through vegetative organs (rhizome, stem, and leaf) with the help of a hand microtome and a botanic razor. The structures of the sections were highlighted by double coloration (iodine green and ruthenium red), the observation was performed on a Novex microscope. The characteristics structures observed by us (epidermis, vascular bundles, trichomes, angular collenchyma, assimilating parenchyma, stomata, mesophyll) correspond with Toma and Rugină (1998) observations and descriptions. Keywords: Tussilago farfara, histo-anatomy, respiratory diseases

Abstract

The effect of ascorbic acid treatment on the seed germination and seedling growth processes in the early ontogenetic stages in the Trigonella foenum-graecum L. was investigated. Four different concentrations of ascorbic acid were used (50 mg/l, 100 mg/l, 150 mg/l and 200 mg/l). The following indicators were considered: the final germination percentage, mean germination time, germination rate index, coefficient of velocity of germination, the length of the root, the length of the hypocotyl, the length of the seedling, the seedling fresh weight and the seedling vigor index. The main effects were: statistically insignificant changes in the indicators associated with germination; obvious increase of the root and seedling length, seedling fresh weight accumulation and enhance of seedling vigor index at 100 mg/l. The immersion of the seeds in ascorbic acid solution may improve germination and growth of Trigonella foenum-graecum seedlings in the early ontogenetic stages

Abstract

The paper presents 10 species of ichneumon wasps, one of them being reported for the first time in the Danube Delta Biosphere Reserve, Romania. Research has been carried out in some invasive plants areas of the Danube Delta Biosphere Reserve. There are being reported representatives of arthropod fauna (Insecta and Araneida) from four stations containing invasive plants: Pătlăgeanca, Beștepe, Plauru and Sabangia. One particular interest was the identification of the ichneumon wasps species in these areas

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The morphological ontogeny of Lopheremaeus mirabilis (Csiszar 1962), the type species of Lopheremaeus Paschoal, 1988 (Plateremaeidae) is described and illustrated. The adult of this species has six pairs of notogastral setae (lp, h(1), h(2) and p-series), epimeral and anogenital hypertrichy, and 4-5 pairs of anal setae. The cuticle of juveniles is plicate, the larva lacks the gastronotal setae c(3), la, lm and h(3), the nymphs are multideficient (they lack also setae of d-series) and eupheredermous, i.e. they carry the exuvial scalps of previous instars on the gastronotum. In the adult, dorsal and ventral crests are present on all femora and trochanter III, and dorsal crest is present on trochanter IV, the nymphs have also dorsal and ventral crests on genua, whereas the larva has no crests. In all instars, seta d on all genua and tibiae is present, and all tarsi have basal bulb (containing muscles) and uniformly narrow distal stalk (with only tendons) at about mid-length. This species was reported only from Bulgaria, Bosnia-Herzegovina and North Macedonia, Romania and Georgia.

Abstract

The morphological ontogeny of Damaeolus ornatissimus Csiszar, 1962 from Romania is described and illustrated. The nymphs of this species are quadrideficient and eupheredermous, i.e. they carry the exuvial scalps of previous instars on the gastronotum. The bothridial seta of juveniles is fusiform, with long apical flagellum, and all prodorsal and gastronotal setae are smooth, and some of them are covered with thick layer of granular cerotegument. The larva has polygonal pattern on the gastronotum, two pairs of setae on paraproctal valves and 12 pairs of gastronotal setae, the nymphs have 12 pairs of latter setae. In all instars, seta d on all genua and tibiae is present.

Abstract

The morphological ontogeny of Oribatella hungarica Balogh, 1943 is described and illustrated. The nymphs of this species are apopheredermous, i.e. they carry the exuvial scalps of previous instars away from the dorsal integument, using modified setal pair da. The larva has 12 pairs of gastronotal setae, most of them have long barbs, except for c(3) with short barbs and minute h(3), whereas the nymphs have 13 pairs with short barbs (dm and dp are lacking). In the tritonymph, leg seta d on all femora and genu IV, l on femora I and II and pair l on genua and tibiae I and II, l' on femur III and genua and tibiae III and IV, and ft on tarsi II-IV are thicker than other setae on these segments. In all juveniles, a humeral organ is present. In the adult, epimeral seta 4c and leg seta l on genua and tibiae I and II. and l'on gem III and genu and tibia IV are thickened.

Abstract

The morphological ontogeny of Punctoribates astrachanicus Shaldybina, 1973 is described and illustrated. This species was rarely investigated since its description, including its ecological and biological aspects. The adult has fusiform bothridial seta, straight anterior tectum of notogaster, four pairs of saccules, and notogastral setae mostly alveolar, except for short p-series. Tarsus I has a short dorsal projection. Juveniles of this species are light brown, with short and thin prodorsal seta ex, and relatively large humeral organ. The gastronotal setae of larva are of medium size and pointed, in nymphs they are short.

Abstract

The secondary metabolites of lichens have proven to be promising sources of anticancer drugs; one of the most important of these is usnic acid, which is a phenolic compound with dibenzofuran structure that is responsible for the numerous biological actions of lichens of genus Usnea. As a result, in this study, we related to this phenolic secondary metabolite. The aim of the present study is the evaluation of the cytotoxic activity of Usnea barbata (L.) F. H. Wigg dry acetone extract (UBE). In advance, the usnic acid content was determined in various extracts of Usnea barbata (L.) F. H. Wigg: the liquid extracts were found in water, ethanol, acetone, and the dry acetone extract; the highest usnic acid quantity was found in the dry acetone extract. First, the cytotoxic action of UBE was assessed using Brine Shrimp Lethality (BSL) test; a significant lethal effect was obtained after 24 h of treatment at high used concentrations of UBE, and it was quantified by the high mortality rate of the Artemia salina (L.) larvae. Secondly, in vitro cytotoxicity of UBE was evaluated on human tongue squamous cells carcinoma, using CAL 27 (ATCC (R) CRL-2095 (TM)) cell line. The most intense cytotoxic effect of UBE on CAL 27 cells was registered after 24 h; this response is directly proportional with the tested UBE concentrations. The obtained results have been reported regarding usnic acid content of UBE, and the data show that CAL 27 cells death was induced by apoptosis and high oxidative stress.

Abstract

The morphological ontogeny of Ctenobelba brevipilosa Mahunka, 1964 is described and illustrated for the first time; this species was investigated since its description mainly in ecological aspect. The adult instar is of medium size, with setiform bothridial seta, bearing 11-17 anterior branches, and with short notogastral setae. Seta d is absent from genua I-III and all tibia. The bothridial seta of juveniles is setiform and barbed, and prodorsal and gastronotal setae are short, except of medium sized ro. Nymphs are quadrideficient and eupheredennous, i.e. they carry the exuvial scalps of previous instars. Anal valves of protony mph and deutonymph have two pairs of alveolar setae, which is rare in Brachypylina. In the juvenile instars, seta d on all genua and tibiae is present.

Abstract

Viral infections are a major global health problem, representing a significant cause of mortality with an unfavorable continuously amplified socio-economic impact. The increased drug resistance and constant viral replication have been the trigger for important studies regarding the use of nanotechnology in antiviral therapies. Nanomaterials offer unique physico-chemical properties that have linked benefits for drug delivery as ideal tools for viral treatment. Currently, different types of nanomaterials namely nanoparticles, liposomes, nanospheres, nanogels, nanosuspensions and nanoemulsions were studied either in vitro or in vivo for drug delivery of antiviral agents with prospects to be translated in clinical practice. This review highlights the drug delivery nanosystems incorporating the major antiviral classes and their transport across specific barriers at cellular and intracellular level. Important reflections on nanomedicines currently approved or undergoing investigations for the treatment of viral infections are also discussed. Finally, the authors present an overview on the requirements for the design of antiviral nanotherapeutics.

Abstract

Non-aqueous dispersions (NAD) with two types of polymeric nanoparticles (NPs), such as hydrophobic poly(epsilon -caprolactone) (PCL) and hydrophilic cross-linked poly(vinylpyrrolidone) (PNVP), were synthesized in the present study starting from monomer-in-silicone oil (PDMS) polymerizable non-aqueous emulsions stabilized with the same tailor-made PDMS-based block copolymer. These NPs were loaded with CCisplatin, an antitumoral model drug, directly from the emulsion polymerization step, and it was observed that the presence of the drug leads only to a slight increase of the NPs size, from 120 to 150 nm. The drug release kinetics was evaluated at 37 degrees C in phosphate buffer at pH = 7.4 and it appeared that the drug release rate from the hydrophilic cross-linked PNVP-based NPs is higher than that from the hydrophobic PCL-based NPs. Moreover, haemolysis tests revealed the fact that these two types of NPs have a good compatibility with the blood. Furthermore, for both the free and drug-loaded NPs, the in vitro cytotoxicity and apoptosis was studied on two types of cancer cell lines, such as MCF-7 (breast cancer cell line) and A-375 (skin cancer cell line). Both types of NPs had no cytotoxic effect but, at a concentration of 500 mu g/mL, presented an apoptotic effect similar to that of the free drug.

Abstract

Assessing global variation in phenotypic traits and linking that variation to colonization and range expansion is notably rare in invasion biology. Here, we studied variation in seed size in Centaurea solstitialis, a weed with worldwide distribution. Additionally, we explored how seed size variation affects seedling survival of C. solstitialis under favorable precipitation conditions in Anatolia, an ancestral region, and unfavorable precipitation conditions in Argentina, a non-native region. To that end, we conducted seed collections following dispersal pathways of C. solstitialis in ancestral, expanded, and non-native ranges. Locally, collections followed elevation gradients. Also, we performed a greenhouse experiment with C. solstitialis populations varying in seed size and water additions simulating precipitation patterns in Anatolia and Argentina. Seeds from ancestral populations at low elevation were smaller than those from the rest of study populations. Also, seed size in populations at high elevation in the expanded range, the main source of non-native populations, was similar to that in all, but one non-native population, where seeds exhibited further increase. Increments in seed size thus track range expansion in C. solstitialis. Locally, seed size increased with elevation in all three ranges, suggesting convergent responses to that gradient. Seedlings from larger seeds displayed greater survival than those from smaller seeds only under Argentinean conditions. Consequently, populations with large seeds may have been instrumental for colonizing that non-native region. Our findings suggest that ancient and recent dispersal of large-seeded populations contribute to explain the reported global pattern of seed size divergence and worldwide distribution of C. solstitialis.

Abstract

Cancer remains one of the world's most devastating diseases and is responsible for more than 20% of all deaths. It is defined as uncontrolled proliferation of cells and spreads rapidly to healthy tissue. Controlled drug delivery systems offers great opportunities for the development of new non-invasive strategies for the treatment of cancers. The main advantage of these systems is their capacity to accumulate in tumors via enhanced permeability and retention effects. In the present study, an innovative hybrid drug delivery system based on nanocapsules obtained from the interfacial condensation between chitosan and poly(N-vinyl pyrrolidone-alt-itaconic anhydride) and containing both magnetic nanoparticles and an antitumoral drug was developed in order to improve the efficiency of the antitumoral treatment. Using dynamic light scattering, it was observed that the mean diameter of these hybrid nanocapsules was in the range of 43 to 142 nm. SEM confirmed their nanometric size and their well-defined spherical shape. These nanocapsules allowed the encapsulation of an increased amount of 5-fluorouracil and provided controlled drug release. In vitro studies have revealed that these drug-loaded hybrid nanocapsules were able to induce a cytostatic effect on breast carcinoma MCF-7 cell lines (Human Caucasian breast adenocarcinoma - HTB-22) comparable to that of the free drug.

Abstract

The catalytic behavior of iron phthalocyanine (FePc)-sensitized magnetic nanocatalysts was evaluated for their application in the oxidative treatment of Bisphenol A (BPA) under mild environmental conditions. Two types of FePc (Fe(II)Pc and Fe(III)Pc), which are highly photosensitive compounds, were immobilized on the surface of functionalized magnetite. The nanomaterials were characterized by high resolution transmission electron microscopy (HR-TEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analyses (TGA). The generation of singlet oxygen by nanomaterials was also investigated. In the presence of UVA light exposure (365 nm) and 15 mM H2O2, the M@Fe(III)Pc photocatalyst gave the best results; for a catalyst concentration of 2.0 g L-1, around 60% BPA was removed after 120 min of reaction. These experimental conditions were further tested under natural solar light exposure, for which also M@Fe(III)Pc exhibited enhanced oxidative catalytic activity, being able to remove 83% of BPA in solution. The water samples were less cytotoxic after treatment, this being confirmed by the MCF-7 cell viability assay.

Abstract

The paper presents 14 species of ichneumon wasps, three of them being reported for the first time from the Danube Delta Biosphere Reserve. Research has been carried out in some agricultural areas of the Danube Delta Biosphere Reserve. There are being reported representatives of arthropod fauna (Insecta and Araneida) from four stations: Murighiol (barley culture), Sarinasuf (alfalfa culture, the shores of Lake Razelm), Beștepe (sunflower culture) and Beștepe (colza culture). One particular interest was the identification of the ichneumon wasps species in these areas.

Abstract

Usnea barbata (L.)F.H.Wigg. - known as “old man’s beard”, “tree moss”, “songluo” is a lichen in the family Parmeliaceae, genus Usnea. Usnea species have recorded history of therapeutic use dating back over three thousand years in Chinese medicine. The lichen secondary metabolites have shown an impressive range of biological proprieties, including antibiotic, antifungal, antiviral, anti-inflammatory, or anticancer activities. In this study, the antitumor activity of Usnea barbata extract was evaluated by observing the morphological changes on squamous cells carcinoma cell-line CAL 27 (ATCC® CRL-2095 ™) in contact with different concentrations of extract, ranged between 12.5–400 μg/mL. The results obtained were quantified by the intensity of morphological changes of the tumor cells after 24 hours of contact. The most significant activity were recorded for 400 μg/mL extract. This study shows that Usnea barbata (L.)F.H.Wigg. extract has antitumor activity. The analysis of the obtained results showed that the cytotoxicity of lichen extract on CAL 27 tumor cells is directly related to the concentration of the applied solution.

Abstract

Thymus species are important aromatic, medicinal and culinary plants with a significant economic value. This study evaluated for the first time the chemical composition and in vitro bioactivities of the essential oils (EOs) from five Moldavian Thymus species (T. vulgaris, T. x citriodorus, T. calcareus) and cultivars (T. vulgaris 'Faustini', T. citriodorus 'Aureus'). The main compounds in Thymus EOs were: thymol in T. vulgaris and T. calcareus EOs (55.44% and 55.45%, respectively), lavandulol in T. x citriodorus EO (54.27%), and geraniol in T. citriodorus 'Aureus' and T. vulgaris 'Faustini' EOs (60.31% and 31.45%, respectively). T. vulgaris and T. calcareus EOs showed the most potent antioxidant activities (EC50=0.003 mg/mL in ABTS radical cation scavenging assay) and exhibited significant inhibitory effects against aflatoxin-producing Aspergillus flavus fungus (MIC=0.25 mu L/mL). At doses that provided micromolar concentrations of thymol, T. vulgaris and T. calcareus EOs acted genoprotective at preventive and interventional levels against H2O2-induced genomic damage in V79 cells, the former being more active (6.21% and 5.52% vs. 25.13% and 7.26% tail DNA in pre- and post-treatment protocols, respectively). The genoprotective effects may be ascribed to antioxidant potential and, possibly, to stimulation of DNA repair processes. The Moldavian Thymus species are valuable resources of bioactive EOs for pharmaceutical and food industries (T. vulgaris, T.calcareus) but also for flavor industry and perfumery (T. x citriodorus, T. citriodorus 'Aureus', T. vulgaris 'Faustini').

Abstract

Abstract

The aim of this study was to evaluate the performances of Pichia kudriavzevii CR-Y103 yeast strain for the decolorization, biodegradation, and detoxification of cationic dye C.I. Basic Blue 41, a toxic compound to aquatic life with long-lasting effects. Under optimized cultural conditions (10.0-gL(-1) glucose, 0.2-gL(-1) yeast extract, and 1.0-gL(-1) (NH4)(2)SO4), the yeast strain was able to decolorize 97.86% of BB41 (50mgL(-1)) at pH 6 within 4h of incubation at 30 degrees C under shaken conditions (12,238.00-gh(-1) average decolorization rate) and 100% within 12h. The UV-Vis spectral analysis, high-performance liquid chromatography (HPLC), and Fourier-transform infrared spectroscopy (FTIR) analysis confirmed the complete decolorization and degradation of the BB41 dye by P. kudriavzevii CR-Y103. Also, other seven yeast strains, isolated from soil, as P. kudriavzevii (CR-Y108, CR-Y119, and CR-Y112), Candida tropicalis CR-Y128, Cyberlindnera saturnus CR-Y125, and Candida solani CR-Y124 have shown a promising decolorizing potential of azo-dye BB41 (99.89-76.09% decolorization). Phytotoxicity, cytotoxicity, and genotoxicity assays on Trifolium pratense and Triticum aestivum seedlings confirmed the high toxicity of BB41 dye (500ppm), with inhibition on germination rate (%), root and shoot elongation, decreasing of mitoxic index value (with 34.03% in T. pratense and 40.25% in T. aestivum), and increasing the frequency of chromosomal aberrations (6.87 times in T. pratense and 6.25 times in T. aestivum), compared to control. The same biomarkers indicated the nontoxic nature of the BB41 degraded metabolite (500ppm) obtained after P. kudriavzevii CR-Y103 treatment. Moreover, the healthy monkey kidney cells (Vero cells) had a low sensitivity to BB41 biodegraded products (250gmL(-1)) (MTT cell viability assay) and revealed minor DNA damage (comet assay) compared to BB41 dye treatment. These findings show that P. kudriavzevii could be used in eco-friendly bioremediation technologies, applicable for reducing the toxicity of basic azo-dyes containing wastewaters.

Abstract

IntroductionPlant species of Verbascum genus have been intensively investigated in the last decades but most studies focused on evaluation of their biological activities; there are only few studies dealing with their chemical characterisation. ObjectiveDetailed investigation of the qualitative and quantitative chemical composition, antioxidant and cytogenotoxic activities of a previously non-studied Verbascum species (V. ovalifolium Donn ex Sims). MethodsQualitative analysis of secondary metabolites was performed by HPLC-DAD-ESI-Q-TOF-MS/MS, whereas quantitative data were obtained through HPLC-DAD. Antioxidant activity was evaluated using in vitro assays; cytotoxic and genotoxic effects were assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium (MTT) and comet assays, respectively. ResultsMore than 50 secondary bioactive metabolites belonging to various classes (iridoids, phenylethanoids, flavonoids, phenolic acids) were detected in the methanolic extract of V. ovalifolium and its fractions. The fragmentation pathways of acylated catalpol-type iridoid diglycosides are thoughtfully described herein. The extracts showed good free radical scavenging and ferric ion reducing properties correlated with phenolic, flavonoid, chlorogenic acid and verbascoside contents. Moreover, 24h treatment of SK-MEL-2 cells with V. ovalifolium extracts produced significant changes in terms of tumour cell viability. The crude extract and the ethyl acetate fraction showed no important signs of cytogenotoxicity in non-tumour cells. ConclusionThe performed phytochemical and biological analyses contribute to the preclinical knowledge about V. ovalifolium and they could help exploiting it in novel herbal medicinal products. More than 50 bioactive secondary metabolites (iridoids, phenylethanoids, flavonoids, phenolic acids) were detected in the aerial parts of Verbascum ovalifolium Donn ex Sims (Scrophulariaceae) by HPLC-DAD-ESI-Q-TOF-MS/MS. Moreover, the fragmentation pathways of acylated catalpol-type iridoid diglycosides are thoughtfully described herein. Biological activity results showed a significant in vitro antioxidant capacity of the methanolic extract and its fractions, a promising antiproliferative activity in malignant melanoma SK-MEL-2 cells and no important cytotoxicity and genotoxicity in V79 cells.

Abstract

Botrytis cinerea is a necrotrophic fungus that affects over 200 plant species. In vineyards, this pathogen is responsible for one of the most important diseases, commonly known as botrytis bunch rot or grey mould. Keeping infection under control with synthetic fungicides leads to an increased biological resistance of pathogen populations. An alternative way to synthetic products is to obtain natural fungicides by using bioactive compounds of plants. This study focuses on the antifungal properties of a new bioproduct obtained from polymeric proanthocyanidins extracted from grape seeds of 'Feteasca neagra' variety. The bioproduct in solid state presented a total content of polyphenols of 0.625 mg GAE mg(-1), a polyphenolic index of 17.40 and an antioxidant activity of 91.27% scavenged DPPH. The bioproduct with polyphenolic structure showed a moderate effect on the radial growth of fungal mycelium, at EC50 values between 11.23 and 12.15 mg mL(-1). Effective antifungal activity was showed in the inhibition of spore germination, where the EC50 values varied from 1.14 to 1.47 mg mL(-1). These in vitro results sustain the possibility of including the bioproduct in the category of natural fungicides for biological control against Botrytis cinerea fungus.

Abstract

In the category of medicinal plants with remarkable therapeutic properties, lichens are also included. An important representative of this group is the Usnea Adans. genus, with over 650 species spread throughout the globe. Representative species of this genus were also found in Romania, so the studies were conducted on Usnea barbata (L.) EH.Wigg., harvested from the Calimani Mountains, Suceava County. The objectives of this research are: obtaining the dry extract of the Usneae lichen followed by the identification and determination of the usnic acid content by ultra-high performance liquid chromatography (UHPLC) method. Usnic acid content determined in dry extract of Usnea barbata (L.) EH.Wigg. was significant, of 16.53 +/- 6.53% (mean +/- RSD), which supports the continuation of the research with the evaluation of biological effects (antibacterial, antioxidant and antitumoral).

Abstract

New type of nanocapsules based on carboxymethyl chitosan functionalized with AS1411 aptamer and poly(N-vinylpyrrolidone-alt-itaconic anhydride) loaded with 5-Fluorouracil (5-FU) were developed, with the potential to improve the treatment of cancer. Functionalization of nanocapsules with AS1411 aptamer will enhance their recognition by tumor cells, due to the interaction with nucleolin, and subsequent endocytosis. Nanocapsules were prepared by interfacial condensation method in the absence of any toxic crosslinking agents. The condensation reaction took place at the interface between the organic and aqueous phases by opening the anhydride cycles from the copolymer, under the action of the NH2 groups from mixture of chitosan/aptamer-functionalized carboxymethyl chitosan. The nanocapsules diameter varied between 100 and 267 nm as a function of the molar ratio of the polymers. SEM images have revealed that nanocapsules were spherical and presented relatively low dimensional polydispersity. Nanocapsules swelling degree was found between 1000 and 1680% in PBS solution (pH = 7.4) and they allowed the encapsulation of an important amount of 5-Fluorouracil (5-FU). The release efficiency of 5-FU was studied, the processes being controlled by the drug diffusion through the polymeric membrane, as confirmed by the theoretical analysis of the drug release. The cytotoxicity and haemolysis tests performed on the nanocapsules proved their lack of toxicity and their excellent hemocompatibility. The obtained results were encouraging, showing that these original 5-FU-loaded nanocapsules were able to induce a more pronounced cytotoxic effect on neoplastic MCF-7 cells, the occurrence of dead cells being more rapidly than in the case of free 5-FU.

Abstract

The morphological ontogeny of Tectoribates ornatus (Schuster, 1958) based on specimens from Romania is described and illustrated. The juveniles of this species are plicate, unideficient and apheredermous, with leaf-like prodorsal setae in and le and most gastronotal setae. The larva has sclerites on the gastronotum, and some of them bear setae. In the juveniles, seta d is present at solenidion sigma on genua I-III, phi(1) on tibia I and phi on other tibiae, but this seta is lost in the adult. In the nymphs, seta d on all femora and seta l' on genu and tibia I are leaf-like.

Abstract

More than one out of every three new cancers is a skin cancer, and the large majority are basal cell carcinomas (BCC). Targeted therapy targets the cancer's specific genes, proteins, or tissue environment that contributes to cancer growth and survival and blocks the growth as well as the spread of cancer cells while limiting damage to healthy cells. Therefore, in the present study AS1411 aptamer-functionalized liposomes for the treatment of BCC were obtained and characterized. Aptamer conjugation increased liposome size, suggesting that the presence of an additional hydrophilic molecule on the liposomal surface increased the hydrodynamic diameter. As expected, the negatively charged DNA aptamer reduced the surface potential of the liposomes. Vertical Franz diffusion cells with artificial membranes were used to evaluate the in vitro release of 5-fluorouracil (5-FU). The aptamer moieties increase the stability of the liposomes and act as a supplementary steric barrier leading to a lower cumulative amount of the released 5-FU. The in vitro cell viability, targeting capability and apoptotic effects of liposomes on the human dermal fibroblasts and on the basal cell carcinoma TE 354.T cell lines were also evaluated. The results indicate that the functionalized liposomes are more efficient as nanocarriers than the non-functionalized ones.

Abstract

Frequently, in polytunnels are found some nonparasitic or physiological disorders on vegetable plants such as tomatoes, peppers, eggplants, and some melons. The growing stagnation of tomato plants cultivated in polytunnels is evidenced by some symptoms that appear during the vegetation season. These symptoms may be due to biotic (e.g., diseases, pests) and environmental factors such as high fluctuations in soil moisture, low or high values of relative humidity of air, atmospheric drought, high temperatures, soil characteristics, etc. One of the soil attributes that favours physiological disorders, are also the substrate reaction, characterized by pH values lower than 5 or higher than 8. High content of some nutrients such as N, K, Mg, Na, Mn, antagonists to Ca, can also induce physiological disorders. The physiological disorders symptoms are manifested especially on the leaves and fruits. Some physiological disorders can often be misidentified as caused by pathogens, for example Blossom End Rot, can often be misidentified as gray rot caused by pathogen Botrytis cinerea Pers., which is a fungus (Ascomycota). Both disorders manifests by brown spots on fruit skin surface, both are favoured by high humidity and weak airing in polytunnels. One of the objectives of the paper is the environmental factors characterization which influence tomato plants affected by nonparasitic disorders such Blossom-end rot or leaf curling and leaf distorting. Following our investigations, it was found that frequently, some symptoms on tomatoes were not due to a parasite action. The correct identification and differentiation of parasitic disorders caused by the environmental factors, presents a particular practical importance in order to establish prevention measures and cultural recommendations.

Abstract

The research was carried out on the protected crops, greenhouses and solariums investigating the variations of photosynthesis, transpiration, stomatal conductance and also, sugars leaf fractions in different technological systems (organic and conventional) on the varieties of Capsicum annum L. (peppers) and Solanum melongena L. (eggplant). In May-June, photosynthesis, transpiration and stomatal conductance showed higher values for varieties from the conventional system, while in July these parameters are higher for organically grown varieties, especially at the basis and middle part of plant showing an increase in the latter fruiting phenophase. Wue, the A/E ratio, showed in flowering stage high and close values in classic and in organic systems. In ripening stage, Wue have a quite higher value in organic systems and where plants showed a slower growth than in classic. Higher values of substomatal CO2, Ci have been observed in organic systems which involved a slower rate of absorbtion of atmospheric CO2 in photosynthesis process than was observed in classic variants. In ripening phenophasis, in classic technology system, disaccharides fraction is increased because are involved in growth and sugars accumulation in fruit whilst the monosaccharides and polysaccharides decrease but varieties organically grown showed higher values of disaccharides and also of polysaccharides.

Abstract

The purpose of this paper is to highlight certain characteristics of the fruits of Sambucus nigra L. harvested from five different locations in Iasi County (N-E Romania). The following features were considered: histological (epicarp and mesocarp analysis), morphological (number of fruits, fruits fresh weight), physicochemical (refractometric index and total soluble solid substances-TSS) and biochemical (water and dry matter content, total mineral elements). Referring to the histological characteristics, epidermal cells were found to be rectangular, being prominently elongated in sample L2; the size of the hypodermic cells varied according to the analyzed samples and the mesocarp was thick in L1 and thin in L4 and L5. The number of fruits in corymbiform cymes showed value between 126.4±10.66 and 354.4±30.42. The fruits fresh weight showed values between 0.0967 g and 0.1468 g. The fruits harvested from the location 5 are characterized by high values for the parameters number of fruit/ corymbiform cymes, TSS, organic substance, and low values for the content of water and ashes. The above specified parameters indicate specific variations related to the fruits source location, a high content in water and solid matter, a moderate content in mineral elements, and a strong negative correlation between the water content and the soluble solid matter content. © 2019, University of Zagreb. All rights reserved.

Abstract

In consideration of the hazards associated with the presence of the textile azo-dye and their biotransformation products in the environment, the goal of this work was to study bioremediation process by the yeast strain Pichia kudriavzevii CR-Y103 related to the ability to degrade and detoxify the sulfonated Reactive Orange 16 azo-dye. In experimental conditions, the optimal inoculum/dye concentration ratio required for complete decolorization (100%) of culture medium and biomass within 24 h has been 1 g L-1 yeast cell (dry weight)/50 mg L-1 Reactive Orange 16. In the presence of 400 mg L-1 of Reactive Orange 16 (RO16), 95% of the dye was removed after 72 h of incubation. Also, the yeast strain could decolorize other eight textile dyes (56.48-99.98% decolorization within 24 h). NADH-DCIP reductase and azo reductase activities were significantly increased (ca. 5.4 times and ca. 37 times, respectively) during the decolorization process. UV-VIS spectra, high-performance liquid chromatography (HPLC), and Fourier transform infrared spectroscopy (FTIR) analysis confirmed the presence of new biotransformation products in extracted metabolites, highlighting the partial biodegradation of the dye by the new yeast isolate. The phytotoxicity evaluation strongly supported the decreased toxicity of biodegraded products as minor inhibition on germination (%), root and shoots elongation of T. pratense L. and T. aestivum L. seedlings. Increasing of mitotic index value and decreasing the frequency of chromosomal aberrations in tested plant meristem cells treated with biodegraded products, compared with RO16 treatment (500 ppm), confirmed their slightly toxic nature. A cell viability assay also confirmed the reduced toxicity of biodegraded products on healthy monkey kidney cells (Vero cells).

Abstract

The identification and conservation of indigenous rhizobia associated with legume plants and their application as biofertilizers is becoming an agricultural worldwide priority. However, little is known about the genetic diversity and phylogeny of rhizobia in Romania. In the present study, the genetic diversity and population composition of Rhizobium leguminosarum symbiovar trifolii isolates from 12 clover plants populations located across two regions in Romania were analyzed. Red clover isolates were phenotypically evaluated and genotyped by sequencing 16S rRNA gene, 16S-23S intergenic spacer, three chromosomal genes (atpD, glnll and recA) and two plasmid genes (nifH and nodA). Multilocus sequence typing (MLST) analysis revealed that red clover plants are nodulated by a wide genetic diversity of R. leguminosarum symbiovar trifolii sequence types (STs), highly similar to the ones previously found in white clover. Rhizobial genetic variation was found mainly within the two clover populations for both chromosomal and plasmid types. Many STs appear to be unique for this region and the genetic composition of rhizobia differs significantly among the clover populations. Furthermore, our results showed that both soil pH and altitude contributed to plasmid sequence type composition while differences in chromosomal composition were affected by the altitude and were strongly correlated with distance. (C) 2018 Elsevier GmbH. All rights reserved.

Abstract

The symbiotic nitrogen fixing legumes play an essential role in sustainable agriculture. White clover (Trifolium repens L.) is one of the most valuable perennial legumes in pastures and meadows of temperate regions. Despite its great agriculture and economic importance, there is no detailed available information on phylogenetic assignation and characterization of rhizobia associated with native white clover plants in South-Eastern Europe. In the present work, the diversity of indigenous white clover rhizobia originating in 11 different natural ecosystems in North-Eastern Romania were assessed by a polyphasic approach. Initial grouping showed that, 73 rhizobial isolates, representing seven distinct phenons were distributed into 12 genotypes, indicating a wide phenotypic and genotypic diversity among the isolates. To clarify their phylogeny, 44 representative strains were used in sequence analysis of 16S rRNA gene and IGS fragments, three housekeeping genes (atpD, glnII and recA) and two symbiosis-related genes (nodA and nifH). Multilocus sequence analysis (MLSA) phylogeny based on concatenated housekeeping genes delineated the clover isolates into five putative genospecies. Despite their diverse chromosomal backgrounds, test strains shared highly similar symbiotic genes closely related to Rhizobium leguminosarum biovar trifolii. Phylogenies inferred from housekeeping genes were incongruent with those of symbiotic genes, probably due to occurrence of lateral transfer events among native strains. This is the first polyphasic taxonomic study to report on the MLSA-based phylogenetic diversity of indigenous rhizobia nodulating white clover plants grown in various soil types in South-Eastern Europe. Our results provide valuable taxonomic data on native clover rhizobia and may increase the pool of genetic material to be used as biofertilizers.

Abstract

The aim of this study was to carry out a histo-anatomical analysis on the stem, nodal branches and leafs of two Equisetum species, Equisetum pratense and Equisetum sylvaticum, commonly known as horsetail. This study was elaborated using color photographs made by the optical microscopy (OM) and scanning electron microscopy (SEM). Using OM, we could investigate the epidermal wall cells, the outer cortex made of different types of tissues and the center cylinder organized into two regions, an external layer that enclosed vascular bundles and an internal layer represented by a large air cavity. The images achieved by SEM showed some differential characteristics concerning the trichomes and stomata but also the manner in which granules of silicon dioxide are spread throughout the surfaces of Equisetum species. These results may serve as a criterion for differentiation and may have taxonomic value in the Equisetum family because they represent elements of novelty in our country and abroad.

Abstract

The scope of the present work is to study the biodegradability behavior of several novel heterocyclic poly(ether urethanes) when their hard segments are subjected to exposure to the Alternaria tenuissima fungus. The heterocyclic poly(ether urethanes) were chain-extended with various pyridine derivatives that had different functional groups placed in different positions on the pyridine rings. Different ratios of the reactive components were also used. The measurements obtained by the use of the attenuated total reflectance-Fourier transform infrared spectroscopy revealed the different structural changes that occurred after exposure to fungi. The degradation process was analyzed through measurement of the mechanical properties and surface morphology evolution by scanning electron microscopy. The obtained results show that the nature and design of the functional groups on the pyridine ring determine the strength of the cohesion linkages that in turn influence different degradation behaviors under exposure to fungi. (c) 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 46096.

Abstract

The study provides information about separation and identification of natural bioactive compounds from beech (Fagus sylvatica L.) bark with potential therapeutic applications such as antibacterial activity against human pathogens. Beech is a common material used in the wood industry, but its bark is separated from the wood and is considered a by-product. In this study, natural compounds with biological activity were obtained from beech bark by hot water extraction. The high-performance liquid chromatography (HPLC) was used to analyze the phenolic compounds in the beech bark extracts. Spectrophotometric methods were employed for the determination of total phenolic content. Microdilution technique was used for testing the antimicrobial activity of the extract. The following strains were tested: Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa. The yield of extracted polyphenols was of 22.952 mg gallic acid/g dry bark. The compounds identified by HPLC were vanilic acid, catechin, taxifolin and syringin. The extracts were active against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The effect of polyphenolic extract on Gram-negative bacteria was absent at a concentration of 30 mg/mL beach bark extract. Altogether, the use of pure water for extraction of polyphenols from beech bark proved to be an effective eco-friendly method. This method sustains the concept of green chemistry by involving the use of renewable plant resources and also by using water as solvent.

Abstract

Soil morphological, physical and chemical properties are described at four locations along an elevational transect in the northeastern part of the Moldavian Plateau (Romania). These data contribute to the knowledge of the soils of this area and to their classification according to the USDA-Soil Taxonomy, FAO-WRB and the SRTS-Romanian System. The soils were classified as Inceptisols, Alfisols and Mollisols, according to the USDA-Soil Taxonomy; Gleysol, Chernozem and Luvisol, according to the FAO-WRB and Gleiosol, Cernoziom, Preluvosol, Luvosol, according to the SRTS-Romanian System. The selected soils have a range of properties that represent the soilscape of the Moldavian subcarpathian plateau, characterised by a natural forest with oak as the dominant species. The selected soil parameters decreased with increasing elevation; calcium carbonate and clay leaching and accumulation are the main soil formation processes. (C) 2018 International Research and Training Center on Erosion and Sedimentation and China Water and Power Press. Production and Hosting by Elsevier B.V.

Abstract

In this paper two newly recorded species of the family Oppiidae are presented, namely Ramusella alejnicovae (Krivolutsky and Gatilova, 1974) and Lauroppia iranica Akrami and Subias, 2008. Additionally, recent records of two other rare species, i.e., Multioppia (M) perfecta Mahunka and Topercer, 1983 and Ramusella sengbuschi tokyoensis (Aoki. 1974) am given. For each species, a redescription and illustration based on Romanian material is provided and compared with existing descriptions, and data regarding world distribution and ecological requirements are analyzed. In addition, the status of Multioppia (Hammeroppia) insolita Ivan and Vasiliu, 1999, a species considered as a possible synonym of M. (H.) wilsoni laniseta Moritz, 1966 (Subias 2004 updated 2018) is discussed, and reasons to deem insolita as a valid species are pointed out.

Abstract

Our investigation has been focused on establishing the slowing down plant growth at the border of polytunnels. In order to determine the limiting factors of plant growth, we have conducted studies on soil properties, entomofauna and soil mesofauna. The research has been carried out in several polytunnels located in North Eastern Romania (Bacău, Iaşi, Bârlad, Fălticeni, Hârlău, Târgu Frumos, Matca-Tecuci) In the field were developed and characterized soil profiles and samples were taken to determine the states of compactness and humidity. The results of researches revealed a strong growth stagnation of tomatoes, cucumbers, eggplants and peppers. The areas where plants have recorded the lowest growth are found in the lateral parts and at the entrance to the polytunnels, as a result of the larger amounts of rainfall, recorded during the vegetation season. Following the analysis, we found that there is a direct correlation between the width of the over wetted soil strip in the lateral parts of the polytunnel and its constructive parameters (width, height, plastic or glass walls), the distance between the polytunnels and the characteristics of the ploughed and under ploughed soil layers. As a result of a pluvial front, the amount of water fallen on the soil unit between the polytunnels, is 6 to 20 times higher than the land in the vicinity of the polytunnels, due to the water flowing from both sides of the solarium. Compaction of the plough layer between the polytunnels, the presence of the hardpan layer, are the additional factors on which the water infiltration rate depends. In addition to the growing stagnation of cultivated plants, a more pronounced incidence of diseases and a more intense attack of the pests, represented by Thysanoptera (Thrips tabaci) and Homoptera, especially aphids – Myzus persicae and Hemiptera – greenhouse whitefly (Trialeurodes vaporariorum). Mesofauna is known as indicator of edaphic conditions, such as water excess. Our investigation revealed very large differences, both quantitative and qualitative, between samples with excess moisture and dry soil between rows. Thus, the average global density was about 35 times higher in the excessively wet soil, and some groups missing in the dry soil are present here (Astigmata and Trombidiformes among mites), whereas. © SGEM 2018.

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The aim of our proposals for soil sampling regarding mesofauna study in soils from greenhouses or polytunnels is motivated by the high soil heterogeneity and large variability of easy exchangeable soil properties such as water content, soluble salts distribution especially under drip irrigation and local fertilization. The intensive technology for growing vegetables or flowers, high irrigation requirements and the high soil moisture favors, in short time the degradation of morphological, physical, chemical properties and especially biological activity of the greenhouses soils. The large variability of the mesofauna species with different requirements to environmental conditions (e. g., hygrophilous to xerophilous species as regards soil humidity, salt tolerant or sensitive species to soil salinity etc.) has lead us to adapt soil sampling methods to particular conditions of every location. One of particular situation was in greenhouse where practice drip irrigation and local fertilizations of tomato plants. Using the adapted sampling methods, the identified mesofauna species confirmed the large variability of soil properties from plant rows, on the border of wetting strip, at middle distances of the plant rows and on the horticultural substrata used for seedling obtaining. After processing of field and laboratory obtained data we consider that proposed diagram for design location is useful for practical application in order to detect initiation processes of soil degradation and establish the proper measures to prevent decreasing soil quality. © SGEM 2018.

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Ramaria largentii Marr & D. E. Stuntz (orange coral mushroom) is a wild edible mushroom whose chemical composition and bioactivity have not been investigated. Herein, we present a study on the phenolic constituents, antioxidant and antigenotoxic effects of a hydromethanolic extract of the fruiting bodies. Total phenolic content, estimated by Folin-Ciocalteu assay, was found to be 42.33 +/- 0.18 mg GAE/g. Protocatechuic and vanillic acids were detected by HPLC-DAD-ESI-MS. The extract showed good free radical scavenging and reducing capacities (EC50 = 64.3 +/- 0.2 and 61.54 +/- 0.46 mu g/mL, respectively). In normal Vero cells, the extract (100, 200 and 300 mu g/mL) showed no genotoxic potential and moreover, almost completely protected DNA against H2O2-induced damage (2.09-7.91% tail DNA) (24 and 48 h pretreatment). Taken together, the results of our study show that Ramaria largentii extract is devoid of genotoxicity and has a remarkable DNA protective activity against H2O2-induced damage. (C) 2017 Elsevier Ltd. All rights reserved.

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Lepidiumdraba (Brassicaceae) is a major concern for agriculture and biodiversity in the western United States. As current control methods do not provide long-term, sustainable solutions, research has been conducted to find biological control agents. Ceutorhynchusassimilis is one of the currently investigated candidates. Known as oligophagous in the literature, a specialist clade of this root-galling weevil exists in southern Europe. This raised the question of its ability to survive in colder climates in the target range. We investigated the phenology of C.assimilis in the field in southern France (specialist clade) and Romania (generalist clade) and measured various temperature-dependent parameters in the laboratory. In both ranges, weevils were univoltine. Oviposition in autumn started later in France compared to Romania, while mature larvae exited galls (to pupate in the soil) earlier the following year. On average, 25% and 32% of galls from France and Romania were completely below the soil surface, respectively, and this appeared to depend on soil substrate. Weevils transported from France to Romania were able to develop, but at a much lower rate than Romanian weevils. Mortality of overwintering larvae of both clades increased with decreasing temperature and exposure time. At -5 degrees C, lethal times Lt(50) and Lt(95) were 15 and 42days for the specialist clade and 26 and 72days for the generalist clade. A higher proportion of third instar larvae compared to first and second instar larvae survived. Pupation time at different temperatures did not differ between weevils from France or Romania. A climate match model (comparing winter temperatures) indicated that the specialist clade of C.assimilis from France has the potential to establish in some parts of the target range (e.g. Washington, Oregon, California). However, temperature extremes and winters without snow cover will likely limit its establishment unless rapid adaptive evolution takes place.

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This research shows the fungal degradation behavior of two series of composites: a series obtained with silver powder incorporated in sulfadiazine (SD)-based polyurethane urea and another series of composites that incorporates silver through polyurethane chain extension with silver SD. This article reports on the chemical structure variations characterized by Fourier transform infrared (FTIR) spectroscopy and the mechanical properties of these polymers before and after fungal exposure. Although silver SDused as a chain-extenderensures a more orderly dispersion of the silver throughout the hard segment of the polyurethane urea main chain, the FTIR results showed some changes on the surface of these composite films. Both silver composites exhibited higher fungal biodegradation resistance.

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The present study had the aim to evaluate the mining plant activities impact on forest ecosystems taking a common sense view of determination, at foliar level, of photosynthetic pigments content, of catalase's and peroxydase's activity - as enzymes involved in the defense against the reactive species of oxygen, the Krebs cycle dehydrogenases' activity - as a main metabolic path generative of energy, but also of the microbial dehydrogenases' activity from soil as a ecologic biomarker with key role in maintaining the soil's quality. In parallel with the mentioned biochemical indicators were performed a series of histo-anatomic investigations. Were taken in study samples from the foliar tissue from two angiosperms species (Populus tremula_L. and Salix caprea L.) from areas of Suceava's County namely Radauti (the reference area) and Crucea-Botusana (area of uranium mining), but also samples of soil derived from different profiles of deepness (0-15 cm and 15-30 cm). The toxicity impact resulted as a consequence of anthropic activity of uranium holding in the Crucea area it is translated through an inferior content of a and b chlorophyll, the photosynthesis process being largely inhibited, in contrast with the superior net catalase's, peroxidase's, but also of foliar dehydrogenases' activity, these enzymes laying out values even four times higher comparatively with the reference lot. The structural differences evidenced especially at the foliar limb level of the two species could be due to the anthropic pollution resulted as a consequence of the mining activity.

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Background: Haloalkane dehalogenases (EC 3.8.1.5, HLDs) are alpha/beta-hydrolases which catalyze the irreversible cleavage of carbon-halogen bonds of haloalkanes, producing an alcohol, a halide and a hydrogen ion. Haloalkanes are acutely toxic to animals and humans and their toxic effects are mainly observed in the liver, kidneys and central nervous system. Objective: In the present work, the haloalkane dehalogenase from Rhizobium leguminosarum bv. trifolii (DrlA) was characterized. Method: Reverse transcription polymerase chain reaction analysis and enzyme activity assays revealed that the DrlA gene expression in R. leguminosarum bv. trifolii is induced by 1,2dibromoethane (1,2-DBE) during the early exponential phase. The gene of the enzyme was isolated, cloned and expressed in E. coli Rosetta (DE3). Results: Recombinant DrlA displays its high catalytic activity towards 1,2-DBE and the long-chain haloalkane 1-iodohexane. Limited activity was observed for other aliphatic and cyclic haloalkanes, indicating that the enzyme displays restricted substrate specificity, compared to other bacterial HLDs. Homology modelling and phylogenetic analysis suggested that the enzyme belongs to the HLD-II subfamily and shares the same overall fold and domain organization as other bacterial HLDs, however major variations were identified at the hydrophobic substrate-binding cavity, the cap domain and the entrance of the main tunnel that affect the size of the active site pocket and the substrate recognition mechanism. Conclusion: This work sheds new light on the environmental fate and toxicity of 1,2-DBE and provides new knowledge on the structure, function and diversity of HLDs for developing applications in toxicology.

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Ramaria largentii Marr & D. E. Stuntz (orange coral mushroom) is a wild edible mushroom whose chemical composition and bioactivity have not been investigated. Herein, we present a study on the phenolic constituents, antioxidant and antigenotoxic effects of a hydromethanolic extract of the fruiting bodies. Total phenolic content, estimated by Folin-Ciocalteu assay, was found to be 42.33 ± 0.18 mg GAE/g. Protocatechuic and vanillic acids were detected by HPLC-DAD-ESI-MS. The extract showed good free radical scavenging and reducing capacities (EC50 = 64.3 ± 0.2 and 61.54 ± 0.46 μg/mL, respectively). In normal Vero cells, the extract (100, 200 and 300 μg/mL) showed no genotoxic potential and moreover, almost completely protected DNA against H2O2-induced damage (2.09–7.91% tail DNA) (24 and 48 h pre-treatment). Taken together, the results of our study show that Ramaria largentii extract is devoid of genotoxicity and has a remarkable DNA protective activity against H2O2-induced damage. © 2017 Elsevier Ltd

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The paper present histo-anatomic aspects, recorded in seedlings of maize (Zea mays L.) grown under the influence of spruce bark polyphenolic extract (SBPE). Spruce barck polyphenolic extract was characterized in terms of the total content of polyphenols, tannins, flavonols, flavonoids and anthocyanins in a previous paper. It is known that, natural polyphenols are essential compounds in stimulating plant growth and development. The aim of this study was to identify the internal structure changes in maize seedlings, under the influence of SBPE, applied in two concentrations (SB1 - 190 mg GAE / L and SB2 - 130 mg GAE / L). It was observed, compared with the control, that SBPE determines a better development of the absorbents bristle and increase leading bundles number and central vessels metaxylem. SBPE in lower concentrations has positive effects that can be attributed to antioxidant properties. The results recorded in this study open new pathways of research in the field.

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The aim of the present study was to investigate the antigenotoxic and antioxidant activities of the crude hydromethanolic extract from the aerial parts of European Dracocephalum moldavica L. (Moldavian dragonhead). The total phenolic content estimated by the Folin-Ciocalteu assay was 289.55 +/- 2.63 mg of GAE/g of dry extract, and rosmarinic acid was the major polyphenol of Dracocephalum moldavica extract (107.11 +/- 0.83 mg/g of dry extract). In vitro antioxidant assays revelead remarkable scavenging effects against DPPH (EC50 = 23.10 +/- 0.10 mu g/mL), ABTS (EC50 = 8.0 +/- 0.10 mu g/mL) and superoxide anion radicals (EC50= 445.5 +/- 2.3 mu g/mL). The extract showed a high ferrous ion chelating activity (EC50 = 35.70 +/- 0.40 mu g/mL), a considerable reducing capacity, and good hydroxyl radical scavenging properties. Dracocephalum moldavica extract reduced, in a concentration-dependent manner, DNA damage induced by bleomycin in normal human dermal fibroblasts as measured by comet assay and micronucleus test. Exposure of dermal fibroblasts to Dracocephalum moldavica extract (100 mu g/mL) after preincubation with bleomycin (10 mu g/mL) resulted in the most significant antigenotoxic activity. The protective effects may be due to the free radical scavenging activity, iron-chelating properties and the possible intervention on DNA repair processes. (C) 2015 Elsevier B.V. All rights reserved.

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EU legislation restricted many chemicals for root-knot nematodes (Meloidogyne spp., RKNs) control. Alternative ways of plant protection were hence investigated, based on the use of microbial formulations exploiting plant parasites antagonism. RKNs are severe and widespread pests causing extensive damage to crops in greenhouse and field. Several antagonistic microorganisms are suitable for biocontrol, including the nematophagous hyphomycete Pochonia chlamydosporia that parasitizes eggs to acquire additional nourishment and face competition with other soil microorganisms. A commercial product (POCHAR) was developed by Microspore based on P. chlamydosporia with other microbial inocula that can be applied through irrigation. The aim of this study was to test POCHAR's efficacy against RKNs on potato and its promotion effect on tomato in two field trials. Moreover, the research included trials to evaluate the best method for open-field application in crops not managed through drip irrigation, opening up the possibility to treat large areas without major technological needs. RKNs control with POCHAR represented a viable alternative to chemicals. In conclusion, the organic approach developed through the bioformulated product highlighted effective RKNs management, with a potential to sustain both plant nutrition and the related root protection needs.

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Nanoparticulate matter, mainly nanoparticles containing metal ions could represent a promising biotechnological tool when preliminary testing could indicate the ability of inducing chromosomal changes in the plant tissues. Chemical route was applied as synthesis method of ZnFe2O4 powder with nanometric size, thus enabling plants to internalize the particles through endocytosis for further lysosomal digestion. Surface modification with long chain molecular shell was chosen to ensure nanoparticle stability in water dispersion. We present the results of some genetic effects induced by such prepared zinc ferrite nanoparticles as diluted suspension when supplied to agrotechnical plant species of large interest namely sunflower. Various and numerous chromosomal aberrations were identified that could be perpetuated in the form of genetic mutations through adequate biotechnology. Quantitative analysis emphasized remarkable influence of ZnFe2O4 nanoparticles on meristem tissues of freshly germinated seeds which suggested possible new biotechnological tool based on nanotoxicity application in genetics research.

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It is known that the genetic diversity of conspecific rhizobia present in root nodules differs greatly among populations of a legume species, which has led to the suggestion that both dispersal limitation and the local environment affect rhizobial genotypic composition. However, it remains unclear whether rhizobial genotypes residing in root nodules are representative of the entire population of compatible symbiotic rhizobia. Since symbiotic preferences differ among legume populations, the genetic composition of rhizobia found within nodules may reflect the preferences of the local hosts, rather than the full diversity of potential nodulating rhizobia present in the soil. Here, we assessed whether Vicia cracca legume hosts of different provenances select different Rhizobium leguminosarum genotypes than sympatric V. cracca hosts, when presented a natural soil rhizobial population. Through combining V. cracca plants and rhizobia from adjacent and more distant populations, we found that V. cracca hosts are relatively randomly associated with rhizobial genotypes. This indicates that pre-infection partner choice is relatively weak in certain legume hosts when faced with a natural population of rhizobia. (C) 2016 Elsevier GmbH. All rights reserved.

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The main objective of this study was to compare soil mesofauna communities in natural and anthropogenic forests from Danube Delta Biosphere Reserve and establish a baseline data in monitoring the disturbed sites. The abundance and diversity of edaphic microarthropods were analyzed in five plots, three of them being natural forests and two plantations (Canada poplar, and respectively willow). The mites from Trombidiformes and Oribatida were closely investigated being identified at family level. Qualitative analysis of edaphic microarthropods evidences numerical dominance of mites, excepting only one stand (Canada poplar plantation). Among mites Oribatida owns the biggest weight (76.6 - 94.1% of the total mites), followed by Trombidiformes or Mesostigmata. Among insects the collembolans hold higher density in the poplar plantation, and the lowest one in the willow plantation. Structure of mites communities differs between the investigated ecosystems both in quantitative and qualitative aspects depending on particular conditions of each plot. © 2016 Vasile Goldis University Press.

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We determined the host range of the parasitoid Trichomalus perfectus (Walker), a candidate for classical biological control of cabbage seedpod weevil, Ceutorhynchus obstrictus (Marsham), an important pest of canola in Canada. Studies were conducted in Europe and in North America. In laboratory experiments, the levels of parasitism (acceptance) of Ceutorhynchus turbatus Schultze, C. cardariae Korotyaev, C. omissus Fall and C. querceti (Gyllenhal) by T. perfectus were not significantly different than of the target host C. obstrictus. Although C. typhae (Herbst), C. pallidactylus (Marsham), C. americanus Buchanan, C. neglectus Blatchely and Ceutorhynchus sp. nr. nodipennis were parasitised by T. perfectus, the levels of parasitism were significantly lower on these species than on C. obstrictus. Ceutorhynchus peyerimhoffi Hustache, C. erysimi (Fabricius), C. alliariae H. Brisout, C. roberti Gyllenhal, Mogulones borraginis (Fabricius), Mononychus vulpeculus (Fabricius) and the leaf-mining fly Scaptomyza flava (Fallen) were not attacked. Ecological host range surveys in Europe corroborated the prediction that T. perfectus would attack C. cardariae at similar rates to C. obstrictus. In North America, the recent discovery of T. perfectus in a C. omissus population suggests that laboratory findings predicting that C. omissus is a preferred host may be the case in the field. We found that T. perfectus attacks larvae of some Ceutorhynchus spp. feeding on Brassicaceae and does not attack species outside of that host range. Thus, the parasitoid can be defined as narrowly oligophagous. These results demonstrate the value of ecological host range studies in the area of origin to validate hypotheses generated through laboratory host range experiments.

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Lepidium draba (Brassicaceae) is a clonal herb, originating from Eurasia, which is invasive in North America. A classical biological control project was initiated in 2001, and the gall-forming weevil Ceutorhynchus cardariae was prioritized as a candidate agent. We studied its biology and field host range between 2003 and 2014 in the laboratory and a common garden in Switzerland and in the field in Romania. Ceutorhynchus cardariae is a univoltine to bivoltine species. In Switzerland, oviposition usually started at the beginning of March and can occur at temperatures as low as 2.5 degrees C. Galls are formed on stems, leaf stalks and midribs of L.draba rosettes and bolting plants. Gall size increased with an increasing number of larvae per gall. The three larval instars feed inside the galls and leave the plant to pupate in the soil once mature. In Switzerland, development from egg to adult took about 12weeks in spring. Adults emerged from May to July. After a brief feeding period, adults aestivate. From late summer, feeding recommenced and females may oviposit, forming a partial second generation. Eggs and all larval instars can be found in galls throughout winter. The rate of larval ectoparasitism reached 78%, while endoparasitism was low with a maximum of 2.3%. Lepidium draba populations differed in their suitability for development (number of C.cardariae produced), indicating that effectiveness of C.cardariae - in case released - may be variable. In the field, we observed that gall formation by C.cardariae can severely stunt or even kill shoots. Investigations on the field host range of C.cardariae indicated that only the closely related Lepidium campestre may act as an alternative host for the weevil in Europe. Host-specificity tests are underway to determine its environmental safety before field release in North America is being considered.

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The application of commercial rhizobial inoculants to legume crops is proving to be an alternative to synthetic fertilizer use. The challenge for sustainable agriculture resides in the compatibility between crop, inoculants and environmental conditions. The evaluation of symbiotic efficiency and genetic diversity of indigenous rhizobial strains could lead to the development of better inoculants and increased crop production. The genetic variability of 32 wild indigenous rhizobial isolates was assessed by RAPD (Random Amplified Polymorphic DNA). The strains were isolated from red clover (Trifolium pratense L.) nodules from two distinct geographical regions of Northern and Eastern Romania. Three decamer primers were used to resolve the phylogenetic relationships between the investigated isolates. Cluster analysis revealed a high diversity; most strains clustered together based on their geographical location.

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The use of extractive mining technologies in various areas of Suceava, Romania (Calimani Mountains - sulfur, Tarnita-Ostra - copper and barite, and Crucea-Botusana - uranium) has resulted in the accumulation of surplus chemicals with direct action on the ecosystem. The aim of this study was to analyze the possible damage on some woody species (Picea abies L., Populus tremula L., Salix alba L., Betula verrucosa Ehrh., and Larix decidua Mill.) and their adaptive and phytoremediation capacity through the biomarkers of oxidative stress and histoanatomical observations. Thus, we determined the total soluble protein concentration and superoxide dismutase, catalase, and peroxidase activities. Cross-sections of the leaves were also made. The results proved the influence of anthropogenic pollution in all studied areas, but the greatest antioxidant defense system reactivity (compared to the control, the unpolluted area of Putna) was detected under the action of uranium on all woody species from Crucea-Botusana. Structural changes, such as the thickening of epidermal cuticle, the thinning of hypodermic cell walls, and the reduction of the phloem, were also observed and attributed to anthropogenic pollution.

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Soil salinity is a major environmental constraint to plant growth and productivity. It was investigated the effects of different sodium chloride concenfrations on Calendula officinalis seedling growth, pigments, proline and protein contents as well as antioxidant responses. Physiological and biochemical parameters were assessed in the seedling after 9 and 36 days of salt freafinent with 50 and 100 mill NaCl. Salinity affected most of the considered parameters. Results showed that the growth and pigment (chl a, chl b, and Car) contents was not negatively affected by the concenfration salt. After 9 days of salinity at marigold seedling it was observed a significant increase of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities at 50 mill NaCl as compared to con frol plant but these were generally decreased at 100 mill NaCl. Moreover, after a prolonged stress (36 days) a decrease of these enyzmes activity was observed with increase of salinity. The saline sfress increased proline content of marigold seedling with the increase of NaCl concenfration at 36-days old.

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We have synthesized a series of diazinium dihydroxyacetophenone derivatives using an efficient reaction pathway, in three steps: O-alkylation, alpha-bromination and N-alkylation reactions. The synthesized derivatives have been evaluated for their cytotoxic activities in vitro against HeLa cell line. Most of the obtained compounds showed a good cytotoxicity against cancer cell line. Compounds 3a-3e and 5a displayed more potent cytotoxic activities against human cancer cell line in comparison with etoposide, 5-fluoro-uracil and methotrexate. The brominated compounds (3a-3e) may possibly be used as lead compounds for developing new anticancer agents. The structures of compounds were assigned by elemental and spectral analysis, the X-ray data proving unambiguously the structure of compounds.

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Maize is one of model plants useful for genetic investigations and also very important for its agrotechnical utilizations. Here the genotoxic effects of low dose X-rays and accelerated electrons in maize caryopses was carried out with focus on the influence of water content at the moment of seed irradiation. X-ray photon beam as well as accelerated electrons were provided with 2.40 Gy min(-1) dose rate. Pre-soaked and dry maize caryopses were irradiated with 0.5-3.0-6.0 Gy. Cytogenetic investigations were carried out based on microscope observations of chromosomes stained by Feulgen method. The mitotic index was found diminished in hydrated samples indicating the negative influence of indirect effects of water radicals. As known the water radiolysis release free radicals that attack biomolecules in addition to the directly absorbed radiation impact. Slight positive influence of 0.5 Gy radiation dose on cell division was evidenced. Chromosomal aberrations were identified like: vagrand chromosomes, C-metaphases, picnotic chromosomes, chromatide bridges. General tendency of aberrant mitoses enhancing was recorded in watered samples - with up to the twice increase for 6.0 Gy radiation dose. The results evidenced the hydration role in monitoring cytogenetic effects of low dose radiations in plant systems -with possible biotechnological applications.

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The natural history of introduced species is often unclear due to a lack of historical records. Even when historical information is readily available, important factors of the invasions such as genetic bottlenecks, hybridization, historical relationships among populations and adaptive changes are left unknown. In this study, we developed a set of nuclear, simple sequence repeat markers and used these to characterize the genetic diversity and population structure among native (Eurasian) and non-native (North and South American) populations of Centaurea solstitialis L., (yellow starthistle). We used these data to test hypotheses about the invasion pathways of the species that were based on historical and geographical records, and we make inferences about historical relationships among populations and demographic processes following invasion. We confirm that the center of diversity and the native range of the species is likely the eastern Mediterranean region in the vicinity of Turkey. From this region, the species likely proceeded to colonize other parts of Europe and Asia via a slow, stepwise range expansion. Spanish populations were the primary source of seed to invade South America via human-mediated events, as was evident from historical records, but populations from the eastern Mediterranean region were also important. North American populations were largely derived from South America, but had secondary contributors. We suggest that the introduction history of non-native populations from disparate parts of the native range have allowed not just one, but multiple opportunities first in South America then again in North America for the creation of novel genotypes via intraspecific hybridization. We propose that multiple intraspecific hybridization events may have created especially potent conditions for the selection of a noxious invader, and may explain differences in genetic patterns among North and South America populations, inferred differences in demographic processes, as well as morphological differences previously reported from common garden experiments.

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Experimental investigation focused on electromagnetic pollution influence on fungi degrading wood is presented in this paper. 970 MHz low power microwaves were used to irradiate Phanerochaete chtysosporum cells combining two exposure times and two specific absorption rates experimentally determined by the scattering parameter method. Total protein content estimation and electrophoretic fractions assay have evidenced diminished biosynthesis with up to 25%, following energy absorption from microwave source, but no qualitative modifications in protein fraction array.

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Heavy metals are determinant factors in increasing environmental pollution, and chromium is considered to be of highest concern because of its genotoxicity in microorganisms, animals, and humans. Relatively few studies are focused on the injury induced in plant genetic material. Therefore, the main objective of this work was to evaluate the extent of the cytogenetic damage induced in root meristems of barley (Hordeum vulgare L.) after short-term seed exposure to 10, 100, 250, and 500 mu M K2Cr2O7 (Cr(VI) concentration is 1.04, 10.39, 25.99, and 51.99 mu g ml(-1)) and 10, 100, 250, and 500 mu M CrCl3 (Cr(III) concentration is 0.52, 5.19, 12.99, and 25.99 mu Mg ml(-1)). Chromium genotoxic potential was proved by significant increases in the rates of the ana-telophase chromosomal aberrations (1.3-2.3 times higher for K2Cr2O7 and 1.7-2.2 times higher for CrCl3, as compared to the control; p<0.05, p<0.01) and of metaphase disturbances (5.0-7.5 times more numerous in chromium-treated groups than in control; p<0.001). The pattern of the chromosomal aberrations is constituted by chromatid bridges, complex aberrations, lagging, and vagrant chromosomes, while the abnormal metaphases are c-like metaphases, sticky metaphases, and metaphases with chromosomes expulsed from equatorial plate. The mitotic indices and the growth of the barley plantlets in the early ontogeny were stimulated by chromium. The changes induced in the frequency of division stages mainly consisted in prophase and telophase accumulation and diminution of metaphase and anaphase proportion.

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Herein we report a feasible study concerning the design, syntheses and in vitro antimicrobial and antitumoral activities of some novel compounds with dihydroxyacetophenone (DA) moiety. An efficient and general method for the preparation of diazine with dihydroxyacetophenone (DDA) skeleton under conventional thermal heating (TH), microwave (MW) and ultrasounds (US) irradiation is presented. Antimicrobial and antitumoral tests prove that some dihydroxyacetophenone compounds (the brominated derivatives BrDA 3) have a significant biological activity. It is also to be pointed out that, basically all the dihydroxyacetophenone derivatives proved to have a powerful antibacterial activity against drug resistant Gram-negative strain Pseudomonas aeruginosa ATCC 27853. Of particular interest could be the excellent antibacterial activity of our dihydroxyacetophenone compounds against drug resistant Gram-negative strain Pseudomonas aeruginosa.

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Background: Extremely low frequency electromagnetic fields aren't considered as a real carcinogenic agent despite the fact that some studies have showed impairment of the DNA integrity in different cells lines. The aim of this study was evaluation of the late effects of a 100 Hz and 5.6 mT electromagnetic field, applied continuously or discontinuously, on the DNA integrity of Vero cells assessed by alkaline Comet assay and by cell cycle analysis. Normal Vero cells were exposed to extremely low frequency electromagnetic fields ( 100 Hz, 5.6 mT) for 45 minutes. The Comet assay and cell cycle analysis were performed 48 hours after the treatment. Results: Exposed samples presented an increase of the number of cells with high damaged DNA as compared with non-exposed cells. Quantitative evaluation of the comet assay showed a significantly (< 0.001) increase of the tail lengths, of the quantity of DNA in tail and of Olive tail moments, respectively. Cell cycle analysis showed an increase of the frequency of the cells in S phase, proving the occurrence of single strand breaks. The most probable mechanism of induction of the registered effects is the production of different types of reactive oxygen species. Conclusions: The analysis of the registered comet indices and of cell cycle showed that extremely low frequency electromagnetic field of 100 Hz and 5.6 mT had a genotoxic impact on Vero cells.

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Differences in morphological or ecological traits expressed by exotic species between their native and non-native ranges are often interpreted as evidence for adaptation to new conditions in the non-native ranges. In turn this adaptation is often hypothesized to contribute to the successful invasion of these species. There is good evidence for rapid evolution by many exotic invasives, but the extent to which these evolutionary changes actually drive invasiveness is unclear. One approach to resolving the relationship between adaptive responses and successful invasion is to compare traits between populations from the native and non-native ranges for both exotic invaders and congeners that are exotic but not invasive. We compared a suite of morphological traits that are commonly tested in the literature in the context of invasion for three very closely related species of Centaurea, all of which are sympatric in the same native and non-native ranges in Europe and North America. Of these, C. solstitialis is highly invasive whereas C. calcitrapa and C. sulphurea are not. For all three species, plants from non-native populations showed similar shifts in key traits that have been identified in other studies as important putative adaptive responses to post-introduction invasion. For example, for all three species plants from populations in non-native ranges were (i) larger and (ii) produced seeds that germinated at higher rates. In fact, the non-invasive C. calcitrapa showed the strongest trait shift between ranges. Centaurea solstitialis was the only species for which plants from the non-native range increased allocation to defensive spines, and allocated proportionally less resources to reproduction, patterns contrary to what would be predicted by theory and other empirical studies to enhance invasion. Our results suggest caution when interpreting the commonly observed increase in size and reproductive capacity as factors that cause exotics to become invaders.

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Three polyphenolic biopreparates from the seeds of three Vitis vinifera variations were obtained, hydroxybenzoic and hydroxycinnamic acids, trans-resveratrol, non-hydrolisable tannins and flavones being identified. The testing was done in vitro, on HeLa neoplasic human cells, and underlines the disturbance of protein synthesis and cellular proliferation, as well as alteration of cellular viability. The impact of the inhibitor of the protein synthesis, mito-inhibitor and of diminishing the cellular viability, with consequences on the development degree of cellular cultures, demonstrates the fact that polyphenolic bio-preparates act like cytostatic and cytotoxic agents.

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In the apple plantations, codling moth (Cydia pomonella L.) is one of the most important pests in Romania, which causes large and constant damages, far above the economic threshold of damage, frequently being about 25-50%, and sometimes even larger. The main aim of this study was the control of codling moth in accordance with the special evolution of its biology and ecology related to local biotope conditions. The purpose of this paper was to underline the dynamics of species evolution of Cydia pomonella L., due to the application of some practical and technical measures meant to improve integrated control system. Latest researches regarding control of codling moth, have revealed some changes in pest biology and also the efficacy of new generation of active substances. During the period 2009-2011, at S. C. D. P. (Station for Research and Development of Fruit-Growing), Iasi, there have been developed experimentations regarding the control of codling moth within the program of integrated control of pathogens and pests from apple orchards by using plant protection active substances which were recently introduced in the phytosanitary treatments. The biological cycle of the species Cydia pomonella L. in the weather conditions of Iasi County, presented two generations/year, a complete generation and a second incomplete one. During the three years of study, the codling moth has recorded three curves of the flight during the vegetative season (two for the first generation and one for the second generation) and with respect to control, this was realized with plant protection substances of latest generation which recorded a very good efficiency. By using these active substances it aimed to improve fruit quality and environmental protection.

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Haloalkane dehalogenases catalyze the hydrolytic cleavage of carbon-halogen bonds in a broad range of environmental pollutants such as aliphatic mono-, di-, and polyhalogenated alkanes. From the biotechnology point of view haloalkane dehalogenases attract attention because of many potential uses for the bioremendation of soil, water and air. In the present study, different Rhizobium strains (Sinorhizobium meliloti 1021, Rhizobium leguminosarum by. trifolii, Mesorhizobium loti MAFF, Bradyrhizobium japonicum usda 110) were screened for their ability to produce stable and active 1,2-dibromoethane-degrading dehalogenase. The results showed that B.japonicum produces the most potent dehalogenase. This enzyme was cloned, expressed in Escherichia coli BL21(DE3), purified and was entrapped in tetraethylorthosilicate derived sol-gel. The tetraethyl orthosilicate sal-gel entrapped haloalkane dehalogenases exhibited higher storage and operational stability at 4 degrees C and 25 degrees C, compared to the free enzyme. Kinetic analysis of the entrapped enzyme using 1,2-dibromoethane showed that substrate turnover was limited by partitioning effects or diffusion through the sal-gel matrix. The biocatalyst was used in a packed bed bioreactor for the biodegradation of 1,2-DBE. Under selected conditions the sal-gel entrapped dehalogenase was able to hydrolyze 91.8% of the loaded 1,2-DBE, within 16.7 h. The results of the present study suggest that the use of HLD biocatalysis may provide a 'green chemistry' tool for sustainable remediation of 1,2-DBE. (C) 2013 Elsevier B.V. All rights reserved.

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The changes in superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities, and in soluble protein level were analysed in Triticum aestivum cv Beti seedlings, after seed exposure to copper, provided as copper acetate and copper citrate, at four concentrations (10, 25, 50, and 100 mu M) containing 0.64, 1.59, 3.18, 6.35 mu g ml(-1) Cu2+, and 1.91, 4.77, 9.53, 19.06 mu g ml(-1) Cu2+, respectively. SOD and POD showed similar patterns, with relatively small fluctuations compared to control. Considerable rise of activity was registered only in 25 mu M copper citrate (increase rate of +152.90% for SOD, and +70.51% for POD). CAT activity was lower than control in all variants, the smallest level being in 25 mu M copper citrate-treated variant (decrease rate of -65.41%). Generally, copper had negative repercussions on soluble protein level. Higher SOD and POD activities and CAT decline in all copper-treated variants indicate that SOD and POD play a more important role than CAT in preventing copper-induced oxidative stress, in the studied wheat cultivar.

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Large morpho-anatomical and biochemical variability was evidenced in all Lonicera caerulea L. provenances. The knowledge of chromosome constitution is of basic importance in this context of elevated phenotype heterogeneity and represents a necessary step in breeding programs. The Romanian selections are tetraploid (2n = 4x = 36). Chromosomes are small-sized (<3.5 mu m), and lengths of haploid complements are close (28.97-29.05 mu m). Only the Lc-SL15 complement is larger (33.89 pm), possibly due to lower chromatin condensation. Karyotypes contain metacentric (85%-100%) and submetacentric chromosomes. According to asymmetry indexes (AsI% = 55.01-59.89, TF% = 40.13-44.98, A(1) = 0.17-0.29, A(2) = 0.23-0.30) and to Stebbins' classification, the karyotypes fall into 1B and 2B categories, considered relatively symmetric and primitive in this system. In the idiogram constructed on average values of cytogenetic parameters of all analyzed selections, the mean chromosome length is <(x)over bar> +/- SE = 1.66 +/- 0.07 mu m, ranging from 2.73 +/- 0.20 to 1.10 +/- 0.02 mu m. The mean relative length is between 9.05 +/- 0.30 and 3.70 +/- 0.12. One submetacentric chromosome pair is present. As a result, the general karyotype formula is K(2n) = 36 = 34 m + 2 sm.

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Copper is one the most abundant heavy metals in agricultural soils and its excess in soil comes from the largely use of this heavy metal in industry and agriculture (as fungicide). Mitotic index, rate and categories of ana-telophase chromosome aberrations, as well as the frequency and types of metaphase disturbances were scored in root tip meristems of Triticum aestivum L. cv. Beti after seed exposure to copper, provided as copper acetate and copper citrate, at four concentrations (10, 25, 50, and 100 mu M) containing 0.64, 1.59, 3.18, 6.35 mu g Cu2+, and 1.91, 4.77, 9.53, 19.06 mu g ml(-1) Cu2+, respectively. Except the mitostimulant effect of 25 mu M concentration, all the other concentrations of copper acetate and copper citrate showed mitodepressive action. The copper genotoxicity is expressed in the increased level (1.5 - 5-fold higher than in control) of the rate of chromosome aberrations in mitotic ana-telophases of copper-treated variants. Chromosome bridges, laggards and complex aberrations are the most numerous, although multipolarity, fragments and micronuclei are present, but with lower frequency and not in all copper-treated variants. Concerning the rate of metaphase disturbances, copper acetate augmented 2 3 times the rate of abnormalities in all variants, whereas only variant treated with 25 mu M copper citrate exceeded the control in a substantial manner. Metaphases with chromosomes expulsed from equatorial plate are numerically preponderant, followed by C-metaphases. These observations constitute a signal about the risks of the widespread and increasing presence of some heavy metals into environment. The results reported here could be considered in a future evaluation of copper effects on other organisms, even on human health, due to large use of copper compounds, inclusively as fungicides.

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The pollution increase, as a result of the release into environment of genotoxic chemicals, including heavy metals, largely affects the ecosystems and the health of living organisms. Although zinc is not considered highly phytotoxic, its excess becomes noxious. In literature, the reports on zinc genotoxicity are equivocal. Therefore, the objective of this work was to evaluate the amplitude of cyrogenetic damage induced in Hordeum vulgare L. cv. 'Madalin' after seed treatment with different concentrations (10, 100, 250, 500 mu M) of Zn+2, provided as zinc sulphate and zinc acetate. The mitostimulatory effect was present at all concentrations of both zinc compounds. The rate of ana-telophase aberrations exceeded by 2-3 times the control, and the frequency of metaphase disturbances was 5.0-10.0 times higher than the control. The results indicate the clastogenic and aneugenic potential of zinc in barley and constitute a signal about the risks of its increasing presence into the environment, with repercussions on living systems, even on human health, due to the extensive use of zinc compounds including as pesticides.

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This study aims to review the species of Oribatula s. str. Berlese recorded in Romanian fauna, according to the most recent monographs and catalogues, but taking the original description and/or subsequent mentions into account, as well. Rich faunistic material was examined for this purpose, collected from various ecosystem types and from most regions of the country. Six species are analyzed from a morphological and biometrical point of view. In this context, reasons to consider Oribatula amblyptera Berlese, 1916 and O. sitnikovae Iordansky, 1991 as valid species are pointed out. A brief description and illustration of each species are given, along with data on their local distribution and ecology. An identification key is proposed.

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The description of the karyotype features and idiogram in Grindelia squarrosa (Pursh) Dunal (Asteraceae), an invasive plant in Romania, are reported here for the first time. The diploid chromosome number is 2n = 2x = 12, in agreement with the data published for the other species of the genus. The karyomorphological data show that the complements of the studied genotypes have small chromosomes (mean chromosome length is (X) over bar +/- SE = 2.56 +/- 0.10 mu m, and mean length of haploid complements is (X) over bar +/- SE = 15.33 +/- 0.69 mu m, with a range of variability comprised between 12.87-17.51 mu m). The karyotypes are made up of six pairs of metacentric and submetacentric chromosomes, with an identical formula of the diploid complement: KF = 2n = 12 = 8m + 2sm + 2sm-SAT. Satellites are located on the short arms of the chromosomes of pair III. The karyotypes show a relatively high level of intra-specific uniformity as well as similar symmetry patterns (R = 1.29-1.53; TF% = 38.78-41.57%; AsI% = 54.54-57.61%; A1 = 0.24-0.32; A2 = 0.08-0.16), belonging to 1A and 2A classes of symmetry. The small size of the chromosomes, the presence of only two chromosome morphometric types, and the preponderance of metacentrics confer a relatively high degree of symmetry to the karyotypes studied.

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Simulated magnetic contamination of plants during early ontogenetic stages was studied in arranged laboratory conditions using magnetic nanoparticles based on compound metal oxides. Aqueous suspensions of magnetic colloidal nanoparticles of Fe3O4, CoFe2O4, and ZnFe2O4, were administrated to freshly germinated sunflower seeds in the same array of dilutions (v/v): 20-40-60-80-100 mu l/l, the results of their genetic impact in the root tip cells being qualitatively and quantitatively analyzed. Cytogenetic tests carried out by optical microscopy means provided data on the types of abnormal cell divisions as well as on the mitosis rate and total percentage of chromosomal aberrations. Considerable diminished mitosis rate was evidenced in all situations, while remarkably enhanced number of chromosomal aberrations was also evidenced for all three cases with higher nanotoxicity revealed in the case of ZnFe2O4 and CoFe2O4. Chromosome fragments, interchromatidian bridges and micronuclei appeared in most analyzed samples with no noticeable difference for one type of magnetic nanoparticles or other.

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The study of polyphenolic extracts from skins, seeds, grape pomace and lees is justified by the huge amount of information from specialized literature, drawing attention to the many pharmacological effects of these biomolecules. The concentrated vegetal extracts obtained from seeds, skins, grape pomace and lees (yeast deposit after fermentation) were characterized according to their content of total polyphenols, anthocyanins, dry matter, ash and pH. The biological material used in the in vitro resting experiments in order to study cytostatic and cytotoxic effects were stabilized cultures of HeLa cancer cells, uncontaminated with mycoplasma and derived from a human uterine cervix carcinoma. Another action of the cytostatic substances could be exerted upon the cell proliferation process. The cell division process of the HeLa cell cultures treated with seeds by-product showed remarkable quantitative changes. The study has also evidenced a great number of dead cells in the composition of the treated HeLa cell cultures, their existence pointing our that the bioactive agent induced a major decrease of the cells viability. The obtained results in the context of the complex evaluation of the in vitro antitumoral property of the extracts, obtained from seeds, skins, grape pomace and wine lees from the 'Areas' grape variety, on He La cells cultures, have demonstrated the significant cytostatic and cytotoxic potential of the seeds polyphenolic biopreparate. The results obtained show that polyphenolic extracts from Vitis vinifera seeds act as cytostatic and cytotoxic agents.

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The aim of this paper consists in 'non-stressing' process use that, with respect to traditional plants operates under milder temperature conditions. Particular attention was devoted to investigate the possibility to purify and concentrate Viscum album extracts with a microfiltration/ultrafiltration unit, allowing the preservation of thermolabile compounds from the extracts and their antineoplastic properties examination. In vitro investigation of the HeLa cancerous cells interaction with biopreparations (extracted, separated and purified from Viscum album through successive ultrafiltrations on membranes) highlighted and quantified their cytostatic and cytotoxic impact, expressed by the inhibition of protein synthesis and cellular proliferation, by cell viability decrease and by the inhibition of cell cultures development.

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The sclerotia of the pyrenomicete Claviceps purpurea (Fr.) Tul. (Hypocreaceae) contains numerous active compounds of which the most known are ergoline alkaloids. In this paper, the mycelian extracts and the corresponding concentrated supernatants, separated from cultures of nine Claviceps purpurea strains, have been biochemically investigated by determination of alkaloids, glucans and proteins. The cytostatic potential of extracts and concentrated supernatants has been also tested on HeLa tumor cells cultures in the presence of bioactive samples. Generally, the cytostatic potential of these bioactive agents was inversely correlated with the intra - or extracellular ergoline alkaloids content, which seemed not to be the exclusive biochemical substratum for this in vitro pharmacodynamic effect.

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The somatic chromosomes and karyotype traits of three Romanian genotypes (coded as CB-1, S-16, L-4) of Hippophae rhamnoides L. have been studied. Diploid chromosome number is 2n = 24. The karyomorphological data show that the complements have small chromosomes (1.33+/-0.014-2.80+/-0.01 mu m, for CB-1 genotype; 1.13+/-0.029-2.90+/-0.029 mu m, for S-16; 1.15+/-0.099-2.88+/-0.024 mu m, for L-4 genotype) and the length of haploid complements is 23.97 mu m, for CB-1, 22.20 mu m, for S-16, 21.73 mu m, for L-4. In CB-1 and S-16 male genotypes, the putative Y-chromosome has larger sizes than X-chromosome and it is of metacentric type (CI = 45.77, r = 1.18, for CB-1; CI = 46.22, r = 1.15, for S-16). The L-4 female karyotype is constituted by twelve pairs of morphologically similar chromosomes. The karyotype formulae are 2n = 24 = 13m + 11sm (putative male: 22 + XY), for CB-1 genotype, 2n = 24 = 18m + 6sm (putative male: 22 + XY), for S-16 genotype, respectively 2n = 24 = 22m + 2sm (L-4 genotype, considered as female: 22 + XX). The small size of chromosomes (<3 mu m), the presence of only two morphometric chromosome types and preponderance of metacentrics confer a relatively high degree of symmetry to the studied karyotypes. Our results show a low intraspecific karyotype variation.

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There is great evidence regarding the beneficial influence of rose hips on human and animal health. In this respect, fruits of native populations of Rosa spp. collected from the northeastern part of Romania (400-1060 in altitude) were assayed for nutritional and some morphological properties, in order to select the best genotypes. The results show a great quantitative variability of analyzed genotypes, regarding dimensions and biochemical indicators as follow: ascorbic acid: 866.91 mg/100 g fw (R. rubiginosa), 615.98 mg/100 g fw (R. carrier); carotenes: 34.95 mg/100 g fw (R. subcanina), 24.64 mg/100 g fw (R. nitidula); protein content: 0.47 g/100 g fw (R. micrantba), 0.25 g/100 g fw (R, caesia); total sugars: 17.63 g/100 g fw (R. andegavensis), 11.55 g/100g fw (R. caryophyllacea). The fruits length, width and dry weight were: 25 mm (R. canina), 12 mm (R. micrantha), 16.5 mm (R. corymbifera), 8 mm (R. micrantha) and 25% dry weight (R. rubiginosa), 3896 dry weight (R. nitidula), respectively. The present study demonstrates the existence of a rich rose germplasm source in north-east area, with good biotechnological fruit quality, which recommends them as functional foods according to their chemical contents (R. rubiginosa, R. vosagiaca, R. subcanina, R. canister and R. caryophyllacea) and also as sources for natural colorants in food technology.

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The wide utilization of plant growth regulators requires the knowledge of possible unwanted effects induced by uncontrolled administration of these chemicals, inclusively at genetic level. The cytogenetic effects induced by two plant growth regulators, an auxin (2,4-D) and a cytokinin (kinetin), in root meristems of plants belonging to two species of economic importance - Raphanus sativus L. and Phaseolus vulgaris L. - were studied. Mitotic index, rate and categories of ana-telophase chromosome aberrations, as well as the frequency and types of metaphase disturbances were comparatively analyzed after seed exposure to two concentrations (1 mg/L and 10 mg/L) for each plant hormone. 2,4-D had a slight cytotoxic effect in radish, but did not show genotoxic potential, whereas kinetin stimulated cell division. In common bean, the small concentrations of 2,4-D and kinetin enhanced mitosis, but the maximum tested concentrations had inhibitive effect on cell division. Generally, the rate of ana-telophase chromosome aberrations induced by 2,4-D and kinetin, at the tested concentrations and at the respective exposure, was not significant. Auxin caused increase of total abnormal metaphases (metaphases with expulsed chromosomes and colchicine-like metaphases) in both species, indifferently of concentration, whereas all the variants exposed to kinetin action showed values lower than control, in both species. Therefore, 2,4-D have aneugenic potential by acting as spindle poison and by disturbing the correct separation of the chromosomes to cell poles. The most complex pattern of ana-telophase and metaphase modifications was present in variant of common bean exposed to the action of 10 mg/L 2,4-D.

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