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Tungsten is incorporated in many industrial goods, military applications and medical devices due to its ability to impart flexibility, strength and conductance to materials. Emerging evidence has questioned the safety of tungsten exposure as studies have demonstrated it can promote tumour formation, induce pulmonary disease and alter immune function. Although tungsten is excreted from the body it can accumulate in certain organs such as the brain, colon, liver, kidneys, spleen and bones, where most of the bioaccumulation occurs. Whether prolonged tungsten exposure leads to accumulation in other tissues is unknown. The present study demonstrated that mice exposed to 15 ppm sodium tungstate for 4 weeks in their drinking water showed comparable accumulation in both the bony vertebrae and intervertebral discs (IVDs). Lumbar IVD height was significantly reduced in tungsten-exposed mice and accompanied by decreased proteoglycan content and increased fibrosis. In addition to catabolic enzymes, tungsten also increased the expression of the inflammatory cytokines IL-1β and tumour necrosis factor (TNF)-α as well as the neurotrophic factors nerve growth factor (NGF) and brain-derived nerve factor (BDNF) in IVD cells. Tungsten significantly increased the presence of nociceptive neurons at the endplates of IVDs as observed by the expression of calcitonin gene-related peptide (CGRP) and anti-protein gene product 9.5 (PGP9.5) in endplate vessels. The present study provided evidence that tungsten may enhance disc degeneration and fibrosis as well as increase the expression of markers for pain. Therefore, tungsten toxicity may play a role in disc degeneration disease.

The paper presents the baseline climate of the Cotnari wine growing region, the climate under whose influence were set up the wine grape varieties, wine types and grapevine training systems of this vineyard. It is also presented the baseline climate suitability for wine grape growing and its spatial variation in the vineyard area. The study is based on the climate data for the 1961 to 1980 time period, previous to the beginning of the climate warming and, therefore, considered to be representative for the baseline climate time period. According to study results, the baseline climate of the Cotnari wine growing region was cool, with annual average temperatures by 8.5...10.0°C, with a sum of the effective temperatures by 1081...1382°C, with freezing phenomena at the beginning and towards the end of the growing season, and with very cool nights during the month of September (CI+2). Elements that generate the baseline climate suitability for wine grape growing were: high average temperatures for the month of July (19...21.1°C); long growing season up to 190 days, and high values for the global radiation, up to 93 kcal/cm2/April 1st - September 30th on the sunny slopes within the vineyard area. According to the multicriteria evaluation of the suitability for wine grape growing, 87.9% (1792 ha) of the vineyard surface was characterized by a baseline climate suitable for white quality wines and 11.9% (241.0 ha) by a baseline climate suitable for white table wines, sparkling wines and wines for distillates. [ABSTRACT FROM AUTHOR]

Introduction: Insulin Resistance (IR) can develop into type 2 diabetes mellitus and is closely associated with obesity. However, the non-obese population has also shown a predisposition to the risk of IR due to genetics. Aim: To assess the relationship between IR and obesity in Type 2 Diabetes Mellitus (T2DM) by comparing the proportion of subjects with IR in lean and obese T2DM and to identify the factors predicting IR in T2DM. Materials and Methods: A cross-sectional, hospital-based study was done at Department of Medicine of RL Jalappa hospital, Kolar, Karnataka on 106 T2DM patients aged >18 years. The study population was grouped into lean (BMI30 kg/m2 ). IR was calculated using Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) and was considered as primary outcome variable. Obesity was considered as primary explanatory variable. Age, Gender, fasting insulin, C-peptide, Fasting Blood Sugar, Glycated haemoglobin (GHB or HbA1c) were the other explanatory variables. Descriptive analysis was carried out using mean and standard deviation for quantitative variables, frequency and proportion for categorical variables. Chi-square test was used to test statistical significance between the groups. Univariate logistic regression analysis was done to identify the predictors of IR. IBM Statistical Package for Social Sciences (SPSS) version 22 was used for statistical analysis. The p-value

Background: Coronary artery disease (CAD) is a worldwide health epidemic. Acute coronary syndrome is a potentially life-threatening condition and patient may die or become disabled in the prime of life. The aim of this study was to determine the conventional risk factors of CAD in young and elderly aged patients in Andhra Pradesh. Materials and Methods: Total of 190 CAD patients admitted in ICCU at Santhiram Medical College General Hospital, Nandyal, Andhra pradesh were selected for the study. In this, 90 patients were aged between 18-45 years, and 100 were more than 45 years of age. These patients were evaluated for risk factor contributing to occurrence of CAD. Results: The hypertension (20%), smoking (22%), diabetes mellitus (11%) and dyslipidemia (8%) were the most common risk factors in young patients. Overall risk factors were more likely in males compared to females (18 to

Interfaces between AISI 304 stainless steel screws and cranial bone were investigated after long-term implantation lasting for 42 years. Samples containing the interface regions were analyzed using state-of-the-art analytical techniques including secondary ion mass, Fourier-transform infrared, Raman, and X-ray photoelectron spectroscopies. Local samples for scanning transmission electron microscopy were cut from the interface regions using the focused ion beam technique. A chemical composition across the interface was recorded in length scales covering micrometric and nanometric resolutions and relevant differences were found between peri-implant and the distant cranial bone, indicating generally younger bone tissue in the peri-implant area. Furthermore, the energy dispersive spectroscopy revealed an 80 nm thick steel surface layer enriched by oxygen suggesting that the AISI 304 material undergoes a corrosion attack. The attack is associated with transport of metallic ions, namely, ferrous and ferric iron, into the bone layer adjacent to the implant. The results comply with an anticipated interplay between released iron ions and osteoclast proliferation. The interplay gives rise to an autocatalytic process in which the iron ions stimulate the osteoclast activity while a formation of fresh bone resorption sites boosts the corrosion process through interactions between acidic osteoclast extracellular compartments and the implant surface. The autocatalytic process thus may account for an accelerated turnover of the peri-implant bone.

Background: Orthodontic treatment, particularly involving premolar extractions, has been a subject of ongoing debate within the orthodontic community. The impact of such interventions on the pharyngeal airway, a critical component of the respiratory system, remains a topic of exploration., Objective: This retrospective cephalometric study aims to investigate changes in pharyngeal airway dimensions following orthodontic treatment involving either all four or all five premolar extractions., Methods: A sample of 68 participants, extracted from orthodontic records, underwent cephalometric analysis to quantify changes in pharyngeal airway dimensions. The study compared two groups: those treated with all four premolar extractions (n=34) and those treated with all five premolar extractions (n=34). Cephalometric radiographs taken before and after treatment were analyzed, focusing on airway width, length, and volume., Results: Preliminary findings indicate significant changes in airway dimensions within each group. In the all four premolar extraction group, there was a statistically significant decrease in airway width (p=0.02) and volume (p=0.04). Similarly, the all five premolar extraction group exhibited significant reductions in airway width (p=0.03) and volume (p=0.02). However, the between-group comparisons revealed no significant differences in changes between the two groups., Conclusion: This study sheds light on the intricate relationship between orthodontic interventions, specifically premolar extractions, and changes in pharyngeal airway dimensions. While significant changes were observed within each group, the lack of significant differences between the all four and all five premolar extraction groups raises intriguing questions about the specific impact of premolar extraction patterns on the upper airway., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Badepalli et al.)

Background Temporomandibular joint disorders (TMD) represent a prevalent group of conditions impacting the temporomandibular joint. Among the therapeutic interventions, occlusal splint therapy has gained recognition for its potential to address TMD symptoms, particularly in cases involving joint displacement. Objective This study aims to investigate the effectiveness of occlusal splint therapy in cases of moderate TMD with joint displacement, focusing on changes in condylar position, joint morphology, and patient-reported outcomes. Methods A retrospective analysis was conducted involving 148 participants who underwent occlusal splint therapy between January 2018 and December 2020. Data were collected through cone beam computed tomography (CBCT) imaging for precise assessments of condylar position and joint morphology. Ethical approval was obtained, and participants provided informed consent. Baseline characteristics, medical history, and TMD severity were recorded. Occlusal splint therapy included individualized fabrication, occlusal analysis, adjustments for optimal fit, and prescribed wear schedules. Follow-up included CBCT scans at specified intervals (three months and six months), with participant-reported outcomes collected. The data analysis was conducted using IBM SPSS Statistics for Windows, Version 22.0 (Released 2013; IBM Corp., Armonk, NY, USA). Paired t-tests or nonparametric equivalents were employed to assess changes in condylar position and joint morphology. Subgroup analyses were conducted to explore potential factors influencing treatment outcomes. The significance level was set at p < 0.05 for all statistical tests. Results The entire cohort (n = 148) had a mean age of 32.5 years (± 8.1), with a balanced gender distribution. Changes in condylar position revealed a statistically significant improvement (p = 0.03), with a mean decrease of 0.2 mm posttreatment. Joint morphology changes indicated increased joint space width (p = 0.01), improved disc position (p = 0.02), and nonsignificant alterations in bony structures (p = 0.10). Patient-reported outcomes demonstrated significant improvements in pain levels, jaw functionality, and satisfaction (all p < 0.001). Age and gender subgroup analyses showed consistent improvements in condylar position, joint morphology, and patient-reported outcomes across different groups. Conclusion Occlusal splint therapy demonstrated effectiveness in improving condylar position, joint morphology, and patient-reported outcomes in cases of moderate TMD with joint displacement. The findings underscore the potential of occlusal splint therapy as a viable intervention for managing TMD, providing valuable insights for clinicians and researchers., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Mathew et al.)

Hypothyroidism is an endocrine disorder which occurs due to a deficiency of thyroid hormones. Hoffmann's syndrome is a rare complication of hypothyroidism - presenting as hypothyroid myopathy. We describe the case of a 20-year-old lactating female, known to have hypothyroidism (diagnosed during her pregnancy and having discontinued treatment following delivery), presenting with complaints of pain, swelling of bilateral calf muscles with cramps in bilateral lower limbs. Symptoms of muscle pseudohypertrophy with muscle stiffness are relatively rare in subclinical hypothyroidism and it is important to identify and diagnose this rare condition, and initiate appropriate treatment., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

This study examines Hg distribution in stream sediments impacted by hard coal mining in the Upper Silesian Coal Basin (USCB), Czechia. By means of a comparative analysis, geological samples and samples from stream sediments were used to carry out a comprehensive assessment of the effects of anthropogenic activities on Hg distribution and speciation. Total Hg (THg), total organic and inorganic carbons (TOC and TIC), and total sulphur (TS) were measured in the samples to reveal a potential relationship. In addition, THg and TS species were discussed in order to elucidate their mobility pattern in the environment. The results have shown that there are no correlations between THg, TS, and TOC indicating overlapping Hg sources attributed to industrial processes. Geological samples, particularly coal and associated sedimentary rocks, contained lower Hg concentrations compared to a variety of stream sediments. The main Hg species identified in the samples was a stable β-HgS, which decreases its mobility in the riverine environment. It follows that Hg enrichment and speciation is linked to industrial processes, which are the main origin/cause for Hg enrichment and transformation. Minor proportions of HgO in some samples show Hg oxidation upon diagenesis, while HgCl 2 is attributed to the chemical loads from the former coking plant., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Data of neurocognitive deficits in survivors of acute lymphoblastic leukemia (ALL) is scarce from low middle-income countries (LMICs), and is influenced by biological and cultural variations. The objective of this study was to assess the prevalence and spectrum of neurocognitive deficits in a cohort of survivors from India., Procedure: Seventy survivors of childhood ALL were evaluated for neurocognitive deficits by the Indian adaptation of Wechsler Intelligence Scale for Children - Fourth Edition (WISC-IV INDIA ). The prevalence of neurocognitive deficits was calculated based on the full-scale intelligence quotient (FSIQ), and scores in discrete domains like verbal comprehension, perceptual reasoning, working memory, and processing speed were calculated and compared to demographics, treatment, and sociocultural factors., Results: The mean (SD) current age and time since diagnosis was 10.5 (±3.2) years and 5 (±2.8) years, respectively. The mean FSIQ was 86.1 ± 20.5, with significant neurocognitive deficit (FSIQ <90) being prevalent in 50% (95% CI: 38%-62%) of the cohort. The proportion of survivors with deficits in individual domains of verbal comprehension, perceptual reasoning, working memory, and processing speed were 49%, 50%, 47%, and 44%, respectively. The odds of having neurocognitive deficits were higher when a child belonged to lower socioeconomic strata (OR 5.7, p = .004), parents with lower education attainment (OR 4.3, p = .041), and whose birth order was higher (OR 20.1, p = .005). Age at diagnosis/assessment, chemotherapy received, or dose of radiotherapy did not have a direct impact on neurocognition., Conclusions and Relevance: Rates of neurocognitive deficits are higher in survivors in LMICs, with socioeconomic variables contributing more than the direct neurotoxic effects of treatment., (© 2022 Wiley Periodicals LLC.)

The work presented here provides a complex environmental impact of sediments in vicinity to the area of the former Lazy coal mine site in the Upper Silesian Coal Basin (Czech Republic). The main aim of this work has been to determine the degree of contamination, to describe the organic matter, and to carry out sorption isotherms to see the size and distribution of pores in the monitored sediments that are the crucial parameters to assumption of removal mechanisms of elements carried in mine water. The results show that the greatest enrichment of Mn, Sr, Ba, and was in sediments of the first tens of meters from the mine water discharge sediments. Ba and Sr were precipitated as mineral barite and thus formed a dominant insoluble component in the river sediments, which were further carried by water flow towards the water reservoirs. Predominant amounts of fossil material and smaller quantities of carbonized and recent organic matter were altered by weathering and erosion processes. The coal materials have a relatively beneficial sorption capacity, which increases with the carbon content. The overburden waste should be considered for use in removing heavy metals in-situ., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The composition of lipids in soil offers clues to soil degradation processes due their persistency and selectivity in soil, and close relation to long-term processes in the ecosystem, thanks to their role in cell membranes of organisms. Organic solvent-extractable compounds were recovered from soils collected at two sites differing in the degree of forest damage. Gas chromatography/mass spectroscopy and Fourier transform infrared spectroscopy were applied in order to characterize solvent-extractable lipids. Raman spectroscopy was also applied as it provides distinct advantages for determining the structural order of carbonaceous materials. The organic matter measurement techniques were combined with an established simultaneous multi-element measurement technique. Variations in individual soil horizons from the sites were reflected in the crystallinity of epicuticular waxes, presence of long-chain aliphatic hydrocarbons, concentrations of n-alkanes, saturated and unsaturated fatty acids, dicarboxylic acids, and in the content of aromatic structures, hydroxyl, ester, and carboxylic acid groups. The results are explained by differently transformed organic matter. The concentrations of elements in the soils were also affected by atmospheric depositions, including higher accumulations of arsenic and antimony, and lower contents of natural nutrients. These data have potential to be used as sensitive biogenic indicators of ecosystem damage by long-term atmospheric depositions.

A woody-biochar was added to waste biomass during a composting process. The resulting compost-char was amended to a metal contaminated soil and two plant species, L. perenne and E. sativa, were grown in a pot experiment to determine 1) plant survival and stress factors, 2) uptake of metals to plants and, 3) chemical characteristics of sampled soils and pore waters. Compost supplemented with biochar after the composting process were also tested, as well as a commercially available compost, for comparison. Co-composting with biochar hastened the composting process, resulting in a composite material of reduced odour, increased maturity, circum-neutral pH and increased moisture retention than compost (increase by 3% of easily removable water content). When amended to the soil, CaCl 2 extractable and pore water metals s were reduced by all compost treatments with little influence of biochar addition at any tested dose. Plant growth success was promoted furthest by the addition of co-composted biochar to the test soil, especially in the case of E. sativa. For both tested plant species significant reductions in plant metal concentrations (e.g. 8-times for Zn) were achieved, against the control soil, by compost, regardless of biochar addition. The results of this study demonstrate that the addition of biochar into the composting process can hasten the stability of the resulting compost-char, with more favourable characteristics as a soil amendment/improver than compost alone. This appears achievable whilst also maintaining the provision of available nutrients to soils and the reduction of metal mobility, and improved conditions for plant establishment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Introduction: Neuroblastoma ranks third among pediatric malignancies., Case Report: The case of a 3-year-old child is presented, who suddenly had frequent, unproductive, emetic cough; fever; and weight loss. Lung X-ray showed an opacity situated in the posterior superior mediastinum. Thoracic ultrasound revealed a slightly inhomogeneous, hypoechoic mass located in the posterior superior mediastinum. Computed tomography evidenced a tumor mass with homogeneous appearance in the costo-vertebral groove. Histological examination confirmed the diagnosis of ganglioneuroblastoma., Conclusion: Although history and clinical examination provided few elements, diagnosis was made based on imaging and histopathological examination.

Tumor suppressor p53 is mutated in about 50% of cancers. Most malignant melanomas carry wild-type p53, but p53 activity is often inhibited due to overexpression of its negative regulators Mdm2 or MdmX. We performed high throughput screening of 2448 compounds on A375 cells carrying p53 activity luciferase reporter construct to reveal compounds that promote p53 activity in melanoma. Albendazole and fenbendazole, two approved and commonly used benzimidazole anthelmintics, stimulated p53 activity and were selected for further studies. The protein levels of p53 and p21 increased upon the treatment with albendazole and fenbendazole, indicating activation of the p53-p21 pathway, while the levels of Mdm2 and MdmX decreased in melanoma and breast cancer cells overexpressing these proteins. We also observed a reduction of cell viability and changes of cellular morphology corresponding to mitotic catastrophe, i.e., G2/M cell cycle arrest of large multinucleated cells with disrupted microtubules. In summary, we established a new tool for testing the impact of small molecule compounds on the activity of p53 and used it to identify the action of benzimidazoles in melanoma cells. The drugs promoted the stability and transcriptional activity of wild-type p53 via downregulation of its negative regulators Mdm2 and MdmX in cells overexpressing these proteins. The results indicate the potential for repurposing the benzimidazole anthelmintics for the treatment of cancers overexpressing p53 negative regulators.

Two pieces of studlovite - Eocene amber from Študlov (Southeast Moravia, Czech Republic) were investigated. To arrive at a more detailed description, attenuated total reflection Fourier transform infrared spectroscopy, Raman spectroscopy, pyrolysis-gas chromatography/mass spectrometry, and gas chromatography-mass spectrometry were used. Both studlovite samples revealed signs of the same plant source, with higher polymerisation and a higher degree of maturation of the fossilized matter. Despite their close spectral resemblance, they differed in their detailed chemical composition, and in structure. Layering of one of the pieces studied showed how the resin was built and what impact the process had on the chemical composition of the amber. Characterization of the organic matter was completed with an analysis of trace elements in amber samples using scanning electron microscope combined with elemental distribution analysis (SEM/EDAX). The results demonstrated the paleoenvironmental conditions that occurred in the plant during resin exudation following wounding., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Novel multiresponsive hybrid biocompatible systems of κ-carrageenan-graft-poly(2-isopropyl-2-oxazoline-co-2-butyl-2-oxazoline)s with unique combination of responsiveness to external stimuli were synthesized and studied. The polymer thermoresponsive behavior proved the existence of both lower and upper critical solution temperatures in aqueous milieu, forming gel at lower temperature, a solution at room temperature and cloudy nanophase-separated dispersion at elevated temperature. The limit temperatures can easily be adjusted by the polyoxazoline graft length and grafting density. Moreover, the polymer behavior is additionally dependent on the concentration of potassium ions. The polymers behave similarly as the original κ-carrageenan, and thus, the poly(2-alkyl-2-oxazoline) grafts do not decrease the ability of the κ-carrageenan to form the self-assembled structures. Molecular principles beyond this multistimuli-responsive behavior were elucidated with the use of dynamic light scattering, magnetic resonance and fluorescence measurements as well as atomic force microscopy. These polymers could be used in a wide range of biological applications demanding thermo- and potassium-responsiveness., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The climatic variability hypothesis was applied to the thermal tolerance of edaphic and cave Collembola occupying contrasting environments. Collembola belonged to four categories - trogloxene, subtroglophile, eutroglophile and troglobiont - with a different degree of affinity to subterranean habitats. Altogether, specimens of 17 species were exposed to a one-hour laboratory survival test. The impact of temperature, species and species-temperature interaction on cold and heat survival was statistically significant. There was a decrease trend in cold and heat tolerance from trogloxenes, over subtroglophiles and eutroglophiles to troglobionts. It was shown that obligate cave species, restricted to climatic-stable cave conditions, retain a functional thermal resistance, i.e. the genetically determined ability to tolerate relatively broader temperature ranges. Our results outlined the direct relationship between the thermal tolerances of species and the size of their geographic distributions. It was also observed that cold resistance of Collembola decreased significantly with increasing species body length, indicating that body size plays an important role in temperature tolerances of arthropods inhabiting soil and subterranean habitats., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The digestive tract microbiota (DTM) plays a plethora of functions that enable hosts to exploit novel niches. However, our understanding of the DTM of birds, particularly passerines, and the turnover of microbial communities along the digestive tract are limited. To better understand how passerine DTMs are assembled, and how the composition changes along the digestive tract, we investigated the DTM of seven different compartments along the digestive tract of nine New Guinean passerine bird species using Illumina MiSeq sequencing of the V4 region of the 16S rRNA. Overall, passerine DTMs were dominated by the phyla Firmicutes and Proteobacteria. We found bird species-specific DTM assemblages and the DTM of different compartments from the same species tended to cluster together. We also found a notable relationship between gut community similarity and feeding guilds (insectivores vs. omnivores). The dominant bacterial genera tended to differ between insectivores and omnivores, with insectivores mainly having lactic acid bacteria that may contribute to the breakdown of carbohydrates. Omnivorous DTMs were more diverse than insectivores and dominated by the bacterial phyla Proteobacteria and Tenericutes. These bacteria may contribute to nitrogen metabolism, and the diverse omnivorous DTMs may allow for more flexibility with varying food availability as these species have wider feeding niches. In well-sampled omnivorous species, the dominant bacterial genera changed along the digestive tracts, which was less prominent for insectivores. In conclusion, the DTMs of New Guinean passerines seem to be species specific and, at least in part, be shaped by bird diet. The sampling of DTM along the digestive tract improved capturing of a more complete set of members, with implications for our understanding of the interactions between symbiont and gut compartment functions.

A conceptually new bimodal immunoradiotherapy treatment was demonstrated using thermoresponsive polymer β-glucan-graft-poly(2-isopropyl-2-oxazoline-co-2-butyl-2-oxazoline) bearing complexes of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid with yttrium-90(III) at the graft ends. The behavior of this thermoresponsive polymer in aqueous solutions was studied, and it showed the appropriate cloud point temperature for brachytherapy applications. The polymer was tested in vitro, and it exhibited nontoxicity and active uptake into cancer cells and macrophages with colocalization in the lysosomes and macrophagosomes. Moreover, the observed oxidative burst response of the leukocytes established the immunostimulatory properties of the polymer, which were also studied in vivo after injection into the thigh muscles of healthy mice. The subsequent histological evaluation revealed the extensive immune activation reactions at the site of injection. Furthermore, the production of tumor necrosis factor α induced by the prepared polymer was observed in vitro, denoting the optimistic prognosis of the treatment. The biodistribution study in vivo indicated the formation of the polymer depot, which was gradually degraded and excluded from the body. The radiolabeled polymer was used during in vivo antitumor efficiency experiments on mice with EL4 lymphoma. The immunoradiotherapy group (treated with the radiolabeled polymer) demonstrated the complete inhibition of tumor growth during the beginning of the treatment. Moreover, 7 of the 15 mice were completely cured in this group, while the others exhibited significantly prolonged survival time compared to the control group. The in vivo experiments indicated the considerable synergistic effect of using immunoradiotherapy compared to separately using immunotherapy or radiotherapy.

The efficiency of clinical trials involving transplantation of multipotent mesenchymal stromal cells (MSCs) is often insufficient due to harsh conditions present within the target tissue including hypoxia, low nutrient supply as well as inflammatory reactions. This indicates the necessity for optimization of cell-based therapy approaches which might include either modification of the cell manufacturing process or specific cell pretreatment procedures prior to transplantation. Recent reports confirm evidence that the aggregation of MSCs into three-dimensional (3D) multicellular spheroids results in enhancement of the overall therapeutic potential of cells, by improving the anti-inflammatory and angiogenic properties, stemness and survival of MSCs after transplantation. Such an MSCs spheroid generation approach may open new opportunities for the enlargement of MSCs applications in clinical research and therapy. However, the unification and optimization of 3D spheroid generation techniques, including the selection of appropriate clinical-grade culture conditions and methods for their large-scale production, are still of great importance. The current review addresses questions regarding therapeutic-associated properties of 3D multicellular MSCs spheroids in vitro and during preclinical animal studies, with special attention to the possibilities of translating these research achievements toward further clinical manufacturing and applications.

Off-the-shelf availability of human adipose-derived mesenchymal stromal cells (ASCs) for regenerative medicine application requires the development of nontoxic, safe, and efficient protocols for cryopreservation. Favorably, such cell processing protocols should not contain xenogeneic or toxic components, such as fetal bovine serum (FS) and dimethyl sulfoxide (DMSO). The objective of the study was to assess the sensitivity of ASCs to DMSO-free cryopreservation protocol depending on their expansion conditions: conventional, based on the application of FS or xeno-free, using PL as a medium supplement. ASCs expansion was carried out in α-MEM supplemented either with FS or PL. For DMSO- and xeno-free cryopreservation ASCs were pretreated with different concentrations of sucrose during 24 h of culture. Pretreated ASCs were cryopreserved in α-MEM containing 100-300 mM of sucrose with the cooling rate of 1 degree/min. ASCs were tested for survival (Trypan Blue test), viability (MTT test), recovery (Alamar Blue test), proliferation and ability to multilineage differentiation. The optimal concentrations of sucrose for ASCs pretreatment and as an additive in cryoprotective solution, which provided highest cell survival, comprised 100 and 200 mM, correspondingly. Survival and recovery rates of platelet lysate (PL)-expanded ASCs after DMSO-free cryopreservation comprised 59 and 51%, and were higher than in FS-cultured cells. After DMSO-free cryopreservation PL-processed ASCs had a shorter population doubling time and higher capacity for osteogenic differentiation than FS-processed cultures. The described DMSO- and xeno-free processing may form the basis for the development of safe and efficient protocols for manufacturing and banking of ASCs, providing their off-the-shelf availability for regenerative medicine applications.

KEY MESSAGE : bZIP TF network in pollen. Transcriptional control of gene expression represents an important mechanism guiding organisms through developmental processes and providing plasticity towards environmental stimuli. Because of their sessile nature, plants require effective gene regulation for rapid response to variation in environmental and developmental conditions. Transcription factors (TFs) provide such control ensuring correct gene expression in spatial and temporal manner. Our work reports the interaction network of six bZIP TFs expressed in Arabidopsis thaliana pollen and highlights the potential functional role for AtbZIP18 in pollen. AtbZIP18 was shown to interact with three other pollen-expressed bZIP TFs-AtbZIP34, AtbZIP52, and AtbZIP61 in yeast two-hybrid assays. AtbZIP18 transcripts are highly expressed in pollen, and at the subcellular level, an AtbZIP18-GFP fusion protein was located in the nucleus and cytoplasm/ER. To address the role of AtbZIP18 in the male gametophyte, we performed phenotypic analysis of a T-DNA knockout allele, which showed slightly reduced transmission through the male gametophyte. Some of the phenotype defects in atbzip18 pollen, although observed at low penetrance, were similar to those seen at higher frequency in the T-DNA knockout of the interacting partner, AtbZIP34. To gain deeper insight into the regulatory role of AtbZIP18, we analysed atbzip18/- pollen microarray data. Our results point towards a potential repressive role for AtbZIP18 and its functional redundancy with AtbZIP34 in pollen.

We have developed a biodegradable, biocompatible system for the delivery of the antituberculotic antibiotic rifampicin with a built-in drug release and nanoparticle degradation fluorescence sensor. Polymer nanoparticles based on poly(ethylene oxide) monomethyl ether-block-poly(ε-caprolactone) were noncovalently loaded with rifampicin, a combination that, to best of our knowledge, was not previously described in the literature, which showed significant benefits. The nanoparticles contain a Förster resonance energy transfer (FRET) system that allows real-time assessment of drug release not only in vitro, but also in living macrophages where the mycobacteria typically reside as hard-to-kill intracellular parasites. The fluorophore also enables in situ monitoring of the enzymatic nanoparticle degradation in the macrophages. We show that the nanoparticles are efficiently taken up by macrophages, where they are very quickly associated with the lysosomal compartment. After drug release, the nanoparticles in the cmacrophages are enzymatically degraded, with half-life 88±11 min., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Adiabatic shear banding (ASB) is a unique dynamic failure mechanism that results in an unpredicted catastrophic failure due to a concentrated shear deformation mode. It is universally considered as a material or structural instability and as such, ASB is hardly controllable or predictable to some extent. ASB is modeled on the premise of stability analyses. The leading paradigm is that a competition between strain (rate) hardening and thermal softening determines the onset of the failure. It was recently shown that microstructural softening transformations, such as dynamic recrystallization, are responsible for adiabatic shear failure. These are dictated by the stored energy of cold work, so that energy considerations can be used to macroscopically model the failure mechanism. The initial mechanisms that lead to final failure are still unknown, as well as the ASB formation mechanism(s). Most of all - is ASB an abrupt instability or rather a gradual transition as would be dictated by microstructural evolutions? This paper reports thorough microstructural characterizations that clearly show the gradual character of the phenomenon, best described as a nucleation and growth failure mechanism, and not as an abrupt instability as previously thought. These observations are coupled to a simple numerical model that illustrates them.

The presence of uranium, with a bulk mass fraction of about 1.5 wt% and radiolytic alterations are a feature of Cenomanian amber from Křižany, at the northeastern edge of the North Bohemian Cretaceous uranium ore district. Pores and microcracks in the amber were filled with a mineral admixture, mainly in the form of Zr-Y-REE enriched uraninite. As a result of radiolytic alterations due to the presence of uranium, structural changes were observed in the Křižany amber in comparison with a reference amber from Nové Strašecí in central Bohemia; this was of similar age and botanical origin but did not contain elevated levels of uranium. Structural changes involved an increase in aromaticity due to dehydroaromatization of aliphatic cyclic hydrocarbons, loss of oxygen functional groups, an increase in the degree of polymerization, crosslinking of CC bonds, formation of a three-dimensional hydrocarbon network in the bulk organic matrix, and carbonization of the organic matrix around the uraninite infill., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Photon-upconverting nanoparticles (UCNPs) emit light of shorter wavelength under near-infrared excitation and thus avoid optical background interference. We have exploited this unique photophysical feature to establish a sensitive competitive immunoassay for the detection of the pharmaceutical micropollutant diclofenac (DCF) in water. The so-called upconversion-linked immunosorbent assay (ULISA) was critically dependent on the design of the upconversion luminescent detection label. Silica-coated UCNPs (50 nm in diameter) exposing carboxyl groups on the surface were conjugated to a secondary anti-IgG antibody. We investigated the structure and monodispersity of the nanoconjugates in detail. Using a highly affine anti-DCF primary antibody, the optimized ULISA reached a detection limit of 0.05 ng DCF per mL. This performance came close to a conventional enzyme-linked immunosorbent assay (ELISA) without the need for an enzyme-mediated signal amplification step. The ULISA was further employed for analyzing drinking and surface water samples. The results were consistent with a conventional ELISA as well as liquid chromatography-mass spectrometry (LC-MS).

The effect of dietary administered young barley containing a mixture of phytochemicals to female rats for the prevention of N-methyl-N-nitrosourea-induced mammary carcinogenesis was evaluated. After carcinogen administration (14 wk), mammary tumors were removed and prepared for histopathological and immunohistochemical analysis. Moreover, in vitro evaluation of possible mechanisms in MCF-7 breast cancer cell line was performed. Barley (0.3%) demonstrated mild antitumor effect in mammary carcinogenesis, yet 3% barley did not further improve this effect. Immunohistochemical analysis of rat tumor cells in treated groups showed significant increase in caspase-3 expression and significant reduction in Ki67 expression. In addition, 3% barley significantly decreased dityrosine levels versus control. Barley in higher dose significantly decreased serum low-density lipoprotein-cholesterol in rats. In vitro studies showed that barley significantly decreased survival of MCF-7 cells in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and significantly decreased 5-bromo-20-deoxyuridine incorporation versus control. Barley prevented cell cycle progression and extended incubation with barley showed significant increase in the percentage of annexin V/propidium iodide-positive MCF-7 cells. Our results propose an antitumor effect for the mixture of phytochemicals present in young barley in a breast cancer model.

We determined the total system elastic Helmholtz free energy, under the constraints of constant temperature and volume, for systems comprised of one or more perfectly bonded hard spherical inclusions (i.e. "hard spheres") embedded in a finite spherical elastic solid. Dirichlet boundary conditions were applied both at the surface(s) of the hard spheres, and at the outer surface of the elastic solid. The boundary conditions at the surface of the spheres were used to describe the rigid displacements of the spheres, relative to their initial location(s) in the unstressed initial state. These displacements, together with the initial positions, provided the final shape of the strained elastic solid. The boundary conditions at the outer surface of the elastic medium were used to ensure constancy of the system volume. We determined the strain and stress tensors numerically, using a method that combines the Neuber-Papkovich spherical harmonic decomposition, the Schwartz alternating method, and Least-squares for determining the spherical harmonic expansion coefficients. The total system elastic Helmholtz free energy was determined by numerically integrating the elastic Helmholtz free energy density over the volume of the elastic solid, either by a quadrature, or a Monte Carlo method, or both. Depending on the initial position of the hard sphere(s) (or equivalently, the shape of the un-deformed stress-free elastic solid), and the displacements, either stationary or non-stationary Helmholtz free energy minima were found. The non-stationary minima, which involved the hard spheres nearly in contact with one another, corresponded to lower Helmholtz free energies, than did the stationary minima, for which the hard spheres were further away from one another.

The process of DNA transposition involves the binding, cleavage, and recombination of specific DNA segments (transposable elements, TE) and is catalyzed by special enzymes encoded by the TE transposases. REP-associated tyrosine transposases (RAYTs) are a class of Y1 nucleases related to the IS200/IS605 transposases associated with a bacterial TE known as repetitive extragenic palindrome elements (REPs). Although RAYT has been subject of numerous studies, where DNA binding and cleavage by RAYT have been confirmed for Escherichia coli, the molecular mechanism of DNA insertion has not been fully understood. In this work, it is demonstrated that surface plasmon resonance (SPR) biosensor technology combined with a system of DNA hairpin probes (mimicking the natural REP sequence) and short oligonucleotides (ONs) can provide a rapid and real-time platform for monitoring and quantification of RAYT activity. We utilized RAYT from E. coli (strain MG1655) as a model system, where we evaluated its activity towards both a natural REP sequence as well as REP sequences having modifications targeting specific features of the DNA crucial for the DNA binding and cleavage. The characteristics of the RAYT-DNA interaction obtained by means of the SPR approach were compared with the results of SDS-PAGE analysis.

In mammals, germ cell differentiation is initiated in the Primordial Germ Cells (PGCs) during fetal development. Prenatal exposure to environmental toxicants such as endocrine disruptors may alter PGC differentiation, development of the male germline and induce transgenerational epigenetic disorders. The anti-androgenic compound vinclozolin represents a paradigmatic example of molecule causing transgenerational effects on germ cells. We performed prenatal exposure to vinclozolin in mice and analyzed the phenotypic and molecular changes in three successive generations. A reduction in the number of embryonic PGCs and increased rate of apoptotic cells along with decrease of fertility rate in adult males were observed in F1 to F3 generations. Blimp1 is a crucial regulator of PGC differentiation. We show that prenatal exposure to vinclozolin deregulates specific microRNAs in PGCs, such as miR-23b and miR-21, inducing disequilibrium in the Lin28/let-7/Blimp1 pathway in three successive generations of males. As determined by global maps of cytosine methylation, we found no evidence for prominent changes in DNA methylation in PGCs or mature sperm. Our data suggest that embryonic exposure to environmental endocrine disruptors induces transgenerational epigenetic deregulation of expression of microRNAs affecting key regulatory pathways of germ cells differentiation.

Polybrominated diphenyl ethers (PBDE) were, and in some countries still are, used as flame retardants for plastic materials. When released from plastics, PDBE cause harm to the environment. This creates the incentive for further investigation of the PBDE degradation. This work focused on a formulation of a PBDE photodegradation model based on the PBDE properties obtained by the quantum chemical calculations. The proposed model predicted degradation routes of arbitrary PBDE congener. The routes of selected congeners were validated by the two independently published data sets and showed the high fitting degree. The model can be easily modified for any reactor system if the initial reaction rate constant of one congener is available for the given system.

Background: Sperm proteins are important for the sperm cell function in fertilization. Some of them are involved in the binding of sperm to the egg. We characterized the acrosomal sperm protein detected by a monoclonal antibody (MoAb) (Hs-8) that was prepared in our laboratory by immunization of BALB/c mice with human ejaculated sperms and we tested the possible role of this protein in the binding assay., Methods: Indirect immunofluorescence and immunogold labelling, gel electrophoresis, Western blotting and protein sequencing were used for Hs-8 antigen characterization. Functional analysis of GAPDHS from the sperm acrosome was performed in the boar model using sperm/zona pellucida binding assay., Results: Monoclonal antibody Hs-8 is an anti-human sperm antibody that cross-reacts with the Hs-8-related protein in spermatozoa of other mammalian species (boar, mouse). In the immunofluorescence test, Hs-8 antibody recognized the protein localized in the acrosomal part of the sperm head and in the principal piece of the sperm flagellum. In immunoblotting test, MoAb Hs-8 labelled a protein of 45 kDa in the extract of human sperm. Sequence analysis identified protein Hs-8 as GAPDHS (glyceraldehyde 3-phosphate dehydrohenase-spermatogenic). For this reason, commercial mouse anti-GAPDHS MoAb was applied in control tests. Both antibodies showed similar staining patterns in immunofluorescence tests, in electron microscopy and in immunoblot analysis. Moreover, both Hs-8 and anti-GAPDHS antibodies blocked sperm/zona pellucida binding., Conclusion: GAPDHS is a sperm-specific glycolytic enzyme involved in energy production during spermatogenesis and sperm motility; its role in the sperm head is unknown. In this study, we identified the antigen with Hs8 antibody and confirmed its localization in the apical part of the sperm head in addition to the principal piece of the flagellum. In an indirect binding assay, we confirmed the potential role of GAPDHS as a binding protein that is involved in the secondary sperm/oocyte binding.

The thermal stability of the monosubstituted cationic cyclodextrin (CD) derivatives PEMEDA-β-CD and PEMPDA-β-CD, which differ in their substituent linker length (ethylene and propylene, respectively), was studied via (1)H NMR experiments. PEMPDA-β-CD exhibited higher resistance towards the Hofmann degradation and was chosen as a more suitable host molecule for further studies. Inclusion properties of PEMPDA-β-CD in solution with a series of simple aromatic guests (salicylic acid, p-methoxyphenol and p-nitroaniline) were determined by isothermal titration calorimetry (ITC) and compared to the native β-CD. Permanently charged cationic CD derivatives were successfully deposited on the anionic solid surface of polymeric Nafion(®) 117 membrane via electrostatic interactions. Deposition kinetics and coverage of the surface were determined by ELSD. Finally, the ability of the CD derivatives bound to the solid surface to encapsulate aromatic compounds from aqueous solution was measured by UV-vis spectroscopy. The obtained results are promising for future industrial applications of the monosubstituted β-CD derivatives, because the preparation of cationic CD derivatives is applicable in large scale, without the need of chromatographic purification. Their ionic deposition on a solid surface is simple, yet robust and a straightforward process as well.

The ball pen probe (BPP) technique is used successfully to make profile measurements of plasma potential, electron temperature, and radial electric field on the Mega Amp Spherical Tokamak. The potential profile measured by the BPP is shown to significantly differ from the floating potential both in polarity and profile shape. By combining the BPP potential and the floating potential, the electron temperature can be measured, which is compared with the Thomson scattering (TS) diagnostic. Excellent agreement between the two diagnostics is obtained when secondary electron emission is accounted for in the floating potential. From the BPP profile, an estimate of the radial electric field is extracted which is shown to be of the order ∼1 kV/m and increases with plasma current. Corrections to the BPP measurement, constrained by the TS comparison, introduce uncertainty into the ER measurements. The uncertainty is most significant in the electric field well inside the separatrix. The electric field is used to estimate toroidal and poloidal rotation velocities from E × B motion. This paper further demonstrates the ability of the ball pen probe to make valuable and important measurements in the boundary plasma of a tokamak.

A simple inexpensive light-emitting diode (LED)-based fluorescence detector for detection in capillary separations is described. The modular design includes a separate high power LED source, detector head, designed in the epifluorescence arrangement, and capillary detection cells. The detector head and detection cells were printed using a 3D printer and assembled with commercially available optical components. Optical fibers were used for connecting the detector head to the LED excitation source and the photodetector module. Microscope objective or high numerical aperture optical fiber were used for collection of the fluorescence emission from the fused silica separation capillary. As an example, mixture of oligosaccharides labeled by 8-aminopyrene-1,3,6-trisulfonate (APTS) was separated by capillary zone electrophoresis and detected by the described detector. The performance of the detector was compared with both a semiconductor photodiode and photomultiplier as light sensing elements. The main advantages of the 3D printed parts, compared to the more expensive alternatives from the optic component suppliers, include not only cost reduction, but also easy customization of the spatial arrangement, modularity, miniaturization, and sharing of information between laboratories for easy replication or further modifications of the detector. All information and files necessary for printing the presented detector are enclosed in the Supporting Information.

Rheological properties of chitosan acetate solutions containing chitin nanofibrils (n-chitin) and biocompatible plasticizers intended for preparation of biodegradable films are reported in the steady, oscillatory and transient shear flow. The experiments were performed on slurries with an optimum proportion of 65/35 wt.% between chitosan and n-chitin in the films which was determined from our results of mechanical properties and absorption of water vapor. The time-dependent dynamic experiments revealed the chitin nanofibrils as an effective "gelling agent" of chitosan phase. The phenomenon is explained by a chitosan-like surface of n-chitin and by the interactions inducing orientational cooperativity of chitosan molecules dissolved in close neighborhood of the anisotropic chitin nanofibrils. Additions of glycerol or poly(ethylene glycol), improving mechanical properties of the films, delay significantly the onset of gelation of chitosan/n-chitin slurries. The effect is induced by an increase in viscosity of the slurries and by their enhanced chaotropic character., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We study how the size of spherical gold nanoparticles (AuNPs) influences their ability to enhance the response of optical biosensors based on surface plasmon resonance (SPR). We present a theoretical model that relates the enhancement generated by the AuNPs to their composition, size, and concentration, thus allowing for accurate predictions regarding the SPR sensor response to various AuNPs. The effect of the AuNP size is also investigated experimentally using an SPR biosensor for the detection of carcinoembryonic antigen (CEA) in which AuNPs covered with neutravidin (N-AuNPs) are used in the last step of a sandwich assay to enhance the sensor response to biotinylated secondary antibody against CEA. The experimental data are in excellent agreement with the results of the theoretical analysis. We demonstrate that the sensor response enhancement generated by the N-AuNPs is determined by (i) the sensor sensitivity to N-AuNP surface density (Sσ) and (ii) the ability of the N-AuNPs to bind to the functionalized surface of the sensor. Our results indicate that, while Sσ increases with the size of the N-AuNP, the ability of the functionalized surface of the sensor to bind the N-AuNPs is affected by steric effects and decreases with the size of N-AuNP.