Quantitative analysis of constitutive and 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced cytochrome P450 1B1 expression in human lymphocytes.
Exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD or dioxin) results in a broad spectrum of biological responses, including altered metabolism, disruption of normal hormone signaling pathways, reproductive and developmental effects, and cancer. Cytochrome P450 1B1 (CYP1B1) is a dioxin-inducible gene that is active in the formation of 4-hydroxyestradiol, a potentially genotoxic catechol estrogen. Therefore, the analysis of CYP1B1 in humans may be useful in establishing relationships between dioxin exposure and adverse health effects. In this study, we examined the expression of CYP1B1 in human peripheral blood lymphocytes of unexposed individuals using a quantitative reverse transcription-PCR method. Absolute CYP1B1 RNA levels varied more than 30-fold in uncultured mononuclear cells obtained from 10 individuals. In vitro treatment of mitogen-stimulated lymphocytes with TCDD for 1-5 days of culture resulted in a peak induction of CYP1B1 after 3 days. The induction of CYP1B1 RNA levels after 3 days of culture was dose-dependent, exhibited a maximum response above 10 nM TCDD, and varied greatly among different individuals. However, the half maximal dose required for this induction was similar between individuals and comparable to that observed in the MCF-7 and HepG2 human cell lines. These observations indicate that CYP1B1 exhibits variable constitutive expression and is inducible in vitro by TCDD in human lymphocytes and that the magnitude of induction varies within the population. These data define the suitability of CYP1B1 for use as a mechanistically based biomarker in ongoing molecular epidemiological studies of human populations exposed to dioxins and related chemicals that bind the aromatic hydrocarbon receptor.


The ability of the Dementia Rating Scale to predict everyday functioning.
The Dementia Rating Scale (DRS) is a brief neuropsychological assessment battery designed to assess five areas of cognitive functioning in the elderly. The relationship between DRS performance and everyday functioning was examined for 50 psychogeriatric patients. Everyday functioning was assessed with a standardized performance measure examining self-care, safety, money management, cooking, medication administration, and community utilization. Regression analyses were conducted for each of the six functional domains. In addition, correlations between the DRS subscales and the functional areas were computed. Results revealed significant predictive relationships (p <.01) between performance on the DRS and most functional domains. The multiple Rs ranged from .52 - .70, accounting for 27% to 49% of the variance. The Initiation/Perseveration subscale was most heavily weighted in each analysis. Significant correlations were obtained between subdomains of cognition and most functional areas. The findings provide evidence of relationships between DRS performance and functional ability.


Rational Development of a Small-Molecule Activator of CK1γ2 That Decreases C99 and Beta-Amyloid Levels.
Alzheimer's disease (AD) is a debilitating neurodegenerative disorder characterized by the accumulation of β-amyloid (Aβ), C99, and Tau in vulnerable areas of the brain. Despite extensive research, current strategies to lower Aβ levels have shown limited efficacy in slowing the cognitive decline associated with AD. Recent findings suggest that C99 may also play a crucial role in the pathogenesis of AD. Our laboratory has discovered that CK1γ2 phosphorylates Presenilin 1 at the γ-secretase complex, leading to decreased C99 and Aβ levels. Thus, CK1γ2 activation appears as a promising therapeutic target to lower both C99 and Aβ levels. In this study, we demonstrate that CK1γ2 is inhibited by intramolecular autophosphorylation and describe a high-throughput screen designed to identify inhibitors of CK1γ2 autophosphorylation. We hypothesize that these inhibitors could lead to CK1γ2 activation and increased PS1-Ser367 phosphorylation, ultimately reducing C99 and Aβ levels. Using cultured cells, we investigated the impact of these compounds on C99 and Aβ concentrations and confirmed that CK1γ2 activation effectively reduced their levels. Our results provide proof of concept that CK1γ2 is an attractive therapeutic target for AD. Future studies should focus on the identification of specific compounds that can inhibit CK1γ2 autophosphorylation and evaluate their efficacy in preclinical models of AD. These studies will pave the way for the development of novel therapeutics for the treatment of AD.


Inhibition of focal adhesion kinase (FAK) signaling in focal adhesions decreases cell motility and proliferation.
It has been proposed that the focal adhesion kinase (FAK) mediates focal adhesion formation through tyrosine phosphorylation during cell adhesion. We investigated the role of FAK in focal adhesion structure and function. Loading cells with a glutathione-S-transferase fusion protein (GST-Cterm) containing the FAK focal adhesion targeting sequence, but not the kinase domain, decreased the association of endogenous FAK with focal adhesions. This displacement of endogenous FAK in both BALB/c 3T3 cells and human umbilical vein endothelial cells loaded with GST-Cterm decreased focal adhesion phosphotyrosine content. Neither cell type, however, exhibited a reduction in focal adhesions after GST-Cterm loading. These results indicate that FAK mediates adhesion-associated tyrosine phosphorylation, but not the formation of focal adhesions. We then examined the effect of inhibiting FAK function on other adhesion-dependent cell behavior. Cells microinjected with GST-Cterm exhibited decreased migration. In addition, cells injected with GST-Cterm had decreased DNA synthesis compared with control-injected or noninjected cells. These findings suggest that FAK functions in the regulation of cell migration and cell proliferation.


Safety and preliminary efficacy of the Gam-COVID-Vac vaccine and outcomes of SARS-CoV-2 infection in Russian patients with genitourinary malignancies.
To our knowledge, there is no clinical data pertaining to COVID-19 outcomes and safety of COVID-19 vaccination in Russian patients with genitourinary (GU) malignancies. Aim of our analysis was to describe the characteristics of the COVID-19 infection course as well as preliminary safety and efficacy of Gam-COVID-Vac vaccine in patients with active GU malignancies. Patients were retrospectively identified at nine cancer centers in different regions. Patients were included if COVID-19 was diagnosed by a polymerase chain reaction. Data from additional patients with GU cancers who had no positive SARS-CoV-2 RT-PCR test before vaccination and who received two doses of Gam-COVID-Vac (Sputnik V) between 11 February and 31 August 2021 were collected for safety assessment. Anonymized data were collected through an online registry covering demographics, treatments, and outcomes. The Gam-COVID-Vac vaccine was well tolerated; no grade 3-5 toxicities were reported in 112 vaccinated metastatic GU cancer patients. The most common grade 1 adverse events (81%) were injection site reactions (76%), flu-like illness (68%), and asthenia (49%). Five patients experienced grade 2 chills (4.5%) and 3 patients had grade 2 fever (2.7%). With median follow-up of 6.2 months, two COVID-19 cases were confirmed by RT-PCR test in the vaccine group (of 112 participants; 1.8%). Eighty-eight patients with COVID-19 disease were included in the analysis. The average age as of the study enrollment was 66 (range 39-81) and the majority of patients were male with renal cell carcinoma (RCC). Thirty-six patients (41%) had evidence of metastatic disease, of these 22 patients were receiving systemic therapy. More than half of patients required hospitalization. Fifty-four patients (61%) experienced complications. Sixteen patients who developed COVID-19 pneumonia required mechanical ventilator support. Sixteen patients (18%) died in a median of 23.5 days after the date of COVID-19 diagnosis was established. The 3-month survival rate was 82%. Clinical and/or radiographic progression of cancer during COVID-19 infection or the subsequent 3 months was observed in 10 patients (11.4%). Patients with GU malignancies are at increased risk of mortality from COVID-19 infection when compared to the general population. Vaccination could be safe in GU cancer patients. retrospectively registered.


Experiencing versus contemplating: Language use during descriptions of awe and wonder.
Awe and wonder are theorised to be distinct from other positive emotions, such as happiness. Yet little empirical or theoretical work has focused on these emotions. This investigation explored differences in language used to describe experiences of awe and wonder. Such analyses can provide insight into how people conceptualise these emotional experiences, and whether they conceptualise these emotions to be distinct from other positive emotions, and each other. Participants wrote narratives about experiences of awe, wonder and happiness. There were differences in the language used to describe these positive emotional states, consistent with the theorised functions of each emotion. Awe was related to observing the world, reflected in greater use of perception words. Wonder was related to trying to understand the world, reflected in greater use of cognitive complexity and tentative words. Language use for both emotions reflected an environmental focus, whereas language use for happiness reflected a social/relationship focus.


Assessing Early Atherosclerosis by Detecting and Imaging of Hypochlorous Acid and Phosphorylation Using Fluorescence Nanoprobe.
The assessment of early atherosclerosis (AS) is of great significance for the early diagnosis and mechanism research. Herein, a novel nanoprobe PCN@FL is developed to realize the simultaneous detection and imaging of phosphorylation and hypochlorous acid (HClO). The selective recognition of HClO is achieved through the specific interaction between DMTC and HClO, while the levels of phosphorylation are detected via the specific interaction between Zr (IV) and phosphates. The nanoprobe can be utilized to monitor the fluctuations in HClO and phosphate in early atherosclerosis. It is observed that the levels of HClO and phosphate in the serum of early AS mice are higher than those of the normal mice. Ultimately, the levels of hypochlorous acid and phosphorylation in the inner wall of aortic vessels are imaged by two-photon microscope. The results show that the levels of HClO and phosphorylation in the early atherosclerotic mice are significantly higher than those of in normal mice. The nanoprobe provides a suitable fluorescent tool for simultaneous detection and imaging of HClO and phosphorylation, which holds promise for early atherosclerotic disease assessment.


In vitro evaluation of a soluble Leishmania promastigote surface antigen as a potential vaccine candidate against human leishmaniasis.
PSA (Promastigote Surface Antigen) belongs to a family of membrane-bound and secreted proteins present in several Leishmania (L.) species. PSA is recognized by human Th1 cells and provides a high degree of protection in vaccinated mice. We evaluated humoral and cellular immune responses induced by a L. amazonensis PSA protein (LaPSA-38S) produced in a L. tarentolae expression system. This was done in individuals cured of cutaneous leishmaniasis due to L. major (CCLm) or L. braziliensis (CCLb) or visceral leishmaniasis due to L. donovani (CVLd) and in healthy individuals. Healthy individuals were subdivided into immune (HHR-Lm and HHR-Li: Healthy High Responders living in an endemic area for L. major or L. infantum infection) or non immune/naive individuals (HLR: Healthy Low Responders), depending on whether they produce high or low levels of IFN-γ in response to Leishmania soluble antigen. Low levels of total IgG antibodies to LaPSA-38S were detected in sera from the studied groups. Interestingly, LaPSA-38S induced specific and significant levels of IFN-γ, granzyme B and IL-10 in CCLm, HHR-Lm and HHR-Li groups, with HHR-Li group producing TNF-α in more. No significant cytokine response was observed in individuals immune to L. braziliensis or L. donovani infection. Phenotypic analysis showed a significant increase in CD4+ T cells producing IFN-γ after LaPSA-38S stimulation, in CCLm. A high positive correlation was observed between the percentage of IFN-γ-producing CD4+ T cells and the released IFN-γ. We showed that the LaPSA-38S protein was able to induce a mixed Th1 and Th2/Treg cytokine response in individuals with immunity to L. major or L. infantum infection indicating that it may be exploited as a vaccine candidate. We also showed, to our knowledge for the first time, the capacity of Leishmania PSA protein to induce granzyme B production in humans with immunity to L. major and L. infantum infection.


Using an RNA Aptamer to Inhibit the Action of Effector Proteins of Plant Pathogens.
In previous work, we experimentally demonstrated the possibility of using RNA aptamers to inhibit endogenous protein expression and their function within plant cells In the current work, we show that our proposed method is suitable for inhibiting the functions of exogenous, foreign proteins delivered into the plant via various mechanisms, including pathogen proteins. Stringent experimentation produced robust RNA aptamers that are able to bind to the recombinant HopU1 effector protein of P. syringae bacteria. This research uses genetic engineering methods to constitutively express/transcribe HopU1 RNA aptamers in transgenic A. thaliana. Our findings support the hypothesis that HopU1 aptamers can actively interfere with the function of the HopU1 protein and thereby increase resistance to phytopathogens of the genus P. syringae pv. tomato DC 3000.


Platelet-derived growth factor in chemotactic for fibroblasts.
Chemotaxis assays in modified Boyden chambers were used to detect fibroblast chemoattractants in materials released from early-stage inflammatory cells, namely, mast cells, platelets, and neutrophils. Strong attractant activity was found in substances released from platelets. This activity was accounted for mainly by the platelet-derived growth factor (PDGF), which is released from the platelets and which was active as a chemoattractant at 0.5-1.0 mitogenic units/ml. The mitogenic activity of purified PDGF, measured by [3H]thymidine incorporation, occurs at a similar concentration range. By varying the gradient of PDGF, we demonstrated that PDGF stimulates chemotaxis rather than random motility. Preincubation of suspensions of fibroblasts in the presence of PDGF decreased the subsequent migration of cells to a gradient of PDGF as well as to a gradient of fibronectin, which is also in attractant for fibroblasts. The chemotactic response of fibroblasts to PDGF was not inhibited by hydroxyurea or azidocytidine but was inhibited by actinomycin D and cycloheximide, suggesting that synthesis of RNA and proteins but not of DNA is required for the chemotactic response to occur. Fibroblast growth factor, epidermal growth factor, nerve growth factor, and insulin were not chemotactic for human skin fibroblasts, suggesting that the chemoattractant activity of PDGF for fibroblasts is not a general property of growth factors and mitogens. These results suggest that PDGF could have two functions in wound healing: to attract fibroblasts to migrate into the clot and then to induce their proliferation.


Long-term results following cranial hydroxyapatite prosthesis implantation in a large skull defect model.
A large skull defect may occur after different events such as trauma, tumor resection, and vascular injuries. There is still some doubt about the best material to use for reconstruction. Hydroxyapatite ceramic is one of the materials in use, and its biocompatibility and osteoconductivity are well established. This study evaluated the interaction of a commercial hydroxyapatite custom-made prosthesis implanted in a large skull defect, to assess its osteointegration and its habitability with newly formed bone over time. Ten sheep underwent craniectomy and reconstruction of the skull defect with a porous hydroxyapatite cranial prosthesis. The animals were divided into two groups: animals in group A were euthanized after 6 months and animals in group B were euthanized after 12 months. At the end of the experimental periods, each implant was evaluated macroscopically and radiologically, and analyzed by micro-computed tomography, histology, histomorphometry, and microhardness techniques. During the study, no adverse events occurred, and there was no evidence of inflammation or negative tissue reactions. Histology and histomorphometry showed new bone formation inside the implant in both experimental periods; newly formed bone had increased significantly (p < 0.05) by over 300 percent between 6 and 12 months. Three-dimensional micro-computed tomographic analysis showed new bone formation and material remodeling. Microhardness analysis indicated that the mineralization process and the mechanical properties of newly formed bone were not altered. The hydroxyapatite prosthesis showed its osteoconductivity and good biocompatibility. A low rate of fibrous tissue formation and a high rate of bony regeneration were found.


Preventing HIV outbreaks among people who inject drugs in the United States: plus ça change, plus ça même chose.
: This editorial review covers current trends in the epidemiology of HIV among people who inject drugs (PWID) in the United States, including four recent HIV outbreaks. We discuss gaps in the prevention and treatment cascades for HIV and medications for opioid disorder and propose lessons learned to prevent future HIV outbreaks. Over the last decade, North America has been in the throes of a major opioid epidemic, due in part to over-prescribing of prescription opiates, followed by increasing availability of cheap heroin, synthetic opioids (e.g. fentanyl), and stimulants (e.g. methamphetamine). Historically, HIV infection among PWID in the US had predominantly affected communities who were older, urban and Black. More recently, the majority of these infections are among younger, rural or suburban and Caucasian PWID. All four HIV outbreaks were characterized by a high proportion of women who inject drugs and underlying socioeconomic drivers such as homelessness and poverty. We contend that the US response to the HIV epidemic among PWID has been fractured. A crucial lesson is that when evidence-based responses to HIV prevention are undermined or abandoned because of moral objections, untold humanitarian and financial costs on public health will ensue. Restructuring a path forward requires that evidence-based interventions be integrated and brought to scale while simultaneously addressing underlying structural drivers of HIV and related syndemics. Failing to do so will mean that HIV outbreaks among PWID and the communities they live in will continue to occur in a tragic and relentless cycle.


[Life and working conditions among physicians at a university hospital].
This paper evaluated the life and working conditions of physicians who worked in a university hospital. Two questionnaires, one for occupational data and the Medical Outcomes Study 36 - Item Short-Form Health Survey (SF-36) were applied. 91 surgeons and 96 physicians, mainly of the masculine sex, participated in the inquiry. The average age among surgeons was 44.5 and among physicians, 46. The average of weekly working hours was 51.8 hours among the surgeons and 44.1 among the physicians. 87.9% of the surgeons and 51.6% physicians also mentioned working in other hospitals. Only in Physical Aspects there was a significant statistical difference between the groups, with lower rates among physicians.


Effects of Elevated Temperature on Pisum sativum Nodule Development: II-Phytohormonal Responses.
High temperature is one of the most important factors limiting legume productivity. We have previously shown the induction of senescence in the apical part of nodules of the pea SGE line, formed by Rhizobium leguminosarum bv. viciae strain 3841, when they were exposed to elevated temperature (28 °C). In this study, we analyzed the potential involvement of abscisic acid (ABA), ethylene, and gibberellins in apical senescence in pea nodules under elevated temperature. Immunolocalization revealed an increase in ABA and 1-aminocyclopropane-1-carboxylic acid (ACC, the precursor of ethylene biosynthesis) levels in cells of the nitrogen fixation zone in heat-stressed nodules in 1 day of exposure compared to heat-unstressed nodules. Both ABA and ethylene appear to be involved in the earliest responses of nodules to heat stress. A decrease in the gibberellic acid (GA3) level in heat-stressed nodules was observed. Exogenous GA3 treatment induced a delay in the degradation of the nitrogen fixation zone in heat-stressed nodules. At the same time, a decrease in the expression level of many genes associated with nodule senescence, heat shock, and defense responses in pea nodules treated with GA3 at an elevated temperature was detected. Therefore, apical senescence in heat-stressed nodules is regulated by phytohormones in a manner similar to natural senescence. Gibberellins can be considered as negative regulators, while ABA and ethylene can be considered positive regulators.


Harmonic analysis of DNA dynamics in a viscous medium.
The harmonic dynamics of normal modes of double-stranded DNA in a viscous fluid are investigated. The model DNA consists of two backbone-supported DNA strands coiling around a common helix axis with base stacking, sugar puckering, interstrand hydrogen bonding, and intrastrand sugar-base interactions assigned values based on published data. Assuming that the DNA bases are shielded from direct bombardment by the solvent, analytical solutions are obtained. The dissipation and fluctuation of the normal modes of the bases moving along the spirals display the effect of the medium indirectly through interactions with the backbone. The dynamics of the backbone are found to be overdamped with the characteristic damping times extending to the picosecond region for disturbance in position and to the sub-picosecond region for disturbance in velocity. In addition to the dynamic mode of a rigid rod, the motions of the bases are coupled to the motions of the backbone with comparable amplitudes for disturbance in position. For disturbance in velocity, however, the bases are effectively at rest, not being able to follow the motions of the backbone. The angular frequencies of the underdamped vibrational modes, identified as the ringing modes of the bases with the backbone effectively at rest, are insensitive to the viscosity and lie in the low frequency region of the Raman spectrum. These findings indicate that the backbone of DNA plays a significant role in modulating the dynamics of double-stranded DNA in an overdamping environment. This modulation of the dynamics of the motions of the bases in DNA by environmental impediments to molecular motion is briefly discussed in connection with protein- and drug- DNA interactions as well as gene regulation.


Seroprevalence and incidence of hepatitis E virus infection in the general population of Iwate prefecture, Japan: A retrospective cohort study.
Hepatitis E virus (HEV) causes subclinical or acute self-limiting hepatitis. We surveyed the current seroprevalence and incidence of HEV infection among the general population in Iwate Prefecture, Japan, where the endemic infection is presumed to be low. Between 2014 and 2016, we recruited individuals from Iwate Prefecture, Japan, who visited a general medical work-up program. Serum anti-HEV antibody and HEV RNA were measured twice, with an interval of 2 years. Anti-HEV antibody was measured with enzyme-linked immunosorbent assay and HEV RNA with reverse transcription-polymerase chain reaction. Study participants comprised 1284 Japanese (650 men and 634 women) with age ranging 20-89 years. A total of 90 participants were found to be positive for anti-HEV immunoglobulin G on the first visit, with a prevalence of 7.0% (95% confidence interval [CI] 5.6%-8.4%). Seroprevalence was higher in men than in women (10.1% vs. 3.7%, p < 0.001), and in those aged in their 50s-80s than in those aged in their 20s-40s (p = 0.006). Positive seroconversion indicating new HEV infection was found in seven of 1194 seronegative participants (0.59%; 95% CI 0.15%-1.0%), indicating the incidence of HEV infection to be 272 per 100 000 person-years (95% CI 109-561). Our observations suggest that the incidence of HEV infection is high and that it is a leading cause of hepatitis virus infection in Iwate Prefecture, Japan.


Insulin effects on glucose tolerance, hypermetabolic response, and circadian-metabolic protein expression in a rat burn and disuse model.
Insulin controls hyperglycemia after severe burns, and its use opposes the hypermetabolic response. The underlying molecular mechanisms are poorly understood, and previous research in this area has been limited because of the inadequacy of animal models to mimic the physiological effects seen in humans with burns. Using a recently published rat model that combines both burn and disuse components, we compare the effects of insulin treatment vs. vehicle on glucose tolerance, hypermetabolic response, muscle loss, and circadian-metabolic protein expression after burns. Male Sprague-Dawley rats were assigned to three groups: cage controls (n = 6); vehicle-treated burn and hindlimb unloading (VBH; n = 11), and insulin-treated burn and hindlimb unloading (IBH; n = 9). With the exception of cage controls, rats underwent a 40% total body surface area burn with hindlimb unloading, then IBH rats received 12 days of subcutaneous insulin injections (5 units·kg(-1)·day(-1)), and VBH rats received an equivalent dose of vehicle. Glucose tolerance testing was performed on day 14, after which blood and tissues were collected for analysis. Body mass loss was attenuated by insulin treatment (VBH = 265 ± 17 g vs. IBH = 283 ± 14 g, P = 0.016), and glucose clearance capacity was increased. Soleus and gastrocnemius muscle loss was decreased in the IBH group. Insulin receptor substrate-1, AKT, FOXO-1, caspase-3, and PER1 phosphorylation was altered by injury and disuse, with levels restored by insulin treatment in almost all cases. Insulin treatment after burn and during disuse attenuated the hypermetabolic response, increased glucose clearance, and normalized circadian-metabolic protein expression patterns. Therapies aimed at targeting downstream effectors may provide the beneficial effects of insulin without hypoglycemic risk.


Anti-inflammatory mechanism of galangin in lipopolysaccharide-stimulated microglia: Critical role of PPAR-γ signaling pathway.
Since microglia-associated neuroinflammation plays a pivotal role in the progression of neurodegenerative diseases, controlling microglial activation has been suggested as a potential therapeutic strategy. Here, we investigated the anti-inflammatory effects of galangin (3,5,7-trihydroxyflavone) in microglia and analyzed the underlying molecular mechanisms. Galangin inhibited the expression of inducible nitric oxide synthase (iNOS) and pro-inflammatory cytokines and enhanced the expression of anti-inflammatory interleukin (IL)-10 in lipopolysaccharide (LPS)-stimulated BV2 microglia. Galangin also suppressed microglial activation and the expression of pro-inflammatory markers in LPS-injected mouse brains. The results of mechanistic studies have shown that galangin inhibited LPS-induced phosphorylation of p38 mitogen activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK), phosphatidylinositol 3-kinase (PI3K)/Akt, and nuclear factor (NF)-κB activity. On the contrary, galangin increased the activity of transcription factors, such as nuclear factor-E2-related factor 2 (Nrf2), cAMP response element-binding protein (CREB), and peroxisome proliferator-activated receptor (PPAR)-γ, known to play an anti-inflammatory role. In addition, galangin showed antioxidant effects by suppressing the expression of NADPH oxidase subunits p47phox and gp91phox, and by enhancing hemeoxygenase-1. We then investigated whether PPAR-γ was involved in the anti-inflammatory function of galangin. Pretreatment with a PPAR-γ antagonist or siRNA significantly blocked galangin-mediated upregulation of IL-10 and attenuated the inhibition of tumor necrosis factor (TNF)-α, nitric oxide (NO), and IL-6 in LPS-stimulated microglia. Moreover, the PPAR-γ antagonist reversed the effects of galangin on NF-κB, Nrf2, and CREB. Altogether, our data suggest that PPAR-γ plays a key role in mediating the anti-inflammatory effects of galangin by modulating the NF-κB and Nrf2/CREB signaling pathways.


Development of a gastrointestinal tract microscale cell culture analog to predict drug transport.
Microscale cell culture analogs (microCCAs) are used to study the metabolism and toxicity of a chemical or drug. These in vitro devices are physical replicas of physiologically based pharmacokinetic models that combine microfabrication and cell culture. The goal of this project is to add an independent GI tract microCCA to a multi-chamber chip microCCA representing the primary circulation. The GI tract microCCA consists of two chambers separated by a microporous membrane on which intestinal epithelial cells are cultured. Compounds of interest are pumped through the top chamber, allowing drug to be absorbed through the epithelial layer and circulated into the chip microCCA. The chip and GI tract microCCAs have been used to recreate the toxic effects of acetaminophen. Preliminary results have shown that the GI tract microCCA acts as a barrier to drugs entering the chip, mimicking in vivo function in this regard.


Intrauterine Growth Restriction: Effects on Neural Precursor Cell Proliferation and Angiogenesis in the Foetal Subventricular Zone.
Exposure to adverse prenatal factors can result in abnormal brain development, contributing to the aetiology of several neurological disorders. Intrauterine insults could occur during neurogenesis and gliogenesis, disrupting these events. Here we investigate the effects of chronic placental insufficiency (CPI) on cell proliferation and the microenvironment in the subventricular zone (SVZ). At 30 days of gestation (DG; term ∼67 DG), CPI was induced in pregnant guinea pigs via unilateral uterine artery ligation to produce growth-restricted (GR) foetuses (n = 7); controls (n = 6) were from the unoperated horn. At 60 DG, foetal brains were stained immunohistochemically to identify proliferating cells (Ki67), immature neurons (polysialylated neuronal cell adhesion molecule), astrocytes (glial fibrillary acidic protein), microglia (ionised calcium-binding adaptor molecule-1, Iba-1) and the microvasculature (von Willebrand factor) in the SVZ. There was no overall difference (p > 0.05) in the total number of Ki67-immunoreactive (IR) cells, the percentage of SVZ occupied by blood vessels or the density of Iba-1-IR microglia in control versus GR foetuses. However, regression analysis across both groups revealed that both the number of Ki67-IR cells and the percentage of SVZ occupied by blood vessels in the ventral SVZ were negatively correlated (p < 0.05) with brain weight. Furthermore, in the SVZ (dorsal and ventral) the density of blood vessels positively correlated (p < 0.05) with the number of Ki67-IR cells. Double-labelling immunofluorescence suggested that the majority of proliferating cells were likely to be neural precursor cells. Thus, we have demonstrated an association between angiogenesis and neurogenesis in the foetal neurogenic niche and have identified a window of opportunity for the administration of trophic support to enhance a neuroregenerative response.


Integrating peptides' sequence and energy of contact residues information improves prediction of peptide and HLA-I binding with unknown alleles.
The HLA (human leukocyte antigen) class I is a kind of molecule encoded by a large family of genes and is characteristic of high polymorphism. Now the number of the registered HLA-I molecules has exceeded 3000. Slight differences in the amino acid sequences of HLAs would make them bind to different sets of peptides. In the past decades, although many methods have been proposed to predict the binding between peptides and HLA-I molecules and achieved good performance, most experimental data used by them is limited to the HLAs with a small number of alleles. Thus they are inclined to obtain high prediction accuracy only for data with similar alleles. Because the peptides and HLAs together determine the binding, it's necessary to consider their contribution meanwhile. By taking into account the features of the peptides sequence and the energy of contact residues, in this paper a method based on the artificial neural network is proposed to predict the binding of peptides and HLA-I even when the HLAs' potential alleles are unknown. Two experiments in the allele-specific and super-type cases are performed respectively to validate our method. In the first case, we collect 14 HLA-A and 14 HLA-B molecules on Bjoern Peters dataset, and compare our method with the ARB, SMM, NetMHC and other 16 online methods. Our method gets the best average AUC (Area under the ROC) value as 0.909. In the second one, we use leave one out cross validation on MHC-peptide binding data that has different alleles but shares the common super-type. Compared to gold standard methods like NetMHC and NetMHCpan, our method again achieves the best average AUC value as 0.847. Our method achieves satisfactory results. Whenever it's tested on the HLA-I with single definite gene or with super-type gene locus, it gets better classification accuracy. Especially, when the training set is small, our method still works better than the other methods in the comparison. Therefore, we could make a conclusion that by combining the peptides' information, HLAs amino acid residues' interaction information and contact energy, our method really could improve prediction of the peptide HLA-I binding even when there aren't the prior experimental dataset for HLAs with various alleles.


Electrophysiologic evaluation of psychogenic movement disorders.
Psychogenic movement disorders (PMD) are a group of disorders which are in the border zone between neurology and psychiatry. All necessary laboratory investigations should be done to rule out an underlying organic disorder. While clinical acumen of a trained movement disorder specialist may be sufficient to diagnose most PMD, there are clinical situations where electrophysiological tests are required either to rule out an organic movement disorder or even diagnose a PMD. Current electrophysiological test are most useful for tremor, followed by jerks and least for spasms or dystonia. Commonly used electrophysiologic tests include multichannel surface electromyography (EMG), accelerometry, electroencephalography time locked with EMG, premovement potential (Bereitschaftspotential), and somatosensory evoked potentials. Psychogenic tremor is a low frequency tremor with variable frequency and duration of EMG bursts, entrainable, has a high coherence with voluntary movements, and presence of coactivation sign. Patients with psychogenic jerks have well organized triphasic pattern of activation of agonist and antagonist muscles. The jerks are associated with EMG bursts of long duration (usually > 70 ms), long and variable latencies in stimulus induced jerks, absence of craniocaudal pattern of muscle recruitment in apparent startle response, and often a Breitschaftspotential (premovement potential) precedes the jerk. Electrophysiological characterization of psychogenic dystonia is difficult and the tests are usually performed to rule out organic dystonia with characteristic findings. Finally, caution should be exerted in interpreting the electrophysiological tests as both false positive and false negative diagnosis of PMD may still occur.


Which is the Ideal Breast Size?: Some Social Clues for Plastic Surgeons.
To provide plastic surgeons with more detailed information as to factors affecting the perception of female attractiveness, the present study was aimed to investigate whether the interaction effect of breast and body size on ratings of female attractiveness is moderated by sociodemographic variables and whether ratings of shapeliness diverge from those of attractiveness.A community sample of 958 Italian participants rated the attractiveness and the shapeliness of 15 stimuli (5 breast sizes × 3 body sizes) in which frontal, 3/4, and profile views of the head and torso of a faceless woman were jointly shown.Bigger breast sizes obtained the highest attractiveness ratings, but the breast-by-body size interaction was also significant. Evidence was found of a moderator role of sex, marital status, and age. When the effects of breast and body size and their interaction had been ruled out, sex differences were at best very slight and limited to very specific combinations of breast and body sizes. Ratings of attractiveness and shapeliness were highly correlated and did not significantly differ.Results suggest that to address women's psychological needs, concerns, and expectations about their appearance, plastic surgeons should not simply focus on breast size but should carefully consider the 'big picture': the body in its entirety.


A plant oxylipin, 12-oxo-phytodienoic acid, inhibits proliferation of human breast cancer cells by targeting cyclin D1.
Cyclin D1 overexpression has been associated with poor prognosis and resistance to therapy in human breast cancer. Thus, the development of therapeutic agents that selectively target cyclin D1 activity is of clinical interest. This study demonstrates that 12-oxo-phytodienoic acid (OPDA), a phytohormone with critical functions in growth and development in plants, induces growth arrest in MDA-MB-231 and T47D breast cancer cells. In response to OPDA treatment, the human breast cancer cell lines exhibit a progressive decline in cyclin D1 expression, which is tightly associated with the accumulation of hypophosphorylated form of the retinoblastoma protein (Rb) and G1 arrest. The decrease in cyclin D1 protein expression accompanies a dramatic decline in nuclear but not membranous beta-catenin expression and activation of glycogen synthase kinase-3-beta (GSK3beta) caused by inhibition of its serine-9 phosphorylation. The proteasome inhibitor MG132 blocks OPDA-mediated decrease in cyclin D1. In addition, the overexpression of T286A, a cyclin D1 mutant which is refractory to phosphorylation by GSK3beta and proteosomal degradation, is resistant to OPDA-mediated Rb dephosphorylation as well as G(1) cell cycle arrest. Thus, our results demonstrate that degradation of cyclin D1 protein is a key event in OPDA induced growth inhibition in breast cancer cells. These data provide the basic foundation for future efforts to develop OPDA-based approaches in the prevention and treatment of breast cancer and other types of cancer.


Basic dyes for the staining of DNA in mammalian tissues and absorption spectra of stained nuclei in the visible light.
This communication presents informations on the staining of DNA with basic dyes, such as ethyl violet, Janus green, a new red dye obtained from Janus green, and Giemsa, belonging to different chemical groups. It has been found that DNA of tissue sections from which RNA has been extracted selectively with cold concentrated phosphoric acid can be stained with aqueous solutions of these dyes. Further, DNA of tissue sections from which RNA has been extracted and the sections then hydrolysed in 6 N HCl at 28 degrees C for 15 min can also be stained with aqueous solutions of ethyl violet, Janus green, and Giemsa. Moreover, tissue sections that have been hydrolysed in 6 N HCl and then stained with aqueous solutions of these dyes, including the new red dye obtained from Janus green, reveal well-stained nuclei. The absorption spectra of nuclei stained with aqueous solutions of ethyl violet, Janus green and Giemsa, following the above stated three procedures of staining have been presented. Absorption data of nuclei stained with the new red dye after Feulgen hydrolysis of tissue sections as well as those of nuclei in tissue sections from which RNA has been extracted have also been presented herein. The implications of these findings have been discussed.


Design of the Study of Tranexamic Acid during Air Medical Prehospital Transport (STAAMP) Trial: Addressing the Knowledge Gaps.
Abstract Hemorrhage and coagulopathy remain major drivers of early preventable mortality in military and civilian trauma. The development of trauma-induced coagulopathy and hyperfibrinolysis is associated with poor outcomes. Interest in the use of tranexamic acid (TXA) in hemorrhaging patients as an antifibrinolytic agent has grown recently. Additionally, several reports describe immunomodulatory effects of TXA that may confer benefit independent of its antifibrinolytic actions. A large trial demonstrated a mortality benefit for early TXA administration in patients at risk for hemorrhage; however, questions remain about the applicability in developed trauma systems and the mechanism by which TXA reduces mortality. We describe here the rationale, design, and challenges of the Study of Tranexamic Acid during Air Medical Prehospital transport (STAAMP) trial. The primary objective is to determine the effect of prehospital TXA infusion during air medical transport on 30-day mortality in patients at risk of traumatic hemorrhage. This study is a multicenter, placebo-controlled, double-blind, randomized clinical trial. The trial will enroll trauma patients with hypotension and tachycardia from 4 level I trauma center air medical transport programs. It includes a 2-phase intervention, with a prehospital and in-hospital phase to investigate multiple dosing regimens. The trial will also explore the effects of TXA on the coagulation and inflammatory response following injury. The trial will be conducted under exception for informed consent for emergency research and thus required an investigational new drug approval from the U.S. Food and Drug Administration as well as a community consultation process. It was designed to address several existing knowledge gaps and research priorities regarding TXA use in trauma.


Targeting molecular pathways with camptothecin as novel therapy for gastric cancer.
Novel chemotherapeutic agents are needed to treat gastric cancer for which the prognosis remains dismal. The antitumor alkaloid camptothecin (CPT) may be useful in the treatment of certain solid tumors; however, its effects on gastric cancer are largely undefined. The purpose of our study was to characterize the effects of CPT on human gastric tumors in vivo and to determine the cellular mechanisms involved in CPT-mediated inhibition. Two human gastric cancers, WIL and TOR, were transplanted subcutaneously into athymic nude mice. After tumors reached 50 to 100 mm(2), mice were randomized into three groups to receive injections of either low-dose CPT (5 mg/kg), high-dose CPT (10 mg/kg), or vehicle (control) intraperitoneally 3 days a week for 3 weeks. Tumors were measured and weighed, and protein levels of the cell cycle inhibitor, p21Waf1/Cip1, and the antiapoptotic protein, Bcl-2, were assessed. Both dosages of CPT significantly inhibited growth of WIL and TOR gastric tumors. CPT (10 mg/kg) reduced tumor size compared to baseline, establishing this as a tumoricidal dosage. Treatment with CPT was associated with increased levels of p21Waf1/Cip1 and decreased levels of Bcl-2. CPT effectively kills human gastric cancers associated with increased levels of p21Waf1/Cip1 and decreased levels of Bcl-2. By activating cell cycle withdrawal and cell death through induction of p21Waf1/Cip1 and downregulation of Bcl-2, CPT may be an effective agent for gastric cancer.


Imunovir in the treatment of immunodepression of diverse etiology.
Immunodepression associated with a variety of situations such as cancer or any of its major modalities of treatment (surgery, irradiation, or chemotherapy) has been effectively alleviated with Imunovir (inosine pranobex-BAN), and this has been associated with demonstrable clinical benefit to these patients. One hundred and six immunodepressed patients with solid tumors undergoing radiotherapy were treated with either Imunovir or placebo; 64% of Imunovir-treated patients were immunorestored after 3 months compared to 23% in the placebo group. Imunovir was also effectively used in 75 patients with malignant hematological disorders both as an immunorestorative agent given prophylactically to prevent infection and as a therapeutic agent to treat infections in these immunodepressed patients. In different studies involving surgical patients treated with either Imunovir or placebo, 70-81% of hypoergic or anergic patients in the Imunovir group became normoergic by day 14 of treatment compared to 5-17% of the placebo group, and this enhanced immunorestoration was associated with lower incidence of local sepsis (P less than 0.05), systemic sepsis (P less than 0.025), and postoperative mortality (P less than 0.05).


Microglia induce neurogenic protein expression in primary cortical cells by stimulating PI3K/AKT intracellular signaling in vitro.
Emerging evidence suggests that microglia can support neurogenesis. Little is known about the mechanisms by which microglia regulate the cortical environment and stimulate cortical neurogenesis. We used an in vitro co-culture model system to investigate the hypothesis that microglia respond to soluble signals from cortical cells, particularly following mechanical injury, to alter the cortical environment and promote cortical cell proliferation, differentiation, and survival. Analyses of cortical cell proliferation, cell death, neurogenic protein expression, and intracellular signaling were performed on uninjured and injured cortical cells in co-culture with microglial cell lines. Microglia soluble cues enhanced cortical cell viability and proliferation cortical cells. Co-culture of injured cortical cells with microglia significantly reduced cell death of cortical cells. Microglial co-culture significantly increased Nestin + and α-internexin + cortical cells. Multiplex ELISA and RT-PCR showed decreased pro-inflammatory cytokine production by microglia co-cultured with injured cortical cells. Inhibition of AKT phosphorylation in cortical cells blocked microglial-enhanced cortical cell viability and expression of neurogenic markers in vitro. This in vitro model system allows for assessment of the effect of microglial-derived soluble signals on cortical cell viability, proliferation, and stages of differentiation during homeostasis or following mechanical injury. These data suggest that microglia cells can downregulate inflammatory cytokine production following activation by mechanical injury to enhance proliferation of new cells capable of neurogenesis via activation of AKT intracellular signaling. Increasing our understanding of the mechanisms that drive microglial-enhanced cortical neurogenesis during homeostasis and following injury in vitro will provide useful information for future primary cell and in vivo studies.


Family physicians' management of genetic aspects of a cardiac disease: a scenario-based study from slovenia.
The aim of this study was to find out how Slovenian family physicians (FPs) would manage a hypothetical clinical case, to explore their views about possible ethical dilemmas associated with this clinical case and to determine possible associations with demographic and other characteristics of FPs. This was an observational cross-sectional postal study in the Slovenian FPs' surgeries. The study population consisted of the whole population of Slovenian FPs (n = 950). The main outcome measures were the percentages of the answers of FPs on different questions about the clinical case on the management of patient and his relative with hereditary cardiomyopathy. There were 271 FPs who answered the questionnaire (response rate was 27.1%). A sample included 66 (24.4%) men and the mean age of all respondents was 45.5 ± 10.6 years. When dealing with the clinical case, most FPs expressed willingness to take the patient's family history. Only 34.2% FPs did not believe that ordering genetic tests was part of their job. Additionally, only 50.0% of them felt competent to interpret the genetic risk, 25.0% of them would give information about genetic testing and only 6.0% would interpret the results of the genetic testing. Family physicians in Slovenia were willing to include genetic tasks into routine management of their patients, but they do not feel competent enough to interpret the genetic risks and the results of genetic testing. However, an important part of FPs would not refer patients at risk to genetic counseling. The inclusion of genetic topics to family medicine specialization curriculum is needed.


Is resistant hypertension an independent predictor of all-cause mortality in individuals with type 2 diabetes? A prospective cohort study.
Resistant hypertension is independently associated with an increased risk of death in the general hypertensive population. We assessed whether resistant hypertension is an independent predictor of all-cause mortality in individuals with type 2 diabetes from the Renal Insufficiency And Cardiovascular Events (RIACE) Italian Multicentre Study. On 31 October 2015, vital status information was retrieved for 15,656 of the 15,773 participants enrolled in 2006-2008. Based on baseline blood pressure (BP) values and treatment, participants were categorized as normotensive, untreated hypertensive, controlled hypertensive (i.e., on-target with < 3 drugs), uncontrolled hypertensive (i.e., not on-target with 1-2 drugs), or resistant hypertensive (i.e., uncontrolled with > 3 drugs or controlled with > 4 drugs). Kaplan-Meier and Cox proportional hazards regression analyses were used to assess the association with all-cause mortality. Using the 130/80 mmHg targets for categorization, crude mortality rates and Kaplan-Meier estimates were highest among resistant hypertension participants, especially those with controlled resistant hypertension. As compared with resistant hypertension, risk for all-cause mortality was significantly lower for all the other groups, including individuals with controlled hypertension (hazard ratio 0.81 [95% confidence interval 0.74-0.89], P < 0.0001), but became progressively similar between resistant and controlled hypertension after adjustment for cardiovascular risk factors and complications/comorbidities. Also when compared with controlled resistant hypertension, mortality risk was significantly lower for all the other groups, including controlled hypertension, even after adjusting for cardiovascular risk factors (0.77 [0.63-0.95], P = 0.012), but not for complications/comorbidities (0.88 [0.72-1.08], P = 0.216). BP was well below target in the controlled hypertensive groups (resistant and non-resistant) and values < 120/70 mmHg were associated with an increased mortality risk. Results changed only partly when using the 140/90 mmHg targets for categorization. In the RIACE cohort, at variance with the general hypertensive population, resistant hypertension did not predict death beyond target organ damage. Our findings may be explained by the high mortality risk conferred by type 2 diabetes and the low BP values observed in controlled hypertensive patients, which may mask risk associated with resistant hypertension. Less stringent BP goals may be preferable in high-risk patients with type 2 diabetes. ClinicalTrials.gov, NCT00715481 , retrospectively registered 15 July, 2008.


Blood pressure regulation during the aging process: the end of the 'hypertension era'?
The elderly blood pressure paradigm reflects all of the demographic, technological and therapeutic changes over the past 20-30 years that make it now possible to propose a more integrative approach of 'hypertension'. The aim of the present review was to address what does measured blood pressure really mean and what are its determinants during the aging process. We show that standard blood pressure measurements are not adequate or even misleading for the evaluation of cardiovascular risk especially in the elderly patients and that there is a necessity of a transition to a new approach in determining the arterial risk. Direct arterial measurements including analysis of central and peripheral arterial waveforms and assessment of pulse wave velocity can be reliable and easily performed measurements as an alternative to blood pressure-Korotkoff approach. For these measurements, there are currently sufficient clinical data showing their association with cardiovascular risk. There is also the emergence of reference values and beneficial elements of regression by treatment. This new approach as well as recent knowledge on vascular hemodynamics and biology, represent the swan song for the hypertension concept as defined by blood pressure values.


Translocation of an Anteater (Tamandua tetradactyla) Infected with Rabies from Virginia to Tennessee Resulting in Multiple Human Exposures, 2021.
On August 16, 2021, the Tennessee Department of Health (TDH) was notified of a positive rabies test result from a South American collared anteater (Tamandua tetradactyla) in Washington County, Tennessee. Tamanduas, or lesser anteaters, are a species of anteater in which rabies has not previously been reported. The animal was living at a Tennessee zoo and had been recently translocated from a zoo in Virginia. TDH conducted an investigation to confirm the rabies result, characterize the rabies variant, and ascertain an exposure risk assessment among persons who came into contact with the tamandua. Risk assessments for 22 persons were completed to determine the need for rabies postexposure prophylaxis (rPEP); rPEP was recommended for 13 persons, all of whom agreed to receive it. Using phylogenetic results of the virus isolated from the tamandua and knowledge of rabies epidemiology, public health officials determined that the animal was likely exposed to wild raccoons present at the Virginia zoo. This report describes expansion of the wide mammalian species diversity susceptible to rabies virus infection and summarizes the investigation, highlighting coordination among veterinary and human public health partners and the importance of preexposure rabies vaccination for animal handlers and exotic zoo animals.


Recognition of risk factors and prognostic indicators in Fournier's gangrene.
Fournier's gangrene is a rare but rapidly progressive necrotizing infection that results from the synergistic action of multiple aerobic and anaerobic organisms colonized on the skin and in the anogenital area. This is a case report of a middle-aged man with late presentation of Fournier's gangrene that was treated aggressively by a team of infectious disease specialists, surgeons, intensivists, and intensive care unit specialty nurses over a 3-week period. Despite heroic efforts by the multidisciplinary team, the patient's condition deteriorated and he died of multisystem failure. The purpose of this article was to alert clinicians to the etiology, pathophysiology, risk factors for invasive necrosis, prognostic indicators, and current diagnostic and treatment recommendations. Fournier's gangrene represents a true medical and surgical emergency and requires early recognition and a team approach to management.


Picosecond solvation dynamics--a potential viewer of DMSO-water binary mixtures.
In this work, we have investigated the composition dependent anomalous behavior of dimethyl sulfoxide (DMSO)-water binary mixture by collecting the ultrafast solvent relaxation response around a well known solvation probe Coumarin 480 (C480) by using a femtosecond fluorescence up-conversion spectrometer. Recent molecular dynamics simulations have predicted two anomalous regions of DMSO-water binary mixture. Particularly, these studies encourage us to investigate the anomalies from experimental background. DMSO-water binary mixture has repeatedly given evidences of its dual anomalous nature in front of our systematic investigation through steady-state and time-resolved measurements. We have calculated average solvation times of C480 by two individual well-known methods, among them first one is spectral-reconstruction method and another one is single-wavelength measurement method. The results of both the methods roughly indicate that solvation time of C480 reaches maxima in the mole fraction of DMSO XD = 0.12-0.17 and XD = 0.27-0.35, respectively. Among them, the second region (XD = 0.27-0.35) is very common as most of the thermodynamic properties exhibit deviation in this range. Most probably, the anomalous solvation trend in this region is fully guided by the shear viscosity of the medium. However, the first region is the most interesting one. In this region due to formation of strongly hydrogen bonded 1DMSO:2H2O complexes, hydration around the probe C480 decreases, as a result of which solvation time increases.


First Findings in the Route of the Maize Bushy Stunt Phytoplasma Within Its Vector Dalbulus maidis (Hemiptera: Cicadellidae).
In the pathosystem of Dalbulus madis (DeLong & Wolcott) (Hemiptera: Cicadellidae), a vector of maize bushy stunt phytoplasma (MBSP), the interactions occurring during the passage, invasion, and multiplication of the phytoplasma inside the vector body have been generalized from other pathosystems, with a poor understanding of the specific interactions. With the aim to understand MBSP movement and potential specific interactions with its vector, D. maidis adults were dissected to obtain the intestine and salivary gland of both infected (acquisition access period=4 d; latent period=23 d) and noninfected individuals. The organs were processed for visualization with transmission electronic microscopy. Images of phytoplasma cells were observed in the alimentary canal, epithelium of the mesenteron, hemocele, and salivary gland of the vector, and were confirmed through observation of similar cells in maize roots with advanced disease symptoms. The study of the MBSP movement within its vector shows novel findings between the synergy of the MBSP phytoplasma and D. maidis.


Initial experience with Angioseal™: Safety and efficacy of the endovascular closure device.
Vascular access site management is crucial to safe, efficient, and comfortable diagnostic or interventional percutaneous procedures. The Angioseal™ vascular closure device has been shown to be safe and effective in reducing the time to hemostasis following angiographic or interventional procedures. Relatively few studies have been conducted in the UK to assess the safety and efficacy of the device in a local setting. Data were retrospectively reviewed on 147 patients who underwent either diagnostic angiography or percutaneous interventional procedures from January 2008 to October 2009, and who had the femoral access site closed by 6F VIP Angioseal. A total of 147 patients (F: 49, M: 98), including 80 right femoral punctures, 57 left femoral punctures, and 10 bilateral punctures were reviewed using radiological reports and patients' clinical data. Data on antiplatelet and anticoagulant therapy were recorded. All procedures were carried out by two interventional radiologists at a single institution, under similar operating conditions. There were a total of six complications (4.47%), of which one was a major complication (0.75%), i.e., retroperitoneal bleed. There were five minor complications (3.73%), which included device deployment failure (2), device malfunction (2), and a superficial hematoma (>6 cm). Total complications were 6 out of 157 (3.8%) [95% CI = 0.8-6.8%)]. Successful hemostasis was achieved in less than 5 min in over 97% of patients. Successful device deployment was seen in over 98% of cases. We conclude that in our experience, the Angioseal vascular closure device is a safe and efficient means of achieving hemostasis post antegrade or retrograde puncture for diagnostic and percutaneous intervention procedures.


Predictive model for the size of bubbles and droplets created in microfluidic T-junctions.
We present a closed-form expression that allows the reader to predict the size of bubbles and droplets created in T-junctions without fitting. Despite the wide use of microfluidic devices to create bubbles and droplets, a physically sound expression for the size of bubbles and droplets, key in many applications, did not yet exist. The theoretical foundation of our expression comprises three main ingredients: continuity, geometrics and recently gained understanding of the mechanism which leads to pinch-off. Our simple theoretical model explains why the size of bubbles and droplets strongly depends on the shape of a T-junction, and teaches how the shape can be tuned to obtain the desired size. We successfully validated our model experimentally by analyzing the formation of gas bubbles, as well as liquid droplets, in T-junctions with a wide variety of shapes under conditions typical to multiphase microfluidics.


Biosynthesis of poly(4-hydroxybutyric acid) by recombinant strains of Escherichia coli.
The aim of this study was the production of the homopolyester poly(4-hydroxybutyric acid) (poly(4HB)) with recombinant strains of Escherichia coli. Wild-type strains and other widely used non-recombinant strains of E. coli are not able to produce polyhydroxyalkanoic acids (PHA) as storage compounds and cannot utilize 4-hydroxybutyric acid as sole carbon source. Accordingly, hybrid plasmids of pBluescript vectors were constructed which harbored the Alcaligenes eutrophus PHA synthase gene (phaC) and the Clostridium kluyveri orfZ putatively encoding a 4-hydroxybutyric acid-coenzyme A transferase. A 3.5-kb genomic SmaI/ApaI fragment from A. eutrophus, which comprises phaC, and a 1.8-kb genomic ApaI/EcoRI fragment from C kluyveri, which contained orfZ, were inserted into the SmaI and EcoRI sites of the vectors pKS- and pSK-, respectively. The two resulting plasmids pSKSE5.3 and pKSSE5.3 comprising phaC and orfZ colinear or antilinear to lacZ, respectively, were transformed into E. coli XL1-Blue. Recombinant strains synthesized the homopolyester poly(4HB), when the cells were cultivated in Luria-Bertani broth and if glucose and 4-hydroxybutyric acid were provided as carbon sources. If glucose was omitted, a copolyester of 3-hydroxybutyric acid and 4-hydroxybutyric acid was accumulated. The homopolyester poly(4HB) was also accumulated during cultivation of these strains in M9 mineral salts medium containing glucose plus 4-hydroxybutyric acid as carbon sources. Poly(4HB) could amount up to approximately 80% (w/w) of the cell dry matter if E. coli XL1-Blue harboring pKSSE5.3 was cultivated in M9 mineral salts medium and if the cultures were not sufficiently supplied with oxygen. 4HB was also incorporated into PHA if gamma-butyrolactone was used as carbon source. If levulinic acid, 4-hydroxyvaleric acid or gamma-valerolactone were used as carbon sources, only very low amounts of PHA were accumulated which did not contain 4-hydroxyalkanoic acids as constituents.


[Hygienic evaluation of chronic lymphatic leukemia in Bashkortostan Republic].
Prevalence of chronic lymphatic leukemia increased in Bashkortostan Republic over 1999-2008 with predominance of urban population, male patients, individuals aged 80-84. Reliable correlation was seen between pollutants releases into the ambient air and the morbidity parameters over following 5 years.


Selexipag for the Treatment of Pediatric Pulmonary Hypertension: A Systematic Review.
To systematically evaluate the safety, dosing regimen, and efficacy of selexipag for pediatric patients with pulmonary hypertension (PH). A literature search of the electronic databases of PubMed, Embase, Web of Science, Cochrane Library, and Google Scholar was performed from inception through February 28, 2023. Two reviewers independently searched and evaluated the quality of the studies and pooled data when appropriate. Full-text articles of studies of children diagnosed with PH and treated with selexipag were eligible. Pediatric patients with PH were classified into 2 groups: the add-on therapy group, in which selexipag was used as a third therapy in addition to the baseline treatment, and the transition therapy group, in which patients were switched from parenteral prostacyclin analogs to selexipag. Fourteen studies involving 58 pediatric patients with PH were included. All studies were either case reports or case series. Overall, 30 and 28 patients were in the add-on and transition therapy groups, respectively. In both groups, selexipag was initially administered as 50-200 µg twice daily and titrated to a tolerated dosage of 200-1,600 µg twice daily. Prostacyclin analogs were simultaneously weaned for patients in the transition group. In the add-on therapy group, 16 patients (80.0%) were at low risk of the World Health Organization functional class (WHO FC I/II), 12 (76.9%) were at low risk of the 6-minute walk distance (6MWD; >350 m), and 21 (95.5%) were at low risk of the pulmonary vascular resistance index (PVRi; <20 WU/m2). Furthermore, N-terminal pro-brain natriuretic peptide and mean pulmonary arterial pressure were significantly improved. More than 70% of patients experienced common tolerable side effects, such as headache, nausea, and diarrhea. In the transition therapy group, 5 patients (55.6%) were at low risk according to WHO FC I/II, 6 (66.7%) were at low risk according to 6MWD, and 14 (87.5) were at low risk according to PVRi; however, selexipag had no significant effect on their hemodynamic parameters. Additionally, more than 80% of patients experienced no side effects. Selexipag as add-on therapy or for transition from prostacyclin analogs may have a favorable safety profile and potential efficacy for pediatric patients with PH. Further high-quality evidence of the efficacy and safety of selexipag for the treatment of pediatric PH is warranted.


Trigeminal nerve responses in the rat elicited by chemical stimulation of the tongue.
Epithelial and neural mechanisms underlying trigeminal chemoreception were investigated by recording lingual nerve responses to chemical stimulation of the tongue. The chloride salts, NaCl, KCl, NH4Cl, and CaCl2, each elicited distinctly different, integrated whole-nerve responses (thresholds, 0.5-2.0 M). Incubation of the tongue with lanthanum (2.5 mM), which reduces the permeability of epithelial tight junctions, reversibly attenuated these salts responses. It did not affect neural responses to mechanical and thermal stimuli. Incubation with other established transport inhibitors--amiloride, tetra-ethyl ammonium or tetrodotoxin, had no effect on the salt responses. Acetic and hydrochloric acids (thresholds, 0.1-1.0 M), and MgCl2 and BaCl2 (greater than or equal to 0.5 M), also elicited distinctive responses. Other salts, MgSO4, Na isethionate and LaCl3 (greater than or equal to 1.0 M), and also ethanol (4 M), capsaicin (100 mM), nicotine (29.6 mM) and dextrose (0.5-2.5 M), did not elicit responses. These results indicate that ions of selected salts can diffuse through the tight junctions of the lingual epithelium to activate the trigeminal nerve, and suggest that both the cation and the anion may be important in determining if the nerves are activated and the wave-form of the responses.


An Analytical Approach to Flow-Guided Nanocommunication Networks.
Continuous progress of nanocommunications and nano-networking is opening the door to the development of innovative yet unimaginable services, with a special focus on medical applications. Among several nano-network topologies, flow-guided nanocommunication networks have recently emerged as a promising solution to monitoring, gathering information, and data communication inside the human body. In particular, flow-guided nano-networks display a number of specific characteristics, such as the type of nodes comprising the network or the ability of a nano-node to transmit successfully, which significantly differentiates them from other types of networks, both at the nano and larger scales. This paper presents the first analytical study on the behavior of these networks, with the objective of evaluating their metrics mathematically. To this end, a theoretical framework of the flow-guided nano-networks is developed and an analytical model derived. The main results reveal that, due to frame collisions, there is an optimal number of nano-nodes for any flow-guided network, which, as a consequence, limits the maximum achievable throughput. Finally, the analytical results obtained are validated through simulations and are further discussed.


Changes in Lipid Profile of Keratinocytes from Rat Skin Exposed to Chronic UVA or UVB Radiation and Topical Application of Cannabidiol.
UV radiation is a well-established environmental risk factor known to cause oxidative stress and disrupt the metabolism of keratinocyte phospholipids. Cannabidiol (CBD) is a phytocannabinoid with anti-inflammatory and antioxidant effects. In this study, we examined changes in the keratinocyte phospholipid profile from nude rat skin exposed to UVA and UVB radiation that was also treated topically with CBD. UVA and UVB radiation promoted up-regulation of phosphatidylcholines (PC), lysophosphatidylcholines (LPC), phosphatidylethanolamines (PE) and down-regulation of sphingomyelin (SM) levels and enhanced the activity of phospholipase A2 (PLA2) and sphingomyelinase (SMase). Application of CBD to the skin of control rats led to down-regulation of SM and up-regulation of SMase activity. After CBD treatment of rats irradiated with UVA or UVB, SM was up-regulated and down-regulated, respectively, while ceramide (CER) levels and SMase activity were down-regulated and up-regulated, respectively. CBD applied to the skin of UV-irradiated rats down-regulated LPC, up-regulated PE and phosphatidylserines (PS) and reduced PLA2 activity. In conclusion, up-regulation of PS may suggest that CBD inhibits their oxidative modification, while changes in the content of PE and SM may indicate a role of CBD in promoting autophagy and improving the status of the transepidermal barrier.


Acquired cross resistance to aminoglycosides in gentamicin-sensitive and gentamicin-resistant strains of enterobacteria.
The development of non-specific resistance ot gentamicin, tobramycin and amikacin was studied in 12 clinical isolates of enterobacteria with various patterns of aminoglycoside resistance. Strains were characterised with respect to MIC, transferability of resistance and possession of acetylating and adenylylating enzymes. An increase in aminoglycoside resistance was induced in 10 strains by a single exposure to the concentration of gentamicin, tobramycin or amikacin immediately below the MIC. Such resistance was non-specific; all three aminoglycosides were affected irrespective of which one had been used to induce the increase. Increments in non-specific aminoglycoside resistance were also evoked by exposure of enterobacteria to changing drug concentrations similar to those achieved in plasma during therapy. When strains already resistant to gentamicin or other aminoglycosides were exposed to therapeutically achievable drug concentrations, no further increase in resistance was observed in most cases. This suggests that use of an aminoglycoside to which the organism is resistant, as during "blind" therapy, will not usually compromise subsequent treatment with related antibiotics. The possible relevance of non-specific aminoglycoside resistance to therapy is discussed.


Effects of immunization of rabbits on establishment, survival, and reproductive biology of the tick Hyalomma anatolicum anatolicum.
Four antigenic preparations, viz. salivary gland antigen (SG Ag), whole tick extract antigen (WTE Ag) and 30,000-g supernatant fraction, and pellet of WTE Ag (TES Ag and TEP Ag, respectively), were made from partially fed adult female Hyalomma anatolicum anatolicum. Four groups of 5 rabbits each were immunized with the antigens, and a fifth group was kept as control. Following challenge with adult H. a. anatolicum, a significant decrease in engorgement weight and egg mass weight and an increase in engorgement period and preoviposition period were observed in WTE Ag-immunized rabbits. Similar results were observed with TES Ag and SG Ag, except that change in the engorgement period was insignificant. However, none of the tick parameter measurements showed significant changes with TEP Ag. None of the antigens produced significant changes in the percentage of engorgement or oviposition period of the challenged ticks. Thus, WTE Ag was the most effective in altering tick performances.


Simultaneous Analysis of Elastic and Nonspecific Adhesive Properties of Thin Sample and Biological Cell Considering Bottom Substrate Effect.
A new asymptotically correct contact model has been developed for conical tip based atomic force microscopy (AFM) nanoindentation. This new model provides both elastic and nonspecific adhesion properties of cells and soft gels by taking sample thickness at the point of indentation and its depth of indentation into consideration. The bottom substrate effect (BSE) is the most common source of error in the study of "AFM force maps" of the cellular sample. The present model incorporates an asymptotically correct correction term as a function of depth of indentation to eliminate the substrate effect in the analysis. Later, the model is extended to analyze the unloading portion of the indentation curve to extract the stiffness and adhesive properties simultaneously. A comparative study of the estimated material properties using other established contact models shows that the provided corrections effectively curb the errors coming from infinite thickness assumption. Nonspecific adhesive nature of a cell is represented in terms of adhesion parameter (γa) based on the "work of adhesion," this is an alternative to the peak value of tip-sample attractive (negative) force commonly used as representative adhesion measurement. The simple analytical expression of the model can help in estimating more realistic and accurate biomechanical properties of cells from atomic force microscopy based indentation technique.


A Pilot Study on Data-Driven Adaptive Deep Brain Stimulation in Chronically Implanted Essential Tremor Patients.
Deep brain stimulation (DBS) is an established therapy for Parkinson's disease (PD) and essential-tremor (ET). In adaptive DBS (aDBS) systems, online tuning of stimulation parameters as a function of neural signals may improve treatment efficacy and reduce side-effects. State-of-the-art aDBS systems use symptom surrogates derived from neural signals-so-called neural markers (NMs)-defined on the patient-group level, and control strategies assuming stationarity of symptoms and NMs. We aim at improving these aDBS systems with (1) a data-driven approach for identifying patient- and session-specific NMs and (2) a control strategy coping with short-term non-stationary dynamics. The two building blocks are implemented as follows: (1) The data-driven NMs are based on a machine learning model estimating tremor intensity from electrocorticographic signals. (2) The control strategy accounts for local variability of tremor statistics. Our study with three chronically implanted ET patients amounted to five online sessions. Tremor quantified from accelerometer data shows that symptom suppression is at least equivalent to that of a continuous DBS strategy in 3 out-of 4 online tests, while considerably reducing net stimulation (at least 24%). In the remaining online test, symptom suppression was not significantly different from either the continuous strategy or the no treatment condition. We introduce a novel aDBS system for ET. It is the first aDBS system based on (1) a machine learning model to identify session-specific NMs, and (2) a control strategy coping with short-term non-stationary dynamics. We show the suitability of our aDBS approach for ET, which opens the door to its further study in a larger patient population.


Late outcomes after acute pulmonary embolism: rationale and design of FOCUS, a prospective observational multicenter cohort study.
Acute pulmonary embolism (PE) is a frequent cause of death and serious disability. The risk of PE-associated mortality and morbidity extends far beyond the acute phase of the disease. In earlier follow-up studies, as many as 30 % of the patients died during a follow-up period of up to 3 years, and up to 50 % of patients continued to complain of dyspnea and/or poor physical performance 6 months to 3 years after the index event. The most feared 'late sequela' of PE is chronic thromboembolic pulmonary hypertension (CTEPH), the true incidence of which remains obscure due to the large margin of error in the rates reported by mostly small, single-center studies. Moreover, the functional and hemodynamic changes corresponding to early, possibly reversible stages of CTEPH, have not been systematically investigated. The ongoing Follow-Up after acute pulmonary embolism (FOCUS) study will prospectively enroll and systematically follow, over a 2-year period and with a standardized comprehensive program of clinical, echocardiographic, functional and laboratory testing, a large multicenter prospective cohort of 1000 unselected patients (all-comers) with acute symptomatic PE. FOCUS will possess adequate power to provide answers to relevant remaining questions regarding the patients' long-term morbidity and mortality, and the temporal pattern of post-PE abnormalities. It will hopefully provide evidence for future guideline recommendations regarding the selection of patients for long-term follow-up after PE, the modalities which this follow-up should include, and the findings that should be interpreted as indicating progressive functional and hemodynamic post-PE impairment, or the development of CTEPH.


The First Description of Setaria Tundra (Issaitshikoff & Rajewskaya, 1928) in Roe Deer from Croatia.
Genus Setaria, Viborg 1795, comprises 46 species that parasitize in the peritoneal cavity of Artiodactyla, Perissodactyla and Hyracoidea. The majority of these infections pass unnoticed, but occasionally they can induce severe peritonitis or neurological signs in aberrant hosts and, rarely, even in humans. In this paper we describe for the first time the finding of Setaria tundra in roe deer in Croatia. We examined 45 roe deer and determined the presence of Setaria nematodes in 24.4% of samples, which were subsequently diagnosed as Setaria tundra using molecular methods.


Protracted febrile myalgia of familial Mediterranean fever. Mutation analysis and clinical correlations.
Protracted febrile myalgia (PFM) is a rare form of vasculitic disease that affects patients with familial Mediterranean fever (FMF). Mutation analysis performed in 15 patients who suffered from this disorder showed that 9 of the 15 patients were homozygous for M694V. FMF in these 9 patients was associated with more severe symptoms compared to a group of 30 M694V homozygous FMF patients without PFM.


Analysis of substrate specificity of Trypanosoma brucei oligosaccharyltransferases (OSTs) by functional expression of domain-swapped chimeras in yeast.
N-Linked protein glycosylation is an essential and highly conserved post-translational modification in eukaryotes. The transfer of a glycan from a lipid-linked oligosaccharide (LLO) donor to the asparagine residue of a nascent polypeptide chain is catalyzed by an oligosaccharyltransferase (OST) in the lumen of the endoplasmic reticulum (ER). Trypanosoma brucei encodes three paralogue single-protein OSTs called TbSTT3A, TbSTT3B, and TbSTT3C that can functionally complement the Saccharomyces cerevisiae OST, making it an ideal experimental system to study the fundamental properties of OST activity. We characterized the LLO and polypeptide specificity of all three TbOST isoforms and their chimeric forms in the heterologous expression host S. cerevisiae where we were able to apply yeast genetic tools and newly developed glycoproteomics methods. We demonstrated that TbSTT3A accepted LLO substrates ranging from Man5GlcNAc2 to Man7GlcNAc2 In contrast, TbSTT3B required more complex precursors ranging from Man6GlcNAc2 to Glc3Man9GlcNAc2 structures, and TbSTT3C did not display any LLO preference. Sequence differences between the isoforms cluster in three distinct regions. We have swapped the individual regions between different OST proteins and identified region 2 to influence the specificity toward the LLO and region 1 to influence polypeptide substrate specificity. These results provide a basis to further investigate the molecular mechanisms and contribution of single amino acids in OST interaction with its substrates.


The Effectiveness of Protection Orders in Reducing Recidivism in Domestic Violence: A Systematic Review and Meta-Analysis.
Preventing and reducing domestic violence is a national and international social priority. Civil law protection orders (POs) have been the primary legal response to domestic violence internationally for a number of decades. However, evidence of their effectiveness is mixed due to variations in application within and across countries and variable quality of the research with most studies at high risk of bias. The purpose of this systematic review and meta-analysis was to evaluate the effectiveness of POs in reducing violation rates of domestic violence, compare violation rates reported by victims and police reports, and identify factors that influence violation and reoffense. Two electronic databases were searched; two independent researchers screened abstracts. Data were collected and assessed methodologically, using the Kmet Checklist Appraisal Tool and National Health and Medical Research Council Hierarchy of Evidence. Twenty-five studies that evaluated the effectiveness of POs in reducing recidivism in domestic violence met the eligibility criteria. Meta-analyses of weighted means of violation in the studies were conducted. Violation rates were found to be higher for victim reports compared with police reports. Violation rates were reduced when POs used in combination with arrests. PO violation rates were lower among perpetrators without histories of arrest for committing violence, perpetrators not engaging in stalking, and where couples have had medium to high incomes. There is no consensus among the studies about what the most appropriate methodology is to measure PO effectiveness. Future research should establish a more unified approach to evaluating the effectiveness and violations of POs.


An Eye Simulation for Training in the Use of Ophthalmologic Equipment and Corneal Foreign Body Removal Techniques.
This simulation curriculum is structured to teach the use of common eye assessment equipment and corneal foreign removal techniques using harvested cow eye models, the ophthalmologic burr, and the hollow-bore needle. This curriculum involves using one self-assessment (SA) station and three skills stations. The skill stations teach the use of the iCareTonometer, Woods Lamp, and Slit Lamp, and train corneal foreign body removal using a burr and hollow-bore needle. Sixteen first, second, and third-year (PGY 1-3) residents of the University of Alabama at Birmingham (UAB) Emergency Medicine participated. 100% indicated this should be an annual training with 87% indicating this was highly realistic. There was a 44% Pre-Sim to 94% Post-Sim SA for improvement with iCareTonometer skill. There was a 7% Pre-Sim to 100% post-Sim SA for improvement with Slit-Lamp skill. There was a 32% Pre-Sim to 100% Post-Sim SA improvement with Woods Lamp skill. For CFB removal, there was a 13% Pre-Sim to 94% Post-Sim SA for improvement with burr and hollow-bore needle skill. The self-assessments indicated that this simulation was successful in leading these emergency medicine residents to attaining enhanced self-assessed competencies in the use of common eye assessment equipment and for corneal foreign body removal using the burr and the hollow-bore needle. To establish curriculum globalization, reliability, and competency, larger test groups will have to be studied and objective qualified testers will need to assess competencies.


Personalized oral debriefing versus standardized multimedia instruction after patient crisis simulation.
Simulation experience alone without debriefing is insufficient for learning. Standardized multimedia instruction has been shown to be useful in teaching surgical skills but has not been evaluated for use as an adjunct in crisis management training. Our primary purpose in this study was to determine whether standardized computer-based multimedia instruction is effective for learning, and whether the learning is retained 5 wk later. Our secondary purpose was to compare multimedia instruction to personalized video-assisted oral debriefing with an expert. Thirty anesthesia residents were recruited to manage three different simulated resuscitation scenarios using a high-fidelity patient simulator. After the first scenario, subjects were randomized to either a computer-based multimedia tutorial or a personal debriefing of their performance with an expert and videotape review. After their respective teaching, subjects managed a similar posttest resuscitation scenario and a third retention test scenario 5 wk later. Performances were independently rated by two blinded expert assessors using a previously validated assessment system. Posttest (12.22 +/- 2.19, P = 0.009) and retention (12.80 +/- 1.77, P < 0.001) performances of nontechnical skills were significantly improved in the standardized multimedia instruction group compared with pretest (10.27 +/- 2.10). There were no significant differences in improvement between the two methods of instruction. Computer-based multimedia instruction is an effective method of teaching nontechnical skills in simulated crisis scenarios and may be as effective as personalized oral debriefing. Multimedia may be a valuable adjunct to centers when debriefing expertise is not available.


A tetrapeptide class of biased analgesics from an Australian fungus targets the µ-opioid receptor.
An Australian estuarine isolate of Penicillium sp. MST-MF667 yielded 3 tetrapeptides named the bilaids with an unusual alternating LDLD chirality. Given their resemblance to known short peptide opioid agonists, we elucidated that they were weak (Ki low micromolar) μ-opioid agonists, which led to the design of bilorphin, a potent and selective μ-opioid receptor (MOPr) agonist (Ki 1.1 nM). In sharp contrast to all-natural product opioid peptides that efficaciously recruit β-arrestin, bilorphin is G protein biased, weakly phosphorylating the MOPr and marginally recruiting β-arrestin, with no receptor internalization. Importantly, bilorphin exhibits a similar G protein bias to oliceridine, a small nonpeptide with improved overdose safety. Molecular dynamics simulations of bilorphin and the strongly arrestin-biased endomorphin-2 with the MOPr indicate distinct receptor interactions and receptor conformations that could underlie their large differences in bias. Whereas bilorphin is systemically inactive, a glycosylated analog, bilactorphin, is orally active with similar in vivo potency to morphine. Bilorphin is both a unique molecular tool that enhances understanding of MOPr biased signaling and a promising lead in the development of next generation analgesics.


Health care challenge in coal mines community.
The present paper depicts salient features of environment and living conditions with the comparison of various diseases prevalent among underground coal miners, surface workers, asbestos mine workers and general population of Jharia-Dhanbad coalfield as conducted by CMRS during the past few years. The investigations on coal miners' community comprise of different morbid conditions with respiratory (22%), Pneumoconiosis (11.6%), Skin (35%), Eye (29%), Intestinal parasitic infestation (44.6%), Anaemia (42%), Immunostatus (V.D.R.L. Positive-19.9%), Status of injuries and Blood pressure, Water-borne diseases, housing facilities and excreta disposal. The paper also includes the analysis of disease pattern obtained from hospital records of two coal mines which depicts 19.1%, 24.7% and 16% members of coal miners' families suffering from disorder with respiratory, gastro-intestinal and fever respectively. With speedy industrialization of the country, the mining of coal resource comes first in the chain of socio-economic development. The speedy human industrial activities are based on 80% steam, metallurgical and thermal electrical energy which hinges on coal wings. The coal has also gradually occupied all the phases of social life, our clothes, books, newspapers, cooking gas, chemical paints, dye stuff, oil phenyl, Benzene, Naphthalene, Coal tar, scents and various types of unaccountable products come out from coal derivatives and pushed to serve in the today's market for our daily exigencies. Every day one finds a new coal based industry is coming up in the area. The coal is utilized in two hundred ways in our various walks of social life.(ABSTRACT TRUNCATED AT 250 WORDS)


A multiyear approach to student-driven investigations in exercise physiology.
Many undergraduate institutions offer individual research opportunities for upper-level students in independent study courses and summer undergraduate research programs. These are necessarily limited to a small number of students. Greater numbers of students can benefit from incorporating student-directed investigative experiences into laboratories in standard courses. In human performance investigations, any single course may not offer sufficient numbers of subjects to adequately test hypotheses comparing population groups or to examine longitudinal trends. In this exercise physiology course, exercise testing was conducted in three areas: 1) techniques of body composition analysis, 2) field tests for the estimation of maximal oxygen consumption, and 3) maximal anaerobic and aerobic power. All students enrolled over a 10-yr period participated as subjects and as testers. Working in small research groups, students added their results to those from previous years, generated a variety of hypotheses (correlations between tests, subgroup differences, etc.), and tested them statistically using the complete data set of 217 subjects. They then engaged in collaborative writing and peer review to prepare formal papers on their results. The multiyear approach allowed students to situate their work within and contribute to the accumulation of a large database and to practice essential scientific skills of hypothesis formation, data collection and analysis, collaborative work, and scientific communication. In addition, due to the larger number of subjects available to analyze, students observed statistically significant differences between test groups in the multiyear database that they were unable to demonstrate when conducting analysis on a single course. Finally, the large number of subjects and statistical power offered by the use of the database provides distinct pedagogical advantages.


Gene expression profiling of the tetrapyrrole metabolic pathway in Arabidopsis with a mini-array system.
Tetrapyrrole compounds, such as chlorophylls, hemes, and phycobilins, are synthesized in many enzymatic steps. For regulation of the tetrapyrrole metabolic pathway, it is generally considered that several specific isoforms catalyzing particular enzymatic steps control the flow of tetrapyrrole intermediates by differential regulation of gene expression depending on environmental and developmental factors. However, the coordination of such regulatory steps and orchestration of the overall tetrapyrrole metabolic pathway are still poorly understood. In this study, we developed an original mini-array system, which enables the expression profiling of each gene involved in tetrapyrrole biosynthesis simultaneously with high sensitivity. With this system, we performed a transcriptome analysis of Arabidopsis seedlings in terms of the onset of greening, endogenous rhythm, and developmental control. Data presented here clearly showed that based on their expression profiles at the onset of greening, genes involved in tetrapyrrole biosynthesis can be classified into four categories, in which genes are coordinately regulated to control the biosynthesis. Moreover, genes in the same group were similarly controlled in an endogenous rhythmic manner but also by a developmental program. The physiological significance of these gene clusters is discussed.


The novel EHEC gene asa overlaps the TEGT transporter gene in antisense and is regulated by NaCl and growth phase.
Only a few overlapping gene pairs are known in the best-analyzed bacterial model organism Escherichia coli. Automatic annotation programs usually annotate only one out of six reading frames at a locus, allowing only small overlaps between protein-coding sequences. However, both RNAseq and RIBOseq show signals corresponding to non-trivially overlapping reading frames in antisense to annotated genes, which may constitute protein-coding genes. The transcription and translation of the novel 264 nt gene asa, which overlaps in antisense to a putative TEGT (Testis-Enhanced Gene Transfer) transporter gene is detected in pathogenic E. coli, but not in two apathogenic E. coli strains. The gene in E. coli O157:H7 (EHEC) was further analyzed. An overexpression phenotype was identified in two stress conditions, i.e. excess in salt or arginine. For this, EHEC overexpressing asa was grown competitively against EHEC with a translationally arrested asa mutant gene. RT-qPCR revealed conditional expression dependent on growth phase, sodium chloride, and arginine. Two potential promoters were computationally identified and experimentally verified by reporter gene expression and determination of the transcription start site. The protein Asa was verified by Western blot. Close homologues of asa have not been found in protein databases, but bioinformatic analyses showed that it may be membrane associated, having a largely disordered structure.


[The efficacy of the 1-day treatment of enterobiasis with medamine].
The Soviet anthelminthic agent medamine has been synthesized at the Institute of Medical Parasitology and Tropical Medicine, USSR Ministry of Public Health and the Institute of Plant Chemistry, Academy of Sciences of the Uzbek SSR and is produced at the Tashkent Chemical Pharmaceutical Plant. It is used for one-day treatment of enterobiasis which is to be repeated in 2-3 weeks. A daily drug dose is 10 mg per kg body weight, given 3 times a day after meals. The efficacy of therapy was 92 +/- 4% and 100 +/- 2% after the first and second course of treatment, respectively.


Soybean oligosaccharides alter colon short-chain fatty acid production and microbial population in vitro.
This study was conducted to the determine fermentation characteristics of soybean oligosaccharides (SBOS) in an in vitro system. Digesta collected from the colon of Huanjiang mini-pigs was used as inoculums, and SBOS (0.2 g per 10 mL fermentation broth) was used as substrate during the in vitro fermentation. The inoculum or inoculum + glucose (0.2 g) was used as negative or positive control, respectively. The slurry was fermented in an anaerobic chamber and gas production (GP) recording was taken after 48 h of incubation by referring to the moving scale on the plunger of the glass syringes, and then GP parameters, pH value, NH(3)-N content, short chain fatty acid (SCFA) levels, and microbial community in the fermentation broth were determined. For gas production parameters, pH, and fermentation product determination after 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 28, 32, 36, 42, and 48 h of incubation, SBOS and glucose demonstrated similar responses compared to control including increase maximal gas production, decreased lag time, decreased pH, and accumulation of NH(3) and increased SCFA. The exception was rate of GP, which was higher (P < 0.05) for SBOS compared with glucose. Incubation with SBOS increased (P < 0.05) the microbial diversity and population of Bifidobacterium and Lactobacillus but decreased (P < 0.05) Escherichia and Streptococcus when compared with incubation with glucose. These findings suggested that the SBOS can improve the gut microbiota balance in colon and modulate its metabolism.


Evidence for the presence of P1- and P2-purinoceptors in rat caecum.
Adenosine, 2-chloroadenosine, adenosine triphosphate (ATP) and adenosine tetraphosphate induced concentration-dependent relaxation of rat caecum. Caffeine as well as theophylline, at concentrations (25-100 microM) lower than that known to produce significant phosphodiesterase inhibition, shifted the concentration-response curve for adenosine to the right in a parallel fashion without diminishing the maximal response. The pA2 values for caffeine and theophylline against adenosine were found to be 4.6 +/- 0.26 (slope = 0.97 +/- 0.05) and 4.9 +/- 0.28 (slope = 0.98 +/- 0.07), respectively. Though the relaxant effect of adenosine tetraphosphate was blocked in a non-competitive manner by quinidine (5 microM) and phenoxybenzamine (25 microM), theophylline or caffeine (50 microM) failed to block this response. Similarly, quinidine also blocked the relaxant effect of ATP in a non-competitive manner. Besides, the relaxant effect due to adensine was rapid in onset as compared to a slow relaxant effect of ATP. These observations suggest the presence of P1- and P2-purinoceptors in the caecum. The activation of these receptors by adenosine and ATP, respectively, leads to relaxation of the tissue.


Molecular Analysis of Clinically Defined Subsets of High-Grade Serous Ovarian Cancer.
The diversity and heterogeneity within high-grade serous ovarian cancer (HGSC), which is the most lethal gynecologic malignancy, is not well understood. Here, we perform comprehensive multi-platform omics analyses, including integrated analysis, and immune monitoring on primary and metastatic sites from highly clinically annotated HGSC samples based on a laparoscopic triage algorithm from patients who underwent complete gross resection (R0) or received neoadjuvant chemotherapy (NACT) with excellent or poor response. We identify significant distinct molecular abnormalities and cellular changes and immune cell repertoire alterations between the groups, including a higher rate of NF1 copy number loss, and reduced chromothripsis-like patterns, higher levels of strong-binding neoantigens, and a higher number of infiltrated T cells in the R0 versus the NACT groups.