Physiological and oxidative stress response of carrot (Daucus carota L.) to jumping plant-louse Bactericera trigonica Hodkinson (Hemiptera: Psylloidea) infestation.

BACKGROUND: Carrot is an important vegetable crop grown worldwide. The major economic problem in carrot cultivation is yellow disease caused by Bactericera trigonica, which induces biotic stress and has the greatest impact on crop productivity. Comprehensive studies on the mechanism of carrot defense response to biotic stress caused by B. trigonica infestation have yet to be conducted. METHODS: The changes in photosynthetic pigments, proline, TPC, H2O2 and MDA content, DPPH radical scavenging ability, and antioxidant enzyme activity of SOD, CAT, and POX in carrot leaves in response to insect sex (female and male), rapid response (during the first six hours), and long-term response to B. trigonica infestation were evaluated. RESULTS: The results of our study strongly suggest that B. trigonica infestation causes significant changes in primary and secondary metabolism and oxidative status of carrot leaves. Photosynthetic pigment content, TPC, and DPPH and CAT activities were significantly reduced in carrot leaves in response to insect infestation. On the other hand, proline, H2O2 content, and the activity of the antioxidant enzymes superoxide dismutase and peroxidase were increased in carrot leaves after B. trigonica infestation. The results indicate that B. trigonica attenuates and delays the oxidative stress responses of carrot, allowing long-term feeding without visible changes in the plant. Carrot responded to long-term B. trigonica infestation with an increase in SOD and POX activity, suggesting that these enzymes may play a key role in plant defense mechanisms. CONCLUSIONS: This is the first comprehensive study strongly suggesting that B. trigonica infestation causes significant changes in primary and secondary metabolism and an attenuated ROS defense response in carrot leaves that enables long-term insect feeding. The information provides new insights into the mechanisms of carrot protection against B. trigonica infestation.

Removal of toxic metals from sewage sludge by EDTA and hydrodynamic cavitation and use of the sludge as fertilizer.

Sewage sludge (SS) is rich in plant nutrients, including P, N, and organic C, but often contains toxic metals (TMs), which hinders its potential use in agriculture. The efficiency of removal of TMs by washing with ethylenediamine tetraacetate (EDTA), in combination with hydrodynamic cavitation (HC) and the usability of washed sewage sludge as fertilizer were investigated. The environmental risk was assessed. During 8 wash batches an average 35, 68, 47 and 45 % of Pb, Zn, Cd and Cu, respectively, as well as 22 and 5 % Mn and Fe were removed from the SS. The process solutions and EDTA were recycled at a pH gradient of 12.5-2, which was achieved by adding quicklime (CaO) and then acidification by H2SO4, so that no wastewater was produced, only solid waste (ReSoil® method). The quality of the recycled process solutions (they remained unsaturated with salts) and the efficiency of the washing process were maintained across all batches. On average, 46 % of the EDTA was lost during the process and was replenished. The initial leachability of EDTA-mobilized Pb, Zn, Cu, Cr and Fe remaining in the washed SS increased 6-, 17-, 3-, 11- and 11-fold, respectively, but not to hazardous levels except for Zn. After washing, P and K remained in the SS, plant-available P increased 3.3-fold, while total N and C were reduced by 20.28 and 2.44 %, respectively. Washed SS was used as fertilizer in the pot experiment. The yield of Brassica juncea did not improve, the uptake of TMs by the plants and the leaching of TMs from the soil were minimal. Our study highlighted the drawbacks and potential feasibility of the new SS washing method.

Foliar application of methyl jasmonate affects impatiens walleriana growth and leaf physiology under drought stress.

In this study, the effects of foliar applied methyl jasmonate (MeJA) on drought-stressed Impatiens walleriana growth and leaf physiology parameters: stomatal conductance, chlorophyll, flavonoid, anthocyanin, and nitrogen balance index (NBI), were evaluated. These parameters could serve as indicators of drought tolerance of I. walleriana, a popular horticultural plant worldwide that is very sensitive to drought. The experiment included four treatments: control, drought-stressed plants sprayed with distilled water, drought-stressed plants sprayed with 5 µM MeJA, and drought-stressed plants sprayed with 50 µM MeJA. Foliar spraying with MeJA was performed twice: seven days before and on the day of drought induction. The stressed plant groups were non-irrigated to achieve soil water contents (SWC) of 15 and 5%, while control plants were well-watered throughout the experiment (35-37% SWC). The results of this study showed that drought significantly reduced I. walleriana fresh and dry shoot weight, as well as total leaf area, but did not impact on dry matter content. The foliar application of MeJA improved growth parameters of I. walleriana, depending on the elicitor concentration and drought intensity. Stomatal conductance was slightly reduced at 5% SWC, and foliar applied MeJA at both concentrations. The flavonoid index was slightly reduced at 15 and 5% SWC when 50 µM MeJA was foliar applied, while there were no observed changes in the anthocyanin index in any treatments. The foliar application of 50 µM MeJA increased the chlorophyll index and NBI of I. walleriana at 5% SWC, indicating a contribution of the elicitor to plant drought tolerance at the physiological level.

Alterations in Physiological, Biochemical, and Molecular Responses of Impatiens walleriana to Drought by Methyl Jasmonate Foliar Application.

Drought stress affects plant growth and development through several mechanisms, including the induction of oxidative stress. To cope with drought, plants have drought tolerance mechanisms at the physiological, biochemical, and molecular levels. In this study, the effects of foliar application of distilled water and methyl jasmonate (MeJA) (5 and 50 µM) on the physiological, biochemical, and molecular responses of Impatiens walleriana during two drought regimes (15 and 5% soil water content, SWC) were investigated. The results showed that plant response depended on the concentration of the elicitor and the stress intensity. The highest chlorophyll and carotenoid contents were observed at 5% SWC in plants pre-treated with 50 µM MeJA, while the MeJA did not have a significant effect on the chlorophyll a/b ratio in drought-stressed plants. Drought-induced formation of hydrogen peroxide and malondialdehyde in plants sprayed with distilled water was significantly reduced in plant leaves pretreated with MeJA. The lower total polyphenol content and antioxidant activity of secondary metabolites in MeJA-pretreated plants were observed. The foliar application of MeJA affected the proline content and antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase) in plants that suffered from drought. The expression of abscisic acid (ABA) metabolic genes (IwNCED4, IwAAO2, and IwABA8ox3) was the most affected in plants sprayed with 50 µM MeJA, while of the four analyzed aquaporin genes (IwPIP1;4, IwPIP2;2, IwPIP2;7, and IwTIP4;1), the expression of IwPIP1;4 and IwPIP2;7 was strongly induced in drought-stressed plants pre-treated with 50 µM MeJA. The study's findings demonstrated the significance of MeJA in regulating the gene expression of the ABA metabolic pathway and aquaporins, as well as the considerable alterations in oxidative stress responses of drought-stressed I. walleriana foliar sprayed with MeJA. The results improved our understanding of this horticulture plant's stress physiology and the field of plant hormones' interaction network in general.

Environmental Acceptability of Geotechnical Composites from Recycled Materials: Comparative Study of Laboratory and Field Investigations.

The environmental properties of three geotechnical composites made by recycling wastes were investigated on a laboratory scale and in the field with the use of lysimeters designated for the revitalization of degraded mining sites. Composites were prepared by combining the mine waste with paper-mill sludge and foundry sand (Composite 1), with digestate from municipal waste and paper ash (Composite 2), and with coal ash, foundry slag and waste incineration bottom ash (Composite 3). The results of laboratory leaching tests proved that Composites 1 and 3 are environmentally acceptable, according to the legislative limits, as the potentially hazardous substances were immobilized, while in Composite 2, the legislative limits were exceeded. In the field lysimeters, the lowest rate of leaching was determined for optimally compacted Composites 1 and 3, while for Composite 2 the leaching of Cu was high. This study proved that optimally installed Composites 1 and 3 are environmentally acceptable for use in construction as an alternative to virgin materials, for the revitalization of degraded mining sites or, along with Composite 2, for closure operations with landfills. In this way, locally available waste streams are valorised and channelized into a beneficial and sustainable recycling practice.

Role of Sodium Nitroprusside on Potential Mitigation of Salt Stress in Centaury (Centaurium erythraea Rafn) Shoots Grown In Vitro.

Soil salinity is one of the most common abiotic stressors that affects plant growth and development. The aim of this work was to investigate the influence of sodium nitroprusside (SNP), a donor of nitric oxide (NO), on the physiological response of common centaury (Centaurium erythraea) shoots grown under stress conditions caused by sodium chloride (NaCl) in vitro. Centaury shoots were first grown on nutrient medium containing different SNP concentrations (50, 100 and 250 µM) during the pretreatment phase. After three weeks, the shoots were transferred to nutrient media supplemented with NaCl (150 mM) and/or SNP (50, 100 or 250 µM) for one week. The results showed that salinity decreased photosynthetic pigments, total phenolic content and DPPH (1,1-diphenyl-2-picrylhydrazyl radical) concentration. The activities of antioxidant enzymes, namely superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX), were also reduced under salt stress. However, MDA concentration was decreased, while H2O2 and proline content did not drastically change under the stress conditions caused by NaCl. Exogenous application of SNP altered the biochemical parameters of centaury shoots grown under salt stress. In this case, increased photosynthetic pigment content, total phenolics and proline content were noted, with reduced MDA, but not H2O2, concentration was observed. In addition, the exogenous application of SNP increased the degree of DPPH reduction as well as SOD, CAT and POX activities.

Physiological Response, Oxidative Stress Assessment and Aquaporin Genes Expression of Cherry Tomato (Solanum lycopersicum L.) Exposed to Hyper-Harmonized Fullerene Water Complex.

The rapid production and numerous applications of nanomaterials warrant the necessity and importance of examining nanoparticles in terms to their environmental and biological effects and implications. In this study, the effects of a water-soluble hyper-harmonized hydroxyl-modified fullerene (3HFWC) on cherry tomato seed germination, seedlings growth, physiological response and fruiting was evaluated. Changes in the photosynthetic pigments content, oxidative stress assessment, and aquaporin genes expression in cherry tomato plants were studied after during short- and long-term continuous exposure to 3HFWC nanosubstance (200 mg/L). Increased levels of photosynthetic pigments in leaves, lycopene in fruits, decreased levels of hydrogen peroxide content, activation of cellular antioxidant enzymes such as superoxide dismutase, catalase and peroxidase and increased aquaporin gene expression (PIP1;3, PIP1;5 and PIP2;4) were observed in 3HFWC nanosubstance-exposed plants in comparison to control, untreated cherry tomato plants. The 3HFWC nanosubstance showed positive effects on cherry tomato seed germination, plantlet growth and lycopene content in fruits and may be considered as a promising nanofertilizer.

Molecular Characterization and Expression of Four Aquaporin Genes in Impatiens walleriana During Drought Stress and Recovery.

Aquaporins comprise a large group of transmembrane proteins responsible for water transport, which is crucial for plant survival under stress conditions. Despite the vital role of aquaporins, nothing is known about this protein family in Impatiens walleriana, a commercially important horticultural plant, which is sensitive to drought stress. In the present study, attention is given to the molecular characterization of aquaporins in I. walleriana and their expression during drought stress and recovery. We identified four I. walleriana aquaporins: IwPIP1;4, IwPIP2;2, IwPIP2;7 and IwTIP4;1. All of them had conserved NPA motifs (Asparagine-Proline-Alanine), transmembrane helices (TMh), pore characteristics, stereochemical properties and tetrameric structure of holoprotein. Drought stress and recovery treatment affected the aquaporins expression in I. walleriana leaves, which was up- or downregulated depending on stress intensity. Expression of IwPIP2;7 was the most affected of all analyzed I. walleriana aquaporins. At 15% and 5% soil moisture and recovery from 15% and 5% soil moisture, IwPIP2;7 expression significantly decreased and increased, respectively. Aquaporins IwPIP1;4 and IwTIP4;1 had lower expression in comparison to IwPIP2;7, with moderate expression changes in response to drought and recovery, while IwPIP2;2 expression was of significance only in recovered plants. Insight into the molecular structure of I. walleriana aquaporins expanded knowledge about plant aquaporins, while its expression during drought and recovery contributed to I. walleriana drought tolerance mechanisms and re-acclimation.

Morpho-Physiological and Molecular Evaluation of Drought and Recovery in Impatiens walleriana Grown Ex Vitro.

This study was carried out to examine the drought effect on development, physiological, biochemical and molecular parameters in Impatiens walleriana grown ex vitro. Experiment design included three treatments: Control plants-grown under optimal watering (35%-37% of soil moisture content), drought-stressed plants-non-irrigated to reach 15% and 5% of soil moisture content and recovery plants-rehydrated for four days to reach optimal soil moisture content. Drought reduced fresh weight, total leaf area, as well as dry weight of I. walleriana shoots. Drought up-regulated expression of abscisic acid (ABA) biosynthesis genes 9-cis-epoxycarotenoid dioxygenase 4 (NCED4) and abscisic aldehyde oxidase 2 (AAO2) and catabolic gene ABA 8'-hydroxylase 3 (ABA8ox3) which was followed by increased ABA content in the leaves. Decrement in water potential of shoots during the drought was not accompanied with increased amino acid proline content. We detected an increase in chlorophyll, carotenoid, total polyphenols and flavonols content under drought conditions, as well as malondialdehyde, hydrogen peroxide and DPPH (1,1'-diphenyl-2-picrylhydrazyl) activity. Increased antioxidant enzyme activities (superoxide dismutase, peroxidase and catalase) throughout drought were also determined. Recovery treatment was significant for neutralizing drought effect on growth parameters, shoot water potential, proline content and genes expression.

Three-Dimensional Reconstruction of Erythrocytes Using the Novel Method For Corrective Realignment of the Transmission Electron Microscopy Cross-Section Images.

The detailed kinetics study of erythrocyte deformability is useful for the early diagnosis of blood diseases and for monitoring the blood rheology. Present solutions for a three-dimensional (3D) reconstruction of erythrocytes has a limited potential. This study aimed to use erythrocyte transmission electron images (ETIs) to evaluate the morphological relationship between adjacent ETIs and generate erythrocytes 3D model. First, ultrathin serial sections of skeletal muscle tissue were obtained using an ultramicrotome. Further, the set of ETIs in a capillary were captured by transmission electron microscopy. The images were aligned by translations and rotations using custom software to optimize the morphological relationship between adjacent ETIs. These coordinate transformations exploit the unique principal axis of inertia of each image to define the body coordinate system and hence provide the means to accurately reconnect the adjacent ETIs. The sum of the distances between the corresponding points on the boundary of adjacent ETIs was minimized and, further, was optimized by using physiological relationship between the adjacent ETIs. The analysis allowed to define precise virtual relationship between the adjacent erythrocytes. Finally, extracted erythrocytes' cross-section images allowed to generate 3D model of the erythrocytes.

Collagen-induced arthritis in Dark Agouti rats as a model for study of immunological sexual dimorphisms in the human disease.

Collagen-induced arthritis (CIA) is a frequently used animal model of rheumatoid arthritis, human autoimmune disease that exhibits clear sex bias in incidence and clinical course. Female Dark Agouti rats immunized for CIA showed also greater incidence and higher arthritic score than their male counterparts. The study investigated sex differences in mechanisms controlling the primary immune responses in draining lymph nodes (dLNs), as a factor contributing to this dimorphism. The higher frequencies of CD4 + CD25 + Foxp3- cells, presumably activated effector T (Teff) cells, and IL-17+, IFN-γ + and IL-17 + IFN-γ + T cells were found in female compared with male rat dLNs. However, the frequency of CD4 + CD25 + Foxp3+ T regulatory cells (Treg) did not differ between sexes. Thus, CD4+ Teff cells/Treg ratio, and IL-17+ T cells/Treg and IFN-γ + T cells/Treg ratios were higher in female than in male rats, and among them was found lower frequency of PD-1+ cells. This suggested less efficient control of (auto)immune Th1/Th17 cell responses in female rat dLNs. On the contrary, the frequency of IL-4+ T cells was lower in female than in male rat dLNs. Consistently, the ratio of serum levels of collagen-specific IgG2a (IFN-γ-dependent, with an important pathogenic role in CIA) and IgG1 (IL-4-dependent) was shifted towards IgG2a in female compared with male rats. As a whole, the study suggests that sexual dimorphism in the control of T cell activation/polarization could contribute to sex bias in the susceptibility to CIA. Moreover, the study advises the use of animals of both sexes in the preclinical testing of new drugs for rheumatoid arthritis.

Marginal Gaps between 2 Calcium Silicate and Glass Ionomer Cements and Apical Root Dentin.

INTRODUCTION: The outcome of periapical surgery has been directly improved with the introduction of novel material formulations. The aim of the study was to compare the retrograde obturation quality of the following materials: calcium silicate (Biodentine; Septodont, Saint-Maur-des-Fosses, France), mineral trioxide aggregate (MTA+; Cerkamed Company, Stalowa Wola, Poland), and glass ionomer cement (Fuji IX; GC Corporation, Tokyo, Japan). METHODS: Materials' wettability was calculated concerning the contact angles of the cements measured using a glycerol drop. Cements' porosity was determined using mercury intrusion porosimetry and micro-computed tomographic (µCT) imaging. Extracted upper human incisors were retrofilled, and µCT analysis was applied to calculate the volume of the gap between the retrograde filling material and root canal dentin. Experiments were performed before and after soaking the materials in simulated body fluid (SBF). RESULTS: No statistically significant differences were found among the contact angles of the studied materials after being soaked in SBF. The material with the lowest nanoporosity (Fuji IX: 2.99% and 4.17% before and after SBF, respectively) showed the highest values of microporosity (4.2% and 3.1% before and after SBF, respectively). Biodentine had the lowest value of microporosity (1.2% and 0.8% before and after SBF, respectively) and the lowest value of microgap to the root canal wall ([10 ± 30] × 10-3 mm3). CONCLUSIONS: Biodentine and MTA possess certain advantages over Fuji IX for hermetic obturation of retrograde root canals. Biodentine shows a tendency toward the lowest marginal gap at the cement-to-dentin interface.

Keeping the pieces together: comparison of mass grave excavation methodology.

Mass graves are a complex and confusing mix of bodies, body parts, soils, artifacts, and other feature evidence. Forensic investigations of these complex crime scenes should attempt to maximize the collection of evidence, which includes the mortal remains, in their best possible condition as they were deposited within the graves. Two standard methods of mass grave excavation were examined with the aim of identifying the better approach. Two experienced teams using different methodologies excavated two separate but very comparable mass graves located in the same area. Single disarticulated skeletal elements not associated with a body at the time of removal from the grave were categorized and their counts analyzed to evaluate the efficiency of the differing excavation methodologies. The methodologies used were the 'pedestal' method, which focuses on the body mass, and the 'stratigraphic' method, in which the grave feature and contents are conjointly excavated. The first grave (Grave A), excavated using the 'pedestal' method, was observed to have a disproportionately larger amount of unassociated bones than did the second (Grave B), which used the 'stratigraphic' method. Chi-square (chi2) goodness-of-fit and contingency tests were performed on the total numbers of recorded elements and different categorical groups of bones, based on size and shape, in each grave. Results demonstrate that significantly greater numbers of unassociated elements resulted from the excavation of Grave A using the pedestal method, both in total number of disarticulated bones as well as within 'large' and 'medium' categories. Conversely 'small' skeletal elements were recovered at a higher rate in Grave B. The lower 'large' and 'medium' bone production rates from Grave B indicate that the stratigraphic method better maintained the provenience and articulation of remains than did Grave A, while the higher 'small' bone recovery rate may point to better recovery techniques of Grave B's excavation team.

[Age assessment at the time of death based on panoramic radiography].

BACKGROUND: The determination of age at the time of death is an important method in forensic anthropology and paleodemography. The possible postmortem investigation of the teeth and jaws enables the determination of age at the time of death, as the bones and teeth are both resistant to degradation in soil and characterized by age-related morphological changes. The aim of this study was to determine whether the age-related changes visible on panoramic radiography correlated with age, and enabled the assessment of individual age. METHODS: Seven radiographic parameters were used in the study: tooth loss, occlusal tooth wear, pulp stones, carious teeth, periapical disease, tooth restoration, and alveolar bone loss associated with periodontal disease. RESULTS: The material comprised 314 dental panoramic tomograms of living patients of both sexes with documented age (18 to 77 years). Multiple regression equations were constructed for the age estimation, including four parameters (the number of missing teeth, the number of intact teeth, the distance of cement-enamel junction from the alveolar ridge, the number of abraded teeth). The nature of data treated by regression analysis required the careful choice of parameters, appropriate functional model for each parameter, and the experience of the investigator. With the four parameters included in equations, the error was +/- 2.55 years. CONCLUSION: This preliminary analysis showed that the conventional regression technique could be appropriate for the age estimation based on panoramic radiography, and that an additional study with a larger sample and on wider population was required.

[Reliability of the individual age assessment at the time of death based on sternal rib end morphology in Balkan population].

This paper analyzes the reliability of the Iscan's sternal rib-ends phase method for the assessment of individual age at the time of death in the Balkan population. The method is based on the morphological age changes of the sternal rib ends. The tested samples consisted of 65 ribs from autopsy cases in the Institute for Forensic Medicine, University of Belgrade, during 1999-2002 (23 females, and 42 males of various ages, ranged from 17-91 years), according to the forensic documents. Significant differences between the real chronological age of the individuals and the values established by the Iscan's method was found, especially in the older categories (phases 6 and 7), in both males and females. The results of the discriminative analysis showed the values of the highest diagnostic relevance for the assessment of age in our population: the change of the depth of the articular fossa, the thickness of its walls, and the quality of the bones.

The reliability of sex determination of skeletons from forensic context in the Balkans.

In this study we have tested the applicability of morphological methods for sex assessment, based on seven pelvic and nine cranial traits, using contemporary Balkans population. The material involved in the study comprises 262 pelvic bones and 180 skulls of male individuals from two mass graves in Serbia. The material was examined separately by an experienced and an inexperienced physical anthropologists. Sex was correctly estimated by the experienced anthropologist in 100% of individuals using all of the 16 pelvic and cranial criteria. In fact, sex differences in pelvic morphology were large enough to allow sexing the individuals with 100% accuracy. Among seven features observed on the pelvic bones, the least reliable single sex indicator was the width of the great sciatic notch (with accuracy of 79.15%). Looking at the skull alone, sex was correctly determined in 70.56% cases. It was shown that the most accurate single indicators among cranial methods was the robustness of the mandible (with accuracy of 70.93%), while the sharpness of the supraorbital margins was the least reliable indicator demonstrating accuracy in only 28.75% of crania. Examination of the sample by an individual with training in physical anthropology, but no case experience, suggests that experience is likely to contribute moderately to the accuracy of the sex determination. Namely, the inexperienced anthropologist accurately assessed the sex of the sample 95.04% of the time; 4.06% less accurate than the experienced anthropologist. The two anthropologists showed the least agreement in scoring the ventral arc and composite arc on the pelvic bones.

Factors affecting postmortem tooth loss.

Unassociated human bones are a particular problem during the exhumation of mass graves and a factor that limits anthropological and paleopathological analyses from archaeological contexts. Extensive anthropological literature has focused on the complex taphonomic factors that influences bone assemblages, but little attention has been paid to postmortem tooth loss and factors affecting this process. The following study focuses upon the influence of different factors on postmortem tooth loss. Three samples were investigated in the study: a medieval church cemetery containing 110 individual skeletal remains, and two samples from a series of mass graves made within the same time period in 1999, containing 402 bodies. The frequency of postmortem tooth loss was analyzed relative to postmortem interval for each sample, excavation methods, age distribution, and presence of bone loss associated with periodontal disease. Our results indicate that the degree of alveolar bone loss significantly affected both antemortem and postmortem tooth loss and that the frequency of postmortem tooth loss has the strongest correlation to time since death. These findings suggest that additional care should be taken when exhuming remains from older contexts.

Two cases of joint disease in post-medieval church cemetery of St. Ilija.

Evidence of disease was analyzed from the skeletal remains of 11 individuals dating to the post-Medieval period from church cemetery of St. Ilija in Serbia. Two individuals showed pathological condition affecting joints. It was supposed that first individual had been suffering from Legg-Calvé-Perthes disease. It seems that this condition remained untreated, with extensive bone remodeling, and that the deformity of femoral head and acetabulum caused secondary degenerative joint disease at a relatively early age of this individual. Second case was related to the bony akylosis of the hand finger, probably caused by Dupuytren's disease. In addition, we discussed development of differential diagnosis in both pathological conditions.