The V-ATPase A subunit gene (vma-1) from Giardia lamblia.
The sequence of the gene encoding the A subunit of the vacuolar type ATPase from Giardia lamblia is reported. Comparison of the encoded protein with the homologous subunits of eukaryotic and archaebacterial ATPases reveals high levels of similarity throughout the sequence (e.g., overall 49.1 and 44.6% identity to the homologous subunit from carrot and Halobacterium, respectively). An exception are three regions which are unique to the Giardia subunit. The largest of these regions contains motifs characteristic for eukaryotic spliceosomal introns; however, comparison to the cDNA shows that this region is also present in the mRNA.


The past 40 years' assessment of urban-rural differences in Benzo[a]pyrene contamination and human health risk in coastal China.
China has implemented many environmental regulations to battle against polycyclic aromatic hydrocarbon (PAH) contamination since the 1990s. It remains unclear how the exposure levels of PAHs changed quantitatively since reform and opening up in 1978 in China, whether the human health risks decreased or not, and how about the discrepancy between urban and rural areas. Here, taking Benzo[a]pyrene (BaP) in the rapidly urbanized Bohai region of China as a case, we used the improved Berkeley-Trent-Urban-Rural model to simulate the multimedia concentrations of BaP from 1980 to 2020 based on BaP emissions at a regional scale. The total emission of BaP in 1990 was the highest, with a value of 240 t, while the urban emission peaked in 2010. The BaP emissions from rural areas were two to seven times higher than urban areas, and the differences became smaller over time. Despite this, the average modeled BaP concentrations in urban air and soil were two to tens fold higher than in rural areas, particularly in highly urbanized or industrialized cities. Mostly, the concentrations of BaP in rural areas peaked in 1990, while those in urban areas peaked in 1990 or 2010. Early urbanized Beijing and Tianjin were the hot-spot cities of BaP contamination before 2000, while after 2010, higher concentrations were found in late industrialized Shandong and Hebei. BaP posed potential cancer risks to local residents, and air inhalation accounted for more than 80 % of the total risk. Under the stronger implementation of environmental regulations since the 1990s, it showed great health benefits, particularly for the urban residents in Beijing and Tianjin. The biggest decline in cancer risk was found in the period 2010-2020, and the average decreasing rates were 61.4 % and 57.4 % for urban and rural areas, respectively.


In vivo assays to study histone ubiquitylation.
The importance of histone acetylation, phosphorylation, and methylation in transcription and other DNA-mediated processes is now well established. Histones are also ubiquitylated, but in contrast to the majority of ubiquitylated proteins, ubiquitylated histones are not generally targeted for degradation and may play roles similar to those of other histone modifications. Antibodies against acetylated histones have provided unique insights into the regulation, distribution, and cellular roles of these modified histones. In this report, we describe methods to identify ubiquitylated histones in budding yeast and HeLa cells. We provide protocols to detect ubiquitylated histones that are based on a combination of in vivo genetic and immunological assays. These methods should provide relatively simple and useful tools to study the global regulation of this important but poorly understood histone modification.


Design of a Novel 3D Printed Bioactive Nanocomposite Scaffold for Improved Osteochondral Regeneration.
Chronic and acute osteochondral defects as a result of osteoarthritis and trauma present a common and serious clinical problem due to the tissue's inherent complexity and poor regenerative capacity. In addition, cells within the osteochondral tissue are in intimate contact with a 3D nanostructured extracellular matrix composed of numerous bioactive organic and inorganic components. As an emerging manufacturing technique, 3D printing offers great precision and control over the microarchitecture, shape and composition of tissue scaffolds. Therefore, the objective of this study is to develop a biomimetic 3D printed nanocomposite scaffold with integrated differentiation cues for improved osteochondral tissue regeneration. Through the combination of novel nano-inks composed of organic and inorganic bioactive factors and advanced 3D printing, we have successfully fabricated a series of novel constructs which closely mimic the native 3D extracellular environment with hierarchical nanoroughness, microstructure and spatiotemporal bioactive cues. Our results illustrate several key characteristics of the 3D printed nanocomposite scaffold to include improved mechanical properties as well as excellent cytocompatibility for enhanced human bone marrow-derived mesenchymal stem cell adhesion, proliferation, and osteochondral differentiation in vitro. The present work further illustrates the effectiveness of the scaffolds developed here as a promising and highly tunable platform for osteochondral tissue regeneration.


Remobilization mechanism and release characteristics of phosphorus in saline sediments from the Pearl River Estuary (PRE), South China, based on high-resolution measurements.
The internal loading of phosphorus (P) is commonly considered an essential factor contributing to eutrophication in freshwater bodies. However, investigation of the lability and remobilization characteristics of P in estuarine saline sediments has been limited. In this study, a sequential chemical extraction procedure and high-resolution measurement using the diffusive gradients in thin film (DGT) technique were employed to explore the lability, potential remobilization mechanism and release characteristics of sediment P in the Pearl River Estuary (PRE), South China. The P accumulated significantly in sediments along the west coast of the PRE due to the combined effects of terrestrial P inputs and specific hydrological conditions. The geochemical fractions of sediment P followed the order of organic P (Org-P) (mean: 58.6%) > iron-bound P (Fe-P) (23.4%) > calcium-bound P (Ca-P) (17.4%) > loosely bound P (LS-P) (0.63%). Synchronous vertical variations in DGT-labile Fe and P in the upper and middle parts of the sediment profiles confirmed that Fe-coupled P mobilization occurred in saline sediments. In contrast, sulfate reduction in bottom sediments supposed to decouple the Fe-P cycling relationship. Additionally, the formation of an "iron curtain" (Fe oxyhydroxides) in the oxic surface sediments efficiently prevented upward diffusion of P, leading to relatively low effluxes of P (0.098-6.59 ng cm-2 d-1) across the sediment-water interface.


Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: fragile balance of myocyte survival?
Activation of the caspase cascade through the mitochondrial and/or death receptor pathway was investigated in the failing human myocardium, in which the mode and extent of the cascade activation are unknown. In terminal heart failure, a loss of cardiomyocytes by overload-induced apoptosis is an attractive mechanism, explaining the progressive character of the disease. However, its relevance is unclear, because the specificity of probes for apoptotic deoxyribonucleic acid damage is under debate. Left ventricular specimens from 36 explanted failing and 21 nonfailing donor hearts were used for messenger ribonucleic acid detection by semiquantitative reverse-transcription polymerase chain reaction. From these groups, immunoblot analysis was performed in samples from nine failing and six nonfailing donor hearts. In terminally failing hearts, there was a significant accumulation of cytochrome c in the cytosol, which was associated with activation of caspase-9 and downregulation of its inhibitor, caspase-9S. Similarly, the death receptor-induced pathway revealed activation of caspase-8, combined with downregulation of its inhibitors, flice-like inhibitory protein-L (FLIP(L)) and FLIP(S). The unspecific caspase inhibitors, XIAP, hIAP-1 and hIAP-2, were also downregulated. However, the terminal effector caspase-3 was not activated, and its substrate gelsolin, acting in its uncleaved form as a feedback inhibitor of caspase-3, was not cleaved. In the terminally failing human myocardium, the caspase cascade is partially activated in the presence of a consistent phenotype shift toward enhanced susceptibility to apoptosis. Although the system is still under a fragile control, the partial initiation of the apoptotic program may be of functional relevance also for the surviving cardiomyocytes.


Pancreatic Beta Cell Survival and Signaling Pathways: Effects of Type 1 Diabetes-Associated Genetic Variants.
Type 1 diabetes (T1D) is a complex autoimmune disease in which pancreatic beta cells are specifically destroyed by the immune system. The disease has an important genetic component and more than 50 loci across the genome have been associated with risk of developing T1D. The molecular mechanisms by which these putative T1D candidate genes modulate disease risk, however, remain poorly characterized and little is known about their effects in pancreatic beta cells. Functional studies in in vitro models of pancreatic beta cells, based on techniques to inhibit or overexpress T1D candidate genes, allow the functional characterization of several T1D candidate genes. This requires a multistage procedure comprising two major steps, namely accurate selection of genes of potential interest and then in vitro and/or in vivo mechanistic approaches to characterize their role in pancreatic beta cell dysfunction and death in T1D. This chapter details the methods and settings used by our groups to characterize the role of T1D candidate genes on pancreatic beta cell survival and signaling pathways, with particular focus on potentially relevant pathways in the pathogenesis of T1D, i.e., inflammation and innate immune responses, apoptosis, beta cell metabolism and function.


Colorimetric immunoassay for rapid detection of Staphylococcus aureus based on etching-enhanced peroxidase-like catalytic activity of gold nanoparticles.
A novel colorimetric immunoassay for the detection of Staphylococcus aureus (S. aureus) based on a combination of immunomagnetic separation and signal amplification via etching-enhanced peroxidase-like catalytic activity of gold nanoparticles (AuNPs) was developed. Nanoconjugates composed of gold and iron oxide nanoparticles were synthesized and further modified with antiS. aureus immunoglobulin Y (IgY), which was used for the selective enrichment and rapid separation of target bacteria in complex matrices. AuNPs functionalized with antiS. aureus aptamer were used as an artificial enzyme which has peroxidase-like catalysis activity. Catalytic activity of AuNPs is inhibited by modifying aptamer. However, catalysis of modified AuNPs remarkably enhanced by hydrogen peroxide etching. Based on collecting unbound modified AuNPs in the supernatant and 3,3',5,5'-tetramethylbenzidine-hydrogen peroxide reporting system, the yellow color of solution decreases linearly with increasing the concentration of S. aureus ranging from 10 to 106 cfu/mL. The limit of detection is 10 cfu/mL, and total detection time is 65 min. The recoveries of the S. aureus spiked in food samples are 88.2-119.8%. Schematic illustration of colorimetric method for detection of S. aureus based on the IgY-Fe3O4/Au nanocomposites as capture probes and apt-AuNPs as artificial enzyme with etching-enhanced peroxidase-like catalytic activity.


A comparison of two planning techniques for radiotherapy of high grade astrocytomas.
The purpose of this study was to compare two planning techniques for the delivery of radical localized postoperative radiotherapy in the treatment of high grade astrocytomas. Conventional orthogonal plain film planning (CONP) was compared with CT planning (CTP) in terms of the size of the target volumes treated, the amount of normal brain irradiated and the accuracy of localization. Twenty consecutive adults with high grade astrocytomas, who were treated with radiotherapy between March and October 1996, were planned with CONP and CTP, using postoperative, contrast-enhanced CT scans to define the tumour volume. The planning target areas, volumes and the 100%, 80% and 50% isodose areas produced using the two planning methods were measured and compared using Student's paired t-test. The target volume length was also measured and compared as an independent factor. The difference between entry points of the central axis of the lateral fields was noted. Nineteen of 20 patients had a reduction in planning target volumes using CTP compared with CONP. The difference between the mean volumes was clinically relevant, with 288 cm3 representing a 25% reduction, and statistically significant at the P<0.001 level. The planning target volumes were reduced in 18/20 patients (mean 24 cm2, 23%, P<0.001). Similarly, there were highly significant reductions in the 100%, 80% and 50% isodose areas. The target volume lengths were not found to be significantly different. When considering the accuracy of localization, the entry point of the CONP lateral field deviated by a mean distance of 1.6 cm relative to CTP (superiorinferior 1.3 cm; anteriorposterior 0.8 cm; range 04.1 cm). In two patients, this would have led to a geographical miss of macroscopic disease. In patients with high grade astrocytomas, CTP is preferred to conventional planning. It leads to appreciable reductions in the size of the planning target areas and volumes receiving radical doses of radiation, it significantly reduces the amount of normal brain tissue being irradiated and is more accurate in terms of tumour localization. These differences are likely to lead to a reduction in treatment morbidity. We recommend CTP for all patients receiving radical radiotherapy.


Effects of Polysaccharides From Auricularia auricula on the Immuno-Stimulatory Activity and Gut Microbiota in Immunosuppressed Mice Induced by Cyclophosphamide.
Recently, the immuno-enhancing potential of polysaccharide from Auricularia auricula (AAP) has been an area of research interest. However, the immune-stimulatory activity and mechanisms of AAP in immunosuppressive mice treated with cyclophosphamide (CTX) are still poorly understood. This study aimed to evaluate the immuno-enhancing effects of AAP and mine its possible mechanisms. Firstly, polysaccharides were isolated from A. auricula and purified. Secondly, the immune-stimulatory activities of the first AAP fraction (AAP1) were evaluated in the CTX-treated mice. Results showed that AAP1 significantly enhanced immune organ indexes, remarkably stimulated IFN-γ, IL-2, IL-4, IL-10, and TNF-α levels in the serum, and dramatically up-regulated the mRNA levels of Claudin-1, Occludin and ZO-1. Compared to the CTX group, AAP1 administration restored the gut microbiota composition similar to that of the control group by decreasing the ratio of Firmicutes/Bacteroidetes and increasing the relative abundances of short-chain fatty acid-producing microbiota. This study provides useful information for its further application as an immune-stimulator in foods and drugs.


Ceramic encapsulated latex composites.
This work reports the encapsulation of latex particles in Al2O3-polystyrene (PS) composite films. These films were prepared from PS particles in Al2O3 dispersion at room temperature in various latex contents. Composite films were annealed at elevated temperatures in 10 min time interval above the glass transition temperature (Tg) of polystyrene. Transmitted photon intensities, I(tr) were monitored after each annealing step. AFM micrographs were also used to observe the physical changes of the composite films during annealing. It was observed that latex particles are encapsulated above a critical Al2O3 content of 33 wt% which corresponds to the critical occupation probability of p(c) = 0.33 at which the film obey the site-percolation model with a critical exponent of 0.45. Below p(c), it was seen that complete latex film formation process took place, where transparency of the film was increased by annealing.


Efficacy and Safety of Esketamine Nasal Spray in Patients with Treatment-Resistant Depression Who Completed a Second Induction Period: Analysis of the Ongoing SUSTAIN-3 Study.
Treatment-resistant depression (TRD) is a chronic illness requiring long-term treatment. Esketamine nasal spray (ESK) has been studied in several long-term trials of patients with TRD, including SUSTAIN-1 (NCT02493868) and SUSTAIN-3 (NCT02782104). This subgroup analysis of SUSTAIN-3 evaluated patients with TRD who received a second induction (IND) and maintenance treatment with ESK plus oral antidepressant (AD) after a relapse in SUSTAIN-1. Patients aged 18-64 years who achieved stable remission or response with ESK and subsequently relapsed after randomization to continue ESK or switch to placebo nasal spray (PBO) in SUSTAIN-1 and entered the IND phase of SUSTAIN-3 were included in this interim analysis. Response (≥50% improvement in total score from baseline for Montgomery-Åsberg Depression Rating Scale [MADRS] and Patient Health Questionnaire 9-item [PHQ-9]), remission (MADRS score ≤12; PHQ-9 total score <5), changes in depression rating scores (measured as mean change from baseline), and safety were evaluated (incidence of treatment-emergent and serious adverse events [AE]). Of the 96 eligible patients who entered IND in SUSTAIN-3, 32 (33.3%) were taking ESK+AD at the time of relapse in SUSTAIN-1 and 64 (66.7%) were taking AD+PBO. Substantial improvements in depressive symptoms were observed over the second IND phase in both groups and were maintained over the optimization/maintenance (OP/M) phase. MADRS response rates following a second IND were 71.9% and 73.4% for previously relapsed (PR) ESK+AD and PR-AD+PBO, respectively; remission rates were 62.5% and 60.9%, respectively. During the IND and OP/M phases, 58.3% and 83.3% of patients experienced a treatment-emergent AE, respectively. No patients discontinued due to an AE during the second IND. Patients with TRD benefitted from receiving a second IND and maintenance treatment with ESK and no new safety signals were identified.


Role of lymphovascular invasion and immunohistochemical expression of IMP3 in the risk stratification of superficially invasive pT1 esophageal adenocarcinoma.
A problem in the management of patients with Barrett's esophagus-related pT1 esophageal adenocarcinoma is to distinguish those who should be treated conservatively (endoscopic mucosal resection and/or radiofrequency ablation) from those who require esophago-gastrectomy. Recently, lymphovascular invasion (LVI) has emerged as one of the best predictors of regional lymph node metastasis (LNM) and recurrence-free survival (RFS) in pT1 EAC. However, LVI may be underestimated, both because of interobserver variability and incomplete sampling. The aim of our study was to correlate the presence of LVI, with the immunohistochemical expression of IMP3 in pT1 EAC and assess their role in further stratifying these lesions into high and low risk groups based on the potential for lymph node metastasis and poor outcome. Depth of invasion, assessed in five sublevels (m2, m3, sm1, sm2, and sm3), LVI, and expression of IMP3 were studied in 30 patients who underwent esophagogastrectomy for pT1 EAC (2001-2010) at Hartford Hospital, and correlated with LNM and RFS. IMP3 was considered positive when expressed in >50% of the malignant cells with an intensity of stain of 2-3+. Ten of 18 (55.5%) cases with IMP3 expression demonstrated LVI and 2/10 (20%) showed LNM and died of disease. In contrast, none of the 12 IMP3 negative cases showed LVI (p<0.004; 2-tailed Fisher exact test) or had LNM/DOD. In pT1 EAC, (1) based on IMP3 expression, pT1 EAC may be divided into high risk (LVI+/IMP3+) and low risk (LVI-/IMP3-) categories. (2) Absence of IMP3 expression is associated with a significantly reduced risk of LVI (Negative Predictive Value: 100%). (3) Since identifying lymphovascular invasion and other morphological parameters is prone to significant inter-observer variation, IMP3 may be useful as an ancillary marker especially in these pT1 lesions in predicting their clinical behavior, the risk stratification and potentially on the type of treatment.


Radiosensitivity of pre-irradiated mouse skin to second courses of single and multi-fractionated irradiation--skin shrinkage.
The hind legs of mice were re-irradiated with various gamma-ray doses in single or multi-fractionated exposures 6 or 12 months after conditioning irradiation was administered using a variety of treatment schedules. Dose-response relationships were evaluated to ascertain "residual injuries", permissible doses during the second treatments. Fifty to 60 days after the second treatments, the degrees of shrinkage of the skin on the dorsal aspects of the hind legs were measured and used as endpoints. The residual injuries were affected by the biologically effective doses, or by the treatment schedules of not only the first, but also the second exposure. The higher the biologically effective doses, the greater were the residual injuries. Compared to the previously untreated skin, the pre-irradiated skin was relatively radioresistant to the second course of treatments, regardless of the treatment schedule used. The response of the pre-treated skin to a given test dose (25 or 40 Gy) lessened with increasing dose for each treatment schedule used during the first treatment course.


Detection of TEM & SHV genes in Escherichia coli & Klebsiella pneumoniae isolates in a tertiary care hospital from India.
Extended spectrum beta lactamases (ESBLs) have been observed in virtually all the species of family Enterobacteriaceae. The enzymes are predominantly plasmid mediated and are derived from broad-spectrum beta lactamase TEM-1, TEM-2 or SHV-1 by a limited number of mutations. This study was undertaken to characterize ESBL producers among Escherichia coli and Klebsiella pneumoniae by PCR-RFLP, which were initially screened by phenotypic method. A total of 100 isolates of each species (E. coli and K. pneumoniae) were screened for ESBL production. PCR analysis for β-lactamase genes of the family TEM and SHV was also carried out. PCR products of TEM and SHV genes were subjected to digest with three different restriction enzymes. The digested products were run on 1.5 per cent agarose gel, stained and examined for DNA bands. PCR carried out on plasmid DNA alone detected 30 per cent ESBL positive isolates using TEM primer and 38 per cent using SHV primer, whereas PCR for both plasmid and chromosomal DNA showed 56 per cent positivity for TEM and 60 per cent positivity for SHV. RFLP yielded homogeneous band pattern, suggesting that there may be a point source or a common evolutionary origin for all the ESBL isolates.


Influence of nutritional status on cognitive, functional and neuropsychiatric deficits in Alzheimer's disease.
Nutrition is an important issue for elderly people, especially in patients with dementia. It is also related to clinical outcome, such as cognition, autonomy and behavior. The present study investigated the role of nutritional status on cognitive, functional and neuropsychiatric deficits in patients with Alzheimer's disease (AD). Forty-nine patients underwent neuropsychological, nutritional and neuropsychiatric assessments. The sample was divided into at risk of malnutrition (21 patients) and well-nourished (28 patients) according to the Mini Nutritional Assessment (MNA) score. The groups were similar for clinical and demographic variables, except for MNA score and age. The mean body mass index (BMI) was higher than the normal range. Patients at risk of malnutrition showed greater impairment, both in simple and instrumental activities of daily living (ADL and IADL) and a more severe ideomotor praxis deficit than well-nourished patients. Neuropsychiatric symptoms showed significant differences in hallucination, apathy, aberrant motor behavior and night-time subscales of Neuropsychiatric Inventory (NPI). These symptoms were more severe in patients at risk of malnutrition. Logistic regression analysis showed that malnutrition was an important risk factor for the onset of apathy. We hypothesized that changes in dietary habits and intake and the onset of these disorders reflect the involvement of a common neuroanatomical network.


Age and prognosis of non-small cell lung cancer. Usefulness of a relative survival model.
The aim of our study was the comparative evaluation of a relative survival model and a Cox model to determine the prognostic factors of survival for patients with surgically cured non-small cell lung cancer (NSCLC). We focused particularly on the exact role of age in this survival. 156 patients treated between 1975 and 1988 were studied. Both univariate and multivariate analyses were performed, using the actuarial method and the Cox model for crude survival and the Hakulinen model for relative survival. This study confirmed the poor prognosis of NSCLC, even if a curative surgical procedure has been possible, with a 5-year survival of 48% for stage I tumours but only 6% for stage III tumours. The most significant prognostic factor was the postsurgical TNM staging. The relative survival method of Hakulinen dismissed age as a significant prognostic factor. Our study underlines the usefulness of relative survival methods which should be more frequently employed to allow comparisons between series of different origin and to set up multicentre therapeutic trials.


A comparison of epithelial and neural properties in progenitor cells derived from the adult human ciliary body and brain.
Cells isolated from the ciliary body (CB) of the adult human eye possess properties of retinal stem/progenitor cells and can be propagated as spheres in culture. As these cells are isolated from a non-neural epithelium which has neuroepithelial origin, they may have both epithelial and neural lineages. Since it is the properties of neural progenitor cells that are sought after in a future scenario of autotransplantation, we wanted to directly compare human CB spheres with neurospheres derived from the human subventricular zone (SVZ), which is the best characterized neural stem cell niche in the CNS of adults. The CB epithelium was dissected from donor eyes (n = 8). Biopsies from the ventricular wall were harvested during neurosurgery due to epilepsy (n = 7). CB and SVZ tissue were also isolated from Brown Norwegian rats. Dissociated single cells were cultivated in a sphere-promoting medium and passaged every 10-30 days. Fixed spheres were studied by immunohistochemistry, quantitative RT-PCR and scanning/transmission electron microscopy. We found that both CB and SVZ spheres contained a mixed population of cells embedded in extracellular matrix. CB spheres, in contrast to SVZ neurospheres, contained pigmented cells with epithelial morphology that stained for cytokeratins (3/12 + 19), were connected through desmosomes and tight-junctions and produced PEDF. Markers of neural progenitors (nestin, Sox-2, GFAP) were significantly lower expressed in human CB compared to SVZ spheres, and nestin positive cells in the CB spheres also contained pigment. There was higher expression of EGF and TGF-beta receptors in human CB spheres, and a comparative greater activation of the canonical Wnt pathway. These results indicate that adult human CB spheres contain progenitor cells with epithelial properties and limited expression of neural progenitor markers compared to CNS neurospheres. Further studies mapping the regulation between epithelial and neural properties in the adult human CB spheres are vital to fully utilize them as a clinical source of retinal progenitor cells in the future.


Kinetic study of azole-bridged dinuclear platinum(II) complexes reacting with a hairpin-stabilized double-stranded oligonucleotide.
The cytotoxic dinuclear platinum(II) complexes [[cis-Pt(NH(3))(2)](2)(mu-OH)(mu-pz)](NO(3))(2) (pz=pyrazolate) (1) and [[cis-Pt(NH(3))(2)](2)(mu-OH)(mu-1,2,3-ta-N1,N2)](NO(3))(2) (1,2,3-ta=1,2,3-triazolate) (2), were allowed to react with the hairpin-stabilized double-stranded oligonucleotide d(TATGGCATT(4)ATGCCATA), to determine the amounts of intrastrand and interstrand DNA adducts. The reaction kinetics was investigated by reversed-phase HPLC, and the resulting products were analyzed using mass spectroscopy combined with enzymatic digestion, and Maxam-Gilbert sequencing. The reaction of 1 results in the formation of the 1,2-intrastrand d(GG) adduct as the major final product. The two most abundant products of 2 were identified as isomeric 1,2-intrastrand d(GG) adducts differing probably in platinum coordination to the triazole ring. No GG-interstrand crosslinks were detected with either compound. d(GGC)-d(GCC) sequences of DNA do thus not appear to represent significant targets for forming interstrand crosslinks with either 1 or 2.


Search Strategies of Visually Impaired Persons using a Camera Phone Wayfinding System.
We report new experiments conducted using a camera phone wayfinding system, which is designed to guide a visually impaired user to machine-readable signs (such as barcodes) labeled with special color markers. These experiments specifically investigate search strategies of such users detecting, localizing and touching color markers that have been mounted in various ways in different environments: in a corridor (either flush with the wall or mounted perpendicular to it) or in a large room with obstacles between the user and the markers. The results show that visually impaired users are able to reliably find color markers in all the conditions that we tested, using search strategies that vary depending on the environment in which they are placed.


Effect of the CYP2D6 and APOE Polymorphisms on the Efficacy of Donepezil in Patients with Alzheimer's Disease: A Systematic Review and Meta-Analysis.
Differential responses to donepezil treatment in patients with Alzheimer's disease (AD) have been observed in clinical practice. It remains controversial whether, and to what extent, individual variation in the genes responsible for drug metabolism (CYP2D6) or those associated with AD pathogenesis (APOE) modulate the response to donepezil treatment. The aim of this study was to better understand the potential link between donepezil treatment response and CYP2D6 or APOE polymorphisms. We performed a meta-analysis based on data collected from 1266 donepezil-treated AD patients, and evaluated the association of CYP2D6 or APOE polymorphisms with treatment effectiveness. No significant difference was observed in the responder rate of donepezil treatment between the normal function CYP2D6 alleles group and the decreased/non-functional group [odds ratio (OR) 1.34, 95 % confidence interval (CI) 0.5-3.58; p = 0.56]. However, compared with the increased function CYP2D6 alleles group, the normal function group had a better response to donepezil treatment (OR 1.52, 95 % CI 1.14-2.03; p = 0.005). For the specific CYP2D6 single nucleotide polymorphism rs1080985, patients who carried the G allele had a significantly higher risk of poor response to donepezil treatment. After adjusting the data based on APOE genotype, it was observed that only individuals bearing both the APOE-ε4 allele and the rs1080985-G allele showed a significant increase in the frequency of treatment non-response (OR 1.73, 95 % CI 1.07-2.09; p = 0.03). No independent effect of APOE polymorphism on donepezil clinical responses was found (OR 1.08, 95 % CI 0.85-1.38; p = 0.53). Lastly, in a subgroup analysis based on ethnicity, all results remained consistent. The CYP2D6 genotype may be potentially effective for predicting the response to donepezil treatment in AD patients.


The isolation and identification of some toxic constituents ofAspergillus wentii wehmer.
Extraction of a maize culture of a toxinogenic strain ofA. wentii led to the isolation and characterization of three anthraquinones, three bianthrones, a xanthone and a benzophenone. The structures were derived from spectroscopic data and were supported by chemical degradation. Of these, emodin, 1,6-di-0-methylemodin, 5-0-methylsulochrine and 1,3-di-0-methylemodin bianthrone were mildly toxic to ducklings.


The REThink Online Therapeutic Game: A Usability Study.
Children and adolescents' help-seeking behaviors are often limited by fear, uncertainty, and stigma, as well as challenges with finding the right intervention, help, and a lack of familiarity with the process. A promising direction for the development of therapeutic interventions targeted at children is embedding them into gamified interventions, known as therapeutic or serious games. The aim of this paper is to describe the protocol of the beta REThink therapeutic game and to evaluate the usability of the game in a sample of children and adolescents. The study was delivered in schools, where 137 children and adolescents aged between 9 and 16 years old played the levels of the REThink game, followed by filling in the usability questionnaire. The results show above average evaluations for all levels of the game, for every dimension evaluated, namely presence/immersion, enjoyment, learning effectiveness, narratives, goal clarity, adequacy of learning material, and motivation. This study contributes to the literature on the usability of cognitive behavioral therapy-based therapeutic games for improvements in the emotion regulation abilities of children and adolescents, which can guide researchers interested in testing the REThink game in different protocols, as well to encourage its use by mental health specialists and parents.


Efficacy and Safety of Daprodustat for Treatment of Anemia of Chronic Kidney Disease in Incident Dialysis Patients: A Randomized Clinical Trial.
Daprodustat, a hypoxia-inducible factor prolyl hydroxylase inhibitor, is being evaluated as an oral alternative to conventional erythropoiesis-stimulating agent (ESA) therapy. Few studies of anemia treatment in an incident dialysis (ID) population have been reported. To evaluate the efficacy and safety of daprodustat vs darbepoetin alfa in treating anemia of chronic kidney disease in ID patients. This prospective, randomized, open-label clinical trial was conducted from May 11, 2017, through September 24, 2020, in 90 centers across 14 countries. Patients with advanced CKD were eligible if they planned to start dialysis within 6 weeks from screening or had started and received hemodialysis (HD) or peritoneal dialysis (PD) within 90 days before randomization, had a screening hemoglobin (Hb) concentration of 8.0 to 10.5 g/dL (to convert to grams per liter, multiply by 10) and a randomization Hb of 8.0 to 11.0 g/dL, were ESA-naive or had received limited ESA treatment, and were iron-replete. Randomized 1:1 to daprodustat or darbepoetin alfa. The primary analysis in the intent-to-treat population evaluated the mean change in Hb concentration from baseline to evaluation period (weeks 28-52) to assess noninferiority of daprodustat vs darbepoetin alfa (noninferiority margin, -0.75 g/dL). The mean monthly intravenous (IV) iron dose from baseline to week 52 was the principal secondary end point. Rates of treatment-emergent and serious adverse events (AEs) were also compared between treatment groups to assess safety and tolerability. A total of 312 patients (median [IQR] age, 55 [45-65] years; 194 [62%] male) were randomized to either daprodustat (157 patients; median [IQR] age, 52.0 [45-63] years; 96 [61%] male) or darbepoetin alfa (155 patients; median [IQR] age, 56.0 [45-67] years; 98 [63%] male); 306 patients (98%) completed the trial. The mean (SD) Hb concentration during the evaluation period was 10.5 (1.0) g/dL for the daprodustat and 10.6 (0.9) g/dL for the darbepoetin alfa group, with an adjusted mean treatment difference of -0.10 g/dL (95% CI, -0.34 to 0.14 g/dL), indicating noninferiority. There was a reduction in mean monthly IV iron use from baseline to week 52 in both treatment groups; however, daprodustat was not superior compared with darbepoetin alfa in reducing monthly IV iron use (adjusted mean treatment difference, 19.4 mg [95% CI, -11.0 to 49.9 mg]). Adverse event rates were 76% for daprodustat vs 72% for darbepoetin alfa. This randomized clinical trial found that daprodustat was noninferior to darbepoetin alfa in treating anemia of CKD and may represent a potential oral alternative to a conventional ESA in the ID population. ClinicalTrials.gov Identifier: NCT03029208.


[The occurrence of Treponema in fecal samples from dogs and cats with and without intestinal diseases].
6 (6.9%) of 87 examined dogs without diarrhoea proved to be carriers of Treponema (1x T. hyodysenteriae, 5x T. innocens), whereas in fecal samples from 62 dogs with enteric symptoms no isolation of Treponema succeeded. 5 fecal samples (3.7%) of cats without signs of diarrhoea were found to contain Treponema (1x T. hyodysenteriae, 4x T. innocens), whereas the fecal samples of 31 cats with diarrhoea didn't show any growth of Treponema by cultural investigations. Due to the results of these investigations the conclusion can be drawn that Treponema belong to the usual bacteria of dogs' and cats' intestines and cannot be suspected to cause diarrhoea in these animals primarily.


Application of candidate endophytic fungi isolated from extreme desert adapted trees to mitigate the adverse effects of drought stress on maize (Zea mays L.).
The survival of plants under adverse conditions in desert habitats is related to microbial interactions, which can be an innovative strategy for reducing the effects of drought stress in colonized plants. In this study, two endophytic fungi, Trichoderma harzianum, and Fusarium solani, were recovered from the roots of trees in desert regions of Iran. A greenhouse experiment with two fungal agents (control, T. harzianum, F. solani, and T. harzianum + F. solani) and drought (100, 75, and 50% water-holding capacity) was performed on maize (Zea mays L.). Findings indicate that increasing drought levels negatively affect maize plant growth and physiological traits. However, the symbiotic relationship between fungal endophytes and maize roots increased fresh and dry biomass, root/shoot ratio, leaf area, relative water content, and membrane stability index compared with their control counterparts. Maize plants inoculated with endophytic fungi had 52.07, 40, 33.03, and 55.62% higher total phenolic, proline and soluble sugar concentrations, respectively than uninoculated controls. Photosynthetic parameters, including chlorophyll and carotenoid pigments, chlorophyll fluorescence, and gas exchange, were improved in the endophyte-treated plants. However, with increasing drought stress, maize plants colonized with endophytes, electrolyte leakage, and sub-stomatal CO2 concentrations decreased by 28.93% and 47.62%, respectively, compared to endophyte-free plants. When plants were exposed to higher levels of drought stress, endophytes were more effective in improving most parameters, and inoculation of maize seedlings with a combination of endophytes isolated from plants in harsh regions was more effective in increasing their tolerance to drought stress than individual inoculation of each fungus.


Different efficiencies of attentional orienting in different wandering minds.
This study examined the relations between properties of attentional networks and Mind Wandering (MW) across individuals. For the attentional networks, we measured three components of attention, known as alerting, orienting, and executive control, using the Attention Network Test (ANT). To investigate MW, we measured thought probes embedded in the Sustained Attention to Response Task (SART). Moreover, four performance characteristics of the SART were calculated as behavioral indices of MW. Three of them showed significant associations with probed MW. Most research regarding MW focused on its relation to executive functions, while the present study revealed that MW, as indexed by self-reports and RT variability, was negatively correlated with orienting, specifically the exogenous orienting system. Furthermore, there was a positive association between RT variability and executive control. Our results suggest that individuals with higher tendency of MW are less sensitive to irrelevant external stimuli, supporting the decoupling hypothesis of MW.


Optimized immunohistochemistry in combination with image analysis: a reliable alternative to quantitative ELISA determination of uPA and PAI-1 for routine risk group discrimination in breast cancer.
The determination of the invasion markers urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) has further improved the possibilities for individualized therapy of breast cancer. To date, quantitative measurement by ELISA, that needs large amounts of fresh, frozen material, is the only standardized procedure for diagnostic purposes. Therefore, the aim of this study was the establishment of a reliable alternative method based on immunohistochemistry (IHC) and image analysis requiring only small amounts of fixed tumor tissue. Protein expression of uPA and PAI-1 was analyzed in HOPE-fixed tumor samples using tissue microarrays (TMAs) and semiquantitative image analysis. The results of both methods were significantly correlated and risk assessment showed an overall concordance of 78% (83/107; high- and low-risk) and of 94% (74/79) regarding only high-risk patients. The data demonstrate that optimized IHC in combination with image analysis can provide adequate clinical significance compared to ELISA-derived determination of uPA and PAI-1.


[Breast feeding during methadon- and buprenorphin therapy].
The number of opiate addicted patients treated with opioid replacement therapy is continuously increasing. In Germany, 57.7% of these patients are treated with methadone and 18.6% with buprenorphine. This maintenance therapy provides several advantages while addicted pregnant women and their foetus have a high benefit from appropriate replacement therapy. However, the recommendations concerning breast feeding during an opioid replacement therapy are discussed controversially, because methadone as well as buprenorphine accumulate in breast milk. This accumulation might cause damages to the newborn's health; so, child benefits of breast feeding have to be balanced with possible health risks.This review provides an overview of a selective literature search based on the PubMed-database and german consensus recommendations. Used search terms included: (methadone*) AND (breastfeeding OR lactation), (methadone*) AND (human milk), (buprenorphine*) AND (breastfeeding OR lactation) and (buprenorphine*) AND (human milk).According to the available literature, addicted women, substinated with methadone or buprenorphine are allowed to breast feed their newborns. The advantages of breast feeding prevail the risks of an infant opiate intoxication caused by methadone or buprenorphine.


Correction to: A suite of 19F based relaxation dispersion experiments to assess biomolecular motions.
In the original publication, Figures 3 and 6 were displayed incorrectly due to a mistake made by the publisher. The correct version of Figs. 3 and 6 are given below.


Cash benefits in long-term home care.
This paper tests empirically for differences in prices paid between parts of the cash benefit that clients may and not may keep when it is unspent. In The Netherlands, demand-side subsidies were introduced in 1996. Clients receive a cash benefit to purchase the type of home care (housework, personal care, support with mobility, organisational tasks or social support) they need from the care supplier of their choice (private care provider, regular care agency, commercial care agency or paid informal care provider). Furthermore, they negotiate with the care supplier about price and quantity. Our main findings are the following: (1) the component of the cash benefit that a client may not keep when it is unspent has a positive impact on the price of care. (2) In contrast, the components of the cash benefit a client may keep when it is unspent, have no or a negative impact on the price of care. Both results have important implications for designing health policy. If cash benefits are introduced in long-term home care in an attempt to make consumers more conscious about prices, it is only successful when consumers may keep the unspent part of the cash benefit.


Randomized phase III trial of radiotherapy or chemoradiotherapy with topotecan and cisplatin in intermediate-risk cervical cancer patients after radical hysterectomy.
In cervical cancer patients with intermediate-risk factors, the optimal adjuvant therapy is still controversial. We undertook a randomized trial (ClinicalTrials.gov Identifier: NCT01418859) to compare the efficacy and toxicity of concurrent chemoradiotherapy with topotecan and cisplatin with radiotherapy alone in intermediate-risk cervical cancer patients. Eligible patients were randomly assigned to one of three treatment arms including arm A (radiotherapy only,RT), arm B(concurrent chemoradiotherapy only, CCRT), and arm C (concurrent chemoradiotherapy with following consolidation chemotherapy, CCRT + CT). All eligible patients completed external RT (IMRT or 3D-CRT), receiving 45-50 Gy /25 f uniformly to the pelvis. Concurrent chemotherapy regimen was topotecan 0.75 mg/m(2) for days 1, 2 and 3, followed by cisplatin 25 mg/m(2) for days 1, 2 and 3. Three cycles of consolidation chemotherapy regimen was topotecan 1.5 mg/m(2) for days 1 and 2, and 0.75 mg/m(2) for day 3; followed by cisplatin 25 mg/m(2) for days 1, 2 and 3, repeated every 21 days. Adverse events of each group were investigated and compared. Thirty-nine patients enrolled onto the remaining regimens: 14 to RT, 15 to CCRT and 10 to CCRT + CT. Six patients (15.4%) did not complete the protocol treatment. Hematologic toxicity was more frequent and more severe in the CCRT and CCRT + CT arms compared with the RT arm. The incidence of grade 3-4 neutropenia was significantly different statistically between the RT, CCRT and CCRT + RT groups (15.4%, 46.7% and 100%, respectively; P = 0.002). Specially, three patients in CCRT + CT arm of all six patients who did not complete the protocol treatment discontinued planned therapy because of persistent grade 4 neutropenia. However, there were no significant differences in grade 3-4 non-hematologic toxicities between the three groups(all P > 0.05). Recurrence-free survival and overall survival of each group were not analyzed on account of a median follow-up of only 16 months. Concurrent chemoradiotherapy with topotecan and cisplatin showed severe hematologic toxicity in intermediate-risk cervical cancer patients after radical hysterectomy. Thus, the study was closed ahead of schedule. ClinicalTrials.gov Identifier: NCT01418859 .


Cellular and molecular responses to acute cocaine treatment in neuronal-like N2a cells: potential mechanism for its resistance in cell death.
Cocaine is a highly abused drug that causes psychiatric and neurological problems. Its entry into neurons could alter cell-biochemistry and contribute in the manifestation of early pathological symptoms. We have previously shown the acute cocaine effects in rat C6 astroglia-like cells and found that these cells were highly sensitive to cocaine in terms of manifesting certain pathologies known to underlie psychological disorders. The present study was aimed to discern acute cocaine effects on the early onset of various changes in Neuro-2a (N2a) cells. Whole-cell patch-clamp recording of differentiated cells displayed the functional voltage-gated Na+ and K+ channels, which demonstrated the neuronal characteristics of the cells. Treatment of these cells with acute cocaine (1 h) at in vivo (nM to μM) and in vitro (mM) concentrations revealed that the cells remained almost 100% viable. Cocaine administration at 6.25 μM or 4 mM doses significantly reduced the inward currents but had no significant effect on outward currents, indicating the Na+ channel-blocking activity of cocaine. While no morphological change was observed at in vivo doses, treatment at in vitro doses altered the morphology, damaged the neurites, and induced cytoplasmic vacuoles; furthermore, general mitochondrial activity and membrane potential were significantly decreased. Mitochondrial dysfunction enabled the cells switch to anaerobic glycolysis, evidenced by dose-dependent increases in lactate and H2S, resulting unaltered ATP level in the cells. Further investigation on the mechanism of action unfolded that the cell's resistance to cocaine was through the activation of nuclear factor E2-related factor-2 (Nrf-2) gene and subsequent increase of antioxidants (glutathione [GSH], catalase and GSH peroxidase [GPx]). The data clearly indicate that the cells employed a detoxifying strategy against cocaine. On a broader perspective, we envision that extrapolating the knowledge of neuronal resistance to central nervous system (CNS) diseases could delay their onset or progression.


Double Origin of the Posterior Inferior Cerebellar Artery Diagnosed by MR Angiography: A Report of Two Cases.
The posterior inferior cerebellar artery (PICA) frequently arises from the fenestrated segment of the intracranial vertebral artery (VA), and this common variation can be misinterpreted as or confused with a PICA of double origin. Rarely, a PICA of true double origin occurs when two branches of the PICA arise separately from the intracranial VA and fuse to form an arterial ring. We discovered this rare variation incidentally while interpreting images of magnetic resonance (MR) angiography. This is the first report of MR angiographic findings of this rare variation.


Psychiatric Comorbidities in Pediatric Monogenic Diabetes due to GCK Mutation: Impact on Diabetes-Related Quality of Life.
Monogenic diabetes caused by mutation in the glucokinase gene (GCK-MD) is a rare disorder manifesting in childhood as mild, prevalent hyperglycemia. By consensus, it is managed by dietary supervision and infrequent consultations. However, its impact on the mental health of the affected children is largely unknown. To estimate the prevalence of psychiatric comorbidities in children with monogenic glucokinase-related diabetes (GCK-MD) and evaluate their association with quality of life (QoL). The study invited children with GCK-MD aged 5-18 years identified in the Central National Registry and treated in 3 pediatric diabetes centers in Poland. The control group comprised children with type 1 diabetes (T1D, the most common diabetes type in youth) matched for age and family history of diabetes. Participants underwent a semistructured clinical interview diagnostic for psychiatric comorbidities, questionnaires assessing behavioral problems, depressive symptoms, parental stress, and measuring general and diabetes-related QoL (PedsQl). We included 35 patients with GCK-MDMD and 199 with T1D. Eight (22.9%) GCK-MD patients were diagnosed with psychiatric disorder in their lifetime, compared with 16 (8.1%) in the T1D group (odds ratio 3.4 [95% confidence interval: 1.3-8.7]). Patients with GCK-MD showed better parent-reported general QoL (87.1 ± 11.9 vs 82.0 ± 14.0, P = 0.0060) and higher diabetes-related QoL in both parental (84.5 ± 13.8 vs 74.1 ± 15.2, P < 0.0001) and child's perspective (87.6 ± 10.9 vs 77.3 ± 13.9, P < 0.0001). Psychiatric disorders (+P) were associated with worse child-reported diabetes QoL (T1D+P 66.6 ± 16.7, T1D-P 78.2 ± 13.3, GCK-MD+P 79.6 ± 16.3, GCK-MD-P 90.1 ± 7.5, P = 0.0002). High prevalence of psychiatric disorders in children with GCK-MD and lower QoL emphasizes the need for psychologic surveillance in those otherwise mildly-treated patients.


Cross-strand split tetra-Cys motifs as structure sensors in a beta-sheet protein.
We have designed "split tetra-Cys motifs" that bind the biarsenical fluorescein dye 4',5'-bis(1,3,2-dithioarsolan-2-yl)fluorescein (FlAsH) across strands of a model beta-rich protein. Our strategy was to divide the linear FlAsH binding tetra-Cys sequence such that dye could be fully liganded only when the strands were arranged in space correctly by native protein conformational proximities. We introduced pairs of alternating cysteines on adjacent beta strands of cellular retinoic acid binding protein to create FlAsH binding sites in the native structure. Selective labeling occurred both in vitro and in vivo relative to sites with fewer than four Cys or with inappropriate geometry. Interestingly, two of the split tetra-Cys motif-carrying proteins bound FlAsH whether native or urea unfolded, while one was capable of binding FlAsH only when native. This latter design exemplifies the potential of split motifs as structure sensors.


Psychosocial Factors and Low Back Pain Outcomes in a Pooled Analysis of Low Back Pain Studies.
Assessment of possible relationships between work-related psychosocial measures and self-reported low back pain (LBP) outcomes in a large pooled dataset of 1929 participants from 82 facilities in the United States. Pooled data from three prospective cohort studies were used to calculate odds ratios (OR) and 95% confidence intervals (95% CI) for relationships between psychosocial factors and the LBP outcomes. Personal and occupational confounders were controlled for in adjusted Logistic regression models. Supervisor support and job satisfaction were significantly (P < 0.05) related to all three LBP outcomes. Other psychosocial factors were significantly (P < 0.05) associated with at least one of the LBP outcomes. Adjusted ORs ranged from approximately 1.50 to 3.50 for most associations. There is a significant relationship between work-related psychosocial measures and LBP outcomes.


[Clinical exercise testing and the Fick equation: strategic thinking for optimizing diagnosis].
This article examines the expected exercise-induced changes in the components of the oxygen transport system as described by the Fick equation with a view to enable a critical analysis of a standard incremental exercise test to identify normal and abnormal patterns of responses and generate hypotheses as to potential physiological and/or pathophysiological causes. The text reviews basic physiological principals and provides useful reminders of standard equations that serve to integrate circulatory, respiratory and skeletal muscle functions. More specifically, the article provides a conceptual and quantitative framework linking the exercise-induced increase in whole body oxygen uptake to central circulatory and peripheral circulatory factors with the view to establish the normalcy of response. Thus, the article reviews the exercise response to cardiac output determinants and provides qualitative and quantitative perspective bases for making assumptions on the peripheral circulatory factors and oxygen use. Finally, the article demonstrates the usefulness of exercise testing as an effective integrative physiological approach to develop clinical reasoning or verify pathophysiological outcomes.


Thyroid hormone-induced oxidative stress triggers nuclear factor-kappaB activation and cytokine gene expression in rat liver.
Nuclear factor-kappaB (NF-kappaB) is a redox-sensitive factor responsible for the transcriptional activation of cytokine-encoding genes. In this study, we show that 3,3,5-triiodothyronine (T(3)) administration to rats activates hepatic NF-kappaB, as assessed by electrophoretic mobility shift assay. This response coincides with the onset of calorigenesis and enhancement in hepatic respiration, and is suppressed by the antioxidants alpha-tocopherol and N-acetylcysteine or by the Kupffer cell inactivator gadolinium chloride. Livers from hyperthyroid rats with enhanced NF-kappaB DNA-binding activity show induced mRNA expression of the NF-kappaB-responsive genes for tumor necrosis factor-alpha (TNF-alpha) and interleukin- (IL-) 10, as evidenced by reverse transcription-polymerase chain reaction assay, which is correlated with increases in the serum levels of the cytokines. T(3) also increased the hepatic levels of mRNA for IL-1alpha and those of IL-1alpha in serum, with a time profile closely related to that of TNF-alpha. It is concluded that T(3)-induced oxidative stress enhances the DNA-binding activity of NF-kappaB and the NF-kappaB-dependent expression of TNF-alpha and IL-10 genes.


A neural circuit from the dorsal CA3 to the dorsomedial hypothalamus mediates balance between risk exploration and defense.
An appropriate balance between explorative and defensive behavior is essential for the survival and reproduction of prey animals in risky environments. However, the neural circuit and mechanism that allow for such a balance remains poorly understood. Here, we use a semi-naturalistic predator threat test (PTT) to observe and quantify the defense-exploration balance, especially risk exploration behavior in mice. During the PTT, the activity of the putative dorsal CA3 glutamatergic neurons (dCA3Glu) is suppressed by predatory threat and risk exploration, whereas the neurons are activated during contextual exploration. Moreover, optogenetic excitation of these neurons induces a significant increase in risk exploration. A circuit, comprising the dorsal CA3, dorsal lateral septal, and dorsomedial hypothalamic (dCA3Glu-dLSGABA-DMH) areas, may be involved. Moreover, activation of the dCA3Glu-dLSGABA-DMH circuit promotes the switch from defense to risk exploration and suppresses threat-induced increase in arousal.


Mild kainate toxicity produces selective motoneuron death with marked activation of CA(2+)-permeable AMPA/kainate receptors.
Motoneuron death could be produced by higher sensitivity to excitoxicity during the development and pathological conditions. We report here that in ventral spinal cord cultures mild kainate exposure (12.5 microM, 20 min or 100 microM, 2.5 min) induced selective cobalt stain of motoneurons, indicating a specific Ca2+ entry through the Ca(2+)-permeable AMPA/kainate receptors. This result was associated with a selective motoneuron death as previously described. In these cultures, motoneuron immunoreactivity for the Ca2+ buffering protein, calretinin was negative. These findings suggest that the selective motoneuron death due to a mild excitotoxic insult could be linked to a marked Ca2+ influx associated with the lack of some Ca2+ buffering proteins.


Single-Particle Measurements: A Powerful Method for Investigating Electrochemical Reactions.
The relationship between interface structure (e. g., the facet of the solid phase and the configuration of solvation) and the reactivity of the corresponding electrode is a critical issue in electrochemistry. Compared to macroscopic electrode measurements, electrochemical methods established on the single-particle scale have advantages in establishing the structure-property relationship. In recent years, great achievements have been made in electrochemical energy storage and electrocatalysis that allow the evolution and kinetics of electrodes to be understood by employing single-particle measurements. This concept aims to provide an overview of the update of single-particle measurements in related electrochemical processes. Furthermore, the challenges and prospects for the development and application of single-particle measurements are also discussed.


SYNGAP1-related developmental and epileptic encephalopathy: The impact on daily life.
SYNGAP1-developmental and epileptic encephalopathy (SYNGAP1-DEE) has been recently featured as a distinct genetic disease characterized by global psychomotor delay mainly involving language, moderate-to-severe cognitive impairment, autism spectrum disorder, and a generalized epilepsy with spontaneous and reflex seizures. The severity and variability of function impairment and the impact on patients' and caregivers' daily life are still poorly acknowledged. The SYNGAP1 Italian Family Association developed a survey, shared online with caregivers, exploring several issues, including: epilepsy outcome, comorbidities, daily-living skills, hospitalizations, rehabilitation treatments, economic burden, and COVID-19 pandemic impact. Caregivers of 13 children and adolescents participated in the survey. They most often show a fine and gross-motor impairment and a drug-resistant epilepsy with possibility to experience pluridaily absence seizures that may lead to periods of psychomotor regressions. Eating and sleep problems are reported in the majority. Most parents are concerned about language impairment, behavioral issues and lack of autonomy in daily-living activities. Specific neuropsychological evaluations for autism should be early considered in order to identify intervention strategies involving alternative communication strategies, which can positively affect behavior and quality of life. Rehabilitation treatment should aim to the acquisition and consolidation of personal autonomy.


Investigating the effect of gallium curcumin and gallium diacetylcurcumin complexes on the structure, function and oxidative stability of the peroxidase enzyme and their anticancer and antibacterial activities.
Curcumin has a wide spectrum of biological and pharmacological activities including anti-inflammatory, antioxidant, antiproliferative, antimicrobial and anticancer activities. Complexation of curcumin with metals has gained attention in recent years for improvement of its stability. In this study, the effect of gallium curcumin and gallium diacetylcurcumin on the structure, function and oxidative stability of horseradish peroxidase (HRP) enzyme were evaluated by spectroscopic techniques. In addition to the enzymatic investigation, the cytotoxic effect of the complexes was assessed on bladder, MCF-7 breast cancer and LNCaP prostate carcinoma cell lines by MTT assay. Furthermore, antibacterial activity of the complexes against S. aureus and E. coli was explored by dilution test method. The results showed that the complexes improve activity of HRP and also increase its tolerance against the oxidative condition. After addition of the complexes, affinity of HRP for hydrogen peroxide substrate decreases, while the affinity increases for phenol substrate. Circular dichroism, intrinsic and synchronous fluorescence spectra showed that the enzyme structure around the catalytic heme group becomes less compact and also the distance between the heme group and tryptophan residues increases due to binding of the complexes to HRP. On the whole, it can be concluded that the change in the enzyme structure upon binding to the gallium curcumin and gallium diacetylcurcumin complexes results in an increase in the antioxidant efficiency and activity of the peroxidise enzyme. The result of anticancer and antibacterial activities suggested that the complexes exhibit the potential for cancer treatment, but they have no significant antibacterial activity.


Localization of herpes simplex virus and varicella zoster virus DNA in human ganglia.
Human dorsal root ganglia from 14 randomly autopsied adults and 1 infant (all seropositive for both herpes simplex virus [HSV] and varicella zoster virus [VZV]) were examined for latent HSV-1 and VZV DNA by polymerase chain reaction. Thoracic ganglionic DNA from all subjects and trigeminal ganglionic DNA from 11 adults were analyzed. HSV-1 DNA was detected in trigeminal ganglia from 8 of 11 (73%) adults and in thoracic ganglia from 2 of 14 (14%) adults. VZV DNA was detected in trigeminal ganglia from 10 of 11 (91%) adults and in thoracic ganglia from 12 of 14 (86%) adults. None of the DNA samples were positive with primers specific for HSV-2. These findings indicate the presence of latent HSV-1 and VZV DNA in trigeminal ganglia and latent VZV DNA in thoracic ganglia of most seropositive adults. Furthermore, although HSV-1 latency most commonly develops in trigeminal ganglia, we also show for the first time the presence of HSV-1 latency in thoracic ganglia. Finally, both viruses can become latent in the same trigeminal ganglion.


A SAP domain-containing protein shuttles between the nucleus and cell membranes and plays a role in adhesion and migration in D. discoideum.
The AmpA protein reduces cell adhesion, thereby influencing cell migration in Dictyostelium. To understand how ampA influences cell migration, second site suppressors of an AmpA overexpressing cell line were created by REMI mutagenesis. Mutant candidates were identified by their ability to suppress the large plaques that the AmpA overexpressing cells form on bacterial lawns as a result of their increased rate of migration. One suppressor gene, sma, encodes an uncharacterized protein, which contains a SAP DNA-binding domain and a PTEN-like domain. Using sma gene knockouts and Sma-mRFP expressing cell lines, a role for sma in influencing cell migration was uncovered. Knockouts of the sma gene in a wild-type background enhanced chemotaxis. An additional role for Sma in influencing cell-cell adhesion was also demonstrated. Sma protein transitions between cytosolic and nuclear localizations as a function of cell density. In growing cells migrating to folic acid it is localized to regions of actin polymerization and absent from the nucleus. A role for Sma in influencing ampA mRNA levels is also demonstrated. Sma additionally appears to be involved in ampA pathways regulating cell size, actin polymerization, and cell substrate adhesion. We present insights to the SAP domain-containing group of proteins in Dictyostelium and provide evidence of a role for a SAP domain-containing protein shuttling from the nucleus to sites of actin polymerization during chemotaxis to folic acid and influencing the efficiency of migration.


The Azotobacter vinelandii response regulator AlgR is essential for cyst formation.
Azotobacter vinelandii produces the exopolysaccharide alginate, which is essential for the encystment process. In Pseudomonas aeruginosa, as well as in A. vinelandii, the sigmaE factor encoded by algU is required for transcription of algD, which encodes a key enzyme of the alginate biosynthetic pathway. The P. aeruginosa response regulator AlgR activates transcription of algD. fimS, located upstream algR, is proposed to encode the AlgR cognate sensor kinase. We have cloned and characterized the A. vinelandii algR gene; the deduced amino acid sequence of the protein encoded by this gene shows 79% identity with its P. aeruginosa homolog. Sequence analysis around the algR gene revealed the absence of a fimS homolog. Inactivation of A. vinelandii algR diminished alginate production by 50%, but did not affect algD transcription, and completely impaired the capacity to form mature cysts. Electron microscopy of the cyst structures formed by the algR mutant revealed that the encystment process is blocked at the step of exine formation. The transcriptional regulation of the A. vinelandii algR gene and the role of AlgR in alginate production differ significantly from those of its P. aeruginosa counterparts. These differences could be due to the fact that in A. vinelandii, alginate plays a role in encystment, a function not found in P. aeruginosa.


Acute Limb Ischemia: An Emergency Medicine Approach.
Acute limb ischemia is a medical emergency with significant morbidity and mortality. Rapid diagnosis is required because it is a time-sensitive condition. Timely treatment is necessary to restore blood flow to the extremity and prevent complications. The differential diagnosis of acute limb ischemia is broad. Classification of severity of acute limb ischemia is based on clinical variables. A suspicion of acute ischemia based on history and physical examination warrants heparin administration and vascular surgery consultation. The decision for endovascular thrombolysis or standard surgery depends on etiology, duration, and location of vascular occlusion. This review evaluates the diagnostic approach and management for acute limb ischemia.


Estimating normal optic nerve axon numbers in non-human primate eyes.
The goal of the present study is to develop a semi-automated method to estimate accurately, with minimum variance, the total number of axons by counting a subset of the axons within a primate optic nerve. Using an imaging analysis system, axons in 50% of the area of cross-sections of the retrobulbar optic nerve from five adult Rhesus monkeys were counted and extrapolated as an estimate of total axon number of the optic nerves. Both neural and non-neural areas were sampled. With the coordinates of the counts topographically registered, axon numbers within areas ranging from 1 to 50% were resampled. A Monte Carlo and theoretical estimate of the standard deviation of the total axon count for each sampled area was computed. The mean cross-sectional area of the five optic nerves counted was 7.26 +/- 0.6 mm2, and the mean total axon count of the optic nerve area was 1,304,8168 +/- 89,112. When sampling less than 8% of the optic nerve, the standard deviation within the individual of the total estimated axon number increased sharply. With this technique, the variance within each individual increased only slightly when the counting area was reduced from 50 to 8%, but increased sharply when the counted area became less than 8%. While counting less than 8% of the optic nerve area gives a good estimation of total axon count, the effect of a substantial increase in the standard deviation on the statistical power needed to differentiate group differences will depend on the study design.


Risk analysis of the spread of classical swine fever virus through "neighbourhood infections" for different regions in Belgium.
Risk factors associated with the occurrence of "neighbourhood infections" [Epidemiology of classical swine fever. In: Truszczynski, M. (Ed.), Proceedings of the Workshop on Diagnostic Procedures and Measures to Control Classical Swine Fever in Domestic Pigs and the European Wild Boar. Pulaway, Poland, pp. 119-130] during classical swine fever (CSF) outbreaks were examined based on information collected during a CSF-epidemic, which occurred in the East Flanders Province of Belgium in 1994. The only risk factor that was associated with the occurrence of "neighbourhood infections" was a kernel estimation of the intensity of neighbouring herds (P=0.055) [Interactive spatial data analysis. Pearson Education Limited, Harlow, Essex], i.e. the higher the kernel estimation, the higher the risk for the occurrence of neighbourhood infections. In a second part of the study, the likelihood for the occurrence of neighbourhood infections within an area with a 1 km radius was predicted for every Belgian pig herd, assuming that the herd was infected with CSF-virus. For the prediction of these likelihoods, the model resulting from the risk assessment was used. Finally, the predicted likelihoods were transformed into a raster map after applying a smoothing technique. As a result, different areas in Belgium of higher or lower risk for CSF-virus spread through "neighbourhood infections" could be identified on the map. The areas in Belgium where CSF-outbreaks including "neighbourhood infections" occurred in the past decades were all predicted by the model to be of high risk.


Physical activity levels and weight control status by body mass index, among adults--National Health and Nutrition Examination Survey 1999-2004.
Adequate levels of physical activity can assist with weight control efforts, however, only a modest number of national studies have examined the physical activity patterns by weight control status. This article aims to describe patterns of physical activity among men and women who reported engaging in weight control practices. Data from the National Health and Nutrition Examination Survey (1999-2004) were used. The sample included 14,388 adults (aged >/= 18 years), with measured weights and heights from which body mass index (BMI) (weight/height2; kg/m2) was calculated. Analyses were performed to describe the prevalence of engaging in levels of physical activity (< 150-630 minutes/week) by three levels of weight control status (trying to lose weight, trying to maintain weight, and not trying to lose/maintain weight). We also examined the association between physical activity level and weight control status by BMI. The prevalence of low levels of physical activity (< 150 minutes/week) was highest among those not trying to lose/maintain weight (77.7% men, 81.2% women), than those trying to lose, or maintain weight (64.2%-59.7% men, 68.1%-66.7% women). Significantly more men than women engaged in higher volumes of physical activity (p < 0.001). Among overweight men, those trying to lose weight were more likely to engage in 150-420 minutes/week (OR = 2.2, 95%CI 1.8-2.9) than those not trying to lose/maintain weight. Similarly, overweight women who were trying to lose weight were more likely to engage in 150-420 minutes/week (OR = 2.8, 95%CI 2.1-3.7) than were to those not trying to lose/maintain weight. Despite people's intentions to lose or maintain their weight, the majority of adults do not engage even in the minimum recommended level of physical activity. However, the prevalence of engaging in high levels of physical activity (150-420 minutes/week) was highest among those trying to lose or maintain weight than were with those not trying to lose/maintain weight. Regardless of weight control status, all adults should be encouraged to engage in regular physical activity.


Alterations in T cell-related transcripts at the feto-maternal interface throughout equine gestation.
The tolerance of pregnancy by the maternal immune system is balanced between recognition and protection. In the human this is controlled by balancing helper T cell populations (Th1, Th2) in addition to immune suppression from the regulatory arm (Tregs), but this has not been evaluated in the horse. RNA sequencing was performed on chorioallantois and endometrium of mares at 120, 180, 300 and 330 days of gestation (n = 4/stage), as well as 45-day chorioallantois (n = 4) and diestrus endometrium (n = 3). Transcripts were selected for relativity to Th1, Th2, or Treg-associated. qPCR and immunohistochemistry were used to confirm the results of select differentially expressed genes. In the endometrium, Th1 transcripts were highest in the diestrus mare and decreased as gestational length progressed. In contrast, Th2 transcripts were upregulated in comparison to the diestrus mare and highest in mid gestation. Treg transcripts were found increased in comparison to the diestrus mare, but decreased prepartum. In the chorioallantois no Th1 transcripts changed. The majority of Th2 transcripts increased from 45 to 300 days gestation, and then decreased prepartum. Treg-related transcripts trended down in the chorioallantois from 45 days to 120 days gestation, followed by an upregulation to 300 days and a secondary decline prepartum. The mare experiences a complex and evolving immune profile within the tissues of the feto-maternal interface. This consists of a balance between the Th1 and Th2 response, and a dynamic Treg response that is hypothesized to regulate overall events within the immune system.


Expression of the BRCA1-interacting protein Brip1/BACH1/FANCJ is driven by E2F and correlates with human breast cancer malignancy.
Mutations in the BRCA1-interacting DEAH helicase Brip1 confer an increased risk of breast cancer. In the present study we aimed to unravel the transcriptional control of Brip1 and to determine its expression levels in a set of 101 primary invasive breast carcinomas. Transcription of Brip1 was found to be cell growth-related and controlled by the E2F/retinoblastoma (Rb) pathway through a conserved E2F-responsive site. Repression of Brip1 expression by the cell growth-inhibiting compound 1alpha,25-dihydroxyvitamin D3 depended on this same E2F-responsive site. In spite of its role as a tumor suppressor, both quantitative reverse transcriptase-PCR analyses and immunohistochemical stainings showed significantly elevated Brip1 expression levels in grade 3 tumors as compared to grade 1 or 2 carcinomas. Furthermore, increased Brip1 transcript levels were found in tumors with an estrogen receptor-negative, progesterone receptor-negative or HER-2-positive status. In conclusion, these data show that Brip1 is a genuine target gene for the E2F/Rb pathway and that elevated expression levels of Brip1 are detected in primary invasive breast carcinomas with unfavorable characteristics.


Intestinal microvascular exchange in the rat during luminal perfusion with formyl-methionyl-leucyl-phenylalanine.
Formyl-methionyl-leucyl-phenylalanine (FMLP), a peptide released from bacteria in the gut lumen, is known to both attract and activate neutrophils. The aim of this study was to determine whether luminal perfusion with 1 microM FMLP alters microvascular permeability, blood flow, and neutrophil migration in the small intestine of control rats and rats treated with antineutrophil serum. Microvascular permeability to total plasma proteins was determined from an analysis of lymphatic protein fluxes. Myeloperoxidase activity was used as an index of tissue neutrophil count. Intestinal blood flow was measured using radiolabeled microspheres and the reference blood sample method. In control rats, luminal perfusion with FMLP caused significant increases in blood flow, lymph flow, lymph protein clearance, and microvascular permeability, but it did not alter tissue myeloperoxidase activity. In rats treated with antineutrophil serum, tissue myeloperoxidase levels were reduced by approximately 55%, and the FMLP-induced changes in lymph flow, lymph protein clearance, and microvascular permeability were significantly attenuated. In vitro experiments with isolated rat neutrophils revealed that 1 microM FMLP elicits significant chemotaxis and degranulation yet minimally enhances superoxide production. The results of this study indicate that peptides produced by microorganisms in the gut lumen can increase intestinal microvascular permeability. The FMLP-induced alterations in microvascular exchange appear to be mediated by activated neutrophils.


Megatrends in pathology practice: implications for residency training. Ninth Conference for pathology residency program directors--a summary.
The participants at the Ninth Conference for Pathology Residency Program Directors clearly identified the important trends, the problems, and the opportunities now confronted by the specialty of pathology. Equally important, the speakers identified changes that should be incorporated into residency programs if we are to provide the training and education for practice in a changing medical environment. Evaluations of the program by the participants indicated it had been a valuable conference and it should be maintained as a part of each annual American Society of Clinical Pathologists/College of American Pathologists Fall Meeting.


Gentamicin Vestibulotoxicity: Further Insights From a Large Clinical Series.
To review insights gained from a 21-year experience with gentamicin-induced vestibulotoxicity including differences in vestibulotoxicity between single daily dosing (SDD) and multiple daily dosing (MDD) regimens. Retrospective case series. Tertiary care center. Patients with gentamicin vestibulotoxicity referred to the Hertz Multidisciplinary Neurotology Clinic between January 1993 and September 2014. None. Spectrum of vestibular dysfunction measured using videonystagmography, vestibular evoked myogenic potentials, video head impulse testing, and magnetic scleral search coil testing. Of 53 patients with gentamicin-induced vestibulotoxicity, 24 received SDD and 29 received MDD treatment. The most common indications for treatment were sepsis, endocarditis, and osteomyelitis. Angular acceleration receptor function (semicircular canals) was more commonly affected than linear acceleration receptor function (otolithic organ of the saccule; 100% vs. 62%). A significant proportion of patients (53%) developed vestibulotoxicity in the absence of nephrotoxicity and 40% experienced vestibulotoxicity in a delayed fashion up to 10 days posttreatment cessation (mean 3.9 ± 0.7). Therapeutic monitoring did not necessarily prevent delayed vestibulotoxicity. Nephrotoxicity was less common for SDD compared with MDD (60% vs. 35%, p = 0.01). However, the SDD group experienced vestibulotoxicity at a lower cumulative dose (6.3 vs. 7.0 g, p = 0.04) and shorter duration of therapy (20.7 vs 29.4 d, p = 0.02). Our study further highlights important insights regarding gentamicin-induced vestibulotoxicity. While SDD is associated with decreased risk for nephrotoxicity compared with MDD, it confers a higher risk for vestibulotoxicity.


Methods: A new protocol for in vitro red blood cell glycation.
During diabetes, the characteristic hyperglycemia can induce red blood cell glycation. Several researchers have proposed different protocols to perform an in vitro model to study this phenomenon. In this article, some of the most important in vitro glycation protocols available in the bibliography were compared to each other. The incubation parameters as the suspension medium, glucose concentration, red blood cell concentration, time, and temperature were analyzed. Also, several assays were carried out in our laboratory, and glycated hemoglobin, erythrocyte aggregation and viscoelasticity were determined for the protocol validation. Based on the bibliographic analysis and our experimental results, an optimal protocol for in vitro glycation of red blood cells is presented.


Potential targets for intervention against doxorubicin-induced cardiotoxicity based on genetic studies: a systematic review of the literature.
Cardiotoxicity is a well-known adverse effect of doxorubicin (Dox) administration, but the underlying molecular mechanism of this effect is not fully understood. Over the past two decades, considerable efforts have focused on the potential molecular targets of cardiotoxicity in the hope that novel targeted therapies will be generated to attenuate Dox-induced cardiotoxicity. Here, we provide a comprehensive overview of genetically modified animals that show enhanced or reduced susceptibility to the cardiotoxic effects of Dox. We focused on the process by which the molecules involved in DNA damage, oxidative stress, apoptosis, autophagy and necrosis are affected in the presence of Dox. We also present a protein-protein interaction network and explain the contribution of the components to the process of Dox-induced cardiotoxicity. More importantly, data from the literature have indicated that PI3Kγ and Rac1 are potential targets with therapeutic advantages in cancer therapy; molecules that target these proteins can simultaneously attenuate Dox-induced cardiotoxicity and enhance its anticancer activity. This review highlights the potential molecular targets that are critical regulators involved in Dox-mediated cardiotoxicity, thus providing further insight into the development of potential treatment strategies to prevent the cardiotoxic effects and enhance the anticancer efficiency of Dox in cancer patients.


[Analysis of current fluctuations shunted from small portions of the membrane of a nerve cell soma].
A method was developed to allow the registration of ionic current through the small areas of the membrane (10 mu2). The aim of the method is to obtain information on the properties of single ionic channels by the fluctuation analysis. The current through single Ca channel while transferring Ba (130 mM in the external media) is 0.21 +/- 0.06 X 10(-12) A according to tentative estimation. The corresponding value for Ca transfer is 0.1 X 10(-12) A. The upper limit for the single Ca channel conductance value is gCa less than or equal to 0.6 X 10(-12) PHI-1.


Isolation and characterization of plant and human pathogenic bacteria from green pepper (Capsicum annum L.) in Riyadh, Saudi Arabia.
Forty-three bacterial isolates in five genera were recovered from naturally infected green pepper fruits (38 samples) showing dark brown, irregular-shaped splotches. The pathogenicity test was performed on healthy green pepper fruits and red colonies were from inoculated fruits showing the same symptoms and the infected area developed into soft rot. Their identification was based on phenotypic characters and sequence of the gene fragment coding 16S rRNA. Of 43 isolates, 10 showing splotches on green pepper fruits belonged to genus Serratia on the basis of phenotypic characters. One representative isolate of the genus Serratia has been identified by partial 16S rRNA gene sequencing and phylogenetic analysis as belonging to the Serratia rubidaea and has the potential to cause spot on green pepper. Eleven phytopathogenic bacterial isolates were also obtained at the same time but did not induce any splotch symptoms on artificially infected green pepper. Five out of 11 bacterial isolates were identified as Ralstonia on the basis of biochemical tests. Partial sequencing of 16S ribosomal gene of representative isolate revealed that the isolate is Ralstonia solanacearum. The six remaining isolates were related to Xanthomonas vesicatoria on the basis of biochemical tests. Twenty-two of opportunistic human pathogens were isolated at the same time and related to Proteus and Klebsiella. Opportunistic human pathogens did not produce any symptoms on artificially infected green pepper. One representative isolate for each genus was identified as Klebsiella oxytoca and Proteus mirabilis based on their partial 16S rRNA gene sequences. The virulence of the S. rubidaea, the causal agent of green pepper fruits splotches was attributed to the production and secretion of a large variety of enzymes capable of degrading the complex polysaccharides of the plant cell wall and membrane constituents.


A study on the efficacy of rebamipide for patients with proton pump inhibitor-refractory non-erosive reflux disease.
Reflux symptoms in patients with non-erosive reflux disease (NERD) cannot be easily controlled by treatment with proton pump inhibitors (PPI). The anti-inflammatory function of rebamipide may be effective for protecting the esophageal mucosa. This prospective randomized multicenter placebo-controlled study was performed to clarify the efficacy of rebamipide for NERD patients whose reflux symptoms were refractory to PPI treatment. One hundred forty-nine patients were enrolled on the basis of a QUEST score of over 6 and absence of endoscopically proven esophageal mucosal breaks. All the patients were initially administered 15 mg of lansoprazole for 4 weeks, and the symptoms were then assessed using QUEST and GSRS. PPI-refractory patients were randomly assigned to administration of rebamipide or placebo t.i.d. for 4 weeks. Three of the 149 patients were lost to follow-up, and 60 among the remaining 146 patients were found to be PPI-refractory. Among these PPI-refractory patients, 31 were randomly assigned to a rebamipide group and 29 to a placebo group. At the end of drug administration, the QUEST and GSRS scores did not differ between the rebamipide and placebo groups, although a significantly higher proportion of patients in the rebamipide group showed amelioration of abdominal pain and diarrhea. Administration of rebamipide cannot effectively control reflux symptoms in NERD patients whose symptoms are refractory to PPI therapy.


Race and Ethnicity Differences in Disease Severity and Visual Field Progression Among Glaucoma Patients.
Investigate associations of race/ethnicity and preferred language with baseline glaucoma severity, VF test frequency and disease progression. Retrospective cohort study. Patients receiving VF testing at a tertiary eyecare center between 1998 and 2020 with self-identified race, ethnicity and preferred language were included. Outcome measures were VF MD and age at first visit, VF test frequency, VF MD progression. Among 29,891 patients with VF measurements between 1998 and 2020, 55.1% were female, 71.0% self-identified as White/Caucasian, 14.0% as Black/African American, 7.4% as Asian and 6.4% as Hispanic, and 11.2% preferred a language other than English. Mean VF MD at presentation was worse among Black (-9.3±9.7 dB), Asian (-6.2±7.6 dB) and Hispanic (-8.3±9.3 dB) patients (vs. Whites [-5.5±7.3 dB, p<0.001] or non-Hispanics [-6.2±7.8 dB, p<0.001]). After controlling for age, gender and English proficiency, disparities in glaucoma severity at presentation were reduced, especially among Asian and Hispanic patients. Despite greater severity at presentation, Black patients had lower VF test frequency/person-years (1.07±0.53) compared to Whites (1.12±0.52, p=0.006) and worse VF MD progression (-0.43 dB/year, 95% CI -0.67 to -0.28, p<0.001). In contrast, Hispanics had a higher VF frequency vs. non-Hispanics (1.18±0.64 vs. 1.11±0.52, p<0.001), and no difference in VF progression (p=0.77). Black, Asian and Hispanic patients had greater baseline severity vs. Whites. Unlike other groups, Black patients had a lower VF frequency vs. Whites and greater VF progression. Disparities in baseline severity were partially explained by English proficiency, especially for Asian and Hispanic patients.


Two New Brackish Ciliates, Amphileptus spiculatus sp. n. and A. bellus sp. n. from Mangrove Wetlands in Southern China, with Notes on the Molecular Phylogeny of the Family Amphileptidae (Protozoa, Ciliophora, Pleurostomatida).
Two new brackish pleurostomatid ciliates, Amphileptus spiculatus sp. n. and A. bellus sp. n. were collected from mangrove wetlands of southern China and their morphology and molecular phylogeny were studied. Amphileptus spiculatus sp. n. can be distinguished from congeners by the presence of 11-14 right and 6-8 left kineties, two macronuclear nodules and a conspicuous beak-like anterior body end. Amphileptus bellus sp. n. is characterized by the presence of 2-4 macronuclear nodules, 31-35 right and 6 or 7 left kineties and two types of extrusomes. Phylogenetic analyses based on SSU rDNA sequences data indicate that the family Amphileptidae is paraphyletic.


Oncologic Outcomes of Surgery Versus SBRT for Non-Small-Cell Lung Carcinoma: A Systematic Review and Meta-analysis.
The optimal treatment of stage I non-small-cell lung carcinoma is subject to debate. The aim of this study was to compare overall survival and oncologic outcomes of lobar resection (LR), sublobar resection (SR), and stereotactic body radiotherapy (SBRT). A systematic review and meta-analysis of oncologic outcomes of propensity matched comparative and noncomparative cohort studies was performed. Outcomes of interest were overall survival and disease-free survival. The inverse variance method and the random-effects method for meta-analysis were utilized to assess the pooled estimates. A total of 100 studies with patients treated for clinical stage I non-small-cell lung carcinoma were included. Long-term overall and disease-free survival after LR was superior over SBRT in all comparisons, and for most comparisons, SR was superior to SBRT. Noncomparative studies showed superior long-term overall and disease-free survival for both LR and SR over SBRT. Although the papers were heterogeneous and of low quality, results remained essentially the same throughout a large number of stratifications and sensitivity analyses. Results of this systematic review and meta-analysis showed that LR has superior outcomes compared to SBRT for cI non-small-cell lung carcinoma. New trials are underway evaluating long-term results of SBRT in potentially operable patients.


Diagnostic accuracy of computed tomography coronary angiography according to pre-test probability of coronary artery disease and severity of coronary arterial calcification. The CORE-64 (Coronary Artery Evaluation Using 64-Row Multidetector Computed Tomography Angiography) International Multicenter Study.
The purpose of this study was to assess the impact of patient population characteristics on accuracy by computed tomography angiography (CTA) to detect obstructive coronary artery disease (CAD). The ability of CTA to exclude obstructive CAD in patients of different pre-test probabilities and in presence of coronary calcification remains uncertain. For the CORE-64 (Coronary Artery Evaluation Using 64-Row Multidetector Computed Tomography Angiography) study, 371 patients underwent CTA and cardiac catheterization for the detection of obstructive CAD, defined as ≥50% luminal stenosis by quantitative coronary angiography (QCA). This analysis includes 80 initially excluded patients with a calcium score ≥600. Area under the receiver-operating characteristic curve (AUC) was used to evaluate CTA diagnostic accuracy compared to QCA in patients according to calcium score and pre-test probability of CAD. Analysis of patient-based quantitative CTA accuracy revealed an AUC of 0.93 (95% confidence interval [CI]: 0.90 to 0.95). The AUC remained 0.93 (95% CI: 0.90 to 0.96) after excluding patients with known CAD but decreased to 0.81 (95% CI: 0.71 to 0.89) in patients with calcium score ≥600 (p = 0.077). While AUCs were similar (0.93, 0.92, and 0.93, respectively) for patients with intermediate, high pre-test probability for CAD, and known CAD, negative predictive values were different: 0.90, 0.83, and 0.50, respectively. Negative predictive values decreased from 0.93 to 0.75 for patients with calcium score <100 or ≥100, respectively (p = 0.053). Both pre-test probability for CAD and coronary calcium scoring should be considered before using CTA for excluding obstructive CAD. For that purpose, CTA is less effective in patients with calcium score ≥600 and in patients with a high pre-test probability for obstructive CAD.


[Anterior urethral duplication in a male child. A case report].
Urethra duplication is a rare anomaly with numerous anatomic variations. These fall into four main groups: epispadiac, hypospadiac, fusiform, and Y-type. We report on one case of sagittal incomplete hypospadiac anterior duplication of the urethra. Surgical repair consisted of incision of the septum between the two urethras and hypospadias repair at the same setting. Follow-up was characterized by the occurrence of some complications.


Replication control of plasmid pLS1: efficient regulation of plasmid copy number is exerted by the combined action of two plasmid components, CopG and RNA II.
Two elements, the products of genes copG and rnaII, are involved in the copy-number control of plasmid pLS1. RNA II is synthesized in a dosage-dependent manner. Mutations in both components have been characterized. To determine the regulatory role of the two genes, we have cloned copG, rnaII or both elements at various gene dosages into pLS1-compatible plasmids. Assays of incompatibility towards wild-type or mutant pLS1 plasmids showed that: (i) the rnaII gene product, rather than the DNA sequence encoding it, is responsible for the incompatibility, and (ii) CopG and RNA II act in trans and are able to correct up fluctuations in pLS1 copy number. A correlation between the gene dosage at which the regulatory elements were supplied and the incompatibility effect on the resident plasmid was observed. The entire copG-rnaII circuit has a synergistic effect when compared with any of its components in the correction of pLS1 copy-number fluctuations, indicating that, in the homoplasmid steady-state situation, the control of pLS1 replication is exerted by the co-ordinate action of CopG and RNA II.


Two classes of channel-specific toxins from funnel web spider venom.
1. The paralytic effects and neuromuscular actions of Agelenopsis aperta venom on insects were analyzed biochemically and electrophysiologically. 2. Paralysis caused by Agelenopsis venom is correlated with two effects on neuromuscular transmission: postsynaptic inhibition and presynaptic excitation. These effects are explained by the actions of two classes of toxins purified by RPLC, the alpha- and mu-agatoxins. 3. The alpha-agatoxins are low molecular weight, acylpolyamines which cause rapid, reversible paralysis correlated with use-dependent postsynaptic block of EPSPs and ionophoretic glutamate potentials. The mu-agatoxins are cysteine-rich polypeptides which cause irreversible paralysis and repetitive action potentials originating in presynaptic axons or nerve terminals. 4. The joint actions of the alpha- and mu-agatoxins lead to significantly higher rates of paralysis than are obtained by either toxin class alone, and this may relate to enhancement by excitatory mu-agatoxins of use-dependent block caused by alpha-agatoxins.


Novel silicone dressing in full thickness skin grafts - A retrospective controlled case series.
The right choice of dressing after skin grafting, especially in the anatomically complex and aesthetically important head and neck area, is difficult. It is important to have a dressing which is durable and doesn't need a lot of dressing changes. This study introduces a novel, individually moulded silicon dressing; and investigates the number of dressing changes, durability of the dressing and the aesthetic outcome of patients receiving this new type of dressing, in comparison to a control group. The present retrospective single center controlled case series study reports our experiences using two different types of dressing on patients undergoing full thickness skin grafts in the head during the period 01 May 2016 to 01 May 2020. Data were analysed according to the type of dressing with either a transparent moulded silicone dressing or an established silicone sheet dressing. 52 patients were included in the study. We found no difference in the aesthetic outcome and complication rate (p > 0.05) between the two groups. The number of days until first dressing change after surgery, the number of dressing changes or follow up visits in the outpatient care until complete wound healing, and the overall number of dressing changes were all statistically significantly reduced (p < 0.05). Addition-cured silicone used as a moulded dressing in full thickness skin grafts has longer durability and leads to a significant reduction of dressing changes with equal aesthetic outcome compared to an establishes silicone sheet dressing.