First enantioseparation and circular dichroism spectra of Au38 clusters protected by achiral ligands.
Bestowing chirality to metals is central in fields such as heterogeneous catalysis and modern optics. Although the bulk phase of metals is symmetric, their surfaces can become chiral through adsorption of molecules. Interestingly, even achiral molecules can lead to locally chiral, though globally racemic, surfaces. A similar situation can be obtained for metal particles or clusters. Here we report the first separation of the enantiomers of a gold cluster protected by achiral thiolates, Au(38)(SCH(2)CH(2)Ph)(24), achieved by chiral high-performance liquid chromatography. The chirality of the nanocluster arises from the chiral arrangement of the thiolates on its surface, forming 'staple motifs'. The enantiomers show mirror-image circular dichroism responses and large anisotropy factors of up to 4×10(-3). Comparison with reported circular dichroism spectra of other Au(38) clusters reveals that the influence of the ligand on the chiroptical properties is minor.


Detection of antibodies against Akabane virus in bovine sera by enzyme-linked immunosorbent assay.
An enzyme-linked immunonosorbent assay was established for detection of antibodies to Akabane virus in bovine sera. The assay was shown to be a useful serological tool for studies on Akabane virus infection.


Formation of composite nucleoprotein complexes near the transcription start of the Shrunken gene from maize.
We describe an analysis of protein-DNA interactions detectable with nuclear extracts prepared from maize kernels and DNA fragments from the immediate upstream region of the Shrunken gene from maize. The data demonstrate that sequences from position -235 to the transcription start are recognized by sequence specific nuclear proteins. In footprinting and competition experiments at least six different protein-DNA interactions can be distinguished within this upstream region. Two sequence related inverted repeat structures, 67 and 64 bp in length, cross compete for protein recognition.


Influence of Proteome Profiles and Intracellular Drug Exposure on Differences in CYP Activity in Donor-Matched Human Liver Microsomes and Hepatocytes.
Human liver microsomes (HLM) and human hepatocytes (HH) are important in vitro systems for studies of intrinsic drug clearance (CLint) in the liver. However, the CLint values are often in disagreement for these two systems. Here, we investigated these differences in a side-by-side comparison of drug metabolism in HLM and HH prepared from 15 matched donors. Protein expression and intracellular unbound drug concentration (Kpuu) effects on the CLint were investigated for five prototypical probe substrates (bupropion-CYP2B6, diclofenac-CYP2C9, omeprazole-CYP2C19, bufuralol-CYP2D6, and midazolam-CYP3A4). The samples were donor-matched to compensate for inter-individual variability but still showed systematic differences in CLint. Global proteomics analysis outlined differences in HLM from HH and homogenates of human liver (HL), indicating variable enrichment of ER-localized cytochrome P450 (CYP) enzymes in the HLM preparation. This suggests that the HLM may not equally and accurately capture metabolic capacity for all CYPs. Scaling CLint with CYP amounts and Kpuu could only partly explain the discordance in absolute values of CLint for the five substrates. Nevertheless, scaling with CYP amounts improved the agreement in rank order for the majority of the substrates. Other factors, such as contribution of additional enzymes and variability in the proportions of active and inactive CYP enzymes in HLM and HH, may have to be considered to avoid the use of empirical scaling factors for prediction of drug metabolism.


Coexistence of congeneric native and invasive species: the case of the green algae Codium spp. in northwestern Spain.
We examined the patterns of distribution and abundance, and reproductive traits (presence of gametophytes and size at time of reproduction) in the invasive Codium fragile ssp. fragile and the native C. tomentosum and C. vermilara on intertidal habitats of NW Spain at two dates. All three species coexist in the locations and habitats studied, although abundances were low. We found a greater proportion of C. fragile ssp. fragile towards the east of the Cantabrian coast and on upper levels on the shore, where conditions are more stressful. The proportion of thalli bearing gametangia in C. fragile ssp. fragile was greater than in the native species in all habitats. The presence of gametangia was size-dependent for all species, with the invasive species maturing at a smaller size, which combined with the previous features, might confer competitive advantages to this species over the native species. We also demonstrated that molecular analyses are necessary for the correct identification of C. fragile subspecies.


Comparative Efficacy (DAS28 Remission) of Targeted Immune Modulators for Rheumatoid Arthritis: A Network Meta-Analysis.
The objective of this study was to evaluate the relative efficacy of targeted immune modulators (TIMs) in TIM-naïve/mixed populations (≤ 20% TIM-experienced) and TIM-experienced (> 20% TIM-experienced) adults with moderate-to-severe rheumatoid arthritis with an inadequate response to or intolerance of conventional disease-modifying antirheumatic drugs (cDMARDs). A fixed-effects Bayesian network meta-analysis (NMA) was performed using published study-level data from 41 randomized controlled trials (RCTs) identified from two recent systematic literature reviews conducted by the Institute for Clinical and Economic Review, and two additional phase III trials for filgotinib (FINCH-1, FINCH-2). RCTs that compared TIMs with each other, cDMARD therapy, or placebo were included. Treatments included Janus kinase (JAK) inhibitors, tumor necrosis factor α inhibitors (TNFi), and other non-TNFi therapies. Efficacy was defined as achieving remission with a DAS28 score < 2.6 at 12 and 24 weeks. In the 12-week analysis for the TIM-naïve/mixed population, all TIMs combined with cDMARD therapy were significantly more likely to achieve remission compared with a cDMARD alone, with intravenous tocilizumab showing a substantially greater magnitude of effect (odds ratio 19.36; 95% credible interval 11.01-38.16). Similarly, in the 24-week analysis, intravenous and subcutaneous tocilizumab showed the highest odds ratio of achieving DAS28 remission compared with cDMARD therapy. Similar trends were observed for the analyses on monotherapy or TIM-experienced population. This NMA demonstrated that tocilizumab is associated with a greater likelihood of remission (DAS28 < 2.6) at 12 and 24 weeks compared with most other TIMs, including new JAK inhibitors, when used in combination with a cDMARD or as monotherapy among TIM-naïve/mixed or TIM-experienced populations.


Intermittent urinary retention in first trimester of pregnancy.
Gynecologic diseases are a rare but easily detectable cause of complete urinary retention. Herein we describe a case of intermittent but complete urinary retention secondary to uterine fibroids and prolapse during the first trimester of pregnancy.


Wnt signaling and the regulation of stem cell function.
Canonical Wnt signaling plays a crucial role in controlling cell expansion in many types of stem cells. Recent studies, however, demonstrated that Wnt is not only a general stem cell growth factor but can also influence cell lineage decisions in certain stem cell types by promoting specific fates at the expense of others. Thus, Wnt signaling elicits multiple functions in stem cells. Wnt activity appears to depend on cell-intrinsic properties that might change with time during development, thereby altering the cellular response to Wnt. Moreover, the spatial context of a stem cell also determines how the cell interprets Wnt signal activity, in that synergistic or antagonistic signaling pathways can modulate Wnt signaling. How a stem cell integrates Wnt and other signals and how such signaling networks regulate stem cell function on the molecular level remains to be elucidated.


The functional neural architecture of dysfunctional reward processing in autism.
Functional imaging studies have found differential neural activation patterns during reward-paradigms in patients with autism spectrum disorder (ASD) compared to neurotypical controls. However, publications report conflicting results on the directionality and location of these aberrant activations. We here quantitatively summarized relevant fMRI papers in the field using the anatomical likelihood estimation (ALE) algorithm. Patients with ASD consistently showed hypoactivations in the striatum across studies, mainly in the right putamen and accumbens. These regions are functionally involved in the processing of rewards and are enrolled in extensive neural networks involving limbic, cortical, thalamic and mesencephalic regions. The striatal hypo-activations found in our ALE meta-analysis, which pooled over contrasts derived from the included studies on reward-processing in ASD, highlight the role of the striatum as a key neural correlate of impaired reward processing in autism. These changes were present for studies using social and non-social stimuli alike. The involvement of these regions in extensive networks associated with the processing of both positive and negative emotion alike might hint at broader impairments of emotion processing in the disorder.


Make-up of the epinephrine autoinjector: the effect on its use by untrained users.
Part of the problems related to proper use of the epinephrine autoinjector may be related to the design of the autoinjector itself. We investigated whether minor modifications in the design of the currently available epinephrine autoinjector ease its use and abrogate common use errors. All interns other than those who had previously worked in allergy department in a medical school were invited to the study. Two identical epinephrine autoinjector trainers (Epipen trainer(®) ) were used, one of which was modified by changing the gray safety cap to red and placing a yellow arrow pointing to the black injection tip. A written and visual instruction sheet for each trainer was provided. Participants were asked to demonstrate the use of the Epipen trainer either with the original or with the modified one. They were scored and timed for their demonstration. Out of the 224 interns who were invited to participate, one hundred and sixty-four interns (73.2%) participated in the study. The number of participants correctly demonstrating the use of epinephrine autoinjectors was 22.6% and 65% in unmodified and modified trainer groups, respectively (p < 0.001). The mean time to administer trainers was 26.78 ± 10.6 and 15.88 ± 2.55 s; total median scores were 3.08 ± 1.48 and 4.47 ± 0.84 in unmodified and modified groups, respectively (p < 0.001 for both). Significantly fewer participants had presumptive unintentional injection injury while using modified (5%) compared with unmodified trainer (45.2%) (p < 0.001). Few and simple modifications in the design of epinephrine autoinjector were found effective in increasing its correct use and decreasing common use errors by untrained users. (Clinical trials identifier: NCT01217138).


The role of Nrf2 signaling pathway in the mud crab (Scylla paramamosain) in response to Vibrio parahaemolyticus infection.
The transcription factor Nrf2 plays vital roles in detoxification and antioxidant enzymes against oxidative stress. However, the function of Nrf2 in crustaceans is not well studied. In this study, a novel Nrf2 gene from the mud crab (Sp-Nrf2) was identified. It was encoded 245 amino acids. Sp-Nrf2 expression was ubiquitously expressed in all tested tissues, with the highest expression level in the gill. Sp-Nrf2 protein was mainly located in the nucleus. The expression levels of Sp-Nrf2, and antioxidant-related genes (HO-1 and NQO-1) were induced after Vibrio parahaemolyticus infection, indicating that Nrf2 signaling pathway was involved in the responses to bacterial infection. Over-expression of Sp-Nrf2 could improve cell viability after H2O2 exposure, indicating that Sp-Nrf2 might relieve oxidative stress. Silencing of Sp-Nrf2 in vivo decreased HO-1 and NQO-1 expression. Moreover, knocking down Sp-Nrf2 in vivo can increase malondialdehyde content and the mortality of mud crabs after V. parahaemolyticus infection. Our results indicated that Nrf2 signaling pathway played a significant role in immune response against bacterial infection.


NADH oxidation in phospholipid-enriched cytoplasmic membrane vesicles from Escherichia coli.
NADH oxidation in Escherichia coli cytoplasmic membrane vesicles enriched in anionic phospholipids by de novo synthesis of lipid in the vesicles from acyl-CoA esters and sn-glycerol 3-phosphate has been studied. NADH-oxidase but not NADH-dehydrogenase activity was found to decrease during synthesis and accumulation of phospholipid in the vesicles. Density gradient fractionation showed that NADH-oxidase activity was reduced to approximately 30% in vesicles with a 3-6 fold increase in anionic phospholipid, whereas vesicles with a greater than 10-fold increase in phospholipid had virtually no NADH oxidase activity.


High expression of HIF-1α alleviates benzene-induced hematopoietic toxicity and immunosuppression in mice.
Benzene exposure can cause pancytopenia and immunosuppression, leading to serious diseases such as aplastic anemia (AA) or acute myeloid leukemia (AML), but the underlying mechanism has not been fully elucidated. Hypoxia-inducible factor 1 (HIF-1) is an important transcription factor that regulates many downstream target genes. In this study, we reported a novel mechanism by which high expression of HIF-1α alleviated benzene toxicity. Mice with high expression of HIF-1α (HIF-1α+) were obtained by the Tet-on system and doxycycline induction, and they and wild-type (WT) mice were exposed to 150 mg/kg benzene for 0, 1, 3, 7, 10, 14, and 28 days. Dynamic changes in hematopoietic and immune-related indicators and the role of HIF-1α were explored. The level of white blood cells in mice reached the highest level on the third day, and immunity was activated and then suppressed within 10 days. Significant pancytopenia and immunosuppression occurred at 14 days and were more pronounced at 28 days. The levels of HIF-1α, EPO, VEGF, RORγt, and IL-17 in WT mice gradually decreased with increasing benzene exposure days, while the levels of Foxp3 and IL-10 increased. These changes were alleviated in HIF-1α+ mice. High expression of HIF-1α increased the levels of EPO and VEGF, which helped to maintain the stability of the hematopoietic microenvironment. Simultaneously, it attenuated benzene-induced immunosuppression by alleviating the Th17/Treg imbalance. HIF-1α is expected to be a new target for benzene-induced diseases such as AA and AML.


Both swine and human cells are capable to support the replication of swine hepatitis E virus type 4 in vitro.
Using swine anal swab or liver as inocula, cell-culture systems were developed for swine hepatitis E virus (HEV) in swine cells (IBRS-2) and human cells (A549). Both positive and negative strand of swine HEV RNA were detected continuously. Cytopathic effect appeared from passage 8 in IBRS-2 and passage 22 in A549. Viral antigen was detected by indirect immunofluorescent assay in infected cells. Progenies harbored mutations in the third nucleotide of codon. Amino acid changes occurred in passage 8 in IBRS-2 and rescued in passage 10. Full-length genome sequence of a swine HEV isolate from liver was determined to be genotype 4. Taken together, our data suggest that swine HEV is able to replicate in both swine and human cells in vitro.


External quality assessment for molecular typing of Salmonella 2013-2015: performance of the European national public health reference laboratories.
We report the results of three consecutive External Quality Assessments (EQAs) for molecular subtyping of Salmonella to assess the performance of the European national public health reference laboratories (NPHRLs). The EQA included the molecular typing methods used for European enhanced surveillance of human Salmonella infections: pulsed field gel electrophoresis (PFGE), including gel analysis by the use of the software BioNumerics, and 5-locus multiple locus variable number of tandem repeat analysis (MLVA) for serovar Typhimurium. The participation in the PFGE laboratory part was higher (27/35) than in the gel analysis (19/35) and MLVA (15/35), suggestive of the need for capacity building in methods requiring specialized equipment (MLVA) or software (gel analysis). The majority (25/27) of the participating NPHRLs produced inter-laboratory comparable PFGE gel(s). Two laboratories continued to produce low-quality gels and should have additional technical assistance in the future. In particular, two gel quality evaluation parameters, measuring "image acquisition and running conditions" and "bands", were identified to cause gel quality problems throughout the EQAs. Despite the high number of laboratories participating in the PFGE laboratory part, the participation in gel analysis was low, although increasing. In the MLVA part, the NPHRLs correctly assigned 96% (405/420) allelic profiles according to the nomenclature. In conclusion, the EQAs identified critical parameters for unsuccessful performance and helped to offer assistance to those laboratories that needed it most. The assessments supported the development of quality in molecular typing and promoted the harmonization of subtyping methods used for EU/EEA-wide surveillance of human Salmonella infections.


[Risk Control of UPS Power Supply for Medical Equipment].
Putting forward that the risk of using UPS for medical matching is easy to be ignored by managers. The method of UPS inspection was designed and used in hospital. The qualified rate of UPS for equipment matching was 66.7% and the problems were corrected. Based on the Internet of Things technology, the UPS monitoring technology was prospected. It is effective to control the risk of UPS by recognizing the risk of using UPS and specifying the strategy.


The approach of the European communities to the assessment of the carcinogenic risk of environmental chemicals, in particular, artificial colourings.
A brief outline is given of the rationale of the Environmental Research Programme of the European Economic Community and of its implementation. The major part of the programme is devoted to research on the exposure-effect relationships for mutagens and carcinogens and to the establishment of criteria. As a first step, a battery of test systems for mutagenicity screening is being established. These will be evaluated in a comparative test programme. The second part of the paper describes the specific legislative actions of the European Communities on food colourings. The historical background is given; and the criteria used by the Scientific Committee for Food for classifying and evaluating colouring matters are listed. Proposed regulatory action, on the basis of the report of the Committe, includes lists of colouring matters which are 'unacceptable', 'temporarily acceptable' and 'acceptable'.


Rapid proliferation of the parasitic copepod, Salmincola californiensis (Dana), on kokanee salmon, Oncorhynchus nerka (Walbaum), in a large Colorado reservoir.
Ecologically and economically valuable Pacific salmon and trout (Oncorhynchus spp.) are widespread and susceptible to the ectoparasite Salmincola californiensis (Dana). The range of this freshwater copepod has expanded, and in 2015, S. californiensis was observed in Blue Mesa Reservoir, Colorado, USA, an important kokanee salmon (O. nerka, Walbaum) egg source for sustaining fisheries. Few S. californiensis were detected on kokanee salmon in 2016 (<10% prevalence; 2 adult S. californiensis maximum). By 2020, age-3 kokanee salmon had 100% S. californiensis prevalence and mean intensity exceeding 50 adult copepods. Year and kokanee salmon age/maturity (older/mature) were consistently identified as significant predictors of S. californiensis prevalence/intensity. There was evidence that S. californiensis spread rapidly, but their population growth was maximized at the initiation (the first 2-3 years) of the invasion. Gills and heads of kokanee salmon carried the highest S. californiensis loads. S. californiensis population growth appears to be slowing, but S. californiensis expansion occurred concomitant with myriad environmental/biological factors. These factors and inherent variance in S. californiensis count data may have obscured patterns that continued monitoring of parasite-host dynamics, when S. californiensis abundance is more stable, might reveal. The rapid proliferation of S. californiensis indicates that in 5 years a system can go from a light infestation to supporting hosts carrying hundreds of parasites, and concern remains about the sustainability of this kokanee salmon population.


Effects of GDF5 overexpression on embryonic rat dopaminergic neurones in vitro and in vivo.
Transplantation of embryonic dopaminergic neurones has shown promise for the treatment of Parkinson's disease (PD), but this approach is limited by the poor survival of the transplanted cells. Exogenous dopaminergic neurotrophic factors such as growth/differentiation factor 5 (GDF5) have been found to enhance the survival of transplanted dopaminergic neurones. However, this approach is limited by the rapid degradation of such factors in vivo; thus, methods for long-term delivery of these factors are under investigation. The present study shows, using optimised lipid-mediated transfection procedures, that overexpression of GDF5 significantly improves the survival of dopaminergic neurones in cultures of embryonic day (E) 13 rat ventral mesencephalon (VM) and protects them against 6-hydroxydopamine (6-OHDA)-induced toxicity. In another experiment, E13 VM cells were transfected with GDF5 after 1 day in vitro (DIV), then transplanted into 6-OHDA-lesioned adult rat striata after 2 DIV. The survival of these E13 VM dopaminergic neurones after transfection and transplantation was as least as high as that of freshly dissected E14 VM dopaminergic neurones, demonstrating that transfection was not detrimental to these cells. Furthermore, GDF5-overexpressing E13 VM transplants significantly reduced amphetamine-induced rotational asymmetry in the lesioned rats. This study shows that lipid-mediated transfection in vitro prior to transplantation is a valid approach for the introduction of neurotrophic proteins such as GDF5, as well as lending further support to the potential use of GDF5 in neuroprotective therapy for PD.


[The control of melanin-incorporation into the pupal color-pattern of the cabbage white pieris brassicae L.]
The pupal cuticle of the Cabbage WhitePieris brassicae L. contains a pattern of black patches, the variation of which is controlled by external factors. Their way of influence is investigated.A classification of the melanization of the pupae is based on the variation of the pattern of black patches. The patches vary in their size. In case of weak melanization some of them are absent, but they do not vary in their position and their shape, and they never exceed a certain maximum size.This means that the integument contains a pattern of areas qualified for melanin deposition into the cuticle, and external factors determine to what extent this potency is realized.To a small extent, melanization is temperature-dependent. In total darkness the strongest melanization is achieved between 25° C and 28° C. Above and below that the melanization is weaker.The most important external influence on melanization is that of the light conditions. - It determines the degree of melanization during a sensitive period after the caterpillar has stopped moving about and before it becomes a pre-pupa.The strongest melanization results on dark background in bright illumination, a somewhat weaker melanization in total darkness and the weakest melanization in illumination on light background. Probably a strong contrast between the illumination and the reflection from the ground enhances the melanization.If a red filter is applied to the illumination, the melanization is weakened; although the total intensity of light is smaller, the pupae are lighter. On the other hand, the filtering out of the long-wave range by a blue filter results in stronger melanization. This means that long-waved light suppresses melanization more than short-waved light does.In equal illumination yellow background causes a weaker melanization than white and black background. Evidently, the color of the light reflected by the ground influences the pigmentation in the same way as that of the illumination does.Melanization is influenced not only by the light acting through the eyes, but also by the light which acts on the integument of the thorax and the abdomen. This does, however, not affect the pigmentation of the integument directly, for isolated abdomina do not adapt their color to light conditions.If the body of the pre-pupa is ligatured, only that part of the body anterior to the ligature will adapt its melanization to the light conditions. The posterior parts are uniform in melanization. It is as strong as that of pupae from total darkness. It is concluded that the pigmentation is controlled by a melanization inhibiting factor, which is distributed from the anterior part of the body. - Further ligaturing experiments show that there is a critical period during which this factor is distributed through the body. This period begins while the caterpillar becomes a pre-pupa.Comparison of these results with those obtained from other Rhopalocera show that the incorporation of melanin into the cuticle is always controlled by a melanization inhibiting hormone, while the incorporation of brown pigments is influenced by pigmentation promoting hormone. Therefore, the different results in ligaturing experiments do not necessarily mean species variation in the controlling hormones, but can be explained by a different action of the same hormonal system on the formation of different pigments.


Evaluating the filtration efficiency of commercial facemasks' materials against respiratory aerosol droplets.
Respiratory droplets serve as a viable transmission mechanism for many viruses and other pathogens. Facemasks are commonly used to minimize the risk of this transmission. However, information on the size-resolved filtration efficiency of commonly available commercial facemasks is not readily available in the literature. To fill this gap, the current study performs aerosolized chamber experiments to evaluate the filtration efficiencies of commonly available commercial facemasks' materials in a size range of 0.3-10 µm. Results rank the performance of filtration through commercial facemasks' materials as follows (values in brackets indicate the average filtration efficiencies across 0.3-10 µm): 6-Layer N95 mask (0.918) > N95 mask - without valve (0.88) > KN95 mask (0.84) > N95 mask -with valve (0.834) > Heavy knitted cotton mask (0.808) > Surgical mask (0.778) > Cotton mask-2 layers (0.744) > Nylon fabric mask-2 layers (0.740) > T-shirt fabric mask-2 layers (0.708) > T-shirt fabric mask-1 layer (0.648). The size-resolved filtration efficiencies through the material across the evaluated commercial facemasks ranged from 38-83% in the size range of 0.3-0.5 µm, 55-88% in the size range of 0.5-1 µm, 69-93% in the size range of 1-2.5 µm, 76-96% in the size range of 2.5-5 µm, and 86-99% in the size range of 5-10 µm. Subsequently, the filtration efficiencies of materials post washing (with detergent in warm water and allowing to dry completely) were also evaluated. The average reduction in filtration efficiencies post washing are as follows: 6-Layer N95 mask: 3%, N95 mask - without valve: 2%, KN95 mask: 4%, N95 mask -with valve: 3%, Heavy knitted cotton mask: 4%, Surgical mask: 18%, Cotton mask-2 layers: 11%, Nylon fabric mask-2 layers: 6%, T-shirt fabric mask-2 layers: 6%, T-shirt fabric mask-1 layer: 8%. This decrease in the filtration efficiency was more pronounced for the sub-micron particles than the super-micron ones.Implications: Facemasks are commonly used to minimize the risk of pathogens through ambient air transmission. However, information on the size-resolved filtration efficiency of commonly available commercial facemasks materials is not readily available in the literature. To fill this gap, the current study performs aerosolized chamber experiments to evaluate the filtration efficiencies of commonly available commercial facemasks materials in a size range of 0.3-10 µm. The performance of the commercial facemasks materials as follows in the order of (values in brackets indicate the average filtration efficiencies across 0.3-10 µm): 6-Layer N95 mask (0.918) > N95 mask - without valve (0.88) > KN95 mask (0.84) > N95 mask -with valve (0.834) > Heavy knitted cotton mask (0.808) > Surgical mask (0.778) > Cotton mask-2 layers (0.744) > Nylon fabric mask-2 layers (0.740) > T-shirt fabric mask-2 layers (0.708) > T-shirt fabric mask-1 layer (0.648). The choice of facemask is greatly driven by the size of viable respiratory droplets that need to be eliminated. If droplets with particle size less than 0.5 µm are required to be filtered, N95 masks without the valve or more layers are preferred. If the primary objective is to filter particles between 0.5-1 µm, then N95 (both with or without valves) or KN95 masks are recommended. Surgical masks and heavy knitted cotton masks may also be used for this purpose, but with caution.


Prevalence of Borrelia burgdorferi and diversity of its outer surface protein C (ospC) alleles in blacklegged ticks (Ixodes scapularis) in Delaware.
Characterizing the diversity of genes associated with virulence and transmission of a pathogen across the pathogen's distribution can inform our understanding of host infection risk. Borrelia burgdorferi is a vector-borne bacterium that causes Lyme disease in humans and is common in the United States. The outer surface protein C (ospC) gene of B. burgdorferi exhibits substantial genetic variation across the pathogen's distribution and plays a critical role in virulence and transmission in vertebrate hosts. In fact, B. burgdorferi infections that disseminate across host tissues in humans are associated with only a subset of ospC alleles. Delaware has a high incidence of Lyme disease, but the diversity of ospC in B. burgdorferi in the state has not been evaluated. We used PCR to amplify ospC in B. burgdorferi-infected blacklegged ticks (Ixodes scapularis) in sites statewide and used short-read sequencing to identify ospC alleles. B. burgdorferi prevalence in blacklegged ticks varied across sites, but not significantly so. We identified 15 previously characterized ospC alleles accounting for nearly all of the expected diversity of alleles across the sites as estimated using the Chao1 index. Nearly 40% of sequenced infections (23/58) had more than one ospC allele present suggesting mixed strain infections and the relative frequencies of alleles in single infections were positively correlated with their relative frequencies in mixed infections. Turnover of ospC alleles was positively related to distance between sites with closer sites having more similar allele compositions than more distant sites. This suggests a degree of B. burgdorferi dispersal limitation or habitat specialization. OspC alleles known to cause disseminated infections in humans were found at the highest frequencies across sites, corresponding to Delaware's high incidence of Lyme disease.


Shortened Treatment Regimens for Drug Sensitive TB.
Tuberculosis (TB) has remained a global health challenge despite the availability of effective anti-tubercular drugs and various treatment strategies. Apart from the complications related to TB disease per se, adverse effects of antitubercular therapy (ATT) also contribute to morbidity. In addition to the adverse effects, the long duration of the treatment regimen also reduces the patient's acceptability of ATT. The available "short-course treatment regimens" are still relatively long, thereby adversely affecting treatment compliance. There is a need for effective, safe, short and intensive regimens for TB which can reduce the treatment cost and adverse effects, thereby improving its acceptance. With the emergence of new evidence, the World Health Organization (WHO) has recently endorsed 4 mo short duration ATT regimen for non-severe, drug-sensitive cases of tuberculosis. Even in severe forms of disease like tubercular meningitis (TBM), trials are underway evaluating efficacy and safety of shorter regimens. Inclusion of fluroquinolones and rifapentine help shorten the regimens. These shortened regimens, however, need more close monitoring for adverse effects and may need to be converted to longer course if there is inadequate clinical response. Thus, shorter regimens for pediatric TB are likely to not only decrease the burden on patients and healthcare but also improve compliance and lower the side effects of the drugs due to prolonged exposure. This article reviews the current evidence and the guidelines pertaining to the shortened, intensive regimens for drug-sensitive tuberculosis.


Inhibition of P-glycoprotein stimulates cell death under Hypoxia-mimicking conditions.
The most common drug resistance mechanism in tumor cells is expression on their surface of the energy-dependent pump like P-glycoprotein (P-gp) that expels chemotherapeutic agents from the interior. An imitation of the hypoxic condition by the iron chelator deferoxamine caused Hypoxia-inducible factor 1-alpha (HIF-1α) stabilization and inhibition of doxorubicin-induced apoptosis in colon cancer НСТ116 cells. P-gp blocker verapamil suppressed doxorubicin accumulation leading to cell death induction. Considering these results, P-gp may be used as a potential target to stimulate chemotherapeutic drugs activity that will contribute to more efficient tumor cells elimination.


Determination of the hormonal growth promoter 17alpha-methyltestosterone in food-producing animals: bovine hair analysis by HPLC-MS/MS.
This paper describes the development, validation and application of a confirmatory method to detect 17alpha-methyltestosterone (MT) in bovine hair, to aid in controlling the administration of this growth promoter in meat-producing animals. After cryogenic grinding, MT was removed from the hair matrix using a single step extraction procedure with acetonitrile. Hydroxylamine derivatisation was used to enhance analyte determination with an electrospray ionisation (ESI) source. Determination was carried out using a triple quadrupole liquid chromatography tandem mass spectrometer (LC-MS/MS) in multiple reaction monitoring mode (MRM). The method was validated in accordance with the criteria defined in Commission Decision 2002/657/EC and using deuterated testosterone (T-d(3)) as the internal standard. The decision limit (CCalpha) was 0.07 ng g(-1) and the detection capability (CCbeta) was 0.12 ng g(-1). Repeatability was CV% (7%), within-laboratory reproducibility was CV% (11.0%), and trueness was (87%). Applicability of the method was demonstrated in an animal study. Samples obtained from animal experiments were analyzed and the presence of MT was confirmed.


Microcalorimetric study of Escherichia coli aerobic growth: kinetics and experimental enthalpy associated with growth on succinic acid.
A new batch calorimeter was used to study the aerobic growth of Escherichia coli in a minimal containing a growth-limiting concentration of succinic acid. The shape of the thermograms obtained was simple when the energy source was the only growth-limiting factor and showed a stationary phase when the oxygen transfer was limiting. The experimentally determined value of the heat production during growth was found to be significantly lower than the heat of combustion of the succinic acid corrected for the assimilated-substrate fraction. An interpretation has been given to explain the difference between the experimental and the theoretical values.


Emphasizing conformal avoidance versus target definition for IMRT planning in head-and-neck cancer.
To describe a method for streamlining the process of elective nodal volume definition for head-and-neck (H&N) intensity-modulated radiotherapy (IMRT) planning. A total of 20 patients who had undergone curative-intent RT for H&N cancer underwent comprehensive treatment planning using three distinct, plan design techniques: conventional three-field design, target-defined IMRT (TD-IMRT), and conformal avoidance IMRT (CA-IMRT). For each patient, the conventional three-field design was created first, thereby providing the "outermost boundaries" for subsequent IMRT design. In brief, TD-IMRT involved physician contouring of the gross tumor volume, high- and low-risk clinical target volume, and normal tissue avoidance structures on consecutive 1.25-mm computed tomography images. CA-IMRT involved physician contouring of the gross tumor volume and normal tissue avoidance structures only. The overall physician time for each approach was monitored, and the resultant plans were rigorously compared. The average physician working time for the design of the respective H&N treatment contours was 0.3 hour for the conventional three-field design plan, 2.7 hours for TD-IMRT, and 0.9 hour for CA-IMRT. Dosimetric analysis confirmed that the largest volume of tissue treated to an intermediate (50 Gy) and high (70 Gy) dose occurred with the conventional three-field design followed by CA-IMRT and then TD-IMRT. However, for the two IMRT approaches, comparable results were found in terms of salivary gland and spinal cord protection. CA-IMRT for H&N treatment offers an alternative to TD-IMRT. The overall time for physician contouring was substantially reduced (approximately threefold), yielding a more standardized elective nodal volume. Because of the complexity of H&N IMRT target design, CA-IMRT might ultimately prove a safer and more reliable method to export to general radiation oncology practitioners, particularly those with limited H&N caseload experience.


Preparation and characterization of a dextran-amylase conjugate.
Bacillus amyloliquefaciens alpha-amylase was attached to dextran after activation of the polysaccharide by using a modification of the cyanogen bromide method. The soluble dextran-amylase conjugate was purified by molecular-sieve chromatography. The conjugated enzyme has greater stability than the unmodified enzyme at low pH values, during heat treatment, and on removal of calcium ions with a chelating agent. Attachment of dextran to alpha-amylase did not alter the Michaelis constant of the enzyme acting on starch. The polysaccharide-enzyme conjugate probably consists of a cross-linked aggregate of many dextran and many enzyme molecules, in which a proportion of the enzyme molecules, although not inactivated, are unable to express their activity, except after dextranase treatment.


Investigation of the antitumor effect on breast cancer cells of the electrospun amygdalin-loaded poly(l-lactic acid)/poly(ethylene glycol) nanofibers.
In this study, PLA/PEG nanofibers (NFs) loaded with amygdalin (AMG) and bitter almond kernels extract were produced by electrospinning to prevent local breast cancer recurrence, and the effect of produced NFs on the MCF-7 cell line was investigated in vitro. The electrospun NFs were characterized by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), thermal analysis (DSC) and tensile strength and physical analyzes were performed. Loading of AMG to nanofibers increased fiber diameters from 827.93 ± 174.507 nm to 1855.32 ± 291.057 μm. When drug release results were analyzed, the NFs showed a controlled release profile extending up to 10 h. The encapsulation efficiency of AMG-loaded NFs was calculated at 100 ± 0,01 %, 94 ± 0,02 %, and 88 ± 0,02 %. When in vitro cytotoxicity results were analyzed, showed that all NFs are effective in inducing cytotoxicity against MCF-7 breast cancer cells. Importantly, 20 mg AMG-loaded NFs displayed effectively higher cytotoxic effects against breast cancer cells relative to the other NFs. Considering all the results, AMG-loaded NFs can give sustained release of drugs at the local sites. Therefore, AMG-loaded nanofibers can reduce the risk of local recurrence of cancer after surgery and can be directly implanted into solid tumor cells for treatment.


Life cycle risk assessment of bottom ash reuse.
The life cycle thinking was integrated with risk assessment to develop the life cycle risk assessment (LCRA) methodology in this study. Because LCRA assessed risks from a life cycle perspective of the concerned policies, it was helpful to identify important sources, contaminants, receptors and exposure pathways along the life cycle of reuse activities. The case study showed that different reuse scenarios resulted in risk shift between different life stages and receptors, and using duration of pavement was an essential factor for risk management. When ash reuse strategies were made based on a focus on the stage of reuse, the rank of strategies were shown to be different from the one based on the total population risks over the entire life cycle. This demonstrated the importance of decision criteria used in selecting reuse strategies. The results also showed that when bottom ash was reused, the health risk was shifted to the laborers; the individual risks of laborers were higher than residents through exposure to Cr and Cd via inhalation and dermal contact. Although the population risk at the treatment stage was the highest, the smaller size of exposed population would make it quite effective to reduce the risk of the laborers.


Feasibility study of an active wound dressing based on hollow fiber membranes in a porcine wound model.
A novel active wound dressing (AWD) concept based on a microporous hollow fiber membrane network was investigated in an animal model. It provides a local solution-perfused environment for regenerative cell nutrition, wound irrigation, debris removal, electrolyte balancing, pH regulation, and topical antibiosis. The device is capable of supplying soluble factors, as tested experimentally for the recombinant human growth and differentiation factor-5 (rhGDF-5). Following in vitro studies for rhGDF-5 using primary human keratinocytes and dermal fibroblasts, we employed a porcine partial thickness wound model with five distinct wounds on each back of n=8 pigs. Four wound groups were perfused differently over 9 days and compared with a negative control wound without perfusion: (1) 1% trehalose solution, pH 5.5; (2) rhGDF-5 (150 ng/ml) in 1% trehalose solution, pH 5.5; (3) nutrition solution; and (4) rhGDF-5 (150 ng/ml) in nutrition solution with 1% trehalose, pH 5.5. Promoted wound healing was observed within group 1 and more pronounced within group 2. Groups 3 and 4, with nutrition solution, showed significant adverse effects on wound healing (p<0.05). The investigated AWD concept appears to be an interesting therapeutic tool to study further wound healing support. Additionally, topical application of rhGDF-5 could be promising.


The association between low-density and non-high-density lipoprotein cholesterol with incident cardiovascular disease among low-risk Iranians during 2 decades follow-up.
To examine the associations between low-density and non-high-density lipoprotein cholesterol (LDL-C and non-HDL-C, respectively) with incident cardiovascular disease (CVD) in low-risk subjects. From a total of 2467 non-diabetic aged 40-70 years, free of CVD with LDL-C range 1.81 ≤ LDL-C < 4.91 mmol/L with 10-year atherosclerotic cardiovascular disease (ASCVD) risk < 7.5 %, the associations of LDL-C and non-HDL-C with incident CVD were assessed using multivariable Cox proportional hazard regression analyses adjusted for age, sex, body mass index, waist circumference, HDL-C, triglycerides, chronic kidney disease, current smoking, hypertension, and family history of CVD. During a median follow-up of 18 years, 559 CVD events occurred. Compared to the LDL-C < 2.59 mmol/L as reference, the categories of 2.59 ≤ LDL-C < 3.36, 3.36 ≤ LDL-C < 4.14, and ≥ 4.14 mmol/L were associated with hazard ratios (95 % confidence intervals) of 1.39(0.89-2.18), 1.72(1.11-2.68), and 2.19(1.36-3.51) for incident CVD (P for trend <0.0001), respectively. Compared to the non-HDL-C < 3.36 as reference, the categories of 3.36 ≤ non-HDL-C < 4.14, 4.14 ≤ non-HDL-C < 4.91, and ≥ 4.91 mmol/L were associated with 1.48(0.96-2.30), 1.37(0.89-2.16), and 2.15(1.36-3.39) higher risk for incident CVD (P for trend = 0.001), respectively. Among those with ASCVD score <5 % (n = 2070), even the 2.59 ≤ LDL-C < 3.36 mmol/L increased the risk for CVD [1.73(1.01-2.97)]. Results for non-HDL-C categories remained unchanged compared to those with ASCVD risk < 7.5%. Among Iranian individuals with ASCVD risk as little as < 5 %, LDL-C ≥ 2.59 mmol/L and non-HDL-C ≥ 3.36 mmol/L, independent of traditional risk factors, were associated with a significantly higher risk of incident CVD, individuals that might potentially benefit from pharmacological therapy.


Ontogeny of T-cell mitogen response in Lewis rats: I. Culture conditions and developmental patterns.
Spleen and thymus cells from female Lewis rats 2 to 220 days old were cultured with phytohemagglutinin (PHA) or Concanavalin A (Con A). The optimal dose of Con A for mitogenesis ranged from 1 microgram/ml for suckling rats to 125 micrograms/ml for adult rats. Thymus cells differed from spleen cells in that they required 72 hours for maximum thymidine incorporation; whereas spleen cells attained maximum incorporation within 48 hours. Thymidine uptake in unstimulated spleen cell cultures was high in the neonate and decreased with age. Maximum splenic cell responses to Con A increased from very low levels in the 2-day old to adult levels by 3 weeks coincident with weaning. Maximum PHA responses of splenocytes matured more slowly, reaching adult levels by 8 weeks. Thymus cell responses to Con A and PHA were mature from birth.


Valve-sparing replacement of the noncoronary sinus of Valsalva in patients with a bicuspid aortic valve.
Progressive dilatation of the aortic root and ascending aorta is frequent in patients with bicuspid aortic valve, and isolated dilatation of the noncoronary sinus has been reported. We describe our technique for the selective replacement of the noncoronary sinus of Valsalva in patients with bicuspid aortic valve.


Unsaturated fatty acids repress the expression of ATP-binding cassette transporter A1 in HepG2 and FHs 74 Int cells.
Adenosine triphosphate-binding cassette transporter A1 (ABCA1) plays a critical role in the formation and metabolism of high-density lipoproteins (HDLs). Adenosine triphosphate-binding cassette transporter A1 in the liver and small intestine, in particular, accounts for approximately 90% of plasma HDL cholesterol. Therefore, any alterations in the hepatic and intestinal expression of ABCA1 could have a large impact on HDL biogenesis. We tested the hypothesis that ABCA1 expression is regulated differentially by different types of fatty acids in the liver and small intestine. Human hepatoma HepG2 and human small intestine epithelial FHs 74 Int cells were used as an in vitro model. Cells were incubated with saturated and unsaturated fatty acids in the presence or absence of T0901317, a synthetic agonist of liver X receptor. Unsaturated fatty acids decreased ABCA1 protein levels at 100 μmol/L of concentration regardless of the agonist with a minimal effect on messenger RNA abundance. Incubation of HepG2 and FHs 74 Int cells with rottlerin, a protein kinase C δ (PKCδ) inhibitor, increased ABCA1 protein but did not abolish linoleic acid-induced decrease in ABCA1 protein levels. Depletion of PKCδ using small interfering RNA showed decreased ABCA1 protein levels in control, palmitic acid-, and linoleic acid-treated cells; but the repressive effect of linoleic acid was sustained. In conclusion, our results indicate that unsaturated fatty acids regulate ABCA1 expression in HepG2 and FHs 74 Int cells at the posttranscriptional level and PKCδ is likely to be involved in maintaining ABCA1 protein levels.


Specific PET imaging of xC- transporter activity using a ¹⁸F-labeled glutamate derivative reveals a dominant pathway in tumor metabolism.
(18)F-labeled small molecules targeting adaptations of tumor metabolism possess the potential for early tumor detection with high sensitivity and specificity by positron emission tomography (PET) imaging. Compounds tracing deranged pathways other than glycolysis may have advantages in situations where 2-[¹⁸F]fluoro-2-deoxy-d-glucose (FDG) has limitations. The aim of this study was the generation of a metabolically stable ¹⁸F-labeled glutamate analogue for PET imaging of tumors. Derivatives of l-glutamate were investigated in cell competition assays to characterize the responsible transporter. An automated radiosynthesis was established for the most promising candidate. The resulting ¹⁸F-labeled PET tracer was characterized in a panel of in vitro and in vivo tumor models. Tumor specificity was investigated in the turpentine oil-induced inflammation model in rats. A fluoropropyl substituted glutamate derivative showed strong inhibition in cell uptake assays. The radiosynthesis was established for (4S)-4-(3-[¹⁸F]fluoropropyl)-l-glutamate (BAY 94-9392). Tracer uptake studies and analysis of knockdown cells showed specific transport of BAY 94-9392 via the cystine/glutamate exchanger designated as system x(C)(-). No metabolites were observed in mouse blood and tumor cells. PET imaging with excellent tumor visualization and high tumor to background ratios was achieved in preclinical tumor models. In addition, BAY 94-9392 did not accumulate in inflammatory lesions in contrast to FDG. BAY 94-9392 is a new tumor-specific PET tracer which could be useful to examine system x(C)(-) activity in vivo as a possible hallmark of tumor oxidative stress. Both preclinical and clinical studies are in progress for further characterization.


Ageing and chronic intermittent hypoxia mimicking sleep apnea do not modify local brain tissue stiffness in healthy mice.
Recent evidence suggests that obstructive sleep apnea (OSA) may increase the risk of Alzheimer´s disease (AD), with the latter promoting alterations in brain tissue stiffness, a feature of ageing. Here, we assessed the effects of age and intermittent hypoxia (IH) on brain tissue stiffness in a mouse model of OSA. Two-month-old and 18-month-old mice (N=10 each) were subjected to IH (20% O2 40s - 6% O2 20s) for 8 weeks (6h/day). Corresponding control groups for each age were kept under normoxic conditions in room air (RA). After sacrifice, the brain was excised and 200-micron coronal slices were cut with a vibratome. Local stiffness of the cortex and hippocampus were assessed in brain slices placed in an Atomic Force Microscope. For both brain regions, the Young's modulus (E) in each animal was computed as the average values from 9 force-indentation curves. Cortex E mean (±SE) values were 442±122Pa (RA) and 455±120 (IH) for young mice and 433±44 (RA) and 405±101 (IH) for old mice. Hippocampal E values were 376±62 (RA) and 474±94 (IH) for young mice and 486±93 (RA) and 521±210 (IH) for old mice. For both cortex and hippocampus, 2-way ANOVA indicated no statistically significant effects of age or challenge (IH vs. RA) on E values. Thus, neither chronic IH mimicking OSA nor ageing up to late middle age appear to modify local brain tissue stiffness in otherwise healthy mice.


Transduced Tat-DJ-1 protein protects against oxidative stress-induced SH-SY5Y cell death and Parkinson disease in a mouse model.
Parkinson's disease (PD) is a well known neurodegenerative disorder characterized by selective loss of dopaminergic neurons in the substantia nigra pars compact (SN). Although the exact mechanism remains unclear, oxidative stress plays a critical role in the pathogenesis of PD. DJ-1 is a multifunctional protein, a potent antioxidant and chaperone, the loss of function of which is linked to the autosomal recessive early onset of PD. Therefore, we investigated the protective effects of DJ-1 protein against SH-SY5Y cells and in a PD mouse model using a cell permeable Tat-DJ-1 protein. Tat-DJ-1 protein rapidly transduced into the cells and showed a protective effect on 6-hydroxydopamine (6-OHDA)-induced neuronal cell death by reducing the reactive oxygen species (ROS). In addition, we found that Tat-DJ-1 protein protects against dopaminergic neuronal cell death in 1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine (MPTP)-induced PD mouse models. These results suggest that Tat-DJ-1 protein provides a potential therapeutic strategy for against ROS related human diseases including PD.


[Retinoscopy under cycloplegic and non-cycloplegic conditions in children comparison of measurements of three examiners (author's transl)].
The refractive status of 100 eyes were measured by three ophthalmologists utilizing retinoscopy under non-cycloplegic and cycloplegic conditions. The statistical analysis of the results was done by multiple regression. The retinoscopic measurements of sphere and cylinder of the three examiners show smaller differences under cycloplegic than under non-cycloplegic conditions. The largest differences in the measurements of axis were found in cases of small cylinder power. Especially in children and in patients with strabismus, the authors feel that measurements of the refractive state have to be carried out under cycloplegia.


Understanding the role of oxidative stress in the incidence of metabolic syndrome and obstructive sleep apnea.
Understanding the causes and risk factors of metabolic syndrome is important for promoting population health. Oxidative stress has been associated with metabolic syndrome, and also obstructive sleep apnea. These are two diseases which have common prognostic characteristics for heart disease. The aim of this study was to examine the role of oxidative stress in the concurrent presence of metabolic syndrome and obstructive sleep apnea in a working population. Participants were 163 artisan bakers in Shahroud, Iran, routinely exposed to significant heat stress and other oxidative stress indicators on a daily basis as part of their work. Using a cross-sectional design, data relevant to determining metabolic syndrome status according to International Diabetes Federation criteria, and the presence of obstructive sleep apnea according to the STOP-Bang score, was collected. Analyses included hierarchical binary logistic regression to yield predictors of the two diseases. Hierarchical binary logistic regression showed that oxidative stress - alongside obesity, no regular exercise, and smoking - was an independent predictor of metabolic syndrome, but not obstructive sleep apnea. Participants who were obese were 28 times more likely to have metabolic syndrome (OR 28.59, 95% CI 4.91-63.02) and 44 times more likely to have obstructive sleep apnea (OR 44.48, 95% CI 4.91-403.28). Participants meeting metabolic syndrome criteria had significantly higher levels of malondialdehyde (p < 0.05) than those who did not. No difference in oxidative stress index levels were found according to obstructive sleep apnea status. Our findings suggest that oxidative stress contributes to the onset of metabolic syndrome, and that obstructive sleep apnea is involved in oxidative stress. Whilst obesity, exercise, and smoking remain important targets for reducing the incidence of metabolic syndrome and obstructive sleep apnea, policies to control risks of prolonged exposure to oxidative stress are also relevant in occupations where such environmental conditions exist.


Silane-treated BaTiO3 ceramic powders for multilayer ceramic capacitor with enhanced dielectric properties.
Silane/ceramic combination provides the composites with several advantages from the advancements of new ceramic composite materials with good thermal conductivity, high mechanical and dielectric properties have wide significant applications in electrical and electronic industries. In this study, to enhance the dispersibility of dielectric barium titanate (BaTiO3) ceramic powder and additives for the fabrication of multilayer ceramic capacitors (MLCCs), surface treatment of the precursor of ceramic powder was performed using silane coupling agents. Dielectric ceramic sheets fabricated from ceramic powders that had been surface-treated with different amounts of N-[3-(trimethoxysilyl)propyl]aniline (TMSPA) which increased the surface gloss. In particular, the dielectric properties of the multilayer ceramic sheet fabricated by stacking sheets from the TMSPA-treated ceramic powder sintering at 1200 °C, it was confirmed that the dielectric constant increased from 881 to 2382 and the dielectric loss dropped from 1.96 to 1.34% with utilization of the TMSPA treatment. The physical and dielectric properties of the TMSPA-treated multilayer ceramic sheet were also determined by Fourier-transform infrared spectroscopy, X-ray diffraction, field-emission scanning electron microscopy, glossmetry, and electrochemical impedance analysis. The results revealed that the TMSPA-modified BaTiO3 surfaces considerably increased the dielectric property of the fabricated nanocomposite.


Quantitative reverse transcription PCR to determine the inactivation of Human Rotavirus by chlorine.
Human rotaviruses (HRVs) are the major cause of acute diarrhea in infants and young children. Here, a real-time reverse transcription polymerase chain reaction assay targeting the rotaviral VP4 gene (VP4-RT-qPCR) was established to evaluate the inactivation of HRV upon chlorine disinfection, based on a previous report that damage to the 1227-2354bp region of the VP4 gene was associated with eliminated HRV infectivity by chlorine. In this study, inactivation of HRV by 0.6mg/L free chlorine was assessed in phosphate buffered saline (PBS; pH 7.2), and tap and river water samples, using both TCID50 and RT-qPCR (VP2- and VP4-RT-qPCR) assays, respectively. Among the samples tested, the VP2-RT-qPCR method did not show significant inactivation after chlorine disinfection; however, the reduction in VP4-RT-qPCR signal was correlated with decreased HRV infectivity. Moreover, the higher sensitivity of the VP4-RT-qPCR assay allowed for assessment of chlorine HRV inactivation at longer exposure times compared with the conventional TCID50 assay. Collectively, these results indicated that the VP4-RT-qPCR assay is a rapid, sensitive, and reliable tool to detect infectious HRV following chlorine inactivation, and highlights the potential for further development of qPCR/RT-qPCR assays to provide information regarding viral infectivity from drinking water plants.


Prophylaxis of bladder tumor implantation. Intravesical and systemic chemotherapy.
Several antitumor agents have been evaluated for their effectiveness in reducing the implantation and progressive growth of transitional cell carcinoma in an animal model. This system allows the evaluation of both the topical cytotoxic action as well as the systemic toxicity of the drugs given intravesically. Systemic cyclophosphamide and intravesical epipodophyllotoxin significantly reduced the incidence of tumor cell implantation.


Palladium-Catalyzed syn-Stereocontrolled Ring Opening of Oxabicyclic Alkenes with Arylsulfonyl Hydrazides.
A novel palladium-catalyzed ring-opening reaction of oxabicyclic alkenes with arylsulfonyl hydrazides was first developed. In this work, we provide an efficient one-pot reaction to afford the corresponding cis-2-aryl-1,2-dihydronaphthalen-1-ols and 2-aryl-naphthalenes in moderate to excellent yields (up to 95%) under an open-air condition. Various types of functional groups attached to the substrates were tolerated well in this method. Among them, the cis-1,2-configuration of product 3ag was confirmed by X-ray crystallographic analysis. In addition, a plausible mechanism for ring opening was also proposed.


Structural and functional analysis of the amdR regulatory gene of Aspergillus oryzae.
We have isolated the Aspergillus oryzae homologue of the amdR regulatory gene of Aspergillus nidulans by cross hybridization. Sequence analysis and functional studies have shown that the amdR genes are highly conserved and functionally interchangeable between the two species. The homology between the two genes extends throughout most of the coding sequences, including sequences encoding the DNA-binding domain and putative activation domains. Two regions of nonconserved sequence were also identified. Studies using various amdS::lacZ fusion constructs indicate that the A. oryzae gene product binds similar sequences and responds to inducer in a similar manner to the A. nidulans protein. Inactivation of the A. oryzae gene results in the inability to grow on gamma-amino-butyric acid (GABA) as a carbon and/or nitrogen source indicating that GABA utilization is amdR-dependent in A. oryzae as it is in A. nidulans.


Preterm birth results in alterations in neural connectivity at age 16 years.
Very low birth weight preterm (PT) children are at high risk for brain injury. Employing diffusion tensor imaging (DTI), we tested the hypothesis that PT adolescents would demonstrate microstructural white matter disorganization relative to term controls at 16 years of age. Forty-four PT subjects (600-1250 g birth weight) without neonatal brain injury and 41 term controls were evaluated at age 16 years with DTI, the Wechsler Intelligence Scale for Children-III (WISC), the Peabody Picture Vocabulary Test-Revised (PPVT), and the Comprehensive Test of Phonological Processing (CTOPP). PT subjects scored lower than term subjects on WISC full scale (p=0.003), verbal (p=0.043), and performance IQ tests (p=0.001), as well as CTOPP phonological awareness (p=0.004), but scored comparably to term subjects on PPVT and CTOPP Rapid Naming tests. PT subjects had lower fractional anisotropy (FA) values in multiple regions including bilateral uncinate fasciculi (left: p=0.01; right: p=0.004), bilateral external capsules (left: p<0.001; right: p<0.001), the splenium of the corpus callosum (p=0.008), and white matter serving the inferior frontal gyrus bilaterally (left: p<0.001; right: p=0.011). FA values in both the left and right uncinate fasciculi correlated with PPVT scores (a semantic language task) in the PT subjects (left: r=0.314, p=0.038; right: r=0.336, p=0.026). FA values in the left and right arcuate fasciculi correlated with CTOPP Rapid Naming scores (a phonologic task) in the PT subjects (left: r=0.424, p=0.004; right: r=0.301, p=0.047). These data support for the first time that dual pathways underlying language function are present in PT adolescents. The striking bilateral dorsal correlations for the PT group suggest that prematurely born subjects rely more heavily on the right hemisphere than typically developing adults for performance of phonological language tasks. These findings may represent either a delay in maturation or the engagement of alternative neural pathways for language in the developing PT brain.


Integrated Analysis of Key Pathways and Drug Targets Associated With Vogt-Koyanagi-Harada Disease.
Vogt-Koyanagi-Harada (VKH) disease is a complex disease associated with multiple molecular immunological mechanisms. As the underlying mechanism for VKH disease is unclear, we hope to utilize an integrated analysis of key pathways and drug targets to develop novel therapeutic strategies. Candidate genes and proteins involved in VKH disease were identified through text-mining in the PubMed database. The GO and KEGG pathway analyses were used to examine the biological functions of the involved pathways associated with this disease. Molecule-related drugs were predicted through Drug-Gene Interaction Database (DGIdb) analysis. A total of 48 genes and 54 proteins were associated with VKH disease. Forty-two significantly altered pathways were identified through pathway analysis and were mainly related to immune and inflammatory responses. The top five of significantly altered pathways were termed as "inflammatory bowel disease," "cytokine-cytokine receptor interaction," "allograft rejection," "antigen processing," and "presentation and Herpes simplex infection" in the KEGG database. IFN-γ and IL-6 were identified as the key genes through network analysis. The DGIdb analysis predicted 48 medicines as possible drugs for VKH disease, among which Interferon Alfa-2B was co-associated both with IFN-γ and IL-6. In this study, systematic analyses were utilized to detect key pathways and drug targets in VKH disease via bioinformatics analysis. IFN-γ and IL-6 were identified as the key mediators and possible drug targets in VKH disease. Interferon Alfa-2B was predicted to be a potentially effective drug for VKH disease treatment by targeting IFN-γ and IL-6, which warrants further experimental and clinical investigations.


Relationship between Epicardial and Coronary Adipose Tissue and the Expression of Adiponectin, Leptin, and Interleukin 6 in Patients with Coronary Artery Disease.
Adipose tissue (AT) is an endocrine and paracrine organ that synthesizes biologically active adipocytokines, which affect inflammation, fibrosis, and atherogenesis. Epicardial and perivascular fat depots are of great interest to researchers, owing to their potential effects on the myocardium and blood vessels. The aim of the study was to assess the expression and secretion of adipocytokine genes in the AT of patients with coronary artery disease (CAD) and patients with aortic or mitral valve replacement. This study included 84 patients with CAD and 50 patients with aortic or mitral valve replacement. Adipocytes were isolated from subcutaneous, epicardial (EAT), and perivascular AT (PVAT), and were cultured for 24 h. EAT exhibited the lowest level of adiponectin gene expression and secretion, regardless of nosology, and high expression levels of the leptin gene and interleukin-6 (IL-6). However, EAT adipocytes in patients with CAD were characterized by more pronounced changes in comparison with the group with heart defects. High leptin and IL-6 levels resulted in increased pro-inflammatory activity, as observed in both EAT and PVAT adipocytes, especially in individuals with CAD. Therefore, our results revealed the pathogenetic significance of alterations in the adipokine and cytokine status of adipocytes of EAT and PVAT in patients with CAD.


A review of cerebral palsy for the oral health professional.
Individuals who have cerebral palsy (CP) face many physical challenges throughout their lifetimes in addition to societal barriers that can have an impact on quality of life. The ability to access appropriate dental care has long been an issue for people who have disabilities. Dentists should be integral members of teams of professionals involved in optimizing the health of individuals who have CP. As with all members of this interdisciplinary team, oral health care providers should have a thorough knowledge of the medical, cognitive, and rehabilitative issues associated with CP. With this knowledge the best possible health care can be provided.


Comparison of outpatient services between elderly people with intellectual disabilities and the general elderly population in Taiwan.
This study aims to analyze the ambulatory visit frequency and medical expenditures of the general elderly population versus the elderly with intellectual disabilities in Taiwan, while examining the effects of age, gender, urbanization and copayment status on ambulatory utilization. A cross-sectional study was conducted to analyze data from 103,183 national health insurance claimants aged 65 or older. A total of 1469 had a principal diagnosis of mental retardation (intellectual disability) and claimed medical outpatient services in 2007. The average number of ambulatory visits was 30.1 ± 23.1, which is much higher than in the United States and other developed countries, and the mean annual visits of the elderly with intellectual disabilities was significantly higher than the general population in Taiwan (35.2 ± 28.7 vs. 30.0 ± 23.1). Age and copayment status affected outpatient visit frequency. The mean medical expenditure per visit and the mean annual outpatient cost were 1146.5 ± 4497.7 NT$ and 34,533.7 ± 115,891.7 NT$, respectively. Male beneficiaries tended to have higher average annual medical expenses and mean medical expenses per visit than female beneficiaries. The three most frequent principal diagnoses at ambulatory visits were circulatory system diseases, musculoskeletal system and connective tissue diseases and digestive system diseases. We conclude that the elderly with intellectual disabilities had higher demand than the general population for healthcare services, and the NHI program lowers the barrier to care for populations with special needs.


Effect of Myostatin Gene Mutation on Slaughtering Performance and Meat Quality in Marchigiana Bulls.
The myostatin gene also called Growth Differentiation Factor 8 gene (GDF8) is one of the most investigated loci that can be responsible for several quantitative and qualitative carcass and meat traits in double-muscled beef cattle. The objective of the study was to bring to light the effect of the myostatin polymorphism on slaughtering performance and meat quality in Marchigiana beef cattle. The experiment was carried out on 78 bulls reared according to the "cow-calf" extensive managing system. At the end of the fattening period, in vivo and carcass data were recorded. From each carcass, a steak of Longissimus thoracis was taken and used to determine the meat's analytical composition and colorimetric properties. Finally, from each steak a sample of Longissimus thoracis was collected, then used for DNA extraction and genotyping at the myostatin locus. The heterozygous bulls showed slight superiority in the carcass data (e.g., hot carcass weight: 426.09 kg-heterozygotes vs. 405.32 kg-normal) and meat quality parameters, although not always with statistical significance. Only fat and ashes content were significantly affected by the myostatin genotype (heterozygotes: 2.01%, 1.26%; normal: 3.04%, 1.15%). The greater muscularity of heterozygous animals compared to normal ones could be a starting point to improving productive efficiency in Marchigiana beef cattle.


Targeting angiopoietin-2 as a novel treatment option for kidney fibrosis.
Kidney fibrosis is a hallmark of chronic kidney disease yet is poorly treated. Chang et al. determined that plasma and kidney levels of the vascular growth factor, angiopoietin-2, were elevated in patients with chronic kidney disease and mice with kidney disease. Angiopoietin-2 inhibited the renoprotective effects of angiopoietin-1 and promoted CC chemokine ligand 2-mediated kidney damage, endothelial cell apoptosis, vascular rarefaction, inflammation, fibrosis, and kidney dysfunction. Hence, therapeutically inhibiting angiopoietin-2 may represent a novel means of treating these chronic kidney disease-associated pathologies.


Isoproterenol- and insulin-induced hyperpolarization in rat skeletal muscle.
Using conventional microelectrode techniques, we investigated the combined effects of isoproterenol (Iso) and insulin (Ins) on the resting membrane potential (RMP) of isolated rat skeletal muscles. In soleus muscle, Iso (1 microM) and Ins (4 units/L) separately induced a hyperpolarization of 9.2 mV and 4.8 mV, respectively. Combined administration of Iso and Ins induced a hyperpolarization of 12.4 mV, larger than either one separately. A similar observation was made in Na(+)-loaded rewarming experiments. 8-Br-cAMP (1 mM and 3 mM) and forskolin (10 microM, an adenylate cyclase activator) induced a hyperpolarization of 5.3 mV, 8 mV, and 6.0 mV, respectively. This hyperpolarizing action was blocked by ouabain, indicating that the Na-K pump was involved in the hyperpolarization. 8-Br-cGMP (3 mM) had no effect on RMP; however, it blocked or reversed the hyperpolarization caused by 8-Br-cAMP (1 mM). In addition, 8-Br-cGMP partially inhibited the hyperpolarizing effect of Iso (1 microM) by 40% and completely prevented the effect of Ins. The phorbol ester, PMA, (1 microM, a PKC activator) induced a ouabain-inhibitable hyperpolarization. These results suggest that cAMP and PKC are involved in the Iso- and Ins-induced hyperpolarization and that Iso and Ins influence the RMP presumably through regulation of the electrogenic Na-K pump via different mechanisms.


Effect of tension-free vaginal tape and TVT-obturator on lower urinary tract symptoms other than stress urinary incontinence.
Variable effects on lower urinary tract symptoms (LUTS) other than stress urinary incontinence (SUI) have been reported after tension-free vaginal tape (TVT). We measured the effect of TVT on LUTS using the American Urological Association Symptom Index (AUASI). Patients undergoing TVT completed the AUASI pre- and post-operatively. Total scores (TS), storage scores (SS), and voiding scores (VS) were compared overall and among patients with SUI vs mixed urinary incontinence (MUI) and those who underwent TVT vs TVT-obturator (TVT-O). The mean change in TS and SS was -3.6 and -3.0. Mean reductions in TS and SS were significant in all patient subsets with no change in VS. There was no significant difference in the mean changes in TS between patients with SUI vs MUI or those undergoing TVT vs TVT-O. LUTS are improved after TVT in most patients. In general, voiding symptoms were not adversely affected.


Genome-wide identification and expression profiling of snakin/GASA genes under drought stress in barley (Hordeum vulgare L.).
Gibberellic Acid-Stimulated Arabidopsis (GASA) proteins are present in various plants and have a role in plant growth, stress responses, and hormone crosstalk. GASA coding sequences in barley were discovered in this study. We then investigated gene and protein structure, physicochemical characteristics, evolutionary and phylogenetic relationships, promoter region, post-translational modification, and in silico gene expression. Finally, real-time quantitative PCR (RT-qPCR) was used to examine the expression of GASA genes in root and shoot tissues under drought stress. We found 11 GASA genes spread across six of seven chromosomes in the barley genome. A conserved GASA domain and 12-cysteine residues at the C-terminus were included in the proteins. All GASA genes contained secretory signal peptides. The GASA genes in Hordeum vulgare (HvGASA) have been classified into three subfamilies based on evolutionary analysis. According to synteny analyses, segmental duplications are significant in forming the GASA gene family. According to the cis-elements analyses, GASA genes may be induced by a variety of phytohormones and stresses. Tissue-specific expression analysis indicated that GASA genes had varied expression patterns in different tissues. Contrary to common perception, the expression study of GASA genes under biotic and abiotic stresses revealed that GASA genes are more induced by abiotic stresses than biotic stresses. The qPCR confirmed the response of GASA genes to abiotic stresses and showed different expression patterns of these genes under drought stress. Overall, these results can improve our knowledge about the function of GASA genes and provide data for future researches. The online version contains supplementary material available at 10.1007/s13205-023-03545-8.


The right to enjoy the benefits of scientific progress: in search of state obligations in relation to health.
After having received little attention over the past decades, one of the least known human rights--the right to enjoy the benefits of scientific progress and its applications--has had its dust blown off. Although included in the Universal Declaration of Human Rights (UDHR) and in the International Covenant on Economic, Social and Cultural Rights (ICESCR)--be it at the very end of both instruments -this right hardly received any attention from States, UN bodies and programmes and academics. The role of science in societies and its benefits and potential danger were discussed in various international fora, but hardly ever in a human rights context. Nowadays, within a world that is increasingly turning to science and technology for solutions to persistent socio-economic and development problems, the human dimension of science also receives increased attention, including the human right to enjoy the benefits of scientific progress and its applications. This contribution analyses the possible legal obligations of States in relation to the right to enjoy the benefits of scientific progress and its applications, in particular as regards health.


Vascular endothelial growth factor 165 inhibits pro-fibrotic differentiation of stromal cells via the DLL4/Notch4/smad7 pathway.
Endometrial fibrosis is the main pathological feature of Asherman's syndrome (AS), which is the leading cause of uterine infertility. Much is known about the expression of VEGF165 in luminal/glandular epithelial cells and stromal cells of the endometrium in normal menstrual cycles; however, less is known about the role and mechanism of VEGF165 in endometrial fibrosis. Herein, we report that VEGF165 is a key regulator in endometrial stromal cells to inhibit α-SMA and collagen 1 expression. Compared to human control subjects, patients with AS exhibited decreased VEGF165 expression in the endometrium along with increased fibrotic marker expression and collagen production. A fibrotic phenotype was shown in both mice with conditional VEGF reduction and VEGF165-deleted endometrial stromal cells. Exogenous VEGF165 could suppress TGFβ1-induced α-SMA and collagen 1 expression in human primary endometrial stromal cells. However, this beneficial effect was hindered when the expression of smad7 or Notch4 was inhibited or when Notch signaling was blocked, suggesting that smad7 and Notch4 are essential downstream molecules for VEGFA functioning. Overall, our results uncover a clinical targeting strategy for VEGF165 to inhibit pro-fibrotic differentiation of stromal cells by inducing DLL4/Notch4/smad7, which paves the way for AS treatment.


Elevation of circulating intestinal fatty acid binding protein in a luminal contents-initiated model of NEC.
Necrotizing enterocolitis (NEC) continues to produce significant morbidity and mortality, due in part to the difficulty in detecting its initial manifestations at a stage when compromised intestine may potentially be salvaged. We have previously reported our findings that intestinal fatty acid binding protein (I-FABP) is a sensitive biochemical marker for early intestinal mucosal injury due to mesenteric ischemia. In this study we evaluated the potential of serum I-FABP as a marker for incipient NEC in a nonischemic model of NEC in the rat. Intraluminal instillation of a solution of casein (10 mg/mL) and calcium gluconate (50 mg/mL) in saline acidified to pH 4.0 with propionic acid resulted in a rapid and prolonged increase in serum I-FABP from a baseline of < or = 4.0 ng/mL to 171 +/- 40 ng/mL. Instillation of the same electrolyte solution with either casein or propionic acid alone resulted in a less dramatic elevation of serum I-FABP to 19 +/- 4 ng/mL and 76 +/- 30 ng/mL, respectively. In both cases baseline values of < or = 4.0 ng/mL were reached within 60 minutes. In control animals, which received saline alone, serum I-FABP was undetectable throughout the experiment. Simultaneously, we found that serum hexosaminidase, a putative biochemical marker for intestinal ischemia and NEC, was unchanged in all groups. Light microscopy of the intestinal specimens obtained three hours after instillation of casein and organic acid demonstrated superficial villus necrosis and villus blunting, but no areas of transmural necrosis.(ABSTRACT TRUNCATED AT 250 WORDS)


Sensitive HPLC method for quantitation of paclitaxel (Genexol in biological samples with application to preclinical pharmacokinetics and biodistribution.
A sensitive, specific and reproducible HPLC method has been developed and validated for the quantitative determination of paclitaxel in plasma, tissues and tumor of mice. Tissue specimens including liver, kidneys, spleen, lungs, heart and tumor were separately homogenized in bovine serum albumin (BSA, 40 g/l) in water. Plasma or tissue homogenates (0.1 ml) containing paclitaxel and internal standard (dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylene dioxy biphenyl-2',2' dicarboxylate (DDB), I.S.) were extracted by ethyl acetate (10 ml). A 4.6 mm x 250 mm ODS column was used to separate the components in biological samples with UV detection at 227 nm and gradient system was applied to a quantitation of paclitaxel consisting of acetonitrile-deionized water. The I.S. and paclitaxel were eluted at 13.7 and 18.0 min, respectively, and no interfering peaks were observed. Linear relationships (r(2) > 0.999) were obtained between the peak height ratios and the corresponding biological sample concentrations over the range of 0.1-20 microg/ml. The average intra- and inter-day variations (% R.S.D.s and % deviations) of the assay for biological samples were less than 10%. The LOD and LOQ were 5 and 10 ng/ml, respectively, for paclitaxel using a microsample volume (100 microl) of plasma sample. This HPLC method has been successfully applied for the determination of paclitaxel in pharmacokinetic and biodistribution study in after administration of 50 mg equivalent paclitaxel/kg dose of paclitaxel-loaded polymeric micelle and 20 mg equivalent paclitaxel/kg dose of Taxol to female SPF C57BL/6 mice.


Δ⁹Tetrahydrocannabinol impairs visuo-spatial associative learning and spatial working memory in rhesus macaques.
Cannabis remains the most commonly abused illicit drug and is rapidly expanding in quasi-licit use in some jurisdictions under medical marijuana laws. Effects of the psychoactive constituent Δ⁹tetrahydrocannabinol (Δ⁹THC) on cognitive function remain of pressing concern. Prior studies in monkeys have not shown consistent evidence of memory-specific effects of Δ⁹THC on recognition tasks, and it remains unclear to what extent Δ⁹THC causes sedative versus specific cognitive effects. In this study, adult male rhesus monkeys were trained on tasks which assess spatial working memory, visuo-spatial associative memory and learning as well as motivation for food reward. Subjects were subsequently challenged with 0.1-0.3 mg/kg Δ⁹THC, i.m., in randomized order and evaluated on the behavioral measures. The performance of both vsPAL and SOSS tasks was impaired by Δ⁹THC in a dose and task-difficulty dependent manner. It is concluded that Δ⁹THC disrupts cognition in a way that is consistent with a direct effect on memory. There was evidence for interference with spatial working memory, visuo-spatial associative memory and incremental learning in the latter task. These results and the lack of specific effect of Δ⁹THC in prior visual recognition studies imply a sensitivity of spatial memory processing and/or working memory to endocannabinoid perturbation.


Outcome measures for early phase clinical trials.
As the number of potential neuroprotective agents for ALS increases, the need for early phase trials that screen drugs before proceeding to efficacy trials also grows. However, it is not known which outcome measures perform best and also provide the most meaningful information in brief small trials. We assessed the performance of different outcome measures for use in early phase clinical trials in ALS, and determined what degree of change in the ALSFRS-R that patients could perceive. Thirty patients underwent six monthly ALS Functional Rating Scale (ALSFRS-R), forced vital capacity, manual muscle testing (MMT) and quality of life assessments. Patients rated their perceived level of change with algorithm scales. Linear mixed effects models assessed the associations among variables and Cox proportional-hazards models examined the ability to predict survival. The quantity of missing data was assessed using descriptive statistics. Correlations were found between all variables. The ALSFRS-R provided the most complete data (99.5%), showed a large within-subject correlation (0.91), and best predicted survival (p = 0.002). One-unit change in patient-perceived clinical function paralleled a 9-point decrease in the ALSFRS-R (p = 0.025; 95% CI 8, 10). This trial assessed just 30 patients over six months, but the standard outcome measures each performed dependably; all could be used in short-duration, early phase trials. The ALSFRS-R most strongly predicted survival and provided the most complete data, but large changes may be necessary before patients perceive treatment effects.


Detection of local recurrent disease in musculoskeletal tumors: magnetic resonance imaging versus computed tomography.
A 2 1/2-year prospective study of surgically treated malignant mesenchymal neoplasms showed magnetic resonance imaging (MRI) to be superior to computed tomography (CT) in sensitivity for local recurrent disease measuring less than 15 cm3. Larger masses were detected with similar sensitivity; specificity and predictive values did not differ. The presence of areas of high signal intensity on T2-weighted images proved to be a reliable criterion except in fibrous neoplasms. However, differentiation between non-hemorrhagic fluid collections, cross-sectioned veins or bowel contents and small tumor nodules cannot be made simply by signal intensity, but has to be based upon the evaluation of gross morphologic criteria.


[Induction-resonance energy transfer between the terbium-binding peptide and the red fluorescent proteins Dsred2 and TagRFP].
Two novel FRET-pairs: Tb3+ -binding peptide-DsRed2 and Tb3+ -binding peptide-TagRFP have been produced based on the terbium-binding peptide and the red fluorescent proteins DsRed2 and TagRFP. Two induction-resonance energy transfer processes in both FRET-pairs have been registered, from tryptophan of the terbium-binding peptide to Tb3+ and from sensitized Tb3+ to the acceptor, the chromophore, DsRed2 or TagRFP. The lifetimes of terbium in the presence and absence of the acceptor have been determined. It has been shown that the lifetime of Tb3+ in the presence of 150 mM NaCl decreases in both FRET-pairs. The efficiency of fluorescent resonance transfer from Tb3+ to the acceptor proteins is 28 and 35% for Tb3+ -binding peptide-DsRed2 and Tb3+ -binding peptide-TagRFP, respectively.


Visualizing spatial distribution of small molecules in the rhubarb stalk (Rheum rhabarbarum) by surface-transfer mass spectrometry imaging.
Laser desorption/ionization mass spectrometry imaging (LDI-MSI) with gold nanoparticle-enhanced target (AuNPET) was used for visualization of small molecules in the rhubarb stalk (Rheum rhabarbarum L.). Analysis was focused on spatial distribution of biologically active compounds which are found in rhubarb species. Detected compounds belong to a very wide range of chemical compound classes such as anthraquinone derivatives and their glucosides, stilbenes, anthocyanins, flavonoids, polyphenols, organic acids, chromenes, chromanones, chromone glycosides and vitamins. The analysis of the spatial distribution of these compounds in rhubarb stalk with the nanoparticle-rich surface of AuNPET target plate has been made without additional matrix and with minimal sample preparation steps.


Detection and phylogenetic characterization of human hepatitis E virus strains, Czech Republic.
To determine the origin of hepatitis E virus in the Czech Republic, we analyzed patient clinical samples. Five isolates of genotypes 3e, 3f, and 3g were obtained. Their genetic relatedness with Czech strains from domestic pigs and wild boars and patient recollections suggest an autochthonous source likely linked to consumption of contaminated pork.


[Primary sclerosing cholangitis and primary biliary liver cirrhosis as the causes of intrahepatic cholestasis].
Primary sclerosing cholangitis and primary biliary cirrhosis are rarely diagnosed diseases which should, however, be considered in the diagnosis of cholestasis. The authors present an outline of the modern knowledge on these diseases with particular reference to differential diagnosis.


The complete plastid genome of the brown alga Scytosiphon lomentaria (scytosiphonaceae, phaeophyceae).
The complete plastid genome of Scytosiphon lomentaria was determined in this study. The circular genome was 139,208 bp in length with the GC content of 31.3%. It contained 137 protein-coding genes (PCGs), 27 transfer RNA (tRNA) genes, and 8 ribosome RNA (rRNA) genes. A phylogenetic analysis based on the plastid genomes of Phaeophyceae indicated that S. lomentaria is located in the Ectocarpales lineage, being closely related to Endarachne binghamiae.


Effects of dietary calcium from dairy products on ambulatory blood pressure in hypertensive men.
The effect of consumption of 400 mg vs 1,500 mg of dietary calcium per day was examined in 13 male volunteers who had been diagnosed as hypertensive. Dietary calcium consumption was varied by manipulation of the intake of dairy products over 4-week periods. Caffeine intake (mean = 500 mg/day) was monitored. Neither laboratory blood pressure measured by standard sphygmomanometry nor ambulatory blood pressure monitored by automated sphygmomanometry varied significantly among men consuming baseline, low-calcium, or high-calcium diets (laboratory blood pressure = 136/83, 133/83, and 137/84 mm Hg, respectively; ambulatory blood pressure = 136/86, 138/87, and 138/87 mm Hg, respectively). Serum ionized calcium values did not vary with the three diets (1.25, 1.26, and 1.25 mmol/L, respectively). The parathyroid hormone level decreased (39 mmol/L vs 37 mmol/L) and the urinary calcium:creatinine ratio was elevated (0.41 vs 0.50) in the high-calcium diet. Consumption of a diet containing 1,500 mg calcium per day over 4 weeks did not produce a significant decrease in blood pressure or alterations in calcium metabolic indexes.


Fungal infection profile in critically ill COVID-19 patients: a prospective study at a large teaching hospital in a middle-income country.
Critically ill COVID-19 patients are highly susceptible to opportunistic fungal infection due to many factors, including virus-induced immune dysregulation, host-related comorbidities, overuse and misuse of antibiotics or corticosteroids, immune modulator drugs, and the emergencies caused by the pandemic. This study aimed to assess the incidence, identify the potential risk factors, and examine the impact of fungal coinfection on the outcomes of COVID-19 patients admitted to the intensive care unit (ICU). A prospective cohort study including 253 critically ill COVID-19 patients aged 18 years or older admitted to the isolation ICU of Zagazig University Hospitals over a 4-month period from May 2021 to August 2021 was conducted. The detection of a fungal infection was carried out. Eighty-three (83) patients (32.8%) were diagnosed with a fungal coinfection. Candida was the most frequently isolated fungus in 61 (24.1%) of 253 critically ill COVID-19 patients, followed by molds, which included Aspergillus 11 (4.3%) and mucormycosis in five patients (1.97%), and six patients (2.4%) diagnosed with other rare fungi. Poor diabetic control, prolonged or high-dose steroids, and multiple comorbidities were all possible risk factors for fungal coinfection [OR (95% CI) = 10.21 (3.43-30.39), 14.1 (5.67-35.10), 14.57 (5.83-33.78), and 4.57 (1.83-14.88), respectively]. Fungal coinfection is a common complication of critically ill COVID-19 patients admitted to the ICU. Candidiasis, aspergillosis, and mucormycosis are the most common COVID-19-associated fungal infections and have a great impact on mortality rates.


Integrins in metastatic adenoid cystic carcinoma.
Adenoid cystic carcinoma exhibits a diverse clinical behaviour from very localised invasion to widespread metastatases. Recently two adenoid cystic carcinoma cells lines have been isolated and cultured which distinctly show either a low metastatic tendency (Acc-2) or a highly metastatic behaviour (Acc-M). It was hypothesised that these two types of behaviour may at least in part be explained by the different integrin profile on the cells' surface membrane. The integrins represent the largest known family of cell adhesion molecules and a number of tumour cell processes have been shown to be dependant on their integrin expression. Adenoid cystic carcinoma cells of both types were obtained and successfully cultured. These were then subjected to integrin analysis by a number of monoclonal antibodies to alpha(1), alpha(2), alpha(3), alpha(v), alpha(6), beta(1), alpha(v)beta6, alpha(v)beta4 using a fluorescence activated cell sorter. Although there was an apparent difference of integrin expression in the Acc-M group as compared to the Acc-2 group, this was not statistically significant. It is still possible though, that this might account for the differences in behaviour of the two cell lines, and the relationship between integrins, and invasion and metastases in other tumours is discussed. The pattern of integrin expression in Acc may be prognostically significant and useful for treatment planning in the future.


Therapeutic effects of lisinopril and empagliflozin in a mouse model of hypertension-accelerated diabetic kidney disease.
Hypertension is a critical comorbidity for progression of diabetic kidney disease (DKD). To facilitate the development of novel therapeutic interventions with the potential to control disease progression, there is a need to establish translational animal models that predict treatment effects in human DKD. The present study aimed to characterize renal disease and outcomes of standard of medical care in a model of advanced DKD facilitated by adeno-associated virus (AAV)-mediated renin overexpression in uninephrectomized (UNx) db/db mice. Five weeks after single AAV administration and 4 wk after UNx, female db/db UNx-ReninAAV mice received (PO, QD) vehicle, lisinopril (40 mg/kg), empagliflozin (20 mg/kg), or combination treatment for 12 wk (n = 17 mice/group). Untreated db/+ mice (n = 8) and vehicle-dosed db/db UNx-LacZAAV mice (n = 17) served as controls. End points included plasma, urine, and histomorphometric markers of kidney disease. Total glomerular numbers and individual glomerular volume were evaluated by whole kidney three-dimensional imaging analysis. db/db UNx-ReninAAV mice developed hallmarks of progressive DKD characterized by severe albuminuria, advanced glomerulosclerosis, and glomerular hypertrophy. Lisinopril significantly improved albuminuria, glomerulosclerosis, tubulointerstitial injury, and inflammation. Although empagliflozin alone had no therapeutic effect on renal endpoints, lisinopril and empagliflozin exerted synergistic effects on renal histological outcomes. In conclusion, the db/db UNx-ReninAAV mouse demonstrates good clinical translatability with respect to physiological and histological hallmarks of progressive DKD. The efficacy of standard of care to control hypertension and hyperglycemia provides a proof of concept for testing novel drug therapies in the model.NEW & NOTEWORTHY Translational animal models of diabetic kidney disease (DKD) are important tools in preclinical research and drug discovery. Here, we show that the standard of care to control hypertension (lisinopril) and hyperglycemia (empagliflozin) improves physiological and histopathological hallmarks of kidney disease in a mouse model of hypertension-accelerated progressive DKD. The findings substantiate hypertension and type 2 diabetes as essential factors in driving DKD progression and provide a proof of concept for probing novel drugs for potential nephroprotective efficacy in this model.


Rapid Ag/Sn/Ag transient liquid phase bonding for high-temperature power devices packaging by the assistance of ultrasound.
Rapid transient liquid phase (TLP) bonding process on Ag/Sn/Ag system is achieved in air by the assistance of ultrasonic, which has great potential to be applied to high-temperature power devices packaging. In this study, the influence of ultrasonic effect on the morphology and growth kinetics of Ag3Sn grains, and the joint microstructure, mechanical property and thermal reliability were systematically investigated. Experimental results indicated that the rapid consumption of the "dynamic" transient liquid phase was attributed to the accelerated dissolution of Ag substrate and the extrusion of liquid Sn, which were entirely induced by the complex sonochemical effects on the liquid/solid intermetallic compounds (IMCs) interface. An elongated scallop-like morphology of Ag3Sn grains was developed during Ag/Sn interfacial reaction with ultrasonic, accompanied by widening of grooves between neighbored grains. This phenomenon is called as a strengthening thermal grooving, in which the grooves at grain boundaries provide stable molten channels for Ag atoms diffusion from the substrate. Consequently, the improved elemental diffusion was evaluated through the growth kinetics of Ag3Sn IMCs, with conservative estimation of 6-16.5 times faster than the traditional TLP process. In addition, both excellent mechanical property and thermal reliability of the Ag-Sn intermetallic joint were experimentally verified by shear test and high-temperature storage test, respectively.


Biomimetic sensing layer based on electrospun conductive polymer webs.
The aim of the present study is to combine a bio-inspired nanofibrous artificial epithelium to the electronic nose (e-nose) principles. The sensing device set up was an electronic nose consisting of an array of 9 micro-chemoresistors (Cr-Au, 3×3) coated with electrospun nanofibrous structures. These were comprised of doped polyemeraldine base blended with 3 different polymers: polyethylene oxide, polyvinilpyrrolidone and polystyrene, which acted as carriers for the conducting polymer and were the major responsible of the features of each fibrous overlay (electrical parameters, selectivity and sensitivity ranges). The two sensing strategies here adopted and compared consisted in the use of 2 different textural coatings: a single- and a double-overlay, where the double-overlay resulting from overdeposition of 2 different polymer blends. Such e-nose included a plurality of nanofibres whose electrical parameters were at the same time depending on each polymer exposure to analytes (NO(2), NH(3)) and on the spatial distribution of the interlacing fibres. The morphology of the coating arrangements of this novel e-nose was investigated by scanning electron microscopy (SEM) and its sensor responses were processed by multicomponent data analyses (PCA and PLS) reporting encouraging results for detection and recognition of analytes at ppb levels.