Relation of IL-6, IL-13 and IL-15 gene polymorphisms to the rheumatoid factors, anti-CCP and other measures of rheumatoid arthritis activity.
The aim of the study was to examine the relation between polymorphisms and serum levels of selected cytokines (IL-6, IL-13 and IL-15), production of autoantibodies and factors describing rheumatoid arthritis (RA), such as DAS28 and Total Sharp Score. A total of 156 patients with RA according to the ACR criteria, and 200 control subjects were recruited into the study. The measurements of CRP, anti-CCP, the presence of rheumatoid factors (RFs), radiographs of both hands with calculation of Total Sharp Score (TSS) and DAS28 were obtained from all patients with RA. In total, five polymorphisms in genes coding cytokines (IL-6, IL-13 and IL-15) were detected. The levels of these selected cytokines were measured in serum using ELISA method. A significant difference in allele frequencies between patients with RA and controls was observed for IL-15 -267C/T polymorphism. A higher prevalence of heterozygote variants of IL-15 polymorphisms (14035A/T and -267C/T) in the RF IgG- and RF IgA-negative subgroups was observed. Furthermore, the association of polymorphisms in gene for IL-15 with circulating level of IL-15 (14035A/T and 367G/A) and with total RF and Ig-specific RFs (-267C/T) was found. The relation of IL-15 to RFs IgA, IgM, IgG and the measure of DAS28 was proved. The frequency of the T allele of the IL-13 polymorphism -1112C/T was higher in subgroup with faster progression of the disease (TSS/month ≥ 0.1). In conclusion, we present an association of IL-15 gene polymorphisms with the RFs including subtypes (RF, IgG, IgA) underlined by the relation of increased IL-15 levels in circulation to RFs.


Cytoplasmic dynein-dependent vesicular transport from early to late endosomes.
We have used an in vitro fusion assay to study the mechanisms of transport from early to late endosomes. Our data show that the late endosomes share with the early endosomes a high capacity to undergo homotypic fusion in vitro. However, direct fusion of early with late endosomes does not occur. We have purified vesicles which are intermediates during transport from early to late endosomes in vivo, and analyzed their protein composition in two-dimensional gels. In contrast to either early or late endosomes, these vesicles do not appear to contain unique proteins. Moreover, these vesicles undergo fusion with late endosomes in vitro, but not with each other or back with early endosomes. In vitro, fusion of these endosomal vesicles with late endosomes is stimulated by polymerized microtubules, consistent with the known role of microtubules during early to late endosome transport in vivo. In contrast, homotypic fusion of early or late endosomes is microtubule-independent. Finally, this stimulation by microtubules depends on microtubule-associated proteins and requires the presence of the minus-end directed motor cytoplasmic dynein, but not the plus-end directed motor kinesin, in agreement with the microtubule organization in vivo. Our data strongly suggest that early and late endosomes are separate, highly dynamic organelles, which are connected by a microtubule-dependent vesicular transport step.


Mosquito control exposures and breast cancer risk: analysis of 1071 cases and 2096 controls from the Ghana Breast Health Study.
Epidemiologic data on insecticide exposures and breast cancer risk are inconclusive and mostly from high-income countries. Using data from 1071 invasive pathologically confirmed breast cancer cases and 2096 controls from the Ghana Breast Health Study conducted from 2013 to 2015, we investigated associations with mosquito control products to reduce the spread of mosquito-borne diseases, such as malaria. These mosquito control products were insecticide-treated nets, mosquito coils, repellent room sprays, and skin creams for personal protection against mosquitos. Multivariable and polytomous logistic regression models were used to estimate odds ratios (ORadj) and 95% confidence intervals (CI) with breast cancer risk-adjusted for potential confounders and known risk factors. Among controls, the reported use of mosquito control products were mosquito coils (65%), followed by insecticide-treated nets (56%), repellent room sprays (53%), and repellent skin creams (15%). Compared to a referent group of participants unexposed to mosquito control products, there was no significant association between breast cancer risk and mosquito coils. There was an association in breast cancer risk with reported use of insecticide-treated nets; however, that association was weak and not statistically significant. Participants who reported using repellent sprays were at elevated risks compared to women who did not use any mosquito control products, even after adjustment for all other mosquito control products (OR = 1.42, 95% CI=1.15-1.75). We had limited power to detect an association with repellent skin creams. Although only a few participants reported using repellent room sprays weekly/daily or < month-monthly, no trends were evident with increased frequency of use of repellent sprays, and there was no statistical evidence of heterogeneity by estrogen receptor (ER) status (p-het > 0.25). Our analysis was limited when determining if an association existed with repellent skin creams; therefore, we cannot conclude an association. We found limited evidence of risk associations with widely used mosquito coils and insecticide-treated nets, which are reassuring given their importance for malaria prevention. Our findings regarding specific breast cancer risk associations, specifically those observed between repellent sprays, require further study.


Strong broadband absorption in GaAs nanocone and nanowire arrays for solar cells.
We studied the influence of geometric parameters on the optical absorption of gallium arsenide (GaAs) nanocone and nanowire arrays via finite difference time domain simulations. We optimized the structural parameters of the nanocone and nanowire arrays to maximize the ultimate efficiency across a range of lengths from 100 to 1000 nm. Nanocone arrays were found to have improved solar absorption, short-circuit current density, and ultimate efficiencies over nanowire arrays for a wide range of lengths. Detailed simulations reveal that nanocones have superior absorption due to reduced reflection from their smaller tip and reduced transmission from their larger base. Breaking the vertical mirror symmetry of nanowires results in a broader absorption spectrum such that overall efficiencies are enhanced for nanocones. We also evaluated the electric field intensity, carrier generation and angle-dependent optical properties of nanocones and nanowires. The carrier generation in nanocone arrays occurs away from the surface and is more uniform over the entire structure, which should result in less recombination losses than in nanowire arrays.


Elevation of H3K27me3 level contributes to the radioresistance of nasopharyngeal carcinoma by inhibiting OAS1 expression.
Radiotherapy has long been a main treatment option for nasopharyngeal carcinoma (NPC). However, during clinical treatment, NPC is prone to developing radioresistance, resulting in treatment failure. This study aims to examine the role of histone methylation in the induction of radioresistance. It was found that the radioresistance of NPC cells was related to the increase of the level of H3K27me3. Treatment of cells with histone methyltransferase inhibitor GSK126 increased the radiosensitivity of NPC cells by triggering Bcl2/BAX signaling pathway. Bioinformatics analysis indicated that the expression of OAS1 was reduced in the radioresistant cells but increased in the GSK126-treated cells. Chromatin immunoprecipitation assay confirmed that the decrease of OAS1 expression in radioresistant cells was mainly due to the enrichment of H3K27me3 in its promoter region. Furthermore, downregulation of OAS1 reduced apoptosis due to the inhibition of Bcl2/BAX pathway after irradiation, while OAS1 overexpression increased radiosensitivity. Our findings revealed for the first time that the increase of H3K27me3 level was associated with the decrease of OAS1 expression, leading to the inhibition of apoptosis and ultimately contributing to the radioresistance of NPC cells. Moreover, the histone methyltransferase inhibitor GSK126 could overcome the radioresistance and thus might be a potential therapeutic strategy for NPC.


Ocular motor measures of cognitive dysfunction in multiple sclerosis II: working memory.
Our companion paper documents pervasive inhibitory deficits in multiple sclerosis (MS) using ocular motor (OM) measures. Here we investigated the utility of an OM working memory (WMem) task in characterising WMem deficits in these patients as a function of disease status and disease duration. 22 patients with CIS, 22 early clinically definite MS patients (CDMS: <7 years of diagnosis), 22 late CDMS patients (>7 years from diagnosis), and 22 healthy controls participated. All participants completed the ocular motor WMem task, the paced auditory serial addition test (PASAT), and the symbol digit modalities test (SDMT). Clinical disability was characterised in CDMS patients using the Expanded Disability Severity Scale (EDSS). WMem performance was measured as proportion of errors (WMem errors), saccade latency, and relative sensitivity to WMem loading (WMem effect), an indicator of WMem capacity. All patient groups performed more WMem errors than controls with proportion of WMem errors, and degree of WMem effect increasing with increasing disease duration. A larger WMem effect, reflecting poorer WMem capacity, corresponded to poorer performance on neuropsychological measures, and a higher disability score for CDMS patients with the longest disease duration; an observation that suggests wider implication of WMem executive processes with advancing disease. Conspicuously, performance decrements on standard neuropsychological testing did not similarly increase commensurate with disease duration. The ocular motor WMem task appears to meaningfully dissociate WMem deficit from healthy individuals as well as a function of increasing disease duration. Potentially, this task represents a highly informative and objective method by which to ascertain progressive WMem changes from the earliest inception of MS.


5-Aza-2'-deoxycytidine is chemopreventive in a 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone-induced primary mouse lung tumor model.
Carcinogenesis is a multistep process in which many alterations in both genetic and epigenetic controls lead to a growth advantage for neoplastic cells. Hypermethylation has been established as the basis of genomic imprinting, but recent studies have also shown that alterations in genomic methylation patterns may contribute to tumorigenesis. The chemical 5-aza-2'-deoxycytidine (5-aza-dC) has been used both in vitro and in vivo to inhibit DNA methylation. In this study, we investigated the chemopreventive efficacy of 5-aza-dC in a well-established primary mouse lung tumor model. Five-week-old male (C3H/HeJ x A/J) F1 hybrid mice were treated for 24 consecutive weeks with 5-aza-dC, three times per week i.p. Lung tumors were induced with two consecutive weekly doses of 4-(methyl-nitrosamino)-1-(3-pyridyl)-1-butanone starting 1 week after initial treatment with 5-aza-dC. We demonstrated that 5-aza-dC exhibits a chemopreventive effect in this primary mouse lung tumor model which, like human lung adenocarcinomas, harbors an activating K-ras mutation. Treatment with 5-aza-dC resulted in a 23% reduction in tumor incidence, as well as a 42% reduction in tumor multiplicity. This work supports further investigation of methylation inhibitors likes 5-aza-dC for early intervention, prevention and treatment of lung cancer.


Isolation, characterization, and action of colicin M.
Colicin M was isolated from Escherichia coli K-12 32T 19F/T1. The purified, biologically active protein had a molecular weight of 27,000. It contained phosphatidyl ethanolamine. The molecular weight found for the polypeptide chain by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate was 18,000. Colicin M was found to be firmly integrated in the membrane of the producing strain. The action of the colicin seems to be on the membrane, since cells of the susceptible strain E. coli K-12 ROW/V/22.1 lyse rapidly. Using the phase contrast microscope, lysis was followed by decrease in turbidity of the cell culture and release of protein into the medium. Lysis started at about 15 min after addition of colicin M and was completed after 40 to 60 min. At this time, one-third of the protein had been released from the cells. The number of viable cells dropped within 10 min to 0.01%. Colicin M induced formation of spheroplasts in the presence of 16% sucrose. The electron microscope examination revealed that at first bulges in the cell envelope appear, most frequently occurring equatorially but also occurring at sites all over the cell. In the process of spheroplast formation, the cytoplasmic membrane often retreats from one-half of the outer membrane so that the cytoplasm is confined to one hemisphere. Sucrose did not prevent cells from dying unless cells were pregrown in a sucrose containing medium for several generations before colicin M was added. With cells pregrown in the presence of sucrose, the number of survivors was 100 times higher than in the absence of sucrose.


Mechanism for the Autocatalytic Formation of Optically Active Compounds under Abiotic Conditions.
The bromination of chiral crystalline sam ples of 4,4'-dimethylchalcone was reinvestigated. In the presence of the optically active reaction product, (+)-or (-)-chalcone dibromide, crystallization from solutions of the achiral chalcone is specifically directed toward one-handedness. A feedback mechanism can thus be envisaged where optically active compounds are formed, generate additional material of the same chirality, and communicate this chirality to other regions, simply by cycles of solidification, reaction, and liquefaction.


Effects of in vivo transfection with anti-miR-214 on gene expression in murine molar tooth germ.
MicroRNAs (miRNAs) are an abundant class of noncoding RNAs that are believed to be important in many biological processes through regulation of gene expression. Little is known of their function in tooth morphogenesis and differentiation. MicroRNA-214 (miR-214), encoded by the polycistronic Dnm30os gene, is highly expressed during development of molar tooth germ and was selected as a target for silencing with anti-miR-214. Mandibular injection of 1-100 pmol of anti-miR-214 close to the developing first molar in newborn mice resulted in significant decrease in expression of miR-214, miR-466h, and miR-574-5p in the tooth germ. Furthermore, levels of miR-199a-3p, miR-199a-5p, miR-690, miR-720, and miR-1224 were significantly increased. Additionally, the expression of 863 genes was significantly increased and the expression of 305 genes was significantly decreased. Among the genes with increased expression was Twist-1 and Ezh2, suggested to regulate expression of miR-214. Microarray results were validated using real-time RT-PCR and Western blotting. Among genes with decreased expression were Amelx, Calb1, Enam, and Prnp; these changes also being reflected in levels of corresponding encoded proteins in the tooth germ. In the anti-miR-214-treated molars the enamel exhibited evidence of hypomineralization with remnants of organic material and reduced surface roughness after acid etching, possibly due to the transiently decreased expression of Amelx and Enam. In contrast, several genes encoding contractile proteins exhibited significantly increased expression. mRNAs involved in amelogenesis (Ambn, Amelx, Enam) were not found among targets of miRNAs that were differentially expressed following treatment with anti-miR-214. It is therefore suggested that effects of miR-214 on amelogenesis are indirect, perhaps mediated by the observed miR-214-dependent changes in levels of expression of numerous transcription factors.


Synthetic strategies for the preparation of precursor polymers and of microcapsules suitable for cellular entrapment by polyelectrolyte complexation of those polymers.
The production of microcapsules suitable for the entrapment of mammalian cell by means of polyelectrolyte complexation has, of a necessity, led to the development of novel strategies for the preparation of relatively bioinert polymers which complex efficiently under unique conditions to produce a mechanically resilient membrane with efficient transport properties. In this communication we relate a brief overview of capsule-membrane forming systems for the immunoisolation (or potential immunoisolation) of mammalian cells, which are based upon the complexation of polyelectrolyte (PE) polymers; with emphasis on precursor synthesis and relationships between precursor polymer structure and capsule membrane stability.


Structural Basis for EarP-Mediated Arginine Glycosylation of Translation Elongation Factor EF-P.
Glycosylation is a universal strategy to posttranslationally modify proteins. The recently discovered arginine rhamnosylation activates the polyproline-specific bacterial translation elongation factor EF-P. EF-P is rhamnosylated on arginine 32 by the glycosyltransferase EarP. However, the enzymatic mechanism remains elusive. In the present study, we solved the crystal structure of EarP from Pseudomonas putida The enzyme is composed of two opposing domains with Rossmann folds, thus constituting a B pattern-type glycosyltransferase (GT-B). While dTDP-β-l-rhamnose is located within a highly conserved pocket of the C-domain, EarP recognizes the KOW-like N-domain of EF-P. Based on our data, we propose a structural model for arginine glycosylation by EarP. As EarP is essential for pathogenicity in P. aeruginosa, our study provides the basis for targeted inhibitor design.IMPORTANCE The structural and biochemical characterization of the EF-P-specific rhamnosyltransferase EarP not only provides the first molecular insights into arginine glycosylation but also lays the basis for targeted-inhibitor design against Pseudomonas aeruginosa infection.


Piracetam-induced improvement of mental performance. A controlled study on normally aging individuals.
A double-blind, intra-individual cross-over comparison of the mental performance of 18 aging, non-deteriorated individuals during two 4-week periods of piracetam (1-acetamide-2-pyrrolidone) and placebo administration was performed using conventional and computerized perceptual-motor tasks. In a majority of these tasks the subjects did significantly better when on piracetam than on placebo, a finding consistent with ratings completed by two independent observers. The findings indicate new avenues for the treatment of individuals with reduced mental performance possibly related to disturbed alertness--a neglected group of psychiatric conditions.


The role of exosomes in the promotion of epithelial-to-mesenchymal transition and metastasis.
The progression of a solid cancer from a localised disease to metastatic stages is a key reason for mortality in patients. Amongst the drivers of cancer progression, Epithelial-to-Mesenchymal Transition (EMT) has been shown to be of crucial importance. EMT results in the phenotypic shift of an immotile, treatment-sensitive epithelial cell into an elongated, metastatic and treatment-resistant mesenchymal cell. Depending on the cellular and molecular setting, a myriad of studies have demonstrated that EMT causes increased cancer cell motility, invasiveness, resistance to therapies, dormancy and cancer-stem cell phenotypes, all of which are prerequisites for metastasis. The alteration of non-canonical intercellular signalling events in cancer EMT is a phenomenon that is not completely understood. Recently, extracellular vesicles, especially small vesicles called exosomes, have shown to be involved in cancer cell EMT. Most intriguingly, across different cancer types, cancer-derived exosomes have demonstrated to be capable of transferring a mesenchymal phenotype upon recipient epithelial cells, including epithelial cancer cells. The uptake of EMT-inducing exosomes results in molecular changes, altering miRNA, mRNA, and protein levels, either through direct transfer of these components, or by altering gene expression networks involved in EMT. In this review, we are presenting the current state of research of exosomes in cancer EMT, highlight gaps in our current knowledge and propose strategies for future experiments in this area.


[Short term result of lung cancer treated by photodynamic therapy (PDT)].
Fifty four patients with cancer in the lumen of trachea-bronchus proved by histopathology were treated with PDT. 5 mg/kg hematoporphyrin derivative was given. 24-72 hr after injection, PDT was delivered, usually 1-3 times in density of 200-400 J/cm2. There were 69 cancer foci in 54 patients treated by PDT for 168 times in all. One patient, who received the highest number of treatments, was treated for 12 times intermittently. The treatment result: 14 cancer foci gave a complete response, 32 significant response, 21 minor and 2 no responses. Neither hemorrhage nor other serious complications were found. The results suggest that PDT be an effective method for lung cancer. Only being topical in nature, the implication of PDT is expected to be limited.


Effect of Body Mass Index on the sensitivity of Magnetogastrogram and Electrogastrogram.
Gastric disorders affect the gastric slow wave. The cutaneous electrogastrogram (EGG) evaluates the electrical potential of the slow wave but is limited by the volume conduction properties of the abdominal wall. The magnetogastrogram (MGG) evaluates the gastric magnetic field activity and is not affected as much by the volume conductor properties of the abdominal wall. We hypothesized that MGG would not be as sensitive to body mass index as EGG. We simultaneously recorded gastric slow wave signals with mucosal electrodes, a Superconducting Quantum Interference Device magnetometer (SQUID) and cutaneous electrodes before and after a test meal. Data were recorded from representative pools of human volunteers. The sensitivity of EGG and MGG was compared to the body mass index and waist circumference of volunteers. The study population had good linear regression of their Waist circumference (Wc) and Body Mass Index (BMI) (regression coefficient, R=0.9). The mean BMI of the study population was 29.2 ±1.8 kgm-2 and mean Wc 35.7±1.4 inch. We found that while subjects with BMI≥25 showed significant reduction in post-prandial EGG sensitivity, only subjects with BMI≥30 showed similar reduction in post-prandial MGG sensitivity. Sensitivity of SOBI "EGG and MGG" was not affected by the anthropometric measurements. Compared to electrogastrogram, the sensitivity of the magnetogastrogram is less affected by changes in body mass index and waist circumference. The use of Second Order Blind Identification (SOBI) increased the sensitivity of EGG and MGG recordings and was not affected by BMI or waist circumference.


Robust Student's-t mixture model with spatial constraints and its application in medical image segmentation.
Finite mixture model based on the Student's-t distribution, which is heavily tailed and more robust than Gaussian, has recently received great attention for image segmentation. A new finite Student's-t mixture model (SMM) is proposed in this paper. Existing models do not explicitly incorporate the spatial relationships between pixels. First, our model exploits Dirichlet distribution and Dirichlet law to incorporate the local spatial constrains in an image. Secondly, we directly deal with the Student's-t distribution in order to estimate the model parameters, whereas, the Student's-t distributions in previous models are represented as an infinite mixture of scaled Gaussians that lead to an increase in complexity. Finally, instead of using expectation maximization (EM) algorithm, the proposed method adopts the gradient method to minimize the higher bound on the data negative log-likelihood and to optimize the parameters. The proposed model is successfully compared to the state-of-the-art finite mixture models. Numerical experiments are presented where the proposed model is tested on various simulated and real medical images.


An Evolutionary Perspective of Dyslexia, Stress, and Brain Network Homeostasis.
Evolution fuels interindividual variability in neuroplasticity, reflected in brain anatomy and functional connectivity of the expanding neocortical regions subserving reading ability. Such variability is orchestrated by an evolutionarily conserved, competitive balance between epigenetic, stress-induced, and cognitive-growth gene expression programs. An evolutionary developmental model of dyslexia, suggests that prenatal and childhood subclinical stress becomes a risk factor for dyslexia when physiological adaptations to stress promoting adaptive fitness, may attenuate neuroplasticity in the brain regions recruited for reading. Stress has the potential to blunt the cognitive-growth functions of the predominantly right hemisphere Ventral and Dorsal attention networks, which are primed with high entropic levels of synaptic plasticity, and are critical for acquiring beginning reading skills. The attentional networks, in collaboration with the stress-responsive Default Mode network, modulate the entrainment and processing of the low frequency auditory oscillations (1-8 Hz) and visuospatial orienting linked etiologically to dyslexia. Thus, dyslexia may result from positive, but costly adaptations to stress system dysregulation: protective measures that reset the stress/growth balance of processing to favor the Default Mode network, compromising development of the attentional networks. Such a normal-variability conceptualization of dyslexia is at odds with the frequent assumption that dyslexia results from a neurological abnormality. To put the normal-variability model in the broader perspective of the state of the field, a traditional evolutionary account of dyslexia is presented to stimulate discussion of the scientific merits of the two approaches.


Macrophage migration inhibitory factor engages PI3K/Akt signalling and is a prognostic factor in metastatic melanoma.
Macrophage migration inhibitory factor (MIF) is a widely expressed cytokine involved in a variety of cellular processes including cell cycle regulation and the control of proliferation. Overexpression of MIF has been reported in a number of cancer types and it has previously been shown that MIF is upregulated in melanocytic tumours with the highest expression levels occurring in malignant melanoma. However, the clinical significance of high MIF expression in melanoma has not been reported. MIF expression was depleted in human melanoma cell lines using siRNA-mediated gene knockdown and effects monitored using in vitro assays of proliferation, cell cycle, apoptosis, clonogenicity and Akt signalling. In silico analyses of expression microarray data were used to correlate MIF expression levels in melanoma tumours with overall patient survival using a univariate Cox regression model. Knockdown of MIF significantly decreased proliferation, increased apoptosis and decreased anchorage-independent growth. Effects were associated with reduced numbers of cells entering S phase concomitant with decreased cyclin D1 and CDK4 expression, increased p27 expression and decreased Akt phosphorylation. Analysis of clinical outcome data showed that MIF expression levels in primary melanoma were not associated with outcome (HR = 1.091, p = 0.892) whereas higher levels of MIF in metastatic lesions were significantly associated with faster disease progression (HR = 2.946, p = 0.003 and HR = 4.600, p = 0.004, respectively in two independent studies). Our in vitro analyses show that MIF functions upstream of the PI3K/Akt pathway in human melanoma cell lines. Moreover, depletion of MIF inhibited melanoma proliferation, viability and clonogenic capacity. Clinically, high MIF levels in metastatic melanoma were found to be associated with faster disease recurrence. These findings support the clinical significance of MIF signalling in melanoma and provide a strong rationale for both targeting and monitoring MIF expression in clinical melanoma.


Is Hypertension Diagnostic Testing and Diagnosis Associated with Psychological Distress?
Psychological impacts of hypertension diagnostic testing and new hypertension diagnoses are unclear. BP-CHECK was a randomized diagnostic study conducted in 2017-2019 in an integrated healthcare system. Participants with no hypertension diagnosis or medications and elevated blood pressure (BP) were randomized to one of three diagnostic regimens: (1) Clinic, (2) Home, or (3) Kiosk. Participants completed questionnaires at baseline, after completion of the diagnostic regimens, and at 6 months. Outcomes included changes from baseline in health-related quality of life (HRQOL), BP-related worry, and thoughts about having a stroke or heart attack. Participants (n = 482) were mostly over age 50 (77.0%), and White race (80.3%). HRQOL did not significantly change from baseline to 3 weeks or 6 months. Among all participants, BP-related worry and concerns about having a heart attack or stroke increased significantly from baseline to 3 weeks, with heart attack and stroke concerns significantly higher in the Kiosk compared Clinic and Home groups. At 6 months, thoughts about having a heart attack or stroke returned to baseline overall and in the Kiosk group, however BP-related worry was significantly higher among those with, compared to those without, a new hypertension diagnosis. The hypertension diagnostic process did not lead to short-term or intermediate-term changes in self-reported HRQOL. However, BP-related worry increased short-term and persisted at 6 months among individuals with a new hypertension diagnosis. Results warrant validation in more representative populations and additional exploration of the impacts of this worry on psychological well-being and hypertension control.


Novel model of calcium and inositol 1,4,5-trisphosphate regulation of InsP3 receptor channel gating in native endoplasmic reticulum.
The InsP3R Ca(2+)-release channel has biphasic dependence on cytoplasmic free Ca2+ concentration ([Ca2+]i). InsP3 activates gating primarily by reducing high [Ca2+]i inhibition. To determine whether relieving Ca2+ inhibition is sufficient for activation, we examined single-channels in low [Ca2+]i in the absence of InsP3 by patch clamping isolated Xenopus oocyte nuclei. For both endogenous Xenopus type 1 and recombinant rat type 3 InsP3R channels, spontaneous InsP3-independent activities with low open probability Po (approximately 0.03) were observed in [Ca2+]i < 5 nM, whereas none were observed in 25 nM Ca2+. These results establish the half-maximal inhibitory [Ca2+]i in the absence of InsP3 and demonstrate that the channel can be active when all of its ligand-binding sites are unoccupied. In the simplest allosteric model that fits all observations in nuclear patch-clamp studies, the tetrameric channel can adopt six conformations, the equilibria among which are controlled by two inhibitory, one activating Ca(2+)-binding, and one InsP3-binding sites in a manner similar to the Monod-Wyman-Changeux model. InsP3 binding activates gating by affecting the relative affinity for Ca2+ of one of the inhibitory sites in different channel conformations, transforming it into an activating site. Ca2+ inhibition of InsP3-liganded channels is mediated by an InsP3-independent second inhibitory site.


Canakinumab in gout.
Gout is a painful inflammatory arthritis with a prevalence of approximately 4% in the United States, affecting an estimated 8.3 million adults. The past 20 years have shown significant increases in the number of patients with gout and its incidence may still be increasing. Current treatment options to control the pain and inflammation of acute gout include nonsteroidal anti-inflammatory drugs, colchicine and corticosteroids, although patients are often unresponsive to, intolerant of, or have contraindications for, these therapies. Additional treatment options are therefore needed for this population with difficult-to-treat gout. Currently available and investigational anti-inflammatory agents for acute and chronic gout will briefly be reviewed. Canakinumab , a fully human monoclonal anti-interleukin (IL)-1β antibody that selectively blocks IL-1β and that is being investigated for the treatment of gout, will be discussed in greater detail. Canakinumab has been found to be superior to triamcinolone acetonide in acute gout and to colchicine in gout attack prophylaxis in reducing pain and risk of new gout attacks. Canakinumab's long half-life contributes to its prolonged anti-inflammatory effects.


Detection of mephedrone and its metabolites in fingerprints from a controlled human administration study by liquid chromatography-tandem mass spectrometry and paper spray-mass spectrometry.
The use of synthetic stimulants, including designer cathinones, remains a significant concern worldwide. Thus, the detection and identification of synthetic cathinones in biological matrices is of paramount importance for clinical and forensic laboratories. In this study, distribution of mephedrone and its metabolites was investigated in fingerprints. Following a controlled human mephedrone administration (100 mg nasally insufflated), two mass spectrometry-based methods for fingerprint analysis have been evaluated. The samples deposited on triangular pieces of chromatography paper were directly analysed under ambient conditions by paper spray-mass spectrometry (PS-MS) while those deposited on glass cover slips were extracted and analysed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The LC-MS/MS method was 5-6 times more sensitive than PS-MS but required sample preparation and longer analysis time. Mephedrone was detected in 62% and in 38% of all post-administration samples analysed by LC-MS/MS and PS-MS, respectively. Nor-mephedrone was the only metabolite detected in 3.8% of all samples analysed by LC-MS/MS. A large inter- and intra-subject variation was observed for mephedrone which may be due to several factors, such as the applied finger pressure, angle and duration of contact with the deposition surface and inability to control the 'amount' of collected fingerprint deposits. Until these limitations are addressed, we suggest that the sole use of fingerprints can be a useful diagnostic tool in qualitative rather than quantitative analysis, and requires a confirmatory analysis in a different biological matrix.


Co-design of a question prompt list about pregnancy and childbearing for women with polycystic kidney disease: an exploratory sequential mixed-methods study.
Although women with polycystic kidney disease (PKD) generally have healthy pregnancies and babies, pregnancy is associated with a greater risk of maternal complications and requires planning and management of their condition. Given these possible complications, routine communication about childbearing between women with PKD and their treating team is important. A question prompt list (QPL), a structured list of questions used by patients during consultations with healthcare providers, may be beneficial in assisting women with PKD to discuss their childbearing concerns with, and seek related information from, their treating team. The aims of this study were to co-design a QPL about pregnancy and childbearing for women with PKD, and evaluate its comprehensibility, salience, and acceptability. An exploratory sequential mixed-methods study of women of reproductive age with PKD living in Australia, using an experience-based co-design approach with two phases. Women were recruited from a metropolitan public health service and via social media and invited to complete an anonymous online survey about the development of the PKD QPL (phase one) and participate in an online discussion group about its refinement (phase two). Sixteen women completed the development survey and seven participated in the evaluation discussion group. Participants reported that women with PKD would value and use a QPL to prompt discussions with and seek further information about pregnancy and childbearing from their healthcare providers. Women identified four main topics for the QPL: 'thinking about having a baby', 'pregnancy', 'my medications' and 'after my baby is born'. Within each section a series of questions was developed. Based on the findings, a QPL about pregnancy and childbearing for women with PKD was co-designed. Women with PKD often find it difficult to access information and have discussions with their health care providers about pregnancy and childbearing. The PKD QPL co-designed in this study was perceived to be an acceptable tool which will, from the perspectives of participants, assist women with PKD to access information more easily about pregnancy, childbearing and PKD; ask more targeted questions of their treating team; and make informed childbearing decisions.


What should the optimal target hemoglobin be?
Partial correction of anemia in patients with chronic kidney disease (CKD) improves anemia-related symptoms. However, controversy remains as to whether total correction of anemia provides benefits over and above those afforded by partial correction. There is some evidence showing that normalization of hemoglobin (Hb) concentrations may improve the cardiac hyperdynamic state in CKD patients and reduce the diameter of the left ventricle. Further studies have shown that normalization of Hb improves cognitive function and physical capacities as measured by quality of life (QoL) tests. Large studies have shown that in dialysis patients there is a close inverse relationship between hematocrit (Hct) levels and mortality and morbidity. Moreover, there is evidence suggesting that Hct levels higher than those recommended by European Best Practice Guidelines (EBPG) and the National Kidney Foundation Dialysis Outcome and Quality Initiative (NKF-DOQI provide better outcomes for patients with CKD. However, when Hb concentrations are increased to normal in selected patients with cardiac disease, congestive heart failure, or ischemic cardiopathy, higher mortality rates are evident. Therefore, while the majority of patients with CKD may experience significant benefits when Hb is normalized, it seems prudent to recommend individualized target Hb concentrations for each patient, taking into account factors such as age, sex, employment status, physical activity, and co-morbidities.


Minority undergraduate programs intended to increase participation in biomedical careers.
This article reviews a selection of undergraduate programs intended to increase successful minority participation in science, technology, engineering, and mathematics majors, potentially leading to biomedical careers. The object is to examine their structure, consider how well they address the issues of the target population, and assess the extent to which they have met/meet their goals. As a means of conducting this review, the first step is to examine the concepts used as the building blocks for program design. These concepts are found in a shared, yet often undefined, vocabulary used in most undergraduate programs for minority students. The hypothesis is that a shared vocabulary obscures a broad range of meaning and interpretation that has serious ramifications affecting student success. How these building blocks are understood and implemented strongly reflects the institution where the program is housed. The discussion further considers the nature of a number of programs created by the National Science Foundation and the National Institutes of Health specifically for underrepresented minority students and examines one program in detail, the University of California Berkeley's National Science Foundation Research Experience for Undergraduates Program in Molecular, Cell, and Evolutionary Biology. The characteristics of federally organized programs and the Research Experience for Undergraduates are contrasted with 2 very successful student-centered local programs based on a different conceptual model.


Biotransformation of guttiferones, Symphonia globulifera metabolites, by Bipolaris cactivora, an endophytic fungus isolated from its leaves.
The search for active microorganisms for the biotransformation of guttiferone A (1) and C (6) has been successfully undertaken from a collection of endophytic fungi of Symphonia globulifera. Of the twenty-five isolates obtained from the leaves, three are active and have been identified as Bipolaris cactivora. The products obtained are the result of xanthone cyclisation with the formation of two regioisomers among four possible and corresponding to 1,16-oxy-guttiferone and 3,16-oxy-guttiferone. The biotransformation conditions were studied. Interestingly, both oxy-guttiferones A are present in the plant, and the ratio of 3,16-oxy-guttiferone to 1,16-oxy-guttiferone is 4 : 1, very close to that observed by biotransformation (3.8 : 1). These results are consistent with the involvement of endophytes in their formation pathway from guttiferone A, in planta. Finally, biotransformation made it possible to obtain and describe for the first time oxy-guttiferones C.


Global Phase Diagram of a Spin-Orbital Kondo Impurity Model and the Suppression of Fermi-Liquid Scale.
Many correlated metallic materials are described by Landau Fermi-liquid theory at low energies, but for Hund metals the Fermi-liquid coherence scale T_{FL} is found to be surprisingly small. In this Letter, we study the simplest impurity model relevant for Hund metals, the three-channel spin-orbital Kondo model, using the numerical renormalization group (NRG) method and compute its global phase diagram. In this framework, T_{FL} becomes arbitrarily small close to two new quantum critical points that we identify by tuning the spin or spin-orbital Kondo couplings into the ferromagnetic regimes. We find quantum phase transitions to a singular Fermi-liquid or a novel non-Fermi-liquid phase. The new non-Fermi-liquid phase shows frustrated behavior involving alternating overscreenings in spin and orbital sectors, with universal power laws in the spin (ω^{-1/5}), orbital (ω^{1/5}) and spin-orbital (ω^{1}) dynamical susceptibilities. These power laws, and the NRG eigenlevel spectra, can be fully understood using conformal field theory arguments, which also clarify the nature of the non-Fermi-liquid phase.


pH responsiveness of fibrous assemblies of repeat-sequence amphipathic α-helix polypeptides.
We reported previously that our designed polypeptide α3 (21 residues), which has three repeats of a seven-amino-acid sequence (LETLAKA)3, forms not only an amphipathic α-helix structure but also long fibrous assemblies in aqueous solution. To address the relationship between the electrical states of the polypeptide and its α-helix and fibrous assembly formation, we characterized mutated polypeptides in which charged amino acid residues of α3 were replaced with Ser. We prepared the following polypeptides: 2Sα3 (LSTLAKA)3, in which all Glu residues were replaced with Ser residues; 6Sα3 (LETLASA)3, in which all Lys residues were replaced with Ser; and 2S6Sα3 (LSTLASA)3; in which all Glu and Lys residues were replaced with Ser. In 0.1M KCl, 2Sα3 formed an α-helix under basic conditions and 6Sα3 formed an α-helix under acid conditions. In 1M KCl, they both formed α-helices under a wide pH range. In addition, 2Sα3 and 6Sα3 formed fibrous assemblies under the same buffer conditions in which they formed α-helices. α-Helix and fibrous assembly formation by these polypeptides was reversible in a pH-dependent manner. In contrast, 2S6Sα3 formed an α-helix under basic conditions in 1M KCl. Taken together, these findings reveal that the charge states of the charged amino acid residues and the charge state of the Leu residue located at the terminus play an important role in α-helix formation.


Comprehensive immune monitoring reveals profound immunological changes in pancreas after kidney (PAK) transplant recipients.
Pancreas transplantation is a therapeutic option for the management of complicated Type 1 diabetes mellitus. While standard protocols include use of induction therapy followed by maintenance immunosuppression, the amount, frequency and duration of induction treatment has not been clearly defined. While the effect of various induction regimens on lymphocytes has been demonstrated, a prospective immune monitoring approach is not widely used to determine "immunological titration" of immunosuppression. In this study, we analyzed a patient cohort with pancreas after kidney transplantation and measured a wide range of quantitative and functional T and B cell parameters to identify those that would provide greatest value in personalized prospective immune assessment and design of immunosuppression. While there were significant quantitative differences observed in the 2 groups of PAK patients for various lymphocyte subsets, the notable observation was that lymphocyte subset quantitation was uninformative with regard to T cell function. Patients with normal lymphocyte counts had impaired T cell functional responses and vice versa. The use of immune monitoring to determine optimal IS regimens needs to be studied further to facilitate personalized management of immunosuppression with reduced risk of allograft rejection in PAK, and limited morbidity and mortality related to infection and malignancy.


A case report of fulminant type 1 diabetes mellitus associated with drug-induced hypersensitivity syndrome in an elderly patient with coxsackie B4 virus infection and human leukocyte antigen-A24 haplotype.
Drug-induced hypersensitivity syndrome (DIHS) is a severe systemic adverse drug reaction. Previous studies showed that DIHS is associated with the onset of fulminant type 1 diabetes mellitus (FT1D). Although genetic background and abnormalities in immune response or viral infection are considered to be associated with pathogenesis of FT1D, it remains unclear whether virus infection and specific human leukocyte antigen (HLA) typing are involved in DIHS-associated FT1D. Here, we report a case of a 78-year-old female patient with FT1D after DIHS treatment. She was diagnosed as DIHS caused by carbamazepine, and treatment with predonisolone was initiated. After 46 days from the occurrence of DIHS, she was admitted to our hospital because of type 1 diabetes mellitus and diabetic ketoacidosis. Although her Hemoglobin A1c (HbA1c) was elevated by predonisolone treatment (HbA1c: 9.2%), we diagnosed her as fulminant type 1 diabetes mellitus considering the abrupt onset of the ketoacidosis. Her general condition was improved by treatment with fluid infusion and insulin administration. During her clinical course, the infection of coxsackie B4 virus was observed. In addition, the examination of HLA typing showed HLA-A24 haplotype. These findings suggest that the coxsackie B4 virus infection may be involved in the pathogenesis of DIHS-induced FT1D, and that HLA-A24 haplotype might relate to DIHS-associated FT1D.


Introducing [Mn(CO)3(tpa-κ(3)N)](+) as a novel photoactivatable CO-releasing molecule with well-defined iCORM intermediates - synthesis, spectroscopy, and antibacterial activity.
[Mn(CO)3(tpa-κ(3)N)]Br was prepared as a novel photoactivatable CO-releasing molecule (PhotoCORM) from [MnBr(CO)5] and tris(2-pyridylmethyl)amine (tpa) for the delivery of carbon monoxide to biological systems, with the κ(3)N binding mode of the tetradentate tpa ligand demonstrated by X-ray crystallography. The title compound is a CORM prodrug stable in solution in the dark for up to 16 h. However, photoactivation at 365 nm leads to CO release from the metal coordination sphere and transfer to haem proteins, as demonstrated by the standard myoglobin assay. Different iCORM intermediates could be detected with solution IR spectroscopy and assigned using DFT vibrational calculations. The antibacterial activity of the complex was studied on Escherichia coli. No effects were observed when the cultures were either kept in the dark in the presence of PhotoCORM or illuminated in the absence of metal complex. However, photoactivation of [Mn(CO)3(tpa-κ(3)N)]Br at 365 nm led to the appearance of the spectral signatures of CO-coordinated haems in the terminal oxidases of the bacterial electron transport chain in whole-cell UV/Vis absorption spectra. Significant internalization of the PhotoCORM was demonstrated by ICP-MS measurement of the intracellular manganese concentration. In particular when using medium with succinate as the sole carbon source, a very pronounced and concentration-dependent decrease in the E. coli growth rate could be observed upon illumination in the presence of metal complex, which is attributed to the constrained energy metabolism under these conditions and a strong indicator of terminal oxidase inhibition by carbon monoxide delivered from the PhotoCORM.


Leaflet motion abnormality after TAVI: genuine threat or much ado about nothing?
Symptomatic transcatheter heart valve (THV) thrombosis is noted in up to 1% of patients after transcatheter aortic valve replacement. Recently, hypoattenuated leaflet thickening and reduced leaflet motion of bioprosthetic aortic valves associated with normal transvalvular gradients (and possibly related to subclinical leaflet thrombosis) have been reported. While trans-thoracic echocardiography is a useful initial screening imaging modality for the detection of symptomatic THV thrombosis associated with elevated transvalvular gradients, it has limited utility in the detection of subclinical THV thrombosis. In the present article, we describe the incidence, clinical presentation and strategies for the prevention and treatment of THV thrombosis, as well as the role of imaging modalities in the diagnosis and management of this phenomenon.


Galectin-3 enhances extracellular matrix associations and wound healing in monkey corneal epithelium.
Poor healing of epithelial wounds in cornea is a major clinical problem, leading to persistent epithelial defects and ulceration. The primary cause is poor cell migration over the wound. Carbohydrate-binding protein galectin-3 binds to extracellular matrixes (ECMs) and promotes lamellipodia formation by cross-linking to α3 integrin. Recombinant galectin-3 also facilitates wound healing in the rodent cornea. The purposes of the present experiments were to: (1) establish epithelial wound healing models in monkey corneal explant culture, the models more relevant to human, (2) evaluate the healing effect of galectin-3 in our models, and (3) determine if galectin-3 enhances cell adhesion by interacting with ECMs on corneal surface and their ligand integrins. Monkey corneas with central wounds produced by sodium hydroxide (NaOH) or n-heptanol were incubated with or without recombinant galectin-3. The defected area was stained with sodium fluorescein. Primary isolated corneal epithelial cells from monkey were cultured with or without galectin-3 on plates coated with ECMs or integrins, and the number of adhering cells was counted. Galectin-3 expression in various eye tissues was visualized by immunoblotting. NaOH caused loss of epithelial cells and basement membrane. n-Heptanol removed epithelial cells, but the basement membrane was retained. These corneal defects spontaneously became smaller in a time-dependent manner. Exogenous galectin-3 enhanced wound healing in both NaOH and n-heptanol models. Galectin-3 also enhanced cell adhesion onto the major ECMs found in the basement and Bowman's membranes and onto integrins. Relatively high levels of galectin-3 were detected in corneal and conjunctival epithelium, but tear fluid contained negligible galactin-3. These results suggested that the enhanced binding of epithelial cells to ECMs and integrins caused by galectin-3 might promote cell migration over wounded corneal surfaces. Since tear fluid contained relatively low levels of galectin-3, exogenous galectin-3 may be a beneficial drug to enhance re-epithelialization in human corneal diseases.


Effort-reward imbalance among nurses in stable countries and in countries in transition.
The effort-reward imbalance (ERI) model was used to analyze burnout and intention to leave the nursing profession (ITL) in nurses (n = 21,229) in seven European countries, as part of the NEXT-Study. Data were gathered using the short form of the ERI questionnaire. Burnout was assessed using the Copenhagen Burnout Inventory. Multivariate logistic regression analysis controlled for age and gender. ERI was very high in Poland, Germany, Italy, and Slovakia. High ERI was associated with burnout. Odds ratios (ORs) ranged from 2.6 to 13.2 for ERI and from 3.0 to 5.5 for overcommitment (OC). Nurses with high ERI also had high ORs for ITL (2.6-5.7). The association of OC and ITL was weaker (1.1-1.7). The strong and consistent associations of ERI model results with psychological health and with ITL support the relevance of the model for examining different outcome levels. Nurses in transitional countries, as expected, experienced high ERI. High ERI in stable countries suggests that their economic sectors, particularly health care, may be in transition.


On the ability of consumer electronics microphones for environmental noise monitoring.
The massive production of microphones for consumer electronics, and the shift from dedicated processing hardware to PC-based systems, opens the way to build affordable, extensive noise measurement networks. Applications include e.g. noise limit and urban soundscape monitoring, and validation of calculated noise maps. Microphones are the critical components of such a network. Therefore, in a first step, some basic characteristics of 8 microphones, distributed over a wide range of price classes, were measured in a standardized way in an anechoic chamber. In a next step, a thorough evaluation was made of the ability of these microphones to be used for environmental noise monitoring. This was done during a continuous, half-year lasting outdoor experiment, characterized by a wide variety of meteorological conditions. While some microphones failed during the course of this test, it was shown that it is possible to identify cheap microphones that highly correlate to the reference microphone during the full test period. When the deviations are expressed in total A-weighted (road traffic) noise levels, values of less than 1 dBA are obtained, in excess to the deviation amongst reference microphones themselves.


Improving few-shot relation extraction through semantics-guided learning.
Few-shot relation extraction (few-shot RE) aims to recognize relations between the entity pair in a given text by utilizing very few annotated instances. As a simple yet efficient approach, prototype network-based methods often directly incorporate relation information to enhance prototype representation or leverage contrastive learning to mitigate prediction confusion. Despite achieving good results, the above methods are still susceptible to false judgments of outlier samples and confusion of similar classes. To address these issues, we propose a novel Semantics-Guided Learning (SemGL) method that more effectively utilizes relation information to enhance both the representations of instances and prototypes for improving the performance of few-shot RE. First, SemGL employs the prompt encoder to encode various prompt templates of instances and relation information and obtains more accurate semantic representations of instances, instance prototypes, and concept prototypes via the prompt enhancement from large language models. Then, SemGL introduces a novel technique called relation graph learning, which leverages concept prototypes to cluster homogeneous instances together, emphasizing relation-specific features of concrete instances. Simultaneously, SemGL employs instance-level contrastive learning between instance prototypes and support instances to distinguish between intra-class instances and inter-class instances to promote shared features among intra-class instances. Additionally, prototype-level contrastive learning leverages concept prototypes to pull closer relation-specific features of the concept prototype and shared features of the instance prototype from the same relation. Finally, SemGL utilizes new relation prototypes that integrate interpretable features of concept prototypes and shared features of instance prototypes for prediction. Experimental results on two publicly available few-shot RE datasets demonstrate the effectiveness and efficiency of SemGL in introducing relation information, with particularly promising results for the domain adaptation challenge task.


Feasibility of Different Exercise Modalities for Community-Dwelling Residents With Physical Inactivity: A Randomized Controlled Trial.
Exercise interventions can promote health, but they can be difficult to implement. Moreover, no consensus has been reached regarding which exercise modality promotes the most significant improvement in metabolic health. This feasibility study was conducted to (a) determine the implementation efficacy of supervised and home-based exercise interventions by investigating their respective rates of intervention adherence, adherence to targeted intensity, attrition, and adverse events and (b) explore the preliminary efficacy of 12-week exercise programs among aerobic exercise, aerobic exercise combined with resistance exercise, and high-intensity interval training on body composition, anthropometric parameters, and lipid profiles for community-dwelling residents with physical inactivity. This randomized controlled trial was conducted from April to October 2020. Seventy-two sedentary participants aged 40-70 years were enrolled and randomized into one of four groups: 12-week aerobic exercise, aerobic exercise combined with resistance exercise, high-intensity interval training, and control. The three exercise groups performed at least moderate-intensity supervised exercise twice a week and home-based exercise once a week, whereas the control group maintained their usual daily activities. The target variables, including body composition, anthropometric parameters, and lipid profiles, were measured before and after the 12-week intervention. The intervention adherence rates were 74.01%-87.54% for the supervised exercise group, 64.98%-83.90% for the home-based exercise group, and 82.65%-92.65% for the target exercise intensity group. The attrition rate ranged from 12.50% to 17.65%, and no adverse events were reported in any of the exercise groups. Preliminary efficacy data show the reductions in body weight (95% CI [0.01, 1.20], p = .048) and low-density lipoprotein (95% CI [2.76, 30.32], p = .02) were greater in the exercise groups than the control group, although the intergroup differences were not significant. Body weight and low-density lipoprotein may be efficiently reduced in a 12-week period using any of the three exercise modalities with at least 82.65% adherence to moderate-intensity exercise and 70.84% adherence to exercising 3 times a week.


Randomized Single Blind Sham Controlled Trial of Adjunctive Home-Based tDCS after rTMS for Mal De Debarquement Syndrome: Safety, Efficacy, and Participant Satisfaction Assessment.
Mal de debarquement syndrome is a medically refractory disorder characterized by chronic rocking dizziness that occurs after exposure to passive motion. Repetitive transcranial magnetic stimulation (rTMS) can acutely suppress the rocking dizziness but treatment options that extend the benefit of rTMS are needed. 1) To determine whether transcranial direct current stimulation (tDCS) added after rTMS can extend the benefit of rTMS; 2) to determine whether participants can safely perform tDCS at home. Participants were given five days of rTMS (1 Hz right DLPFC/10 Hz left DLPFC in right-handers, vice versa in left-handers), according to a previously piloted protocol. They received three days of training on tDCS self-administration and were then randomized to either real or sham tDCS for four-weeks (anode left DLPFC/cathode right DLPFC for right-handers, vice versa for left-handers). Twenty-three participants completed the study. Those who received real tDCS after rTMS showed significant improvements in the degree of rocking perception as measured by the MdDS Balance Rating Scale and anxiety ratings by Week 4 of tDCS and a trend for improvement on the Dizziness Handicap Inventory. Two rTMS non-responders responded well to subsequent open-label tDCS. Side effects were mild and not different between real and sham tDCS. There were no episodes of skin burns in a group total of 556 sessions of tDCS. Satisfaction was rated high. Home-based tDCS can be performed safely and may be beneficial in selected individuals. Adequate teaching, automatic device safety features, and a good communications infrastructure are components of successful home therapy.


Changes in the composition and fluidity of membranes in obese (ob/ob) mice: a study of hepatic microsomal NADPH-cytochrome P450 oxidoreductase activity.
The characteristics of microsomal membrane lipids in obese (ob/ob) mice have been probed by studies on the microsomal enzyme NADPH-cytochrome P450 oxidoreductase and by investigation of their fluidity using the fluorescence probe 1,6 diphenylhexatriene. Arrhenius plots of NADPH-cytochrome P450 oxidoreductase activity were biphasic with the break temperature in obese mice reduced from 9.5 to 6.5 degrees C. The activation energy of NADPH-cytochrome P450 oxidoreductase was increased in obese microsomes below the Arrhenius breakpoint. Normalising the body temperature of ob/ob mice did not rectify the Arrhenius plot of NADPH-cytochrome P450 oxidoreductase activity of ob/ob mice. DPH fluorescence polarisation indicated that the gross phospholipids from ob/ob mouse microsomes were more fluid than those from lean mice. Phosphatidylethanolamine (PE) of ob/ob microsomes contains a higher proportion of unsaturated fatty acids resulting mainly from an increase in 22:6 and a fall in 16:0. Phosphatidyl choline (PC) of ob/ob microsomes contains an increased proportion of saturated fatty acids. Phospholipid vesicles of obese microsomal PE were more fluid and obese microsomal PC less fluid than the equivalent phospholipids from lean mice microsomes. The changes in Arrhenius plots of NADPH-cytochrome P450 oxidoreductase activity of obese microsomes are interpreted in relation to the reported phospholipid changes.


Spatially controlled simultaneous patterning of multiple growth factors in three-dimensional hydrogels.
Three-dimensional (3D) protein-patterned scaffolds provide a more biomimetic environment for cell culture than traditional two-dimensional surfaces, but simultaneous 3D protein patterning has proved difficult. We developed a method to spatially control the immobilization of different growth factors in distinct volumes in 3D hydrogels, and to specifically guide differentiation of stem/progenitor cells therein. Stem-cell differentiation factors sonic hedgehog (SHH) and ciliary neurotrophic factor (CNTF) were simultaneously immobilized using orthogonal physical binding pairs, barnase-barstar and streptavidin-biotin, respectively. Barnase and streptavidin were sequentially immobilized using two-photon chemistry for subsequent concurrent complexation with fusion proteins barstar-SHH and biotin-CNTF, resulting in bioactive 3D patterned hydrogels. The technique should be broadly applicable to the patterning of a wide range of proteins.


Hypoglycosylated hFSH Has Greater Bioactivity Than Fully Glycosylated Recombinant hFSH in Human Granulosa Cells.
Previous studies suggest that aging in women is associated with a reduction in hypoglycosylated forms of FSH. Experiments were performed to determine whether glycosylation of the FSHβ subunit modulates the biological activity of FSH in human granulosa cells. Recombinant human FSH (hFSH) derived from GH3 pituitary cells was purified into fractions containing hypoglycosylated hFSH(21/18) and fully glycosylated hFSH(24). The response to FSH glycoforms was evaluated using the well-characterized, FSH-responsive human granulosa cell line, KGN at an academic medical center. Granulosa cells were treated with increasing concentrations of fully- or hypoglycosylated FSH glycoforms for periods up to 48 hours. The main outcomes were indices of cAMP-dependent cell signaling and estrogen and progesterone synthesis. We observed that hypoglycosylated FSH(21/18) was significantly more effective than fully glycosylated FSH(24) at stimulating cAMP accumulation, protein kinase A (PKA) activity, and cAMP response element binding protein (CREB) (S133) phosphorylation. FSH(21/18) was also much more effective than hFSH(24) on the stimulation CREB-response element-mediated transcription, expression of aromatase and STAR proteins, and synthesis of estrogen and progesterone. Adenoviral-mediated expression of the endogenous inhibitor of PKA, inhibited FSH(21/18)- and FSH(24)-stimulated CREB phosphorylation, and steroidogenesis. Hypoglycosylated FSH(21/18) has greater bioactivity than fully glycosylated hFSH(24), suggesting that age-dependent decreases in hypoglycosylated hFSH contribute to reduced ovarian responsiveness. Hypoglycosylated FSH may be useful in follicle stimulation protocols for older patients using assisted reproduction technologies.


Nano-assembly of ursolic acid with platinum prodrug overcomes multiple deactivation pathways in platinum-resistant ovarian cancer.
As the most common cause of gynecological cancer-related deaths worldwide, ovarian cancer requires novel therapy strategies. Pt(ii)-Based antitumor drugs (e.g. cisplatin) are one of the most successful and frequently used drugs in ovarian cancer chemotherapy at present. However, drug resistance and severe side effects are the major problems in cancer treatment. Herein, the design of a reduction responsive platinum(iv) (Pt(iv))/ursolic acid (UA)/polyethylene glycol (PEG) dual prodrug amphiphile (Pt(iv)-UA-PEG) to treat cisplatin-resistant ovarian cancer is reported for the first time. Pt(iv)-UA-PEG could self-assemble into nanoparticles (Pt(iv)-UA NPs) with a fixed and precise Pt/UA ratio, and a constantly high content of drugs. Pt(iv)-UA NPs could be efficiently taken up by cisplatin-resistant ovarian cancer cells and release the drug in intracellular reductive and acidic environments. In vitro studies show that the released UA and cisplatin have different anticancer mechanisms, and their synergistic effects overcome the detoxification and anti-apoptotic mechanisms of cancer cells. Furthermore, the in vivo results indicate that Pt(iv)-UA NPs have a prolonged blood circulation time, enhanced tumor accumulation, and significantly improved antitumor efficacy in A2780/DDP tumor-bearing mice, without causing any side effects. In summary, our results demonstrate that the development of the stimuli-responsive dual prodrug amphiphile nano-assembly provides a new strategy to overcome drug resistance.


Aortic valve area, stroke volume, left ventricular hypertrophy, remodeling, and fibrosis in aortic stenosis assessed by cardiac magnetic resonance imaging: comparison between high and low gradient and normal and low flow aortic stenosis.
Recent works using echocardiography suggested that low gradient (LG), low flow (LF) aortic stenosis (AS) has more pronounced left ventricular (LV) concentric remodeling, smaller LV cavity size, and more interstitial fibrosis compared with high gradient (HG) normal flow (NF) AS. Therefore, we evaluated the accuracy of echocardiographic measurements and compared remodeling and fibrosis in different types of AS by cardiac magnetic resonance (CMR). A total of 128 patients (73±11 years of age; 75 men) with aortic valve area (AVA) <0.6 cm(2)/m(2) and ejection fraction >50% by echocardiography underwent CMR to measure planimetric AVA, phase-contrast indexed stroke volume, LV mass, and focal fibrosis. Using <40 mm Hg and indexed stroke volume <35 mL/m(2) by echocardiography as criteria for LG and LF, 69 (54%) patients were HG/NF, 28 (22%) HG/LF, 17 (13%) LG/NF, and 14 (11%) LG/LF AS. LV outflow tract area, indexed stroke volume, and AVA correlated well between echocardiography and CMR (r=0.7, 0.61, and 0.65, respectively; P<0.001 for all). By CMR, however, planimetric AVA was larger in LF/LG (0.54±0.08 cm(2)/m(2)) and LG/NF (0.61±0.08 cm(2)/m(2)) than in HG/LF (0.46±0.07 cm(2)/m(2); P<0.05) AS, and indexed LV mass was lower in LG/LF (75±12 g/m(2)) and LG/NF (81±18 g/m(2)) than in HG/LF (100±27 g/m(2); P<0.05) AS. All groups of AS had similar LV volumes, predominantly concentric hypertrophy remodeling, and similar amounts of focal fibrosis. CMR confirmed overall accuracy of echocardiographic classification of AS but demonstrated that LG/LF and LG/NF AS have larger AVA, less LV hypertrophy, and similar focal fibrosis compared with HG/LF AS. This challenges the view that LG/LF AS is a more advanced state of AS.


An in vitro study on the effect of free amino acids alone or in combination with nisin on biofilms as well as on planktonic bacteria of Streptococcus mutans.
Free D-amino acids (D-AAs) are one of the most striking features of the peptidoglycan composition in bacteria and play a key role in regulating and disassembling bacterial biofilms. Previous studies have indicated that the antimicrobial peptide nisin can inhibit the growth of the cariogenic bacteria Streptococcus mutans. The present study investigated the effect of free amino acids either alone or in combination with nisin on biofilm and on planktonic S. mutans bacteria. The results of the MIC and MBC analyses showed that D-cysteine (Cys), D- or L-aspartic acid (Asp), and D- or L-glutamic acid (Glu) significantly improve the antibacterial activity of nisin against S. mutans and that the mixture of D-Cys, D-Asp, and D-Glu (3D-AAs) and the mixture of L-Cys, L-Asp, and L-Glu (3L-AAs) at a concentration of 40 mM can prevent S. mutans growth. Crystal violet staining showed that the D- or L-enantiomers of Cys, Asp, and Glu at a concentration of 40 mM can inhibit the formation of S. mutans biofilms, and their mixture generated a stronger inhibition than the components alone. Furthermore, the mixture of the three D-AAs or L-AAs may improve the antibacterial activity of nisin against S. mutans biofilms. This study underscores the potential of free amino acids for the enhancement of the antibacterial activity of nisin and the inhibition of the cariogenic bacteria S. mutans and biofilms.


[Off-label use: update and relevance for urology].
The use of pharmaceuticals beyond the approved indication and conditions (off-label use) is of increasing public interest in times of necessary financial constraints in public health together with the high requirements for drug safety to protect the patient. Remarkably, more than half of the therapies in oncology are performed as off-label use. The discussion on off-label use is controversial and based on different points of interests. Evaluation of therapeutic agents by the pharmaceutical industry is predominantly driven by marketing and business requirements. As a consequence, treatment of rare diseases is often only possible by off-label use, creating more or less an off-label need. Reimbursement by health-care insurance is based on the approval of a pharmaceutical substance for a particular situation, because only the rigorous licensing process assures that the verified efficacy is higher than the, often severe, adverse side effects. It is a well known fact that the sometimes adverse events, which occur on administration of substances in an off-label fashion, are not included in the information on the regular use of a given drug. Finally, physicians request a controlled off-label use, which only allows experienced colleagues and (sub)-specialized oncologists to use pharmaceuticals in an off-label fashion. Up to date no legal documents exist that provide regulations for such an off-label usage.


Characterization and designing of an SAM riboswitch to establish a high-throughput screening platform for SAM overproduction in Saccharomyces cerevisiae.
S-adenosyl- l-methionine (SAM) is a high-value compound widely used in the treatment of various diseases. SAM can be produced through fermentation, but further enhancing the microbial production of SAM requires novel high-throughput screening methods for rapid detection and screening of mutant libraries. In this work, an SAM-OFF riboswitch capable of responding to the SAM concentration was obtained and a high-throughput platform for screening SAM overproducers was established. SAM synthase was engineered by semirational design and directed evolution, which resulted in the SAM2S203F,W164R,T251S,Y285F,S365R mutant with almost twice higher catalytic activity than the parental enzyme. The best mutant was then introduced into Saccharomyces cerevisiae BY4741, and the resulting strain BSM8 produced a sevenfold higher SAM titer in shake-flask fermentation, reaching 1.25 g L-1 . This work provides a reference for designing biosensors to dynamically detect metabolite concentrations for high-throughput screening and the construction of effective microbial cell factories.


The expression profiles of fibroblast growth factor 9 and its receptors in developing mice testes.
An expressional lack of fibroblast growth factor 9 (FGF9) would cause male-to-female sex reversal in the mouse, implying the essential role of FGF9 in testicular organogenesis and maturation. However, the temporal expression of FGF9 and its receptors during testicular development remains elusive. In this study, immunohistochemistry was used to identify the localization of FGF9 and its receptors at different embryonic and postnatal stages in mice testes. Results showed that FGF9 continuously expressed in the testis during development. FGF9 had highest expression in the interstitial region at 17-18 d post coitum (dpc) and in the spermatocytes, spermatids and Leydig cell on postnatal days (pnd) 35-65. Regarding receptor expression, FGFR1 and FGFR4 were evenly expressed in the whole testis during the embryonic and postnatal stages. However, FGFR2 and FGFR3 were widely expressed during the embryonic testis development with higher FGFR2 expression in seminiferous tubules at 16-18 dpc and higher FGFR3 expression in interstitial region at 17-18 dpc. In postnatal stage, FGFR2 extensively expressed with higher expression at spermatids and Leydig cells on 35-65 pnd and FGFR3 widely expressed in the whole testis. Taken together, these results strongly suggest that FGF9 is correlated with the temporal expression profiles of FGFR2 and FGFR3 and possibly associated with testis development.


Endoplasmic reticulum stress does not contribute to steatohepatitis in obese and insulin-resistant high-fat-diet-fed foz/foz mice.
Non-alcoholic fatty liver (steatosis) and steatohepatitis [non-alcoholic steatohepatitis (NASH)] are hepatic complications of the metabolic syndrome. Endoplasmic reticulum (ER) stress is proposed as a crucial disease mechanism in obese and insulin-resistant animals (such as ob/ob mice) with simple steatosis, but its role in NASH remains controversial. We therefore evaluated the role of ER stress as a disease mechanism in foz/foz mice, which develop both the metabolic and histological features that mimic human NASH. We explored ER stress markers in the liver of foz/foz mice in response to a high-fat diet (HFD) at several time points. We then evaluated the effect of treatment with an ER stress inducer tunicamycin, or conversely with the ER protectant tauroursodeoxycholic acid (TUDCA), on the metabolic and hepatic features. foz/foz mice are obese, glucose intolerant and develop NASH characterized by steatosis, inflammation, ballooned hepatocytes and apoptosis from 6 weeks of HFD feeding. This was not associated with activation of the upstream unfolded protein response [phospho-eukaryotic initiation factor 2α (eIF2α), inositol-requiring enzyme 1α (IRE1α) activity and spliced X-box-binding protein 1 (Xbp1)]. Activation of c-Jun N-terminal kinase (JNK) and up-regulation of activating transcription factor-4 (Atf4) and CCAAT/enhancer-binding protein-homologous protein (Chop) transcripts were however compatible with a 'pathological' response to ER stress. We tested this by using intervention experiments. Induction of chronic ER stress failed to worsen obesity, glucose intolerance and NASH pathology in HFD-fed foz/foz mice. In addition, the ER protectant TUDCA, although reducing steatosis, failed to improve glucose intolerance, hepatic inflammation and apoptosis in HFD-fed foz/foz mice. These results show that signals driving hepatic inflammation, apoptosis and insulin resistance are independent of ER stress in obese diabetic mice with steatohepatitis.


Toward critical research ethics: transforming ethical conduct in qualitative health care research.
Based on research conducted with women injection drug users (WIDUs), I discuss the ethical conflicts that researchers and sub-contractors face in gaining access to the life narratives of WIDUs. Foremost among these is the potentially exploitative nature of the study participant-researcher relationship. I suggest that federal and institutional guidelines for human subject research must incorporate additional safeguards to protect study populations such as WIDUs. Moreover, the ethical concerns related to health care research should be addressed in guidelines for ethical conduct with human subjects, research ethics seminars, and required training programs for researchers and subcontractors separately.


Peptic ulcer disease.
This review reports and attempts to place in some context current key observations in the literature, as they relate to peptic ulcer disease. Focus areas in this review are general, the usefulness of symptom-based triage in management decision making and critical review of current practice as it relates to foregut malignancy; Helicobacter pylori, important observations regarding gastric cancer prevention are presented, the role of the organism in "acid rebound" is discussed, and further evidence to support the role of H. pylori eradication in peptic ulcer disease is provided; medication, a number of studies comparing steady state and onset of action efficacy of current proton pump inhibitors at a gastric pH level are reviewed and a new agent is introduced. Problems with and strategies for the safe use of antithrombotic agents are reviewed, and exciting data with regard to a new class of nonsteroidal antiinflammatory drug are provided, now at the "proof of concept" stage. The review period's main "new" information relates to the nonsteroidal antiinflammatory drugs in which a number of exciting agents are being developed. Although inconclusive, the observations with regard to the effect of H. pylori eradication on gastric carcinogenesis is likely to lead to more work in this field.


The effect of gender on clinical outcomes following routine revascularizations with polymer-free sirolimus-eluting stents.
Gender-specific outcomes after percutaneous coronary interventions were studied by a number of research groups with different endpoints and cohorts of different ethnic extractions. The purpose of this report is to use propensity score matching to determine gender-specific differences in clinical outcomes after percutaneous coronary interventions with polymer-free sirolimus-coated stents. The basis for this post hoc analysis was two large all-comers studies with prospectively enrolled patients from Europe and Asia. Data were pooled and analyzed in terms of clinical outcomes to assess the impact of gender in patients with stable coronary artery disease and acute coronary syndrome. The primary endpoint was the accumulated target-lesion revascularization rate whereas secondary endpoints consisted of the event rates for major adverse cardiac events (MACE), myocardial infarction, bleeding events and death from all causes. The purpose of these post hoc analyses was to detect potential differences in clinical outcomes between females and males in unselected and propensity-score-matched cohorts. Overall, in the unmatched cohorts, accumulated target-lesion revascularization rates did not differ between both genders (2.7% vs. 2.0%; P = 0.101), however, accumulated MACE rates were higher in females than in males (5.2% vs. 3.9%; P = 0.020). After propensity-score-matching, primarily adjusting for age, hypertension and diabetes, our data revealed similar accumulated MACE in women and men (5.5% vs. 5.2%; P = 0.749). In the unmatched STEMI subgroup, all-cause mortality was significantly higher in females driven by older age ( P < 0.001). In the propensity-score-matched real-world cohorts, female gender was not a predictor for increased rates of accumulated MACE. In the unmatched STEMI subgroup, all-cause mortality was significantly higher in females due to older age. Age seems to be the determining factor for increased clinical event rates and not gender.


Platelet-Rich Plasma for Knee Osteoarthritis: Internet Marketing and Patient Education-An Appraisal of Content for Websites with the Greatest Search Engine Visibility.
The internet is increasingly being used as a resource for health-related information by the general public. We sought to establish the authorship, content, and accuracy of the information available online regarding platelet-rich plasma (PRP) therapy for knee osteoarthritis. Top 200 search results from each of the 3 leading search engines available online (Google, Yahoo!, Bing) were screened, and 181 websites were finally reviewed for content with emphasis on specific claims, comparing between websites authored by private physicians/groups and other authorship types. Nearly 80% of the websites claimed that PRP injections for osteoarthritis of the knee improve patients' pain. A total of 42.8% of the private websites and 27.6% of nonprivate websites have stated that the procedure can delay or eliminate the need for future surgery. Costs were only mentioned by few (11.6%), and mainly by the nonprivate websites. Both website groups were unlikely to mention that PRP therapy is not the treatment of choice for end-stage knee osteoarthritis (7.9% of private and 17.2% of the nonprivate sites), or to state that patients with less advanced disease may benefit more from the treatment (11.8% and 20.6%, respectively). Private websites were less likely to refer to peer-reviewed literature (18.4% vs. 41.4%) and were more than 3 times less likely to mention lack of adequate evidence (13.2% vs. 48.2%). Patients seeking online information regarding PRP therapy are vulnerable to websites presenting a narrow viewpoint of this treatment modality, putting emphasis on unsubstantiated benefits while disregarding potential drawbacks and concerns.


Deleting key autophagy elongation proteins induces acquirement of tumor-associated phenotypes via ISG15.
Autophagy is a cellular catabolic process that maintains intracellular homeostasis using lysosomal degradation systems. We demonstrate that inhibiting autophagy by depleting essential autophagy elongation proteins, Atg5 or Atg7, induces ISG15 expression through STING-mediated cytosolic dsDNA response. Genome stability is impaired in ATG5- or ATG7-depleted cells, and thus, double-strand breakages of DNA increase and cytosolic dsDNA accumulates. Accumulated cytosolic dsDNA induces the STING pathway to activate type I IFN signals which induce STAT1 activity and downregulate ATF3. When depletion of ATG5 or ATG7 inhibits autophagy, ATF3 is downregulated and STAT1 is upregulated. Furthermore, inhibiting autophagy induces ISG15 expression through STAT1 activation, which promotes acquisition of tumor-associated phenotypes such as migration, invasion, and proliferation. In conclusion, it appears that via the STING-mediated cytosolic dsDNA response, the STAT1-ISG15 axis mediates the relationship between autophagy and the immune system in relation to tumor progression. Moreover, combined with autophagy control, regulating ISG15 expression could be a novel strategy for cancer immunotherapy.


Effects of methotrexate on trophoblast proliferation and local immune responses.
Methotrexate is a folic acid analogue that has been used successfully for the treatment of ectopic pregnancy and, in conjunction with misoprostol, for medical abortions of early intrauterine pregnancies. To administer the most efficacious treatment requires knowledge of the mechanism underlying the induction of methotrexate-induced abortion. This study was designed to ascertain trophoblast integrity, proliferation and differentiation following administration of methotrexate. In addition, to determine if methotrexate affects the local uterine immune response, we ascertained the numbers and identities of decidual leukocytes following treatment. Ten women with undesired intrauterine pregnancies of 42-49 days gestation were recruited to receive methotrexte 50 mg/m2 i.m. A suction aspiration was performed 7 days later. Tissues from gestational age-matched elective surgical abortions were used as controls. Additionally, specimens from women who received methotrexate and misoprostol for abortion in a clinical trial of oral methotrexate in combination with misoprostol, who had a suction abortion because of continued embryonic cardiac activity 14 days after the methotrexate, were evaluated. Immunoreactivity to proliferating cell nuclear antigen and cyclin D3 antibodies was used to demonstrate a marked reduction in the proliferation index of cytotrophoblasts from methotrexate-treated abortions. Methotrexate treatment failures and non-treated pregnancies had a much higher proliferation index. There was no direct destruction of the syncytiotrophoblast, as indicated by the continued presence of human placental lactogen and beta-human chorionic gonadotrophin proteins. A decrease in the total number of leukocyte cells was observed in the decidua of methotrexate-treated samples, with the large granular lymphocyte (LGL) cells showing the greatest decline in numbers. Our conclusions from this study are that methotrexate acts primarily to derail the normal developmental programme of the trophoblast stem cell population, as well as to decrease LGL cell numbers in the decidua.


Assessing the impact of the Gamma variant on COVID-19 patient admissions in a southern Brazilian tertiary hospital-A comparison of dual pandemic phases.
Since the first case of COVID-19, Brazil has undergone infection waves with distinct characteristics. The description of new variants has alerted the emergence of more contagious or virulent viruses. The variant of concern Gamma emerged in Brazil and caused an epidemic wave, but its spread outside the country was limited. We report the clinical-epidemiological profile of hospitalized patients with COVID-19 by comparing two periods. A retrospective cohort study was performed. The primary outcome was to assess individuals with COVID-19 admitted in wards and intensive care units at the academic hospital of the Federal University of Parana (CHC-UFPR) between March 2020 and July 2021, correlating demographic, clinical-epidemiologic, and survival data with the most prevalent viral variant found in each period. We used Kaplan-Meier analysis to estimate the probability of survival and ROC curves to evaluate laboratory tests to find a cutoff point for poor outcomes. Data from 2,887 individuals were analyzed, 1,495 and 1,392 from the first and second periods, respectively. Hospitalization predominated among males in both periods, and the median age was significantly lower in the second one. The frequency of comorbidities was similar. Various demographic factors, clinical assessments, and laboratory tests were examined in relation to greater severity. When comparing the two periods, we observed predominance of the Wild virus during the first wave and the Gamma variant during the second, with no significant difference in outcomes. The findings suggest that despite the association of many factors with increased severity, the temporal variation between the two periods did not result in a notable divergence in the measured outcomes. The COVID-19 pandemic has lasted for a long time, with periods marked by peaks of cases, often caused by the emergence of viral variants, resulting in higher infection rates and rapid dissemination but, for variant Gamma, no apparent greater virulence.


Soft Tissue Aneurysmal Bone Cyst in the Sartorius Muscle of a 13-Year-Old Boy Mimicking Myositis Ossificans: Case Report.
A soft tissue aneurysmal bone cyst is an extremely rare tumor. The objective of the article is to present the clinical, radiological, and histopathological features of a very unusual neoplasm of soft tissues. A 13-year-old male patient presented a painful, mobile, and rapidly growing mass on the posteromedial aspect of his left knee. Imaging studies revealed a mass that arose from the medial surface of the distal sartorius muscle, with extension to the subcutaneous fat tissue. It was a well-circumscribed solid tumor with a peripheral rim calcification on plain film, computerized tomography, and ultrasound (zonal phenomenon). On magnetic resonance imaging, a heterogenous mass on T1-weighted images (WI) and T2-WI was seen, with a peripheral hypointense rim in both sequences. An outstanding edema on T2-WI extending to the soft tissue and muscles of the medial compartment of the knee was detected. The mass was resected, and the "tumoral mimickers" histopathological and molecular (next-generation sequencing) diagnoses confirmed a soft tissue aneurysmal bone cyst. A follow-up showed that the patient was free of disease 12 months after surgery. Soft tissue aneurysmal bone cyst is a rare tumor. Appropriate clinical and radiological correlation should be performed to differentiate it from other tumor mimickers.


Novel toxic effects associated with a tropical Limnothrix/Geitlerinema-like cyanobacterium.
The presence of a toxic strain of a fine filamentous cyanobacterium belonging to the Oscillatorialean family Pseudanabaenacea was detected during a survey of cyanobacterial taxa associated with the presence of cylindrospermopsin in dams in Central Queensland (Australia). The strain, AC0243, was isolated and cultured, its genomic DNA extracted and 16S RNA gene sequenced. Phylogenetic analysis placed AC0243 with Limnothrix species, although this genus appears polyphyletic. Moreover, not all morphological characters are consistent with this genus but more closely fit the description of Geitlerinema unigranulatum (R.N. Singh) Komárek and Azevedo. The potential toxic effects of AC0243 extract were assessed chemically and biologically. Cell free protein synthesis was inhibited by the extract. Exposure of Vero cells to the extract resulted in a significant reduction in cellular ATP levels following 24-72 h incubation. The presence of cylindrospermopsin was excluded based on the nature of responses obtained in cell and cell-free assays; in addition, (i) it could not be detected by HPLC, LC-MS, or immunological assay, and (ii) no genes currently associated with the production of cylindrospermopsin were found in the genome. Other known cyanobacterial toxins were not detected. The apparent novelty of this toxin is discussed.


Cervical analgesic discography. A test for diagnosis of the painful disc syndrome.
The cervical discogenic (painful disc) syndrome consists of scapular pain radiating to the head, shoulder and upper arm, often associated with paraesthesiae but without neurological deficit. Analgesic discography confirms the diagnosis and more accurately locates the pain-producing disc. In this series of patients analgesic discography followed by anterior cervical fusion with or without discectomy resulted in an 81% excellent or good result. Analgesic discography is the most effective test for location of the lesion in the painful disc syndrome.


GIS-based quantification of future nutrient loads into Lake Peipsi/Chudskoe using qualitative regional development scenarios.
This study aims at the quantification of possible future nutrient loads into Lake Peipsi/Chudskoe under different economic development scenarios. This drainage basin is on the borders of Russia, Estonia and Latvia. The sudden disintegration of the Soviet Union in 1991 caused a collapse of agricultural economy, and consequently, a substantial decrease of diffuse and point-source nutrient emissions. For the future, uncertainties about economic development and the priorities that will be set for this region make it difficult to assess the consequences for river water quality and nutrient loads into the lake. We applied five integrated scenarios of future development of this transboundary region for the next twelve to fifteen years. Each scenario consists of a qualitative story line, which was translated into quantitative changes in the input variables for a geographical information system based nutrient transport model. This model calculates nutrient emissions, as well as transport and retention and the resulting nutrient loads into the lake. The model results show that the effects of the different development scenarios on nutrient loads are relatively limited over a time span of about 15 years. In general, a further reduction of nutrient loads is expected, except for a fast economic development scenario.


Cooperative effects in the oxidation of CO by palladium oxide cations.
Cooperative reactivity plays an important role in the oxidation of CO to CO(2) by palladium oxide cations and offers insight into factors which influence catalysis. Comprehensive studies including guided-ion-beam mass spectrometry and theoretical investigations reveal the reaction products and profiles of PdO(2)(+) and PdO(3)(+) with CO through oxygen radical centers and dioxygen complexes bound to the Pd atom. O radical centers are more reactive than the dioxygen complexes, and experimental evidence of both direct and cooperative CO oxidation with the adsorption of two CO molecules are observed. The binding of multiple electron withdrawing CO molecules is found to increase the barrier heights for reactivity due to decreased binding of the secondary CO molecule, however, reactivity is enhanced by the increase in kinetic energy available to hurdle the barrier. We examine the effect of oxygen sites, cooperative ligands, and spin including two-state reactivity.


Natural combinatorial genetics and prolific polyamine production enable siderophore diversification in Serratia plymuthica.
Iron is essential for bacterial survival. Bacterial siderophores are small molecules with unmatched capacity to scavenge iron from proteins and the extracellular milieu, where it mostly occurs as insoluble Fe3+. Siderophores chelate Fe3+ for uptake into the cell, where it is reduced to soluble Fe2+. Siderophores are key molecules in low soluble iron conditions. The ability of bacteria to synthesize proprietary siderophores may have increased bacterial evolutionary fitness; one way that bacteria diversify siderophore structure is by incorporating different polyamine backbones while maintaining the catechol moieties. We report that Serratia plymuthica V4 produces a variety of siderophores, which we term the siderome, and which are assembled by the concerted action of enzymes encoded in two independent gene clusters. Besides assembling serratiochelin A and B with diaminopropane, S. plymuthica utilizes putrescine and the same set of enzymes to assemble photobactin, a siderophore found in the bacterium Photorhabdus luminescens. The enzymes encoded by one of the gene clusters can independently assemble enterobactin. A third, independent operon is responsible for biosynthesis of the hydroxamate siderophore aerobactin, initially described in Enterobacter aerogenes. Mutant strains not synthesizing polyamine-siderophores significantly increased enterobactin production levels, though lack of enterobactin did not impact the production of serratiochelins. Knocking out SchF0, an enzyme involved in the assembly of enterobactin alone, significantly reduced bacterial fitness. This study shows the natural occurrence of serratiochelins, photobactin, enterobactin, and aerobactin in a single bacterial species and illuminates the interplay between siderophore biosynthetic pathways and polyamine production, indicating routes of molecular diversification. Given its natural yields of diaminopropane (97.75 μmol/g DW) and putrescine (30.83 μmol/g DW), S. plymuthica can be exploited for the industrial production of these compounds.


Designed semisynthetic protein inhibitors of Ub/Ubl E1 activating enzymes.
Semisynthetic, mechanism-based protein inhibitors of ubiquitin (Ub) and ubiquitin-like modifier (Ubl) activating enzymes (E1s) have been developed to target E1-catalyzed adenylation and thioesterification of the Ub/Ubl C-terminus during the processes of protein SUMOylation and ubiquitination. The inhibitors were generated by intein-mediated expressed protein ligation using a truncated Ub/Ubl protein (SUMO residues 1-94; Ub residues 1-71) with a C-terminal thioester and synthetic tripeptides having a C-terminal adenosine analogue and an N-terminal cysteine residue. SUMO-AMSN (4a) and Ub-AMSN (4b) contain a sulfamide group as a nonhydrolyzable mimic of the phosphate group in the cognate Ub/Ubl-AMP adenylate intermediate in the first half-reaction, and these constructs selectively inhibit SUMO E1 and Ub E1, respectively, in a dose-dependent manner. SUMO-AVSN (5a) and Ub-AVSN (5b) contain an electrophilic vinyl sulfonamide designed to trap the incoming E1 cysteine nucleophile (Uba2 Cys173 in SUMO E1; Uba1 Cys593 in Ub E1) in the second half-reaction, and these constructs selectively, covalently, and stably cross-link to SUMO E1 and Ub E1, respectively, in a cysteine nucleophile-dependent manner. These inhibitors are powerful tools to probe outstanding mechanistic questions in E1 function and can also be used to study the biological functions of E1 enzymes.