Dissociation and re-association studies on the interaction domains of mannan-binding lectin (MBL)-associated serine proteases, MASP-1 and MASP-2, provide evidence for heterodimer formation.
Activation of the lectin pathway of complement begins with the activation of mannan-binding lectin (MBL)-associated serine proteases, MASP-1 and MASP-2, which are bound to the recognition molecules, MBL and ficolins. MASPs are Ca(2+)-dependent dimers. Dimerization and Ca(2+)-dependent association with the recognition molecules occurs via the first 3 domains, the CUB1-EGF-CUB2 region. The CUB1-EGF-CUB2 (D1-3) regions of MASP-1 and MASP-2, and also their tagged versions, were expressed in E. coli, refolded and purified. The first three domains of MASP-1 are identical with the respective regions of MASP-3 and MAp44, which are also associated with MBL and ficolins. The functionality of the fragments was checked by inhibition of C3 deposition from human serum. Time-course of the dissociation and re-association was examined by size exclusion chromatography. Both refolded proteins are tight Ca(2+)-dependent dimers, as expected. In buffer containing EDTA MASP-1_D1-3 dissociated to monomers, however it took about 1h to reach an equilibrium. Upon re-calcification dimers were re-formed, but this process was even slower; only after overnight incubation was the dimerization completed. MASP-2_D1-3 showed a somewhat different behavior: dissociation by EDTA was even slower, less complete, and higher MW aggregates also appeared. Heterodimer formation was detected by native PAGE. As modeled by the D1-3 fragments, MASP-1 and MASP-2 can readily form heterodimers after dissociation and re-association, however, in the presence of Ca(2+) exchange of subunits is slow between the homodimers. MASP-1:MASP-3 heterodimer formation was modeled by the tagged and untagged D1-3 fragments, and data indicate that subunits of these proteins are readily exchanged even in the presence of Ca(2+). The existence of heterodimers influences the current view on the composition of lectin pathway complexes and their activation.


CD44 is a metastasis suppressor gene for prostatic cancer located on human chromosome 11p13.
We have used microcell fusion-mediated chromosomal transfer to introduce normal human chromosomes into highly metastatic rodent prostatic cancer cells to map the location of a metastasis suppressor gene(s). Using this approach, several chromosomal regions have been identified that harbor such metastatic suppressor genes, including human chromosome 11 between p11.2-13 (T. Ichikawa et al., Cancer Res., 52: 3486-3490, 1992, 54: 2299-2302, 1994; N. Nihei et al., Genes Chromosomes & Cancer, 14: 112-119, 1995; C. W. Rinker-Schaeffer et al., Cancer Res., 54: 6249-6256, 1994). Using positional cloning, a metastatic suppressor gene, termed KAI1, was identified, which is located at human chromosome 11p11.2 (5). Overexpression of KAI1 results in metastasis suppression in certain highly metastatic Dunning R-3327 rat prostatic cancer sublines, such as AT6.1, without metastasis suppression in other highly metastatic sublines, such as AT3.1. This suggests that an additional metastasis suppressor gene is located within the human chromosome 11p11.2-13 region. The CD44 gene is located on human chromosome 11p13 and encodes an integral membrane glycoprotein that participates in specific cell-cell and cell-extracellular matrix interactions. Down-regulation of CD44 expression both at the mRNA and protein levels correlates with metastatic potential within the Dunning system of rat prostatic cancer sublines. Transfection-induced enhanced expression of the Mr 85,000 standard form of CD44 in the highly metastatic AT3.1 rat prostatic cells greatly suppresses their metastatic ability to the lungs without suppression of their in vivo growth rate or tumorigenicity. These results suggest that CD44 is a metastasis suppressor for prostatic cancer and that decreased expression of the standard form of CD44 is involved in the progression of prostatic cancer to a metastatic state.


Direct and indirect effects of perceived social support on health-related quality of life in persons living with HIV/AIDS.
Research has established a link between perceived social support and health-related quality of life (HRQOL) among persons living with HIV/AIDS. However, little is known about the ways through which social support influences HRQOL. This study examined the direct and indirect effects of perceived social support on physical and mental HRQOL in a sample of 602 adults living with HIV in Ontario, Canada. Participants completed the Medical Outcomes Study-HIV (MOS-HIV) health survey, the MOS-HIV Social Support Scale (MOS-HIV-SSS), and the Center for Epidemiological Studies Depression-Revised scale. Data on demographic and clinical characteristics were also collected. The direct and indirect effects of social support on the two MOS-HIV HRQOL summary measures, that is, physical health summary (PHS) and mental health summary (MHS), were estimated in multiple linear regression analyses. Perceived social support had significant direct effects on PHS (B=0.04, p<0.01) and MHS (B=0.05, p<0.01). It also had significant indirect effect on both PHS (B=0.04, p<0.01) and MHS (B=0.11, p<0.01), mediated by depressive symptoms. Interventions that enhance social support have the potential to contribute to better HRQOL either directly or indirectly by decreasing the deleterious effect of depressive symptoms on HRQOL.


[Schizophrenia and the quality of life].
This paper discusses opportunities and constraints for psychiatry to exert an influence on the quality of life of people suffering from schizophrenia. Using the example of neuroleptic drug treatment, the opportunities to both positively and adversely affect quality of life by means of psychiatric treatment are illustrated. De-institutionalisation was used as an example for demonstrating the effect of the quality of mental health services. The fact that efforts on the part of professional helpers to achieve improvements in their patients' quality of life are constrained by existing economic and socio-cultural conditions is shown by discussing the stigmatization of people with schizophrenia and their situation on the labour market.


Condensin phosphorylated by the Aurora-B-like kinase Ark1 is continuously required until telophase in a mode distinct from Top2.
Condensin is a conserved protein complex that functions in chromosome condensation and segregation. It has not been previously unequivocally determined whether condensin is required throughout mitosis. Here, we examined whether Schizosaccharomyces pombe condensin continuously acts on chromosomes during mitosis and compared its role with that of DNA topoisomerase II (Top2). Using double mutants containing a temperature-sensitive allele of the condensin SMC2 subunit cut14 (cut14-208) or of top2, together with the cold-sensitive nda3-KM311 mutation (in β-tubulin), temperature-shift experiments were performed. These experiments allowed inactivation of condensin or Top2 at various stages throughout mitosis, even after late anaphase. The results established that mitotic chromosomes require condensin and Top2 throughout mitosis, even in telophase. We then showed that the Cnd2 subunit of condensin (also known as Barren) is the target subunit of Aurora-B-like kinase Ark1 and that Ark1-mediated phosphorylation of Cnd2 occurred throughout mitosis. The phosphorylation sites in Cnd2 were determined by mass spectrometry, and alanine and glutamate residue replacement mutant constructs for these sites were constructed. Alanine substitution mutants of Cnd2, which mimic the unphosphorylated protein, exhibited broad mitotic defects, including at telophase, and overexpression of these constructs caused a severe dominant-negative effect. By contrast, glutamate substitution mutants, which mimic the phosphorylated protein, alleviated the segregation defect in Ark1-inhibited cells. In telophase, the condensin subunits in cut14-208 mutant accumulated in lumps that contained telomeric DNA and proteins that failed to segregate. Condensin might thus serve to keep the segregated chromosomes apart during telophase.


Stromal-mediated protection of tyrosine kinase inhibitor-treated BCR-ABL-expressing leukemia cells.
Clinical studies of patients with chronic myeloid leukemia revealed that a common pattern of response is a dramatic fall in the circulating population of blast cells, with a minimal or delayed decrease in marrow blasts, suggesting a protective environment. These observations suggest that a greater understanding of the interaction of stromal cells with leukemic cells is essential. Here, we present an in vivo system for monitoring relative tumor accumulation in leukemic mice and residual disease in leukemic mice treated with a tyrosine kinase inhibitor and an in vitro system for identifying integral factors involved in stromal-mediated cytoprotection. Using the in vivo model, we observed high tumor burden/residual disease in tissues characterized as significant sources of hematopoiesis-promoting stroma, with bone marrow stroma most frequently showing the highest accumulation of leukemia in untreated and nilotinib-treated mice as well as partial protection of leukemic cells from the inhibitory effects of nilotinib. These studies, which showed a pattern of leukemia distribution consistent with what is observed in imatinib- and nilotinib-treated chronic myeloid leukemia patients, were followed by a more in-depth analysis of stroma-leukemia cell interactions that lead to protection of leukemia cells from nilotinib-induced cytotoxicity. For the latter, we used the human BCR-ABL-positive cell line, KU812F, and the human bone marrow stroma cell line, HS-5, to more closely approximate the bone marrow-associated cytoprotection observed in drug-treated leukemia patients. This in vitro system helped to elucidate stromal-secreted viability factors that may play a role in stromal-mediated cytoprotection of tyrosine kinase inhibitor-treated leukemia cells.


[Hemodynamic shifts in humans under the conditions of prolonged microgravity and the role of hypovolemia].
Purpose of this work was to elicit the role of hypovolemia in the development of hemodynamic shifts and loss of orthostatic tolerance in humans during prolonged space flights (PSF). Correlation analysis of the data of in-flight ultrasonic investigations of hemodynamics in resting cosmonauts and bio-impedance measurements of body liquids in different periods during PSF pointed out that changes in the left ventricle filling and renal artery resistance (Rren) reflect the degree of hypovolemia with r = 0.95. The dependence of orthostatic tolerance on hypovolemia fell short of this ambiguity. Lower body negative pressure tests (LBNP) performed by 15 cosmonauts in different periods of PSF typically correlated with the body hydration status at r = 0.94; however, several cosmonauts showed a substantial loss of LBNP tolerance despite a moderate hypovolemia. Therefore, we may hypothesize that hypovolemia is a significant yet not the sole factor in orthostatic disorders. Deconditioning, altered control of hemodynamics, decreased vasoconstriction of arterial vessels and increased compliance of the leg veins have an important, in some cases decisive role in degradation of orthostatic tolerance.


Large-Scale Structural Characterization of Drug and Drug-Like Compounds by High-Throughput Ion Mobility-Mass Spectrometry.
Ion mobility-mass spectrometry (IM-MS) can provide orthogonal information, i.e., m/z and collision cross section (CCS), for the identification of drugs and drug metabolites. However, only a small number of CCS values are available for drugs, which limits the use of CCS as an identification parameter and the assessment of structure-function relationships of drugs using IM-MS. Here, we report the development of a rapid workflow for the measurement of CCS values of a large number of drug or drug-like molecules in nitrogen on the widely available traveling wave IM-MS (TWIM-MS) platform. Using a combination of small molecule and polypeptide CCS calibrants, we successfully determined the nitrogen CCS values of 1425 drug or drug-like molecules in the MicroSource Discovery Systems' Spectrum Collection using flow injection analysis of 384-well plates. Software was developed to streamline data extraction, processing, and calibration. We found that the overall drug collection covers a wide CCS range for the same mass, suggesting a large structural diversity of these drugs. However, individual drug classes appear to occupy a narrow and unique space in the CCS-mass 2D spectrum, suggesting a tight structure-function relationship for each class of drugs with a specific target. We observed bimodal distributions for several antibiotic species due to multiple protomers, including the known fluoroquinolone protomers and the new finding of cephalosporin protomers. Lastly, we demonstrated the utility of the high-throughput method and drug CCS database by quickly and confidently confirming the active component in a pharmaceutical product.


Prognostic factor study of survival after enucleation for juxtapapillary melanomas.
We reviewed 242 cases of choroidal and ciliary body melanoma that were treated by enucleation to determine the importance of juxtapapillary location as a prognostic factor for tumor-related deaths. Analysis involved Kaplan-Meier survival curves and Cox proportional hazards analysis. Patients with juxtapapillary tumors had a worse prognosis than those with tumors in other locations (64% vs 78% five-year survival), but the difference was not statistically significant. Juxtapapillary location was not an independent risk factor on Cox multivariate analysis. Risk factors included number of epithelioid cells per high-power field, largest tumor dimension, location of the tumor's anterior margin, and invasion to the line of transection. In 63 cases (26%), the tumor was in contact with the optic disc. For these juxtapapillary tumors, we also evaluated the prognostic importance of six variables relevant to juxtapapillary location. On univariate analysis, tumor height at the disc margin, subarachnoid space invasion, and postlaminar optic nerve invasion were significantly related to survival, while disc compression by tumor, prelaminar optic nerve invasion, and extent of disc perimeter surrounded by tumor were not. On Cox multivariate analysis, however, none of these six variables was an independent risk factor. Risk factors for juxtapapillary tumors included number of epithelioid cells per high-power field and location of the tumor's anterior margin.


Hexavalent molybdenum reduction to Mo-blue by Acinetobacter calcoaceticus.
A local molybdenum-reducing bacterium was isolated and tentatively identified as Acinetobacter calcoaceticus strain Dr.Y12 based on carbon utilization profiles using Biolog GN plates and 16S rDNA comparative analysis. Molybdate reduction was optimized under conditions of low dissolved oxygen (37 degrees C and pH 6.5). Of the electron donors tested, glucose, fructose, maltose and sucrose supported molybdate reduction after 1 d of incubation, glucose and fructose supporting the highest Mo-blue production. Optimum Mo-blue production was reached at 20 mmol/L molybdate and 5 mmol/L phosphate; increasing the phosphate concentrations inhibited the production. An increase in an overall absorption profiles, especially at peak maximum at 865 nm and the shoulder at 700 nm, was observed in direct correlation with the increased in Mo-blue amounts. Metal ions, such as chromium, cadmium, copper, mercury and lead (2 mmol/L final concentration) caused approximately 88, 53, 80, 100, and 20 % inhibition, respectively. Respiratory inhibitors, such as antimycin A, rotenone, sodium azide and cyanide showed in this bacterium no inhibition of the Mo-blue production, suggesting that the electron transport system is not a site of molybdate reduction.


Signatures of Long-Term Balancing Selection in Human Genomes.
Balancing selection maintains advantageous diversity in populations through various mechanisms. Although extensively explored from a theoretical perspective, an empirical understanding of its prevalence and targets lags behind our knowledge of positive selection. Here, we describe the Non-central Deviation (NCD), a simple yet powerful statistic to detect long-term balancing selection (LTBS) that quantifies how close frequencies are to expectations under LTBS, and provides the basis for a neutrality test. NCD can be applied to a single locus or genomic data, and can be implemented considering only polymorphisms (NCD1) or also considering fixed differences with respect to an outgroup (NCD2) species. Incorporating fixed differences improves power, and NCD2 has higher power to detect LTBS in humans under different frequencies of the balanced allele(s) than other available methods. Applied to genome-wide data from African and European human populations, in both cases using chimpanzee as an outgroup, NCD2 shows that, albeit not prevalent, LTBS affects a sizable portion of the genome: ∼0.6% of analyzed genomic windows and 0.8% of analyzed positions. Significant windows (P < 0.0001) contain 1.6% of SNPs in the genome, which disproportionally fall within exons and change protein sequence, but are not enriched in putatively regulatory sites. These windows overlap ∼8% of the protein-coding genes, and these have larger number of transcripts than expected by chance even after controlling for gene length. Our catalog includes known targets of LTBS but a majority of them (90%) are novel. As expected, immune-related genes are among those with the strongest signatures, although most candidates are involved in other biological functions, suggesting that LTBS potentially influences diverse human phenotypes.


Multiple Ribosomal RNA Operons in Bacteria; Their Concerted Evolution and Potential Consequences on the Rate of Evolution of Their 16S rRNA.
Bacterial species differ greatly in the number and location of the rRNA operons which may be present in the bacterial chromosomes and plasmids. Most bacterial species contain more than one ribosomal RNA operon copy in their genomes, with some species containing up to 15 such copies. We review the number and location of the rRNA operons and discuss evolution of 16S rRNA (rrs) genes -which are considered as ultimate chronometers for phylogenetic classification- in bacteria with multiple copies of these genes. In these bacterial species, the rrs genes must evolve in concert and sequence changes generated by mutation or horizontal gene transfer must be either erased or spread to every gene copy to avoid divergence, as it occurs when they are present in different species. Analysis of polymorphic sites in intra-genomic rrs copies identifies putative conversion events and demonstrates that sequence conversion is patchy and occurs in small conversion tracts. Sequence conversion probably arises by a non-reciprocal transfer between two or more copies where one copy contributes only a small contiguous segment of DNA, whereas the other copy contributes the rest of the genome in a fairly well understood molecular process. Because concerted evolution implies that a mutation in any of the rrs copies is either eliminated or transferred to every rrs gene in the genome, this process should slow their evolution rate relative to that of single copy genes. However, available data on the rrs genes in bacterial genomes do not show a clear relationship between their evolution rates and the number of their copies in the genome.


A morphological study of variations in the branching pattern and termination of the radial artery.
Coronary artery bypass grafting is an established means of treating advanced coronary artery disease. In recent years, there has been an increased interest in the radial artery as an entry route during coronary angiography. Accurate knowledge of the branching pattern of this artery and its relation to surrounding structures is of great importance in the care of surgical patients. This study was conducted on 75 formalin-fixed upper limbs in order to note the variations in the branching pattern and termination of the radial artery. The radial artery divided into three branches in 2.7% of cases and into two branches in 52.0% of cases. The radial recurrent artery originated from the brachial artery instead of the radial artery in 12.0% of cases. The radial recurrent artery, palmar carpal artery, first dorsal metacarpal artery and superficial palmar artery were absent in 1.3%, 26.7%, 9.3% and 5.3% of cases, respectively. 6.7% of cases had a high origin of the superficial palmar artery. The rich photographic documentation of the variation of branching pattern and termination of radial artery is not only of academic interest but also useful to surgeons and radiologists working in the same area.


Aligning physical learning spaces with the curriculum: AMEE Guide No. 107.
This Guide explores emerging issues on the alignment of learning spaces with the changing curriculum in medical education. As technology and new teaching methods have altered the nature of learning in medical education, it is necessary to re-think how physical learning spaces are aligned with the curriculum. The better alignment of learning spaces with the curriculum depends on more directly engaged leadership from faculty and the community of medical education for briefing the requirements for the design of all kinds of learning spaces. However, there is a lack of precedent and well-established processes as to how new kinds of learning spaces should be programmed. Such programmes are essential aspects of optimizing the intended experience of the curriculum. Faculty and the learning community need better tools and instruments to support their leadership role in briefing and programming. A Guide to critical concepts for exploring the alignment of curriculum and learning spaces is provided. The idea of a networked learning landscape is introduced as a way of assessing and evaluating the alignment of physical spaces to the emerging curriculum. The concept is used to explore how technology has widened the range of spaces and places in which learning happens as well as enabling new styles of learning. The networked learning landscaped is explored through four different scales within which learning is accommodated: the classroom, the building, the campus, and the city. High-level guidance on the process of briefing for the networked learning landscape is provided, to take into account the wider scale of learning spaces and the impact of technology. Key to a successful measurement process is argued to be the involvement of relevant academic stakeholders who can identify the strategic direction and purpose for the design of the learning environments in relation to the emerging demands of the curriculum.


The thermogenic properties of ephedrine/methylxanthine mixtures: animal studies.
An over-the-counter preparation containing ephedrine, caffeine, and theophylline was examined for thermogenic anti-obesity properties. Administration of the methylxanthines to MSG-induced obese mice for 6 wk had no effect on energy balance or body composition. In contrast, treatment with ephedrine alone caused losses of 14% in body weight and 42% in body fat, effects brought about mainly by a 10% increase in energy expenditure. These changes were accentuated when ephedrine was administered together with one or both methylxanthines: energy expenditure was increased by a further 10%, and led to a reduction of about 25% in body weight and 75% in body fat, while the total food intake and body protein were unaltered. These results indicate that dietary methylxanthines like caffeine and theophylline, although alone have little effect on energy balance, can nevertheless markedly potentiate the thermogenic anti-obesity effect of ephedrine and normalize the body composition of the obese to the lean levels.


Fault Detectability of Asynchronous Switched Boolean Networks: A Set Reachability Approach.
This paper addresses the fault detectability problem of asynchronous switched Boolean networks, which is focused on whether occurrence of the faults would have an impact on the outputs of the considered network. By applying the semi-tensor product method, the asynchronous switching scheme of the considered system is converted into multiple switching signals. Based on them, an augmented system is established to transform the fault detectability issue of the original system into a set reachability problem. Moreover, some criteria for the fault detectability of the asynchronous switched Boolean control networks are also proposed. Additionally, two algorithms are given to design feasible control sequences after explicitly analyzing the considered augmented system. Finally, an example is provided to illustrate the significance of the obtained theoretical results.


Secretory antibodies to citrullinated peptides in plasma and saliva from rheumatoid arthritis patients and their unaffected first-degree relatives.
The aim of this study was to evaluate secretory antibodies to citrullinated proteins (ACPA) in plasma and immunoglobulin (Ig)A ACPA in saliva from patients with rheumatoid arthritis (RA) and their unaffected first-degree relatives (FDRs). Patients with RA (n = 194) and first-degree relatives unaffected by RA (n = 191) were recruited for analysis of secretory antibodies to second-generation cyclic citrullinated peptides (anti-CCP) in plasma. From a subpopulation (25 RA patients, 21 first-degree relatives and 11 controls), saliva samples were obtained for IgA anti-CCP analysis. The presence of secretory ACPA was compared between subject categories, and related to genetic and environmental risk factors. Secretory ACPA occurred in 37 (19%) plasma samples from patients with RA, but only in two (1%) of FDRs. IgA ACPA in saliva was found in three of 25 (12%) patients with RA, but not in any of the 21 FDRs (< 5%). No significant associations were seen between the presence of secretory ACPA and SE or smoking, either among RA patients or among FDRs. Despite occurring in 19% of RA plasma, secretory ACPA was rare in both saliva and plasma among FDRs, even among those positive for conventional ACPA of non-mucosal origin. Longitudinal studies are warranted to determine whether circulating secretory ACPA occurs before or in parallel with the development of clinical arthritis.


Enrichment of SNPs in Functional Categories Reveals Genes Affecting Complex Traits.
Genome-wide association studies (GWAS) have indicated potential to identify heritability of common complex phenotypes, but traditional approaches have limited ability to detect hiding signals because single SNP has weak effect size accounting for only a small fraction of overall phenotypic variations. To improve the power of GWAS, methods have been developed to identify truly associated genes by jointly testing effects of all SNPs. However, equally considering all SNPs within a gene might dilute strong signals of SNPs in real functional categories. Here, we observed a consistent pattern on enrichment of significant SNPs in eight functional categories across six phenotypes, with the highest enrichment in coding and both UTR regions while the lowest enrichment in the intron. Based on the pattern of SNP enrichment in functional categories, we developed a new approach for detecting gene associations on traits (DGAT) by selecting the most significant functional category and then using SNPs within it to assess gene associations. The method was found to be robust in type I error rate on simulated data, and to have mostly higher power in detecting associated genes for three different diseases than other methods. Further analysis indicated ability of the DGAT to detect novel genes. The DGAT is available by http://sparks-lab.org/server/DGAT.


Suspected flunixin poisoning of a wild Eurasian Griffon Vulture from Spain.
Exposure to residues of the nonsteroidal anti-inflammatory drug (NSAID) diclofenac present in livestock carcasses has caused extensive declines in 3 Gyps vulture species across Asia. The carcass of a wild Eurasian Griffon Vulture (Gyps fulvus) was found in 2012 on an Andalucian (Spain) game hunting reserve and examined forensically. The bird had severe visceral gout, a finding consistent with Gyps vultures from Asia that have been poisoned by diclofenac. Liver and kidney samples from this Eurasian Griffon Vulture contained elevated flunixin (an NSAID) levels (median = 2.70 and 6.50 mg/kg, respectively). This is the first reported case of a wild vulture being exposed to and apparently killed by an NSAID outside Asia. It is also the first reported instance of mortality in the wild resulting from environmental exposure to an NSAID other than diclofenac.


Leukemia inhibitory factor, a potent cardiac hypertrophic cytokine, enhances L-type Ca2+ current and [Ca2+]i transient in cardiomyocytes.
This study investigates whether leukemia inhibitory factor (LIF), a potent cardiac hypertrophic cytokine, affects the L-type Ca2+ current (I(Ca,L)) and intracellular Ca2+ concentrations ([Ca2+]i) in cardiomyocytes. I(Ca,L) was recorded using a whole cell patch clamp configuration in guinea pig cardiomyocytes, and the [Ca2+]i transient was detected by use of Fluo-3 in rat cardiomyocytes. Cells were preincubated with LIF (1000 U/ml) for 15 min before whole cell recording. LIF increased I(Ca,L) by 41.8%. LIF synergistically increased I(Ca,L) with isoproterenol. Preincubation with H89 did not inhibit the LIF-induced increase in I(Ca,L), indicating that this phenomenon is PKA-independent. PD98059 completely inhibited the increase in I(Ca,L), and this effect was dose-dependent (IC50=3.6 micromol/l). Other signal transduction inhibitors including AG490, SB203580, chelerythrine, genistein, and KN62 did not affect the LIF-induced increase in I(Ca,L). Perforated patch clamp recording revealed that LIF maximally increased the I(Ca,L) by 25% at 15 min. LIF also increased the peak [Ca2+]i transient level by 63% at 15 min. PD98059 fully inhibited the increase in the [Ca2+]i transient. In conclusion, LIF increased I(Ca,L) and the [Ca2+]i transient in cardiomyocytes, and the Raf-1/MEK/ERK pathway might be involved in the modulation of this activation.


Role of the NH2 -terminal fragment of dentin sialophosphoprotein in dentinogenesis.
Dentin sialophosphoprotein (DSPP) is a large precursor protein that is proteolytically processed into a NH2 -terminal fragment [composed of dentin sialoprotein (DSP) and a proteoglycan form (DSP-PG)] and a COOH-terminal fragment [dentin phosphoprotein (DPP)]. In vitro studies indicate that DPP is a strong initiator and regulator of hydroxyapatite crystal formation and growth, but the role(s) of the NH2 -terminal fragment of DSPP (i.e., DSP and DSP-PG) in dentinogenesis remain unclear. This study focuses on the function of the NH2 -terminal fragment of DSPP in dentinogenesis. Here, transgenic (Tg) mouse lines expressing the NH2 -terminal fragment of DSPP driven by a 3.6-kb type I collagen promoter (Col 1a1) were generated and cross-bred with Dspp null mice to obtain mice that express the transgene but lack the endogenous Dspp (Dspp KO/DSP Tg). We found that dentin from the Dspp KO/DSP Tg mice was much thinner, more poorly mineralized, and remarkably disorganized compared with dentin from the Dspp KO mice. The fact that Dspp KO/DSP Tg mice exhibited more severe dentin defects than did the Dspp null mice indicates that the NH2 -terminal fragment of DSPP may inhibit dentin mineralization or may serve as an antagonist against the accelerating action of DPP and serve to prevent predentin from being mineralized too rapidly during dentinogenesis.


Allergic contact dermatitis: a significant environmental and occupational skin disease.
This review article seeks to provide an overview of allergic contact dermatitis (ACD) as a significant environmental and occupational skin disease, the phases of ACD, its causes from the occupational and environmental perspectives, its detection, the effects of ACD with respect to the social, psychological, occupational, and financial perspectives, and its cure and/or prevention. Human skin is very sensitive and as the largest organ in the body, it is highly prone to direct and indirect contact with the substances from its environment. The skin reacts to these substances (xenobiotics) differently depending on the individual's tolerance level or threshold. Allergic contact dermatitis is a significant environmental and occupational skin disease that should not be ignored in our society because it can affect the quality of life of an affected individual. There are multiple causes of ACD, and these causes of ACD have been discussed from two perspectives: environmental and occupational. The effects of ACD can be psychological, social, financial, and occupational. There is need for more public enlightenment on the effects of ACD as well as a precise understanding that it is not a contagious disease so as to significantly reduce the psychological and social effects of ACD on these patients.


Not so ugly after all: when shame acts as a commitment device.
Most psychological theories and research on shame focus on the ugly aspects and negative consequences of this emotion. Theories on moral emotions, however, assume that shame acts as a commitment device motivating prosocial behavior. To solve this apparent paradox, the authors studied the effects of shame on prosocial behavior. Shame was hypothesized to motivate prosocial behavior when it was relevant for the decision at hand (endogenous). In contrast, shame that was not relevant for the decision at hand (exogenous) was hypothesized to have no such effects. Four experiments with three different shame inductions and two different measures of prosocial behavior confirmed that endogenous shame motivated prosocial behavior for proselfs but that exogenous shame did not. Shame is shown to have a clear interpersonal function in the sense that it acts as a commitment device.


Effect of Vibrio alginolyticus on larval survival of the blue mussel Mytilus galloprovincialis.
The effect of increasing concentrations of Vibrio alginolyticus on survival of Mytilus galloprovincialis larvae was studied in a 48 h static bioassay in 1 l glass bottles. Five bacterial densities were tested ranging from 10(2) to 10(6) bacteria ml(-1). Larval survival and normality (veliger larvae with the typical D-shape) were evaluated after 48 h. An inverse relationship between bacterial concentration and larval survival and normality was observed. In spite of high larval survival (79%) under conditions of high bacterial density (10(5) bacteria ml(-1)), the percent of normal larvae was 11%. Besides an irregular shape, abnormal larvae also presented velum reduction. Results from this study suggest that concentrations of V. alginolyticus lower than 10(3) bacteria ml(-1) should be maintained during M. galloprovincialis larval culture.


Induced Circular Dichroism Analysis of Thermally Induced Conformational Changes on Protein Binding Sites Under a Crowding Environment.
Protein-ligand interactions in crowded cellular environments play a crucial role in biological functions. The crowded environment can perturb the overall protein structure and local conformation, thereby influencing the binding pathway of protein-ligand reactions within the cellular milieu. Therefore, a detailed understanding of the local conformation is crucial for elucidating the intricacies of protein-ligand interactions in crowded cellular environments. In this study, we investigated the feasibility of induced circular dichroism (ICD) using 8-anilinonaphthalene-1-sulfonic acid (ANS) for local conformational analysis at the binding site in a crowding environment. Bovine serum albumin (BSA) concentration-dependent measurements were performed to assess the feasibility of ANS-ICD for analyzing protein interior binding sites. The results showed distinct changes in the ANS-ICD spectra of BSA solutions, indicating their potential for analyzing the internal conformation of proteins. Moreover, temperature-dependent measurements were performed in dilute and crowding environments, revealing distinct denaturation pathways of BSA binding sites. Principal component analysis of ANS-ICD spectral changes revealed lower temperature pre-denaturation in the crowded solution than that in the diluted solution, suggesting destabilization of binding sites owing to self-crowding repulsive interactions. The established ANS-ICD method can provide valuable conformational insights into protein-ligand interactions in crowded cellular environments.


Gelation-induced enhanced fluorescence emission from organogels of salicylanilide-containing compounds exhibiting excited-state intramolecular proton transfer: synthesis and self-assembly.
Self-assembly structure, stability, hydrogen-bonding interaction, and optical properties of a new class of low molecular weight organogelators (LMOGs) formed by salicylanilides 3 and 4 have been investigated by field-emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), UV/Vis absorption and photoluminescence, as well as theoretical studies by DFT and semiempirical calculations with CI (AM1/PECI=8) methods. It was found that salicylanilides form gels in nonpolar solvents due to pi-stacking interaction complemented by the presence of both inter- and intramolecular hydrogen bonding. The supramolecular arrangement in these organogels predicted by XRD shows lamellar and hexagonal columnar structures for gelators 3 and 4, respectively. Of particular interest is the observation of significant fluorescence enhancement accompanying gelation, which was ascribed to the formation of J-aggregates and inhibition of intramolecular rotation in the gel state.


LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC.
Dysregulation of transcription and cytokine expression has been implicated in the pathogenesis of a variety inflammatory diseases. The resulting imbalance between inflammatory and resolving transcriptional programs can cause an overabundance of pro-inflammatory, classically activated macrophage type 1 (M1) and/or helper T cell type 1 (Th1) products, such as IFNγ, TNFα, IL1-β, and IL12, that prevent immune switching to resolution and healing. The low molecular weight fraction of human serum albumin (LMWF5A) is a novel biologic drug that is currently under clinical investigation for the treatment of osteoarthritis and the hyper-inflammatory response associated with COVID-19. This study aims to elucidate transcriptional mechanisms of action involved with the ability of LMWF5A to reduce pro-inflammatory cytokine release. ELISA arrays were used to identify cytokines and chemokines influenced by LMWF5A treatment of LPS-stimulated peripheral blood mononuclear cells (PBMC). The resulting profiles were analyzed by gene enrichment to gain mechanistic insight into the biologic processes and transcription factors (TFs) underlying the identified differentially expressed cytokines. DNA-binding ELISAs, luciferase reporter assays, and TNFα or IL-1β relative potency were then employed to confirm the involvement of enriched pathways and TFs. LMWF5A was found to significantly inhibit a distinct set of pro-inflammatory cytokines (TNFα, IL-1β, IL-12, CXCL9, CXCL10, and CXCL11) associated with pro-inflammatory M1/Th1 immune profiles. Gene enrichment analysis also suggests these cytokines are, in part, regulated by NF-κB and STAT transcription factors. Data from DNA-binding and reporter assays support this with LMWF5A inhibition of STAT1α DNA-binding activity as well as a reduction in overall NF-κB-driven luciferase expression. Experiments using antagonists specific for the immunomodulatory and NF-κB/STAT-repressing transcription factors, peroxisome proliferator-activated receptor (PPAR)γ and aryl hydrocarbon receptor (AhR), indicate these pathways are involved in the LMWF5A mechanisms of action by reducing LMWF5A drug potency as measured by TNFα and IL-1β release. In this report, we provide evidence that LMWF5A reduces pro-inflammatory cytokine release by activating the immunoregulatory transcription factors PPARγ and AhR. In addition, our data indicate that LMWF5A suppresses NF-κB and STAT1α pro-inflammatory pathways. This suggests that LMWF5A acts through these mechanisms to decrease pro-inflammatory transcription factor activity and subsequent inflammatory cytokine production.


Development of sludge based stable aqueous Bacillus thuringiensis formulations.
This study focuses on development of aqueous flowable (suspension) formulations for Bacillus thuringiensis (Bt) based biopesticides from wastewater sludge. Different inerts like sorbitol, sodium monophosphate, sodium metabisulphite, sorbic acid, propionic acid, Tween-80, Triton X-100 and glycerol were tested for formulations. Five different formulations for non-hydrolyzed (NH) secondary sludges were tried and the best combination selected on the basis of various physical parameters like viscosity, particle size, suspendibility, entomotoxicity, and microbiological purity tests. F5 formulations (for secondary sludge) comprising sorbitol, sodium monophosphate and sodium metabisulphite gave better physical and biological characteristics with a small effect on entomotoxicity and spore concentration after 120 days at pH 6, 6.5 and temperatures 40 and 50 degrees C and viscosity change at 40 and 50 degrees C. The formulations were more stable at pH 4.0 to 5.0 and temperatures 4 to 30 degrees C whereas at pH 6.0 and 6.5 and temperatures 40 and 50 degrees C, there was degeneration of the product. Lower proteolytic activity and physical factors like ionic strength and surface group changes at pH 6 and 6.5 were responsible for the instability of the formulation.


Development of DNA Aptamers Against Plasmodium falciparum Blood Stages Using Cell-Systematic Evolution of Ligands by EXponential Enrichment.
New biomarkers have to be developed in order to increase the performance of current antigen-based malaria rapid diagnosis. Antibody production often involves the use of laboratory animals and is time-consuming and costly, especially when the target is Plasmodium, whose variable antigen expression complicates the development of long-lived biomarkers. To circumvent these obstacles, we have applied the Systematic Evolution of Ligands by EXponential enrichment method to the rapid identification of DNA aptamers against Plasmodium falciparum-infected red blood cells (pRBCs). Five 70 b-long ssDNA sequences, and their shorter forms without the flanking PCR primer-binding regions, have been identified having a highly specific binding of pRBCs versus non-infected erythrocytes. Structural analysis revealed G-enriched sequences compatible with the formation of G-quadruplexes. The selected aptamers recognized intracellular epitopes with apparent Kds in the μM range in both fixed and non-fixed saponin-permeabilized pRBCs, improving >30-fold the pRBC detection in comparison with aptamers raised against Plasmodium lactate dehydrogenase, the gold standard antigen for current malaria diagnostic tests. In thin blood smears of clinical samples the aptamers reported in this work specifically bound all P. falciparum stages versus non-infected erythrocytes, and also detected early and late stages of the human malaria parasites Plasmodium vivax, Plasmodium ovale and Plasmodium malariae. The results are discussed in the context of their potential application in future malaria diagnostic devices.


Accumulation of advanced glycation end products as a molecular mechanism for aging as a risk factor in osteoarthritis.
Osteoarthritis (OA) is one of the most prevalent and disabling chronic conditions affecting the elderly. Its etiology is largely unknown, but age is the most prominent risk factor. The current study was designed to test whether accumulation of advanced glycation end products (AGEs), which are known to adversely affect cartilage turnover and mechanical properties, provides a molecular mechanism by which aging contributes to the development of OA. The hypothesis that elevated AGE levels predispose to the development of OA was tested in the canine anterior cruciate ligament transection (ACLT) model of experimental OA. Cartilage AGE levels were enhanced in young dogs by intraarticular injections of ribose. This mimics the accumulation of AGEs without the interference of other age-related changes. The severity of OA was then assessed 7 weeks after ACLT surgery in dogs with normal versus enhanced AGE levels. Intraarticular injections of ribose enhanced cartilage AGE levels approximately 5-fold, which is similar to the normal increase that is observed in old dogs. ACLT surgery resulted in more-pronounced OA in dogs with enhanced AGE levels. This was observed as increased collagen damage and enhanced release of proteoglycans. The attempt to repair the matrix damage was impaired; proteoglycan synthesis and retention were decreased at enhanced AGE levels. Mankin grading of histology sections also revealed more-severe OA in animals with enhanced AGE levels. These findings demonstrate increased severity of OA at higher cartilage AGE levels and provide the first in vivo experimental evidence for a molecular mechanism by which aging may predispose to the development of OA.


Fermentation of mucin by bifidobacteria from rectal samples of humans and rectal and intestinal samples of animals.
Bifidobacteria (246 strains in total) were isolated from rectal samples of infants and adult humans and animals, and from intestinal samples of calves. Twenty-five strains grew well on mucin: 20 from infants, two from adults, and three from goatlings. Poor or no growth on mucin was observed in 156 bifidobacterial strains of animal origin. The difference between human and animal isolates in ability to grow on mucin was significant at p < 0.001. Nine human strains with the best growth on mucin were identified as Bifidobacterium bifidum. These strains produced extracellular, membrane-bound, and intracellular mucinases with activities of 0.11, 0.53, and 0.09 μmol/min of reducing sugars per milligram of protein, respectively. Membrane-bound mucinases were active between pH 5 and 10. The optimum pH of extracellular mucinases was 6-7. Fermentation patterns in cultures grown on mucin and glucose differed. On mucin, the acetate-to-lactate ratio was higher than in cultures grown on glucose (p = 0.012). We showed that the bifidobacteria belong to the mucin-fermenting bacteria in humans, but their significance in mucin degradation in animals seems to be limited.


Cognitive behavioral therapy for posttraumatic stress disorder in women: a randomized controlled trial.
The prevalence of posttraumatic stress disorder (PTSD) is elevated among women who have served in the military, but no prior study has evaluated treatment for PTSD in this population. Prior research suggests that cognitive behavioral therapy is a particularly effective treatment for PTSD. To compare prolonged exposure, a type of cognitive behavioral therapy, with present-centered therapy, a supportive intervention, for the treatment of PTSD. A randomized controlled trial of female veterans (n=277) and active-duty personnel (n=7) with PTSD recruited from 9 VA medical centers, 2 VA readjustment counseling centers, and 1 military hospital from August 2002 through October 2005. Participants were randomly assigned to receive prolonged exposure (n = 141) or present-centered therapy (n = 143), delivered according to standard protocols in 10 weekly 90-minute sessions. Posttraumatic stress disorder symptom severity was the primary outcome. Comorbid symptoms, functioning, and quality of life were secondary outcomes. Blinded assessors collected data before and after treatment and at 3- and 6-month follow-up. Women who received prolonged exposure experienced greater reduction of PTSD symptoms relative to women who received present-centered therapy (effect size, 0.27; P = .03). The prolonged exposure group was more likely than the present-centered therapy group to no longer meet PTSD diagnostic criteria (41.0% vs 27.8%; odds ratio, 1.80; 95% confidence interval, 1.10-2.96; P = .01) and achieve total remission (15.2% vs 6.9%; odds ratio, 2.43; 95% confidence interval, 1.10-5.37; P = .01). Effects were consistent over time in longitudinal analyses, although in cross-sectional analyses most differences occurred immediately after treatment. Prolonged exposure is an effective treatment for PTSD in female veterans and active-duty military personnel. It is feasible to implement prolonged exposure across a range of clinical settings. clinicaltrials.gov Identifier: NCT00032617.


Evaluation of Daily Lives of Children and Adolescents with Celiac Disease and Nursing Approaches.
The incidence of celiac disease has been increasing in recent years. Celiac disease is an autoimmune enteropathy that emerges mostly in the childhood period. This disorder is a chronic condition of the small intestine due to gluten intake in individuals with genetic predisposition. In the treatment of celiac disease, gluten-free diet therapy is applied. However, the adaptation process to the diet may create difficulties for patients. Compliance with diet is much more difficult among patients with celiac disease in childhood and adolescence than in adult patients. Children and adolescents face distinct difficulties in their homes, at school, and in their social lives. Besides following dietary treatment, children and adolescents living with celiac disease need peer support to manage their daily routines. In this article, we aimed to discuss the problems experienced by children and adolescents living with celiac disease and nursing approaches to these problems.


National panorama of adolescent pregnancy in Mexico: lessons learned in a six-year period.
Globally, adolescent pregnancy constitutes a serious public health problem of a multifactorial nature. Specifically for women, it entails various educational, economic and social implications that affect their life project and widen the social gaps in this age group. Furthermore, adolescent girls are more vulnerable because of the health risk involved with pregnancy at a younger age. According to the World Health Organization, "the probability of maternal death is twice as high in adolescents in comparison with women aged between 20 and 30 years, and for those younger than 15 years, the risks are five times higher". In general, adolescents are in great need for education on sexual and reproductive health issues, which should be aimed at increasing information and knowledge about correct use and access to modern contraceptive methods, as well as at demystifying fears and beliefs around their possible side effects. Ensuring proper counseling with trained personnel is equally vital. Public institutions have a social responsibility to support efforts aimed at preventing adolescent pregnancy, based on relevant lines of action and health policies.


Limits of rapid log P determination methods for highly lipophilic and flexible compounds.
Lipophilicity is of crucial importance in many fields including pharmaceutical, environmental, cosmetic and food industries. Whereas different experimental strategies have been developed for rapid lipophilicity determination of new chemical entities, log P determination of highly lipophilic compounds is always challenging. In this study, three published chromatographic methods have been compared on a series of phenylalkanoic acids including the pro-perfume HaloscentD (HD-C12). Different log P values were obtained depending on the chromatographic method used for log P estimation. Molecular modelling suggested that log P variations may be due to the chromatographic conditions applied (isocratic or gradient mode, ratio methanol/water in the mobile phase), responsible of specific conformations of the molecule in solution. Thus, for flexible compounds, published methods have to be used with caution and considered as a good tool to estimate a log P range, depending on the molecular conformational state.


Survival of enterohemorrhagic Escherichia coli O157:H7 in bovine feces applied to lettuce and the effectiveness of chlorinated water as a disinfectant.
Bovine feces are a potential vehicle for transmitting enterohemorrhagic Escherichia coli O157:H7 to humans. A study was undertaken to determine survival characteristics of E. coli O157:H7 on iceberg lettuce using 0.1% peptone water and bovine feces as carriers for inocula. Four levels of inoculum, ranging from 10(0) to 10(5) CFU of E. coli O157:H7 per g of lettuce, were applied. Populations surviving on lettuce stored at 4 degrees C were monitored for up to 15 days. Regardless of the type of carrier, viable cells of E. coli O157:H7 were detected on lettuce after 15 days, even when the initial inoculum was 10(0) to 10(1) CFU/g. Spray treatments of lettuce with 200 ppm chlorine solution or deionized water were equally effective in killing or removing E. coli O157:H7 from lettuce. Holding lettuce for 5 min after spray treatment was not more effective in reducing populations than holding for 1 min before rinsing with water. Prevention of contamination of lettuce with bovine feces that may harbor E. coli O157:H7 as well as other infectious microorganisms is essential to minimizing the risk of illness. The development of sanitizers more efficacious than chlorine for the removal of pathogens from raw fruits and vegetable is needed.


HPV-16 viral transcripts in vulvar neoplasia: preliminary studies.
Specific human papillomavirus (HPV) types are strongly associated with intraepithelial neoplasia and invasive cancer of the cervix. In contrast, the role of HPVs in the pathogenesis of invasive carcinoma of the vulva is poorly understood. We have employed in situ hybridization for the detection of subgenomic transcripts in four vulvar specimens to elucidate the role of HPV type 16 in the development and progression of vulvar cancer. These analyses revealed that the transcripts of the E6-E7 region were more abundant than those of the L1-L2 region in vulvar neoplastic tissues. The transcripts from early and late region of HPV-16 continued to increase with the differentiation of the epithelial cells in both the warty and the basaloid types of vulvar precancerous lesions. This pattern persisted in invasive warty carcinoma but not in basaloid invasive carcinoma; the transcripts in basaloid carcinoma were distributed in an even and discrete pattern. In contrast to earlier studies, L1-L2-region transcripts, as well as viral capsid protein, were detected in focal areas of well-differentiated cells of invasive warty carcinoma. These findings suggest that expression of HPV-16 is regulated by the degree of cellular differentiation.


IL-13-induced STAT3-dependent signaling networks regulate esophageal epithelial proliferation in eosinophilic esophagitis.
Basal zone hyperplasia (BZH) and dilated intercellular spaces (DISs) are thought to contribute to the clinical manifestations of eosinophilic esophagitis (EoE); however, the molecular pathways that drive BZH remain largely unexplored. We sought to define the role of IL-13-induced transcriptional programs in esophageal epithelial proliferation in EoE. We performed RNA sequencing, bioinformatics, Western blot, reverse transcriptase quantitative PCR, and histologic analyses on esophageal biopsies from healthy control and patients with EoE, primary esophageal cells derived from patients with EoE, and IL-13-stimulated esophageal epithelial keratinocytes grown at the air-liquid interface (EPC2-ALI). Genetic (shRNA) and pharmacologic (proteolysis-targeting chimera degrader) approaches and in vivo model of IL-13-induced esophageal epithelial remodeling (Krt5-rtTA x tetO-IL-13Tg) were used to define the role of signal transducer and activator of transcription 3 (STAT3) and STAT6 and secreted frizzled-related protein 1 (SFRP1) in esophageal epithelial proliferation. RNA-sequencing analysis of esophageal biopsies (healthy control vs EoE) and EPC2-ALI revealed 82 common differentially expressed genes that were enriched for putative STAT3 target genes. In vitro and in vivo analyses revealed a link between IL-13-induced STAT3 and STAT6 phosphorylation, SFRP1 mRNA expression, and esophageal epithelial proliferation. In vitro studies showed that IL-13-induced esophageal epithelial proliferation was STAT3-dependent and regulated by the STAT3 target SFRP1. SFRP1 mRNA is increased in esophageal biopsies from patients with active EoE compared with healthy controls or patients in remission and identifies an esophageal suprabasal epithelial cell subpopulation that uniquely expressed the core EoE proinflammatory transcriptome genes (CCL26, ALOX15, CAPN14, ANO1, and TNFAIP6). These studies identify SFRP1 as a key regulator of IL-13-induced and STAT3-dependent esophageal proliferation and BZH in EoE and link SFRP1+ esophageal epithelial cells with the proinflammatory and epithelial remodeling response in EoE.


Strand-specific RNA sequencing reveals extensive regulated long antisense transcripts that are conserved across yeast species.
Recent studies in budding yeast have shown that antisense transcription occurs at many loci. However, the functional role of antisense transcripts has been demonstrated only in a few cases and it has been suggested that most antisense transcripts may result from promiscuous bi-directional transcription in a dense genome. Here, we use strand-specific RNA sequencing to study anti-sense transcription in Saccharomyces cerevisiae. We detect 1,103 putative antisense transcripts expressed in mid-log phase growth, ranging from 39 short transcripts covering only the 3' UTR of sense genes to 145 long transcripts covering the entire sense open reading frame. Many of these antisense transcripts overlap sense genes that are repressed in mid-log phase and are important in stationary phase, stress response, or meiosis. We validate the differential regulation of 67 antisense transcripts and their sense targets in relevant conditions, including nutrient limitation and environmental stresses. Moreover, we show that several antisense transcripts and, in some cases, their differential expression have been conserved across five species of yeast spanning 150 million years of evolution. Divergence in the regulation of antisense transcripts to two respiratory genes coincides with the evolution of respiro-fermentation. Our work provides support for a global and conserved role for antisense transcription in yeast gene regulation.


Trait and social anxiety in adults with chronic stuttering: conclusions following meta-analysis.
The relationship between chronic stuttering and anxiety has been a matter of some debate over the past two decades, with a major emphasis of research focused on examining whether people who stutter have abnormally elevated levels of trait or social anxiety. The major goal of this paper was to perform a systematic literature review and perform meta-analyses on research that has assessed (i) trait anxiety and (ii) social anxiety, in adults who stutter. Only studies that met strict inclusion criteria were selected for the meta-analyses. Two meta-analyses were conducted, the first for trait anxiety, and the second for social anxiety. Meta-analysis combines statistically the results of selected studies that meet strict design criteria, thereby clarifying the size of differences in trait and social anxiety between adults who stutter and adults who do not stutter. Meta-analytic results confirmed that adults with chronic stuttering do have substantially elevated trait and social anxiety. The overall effect size for trait and social anxiety was calculated to be .57 and .82, respectively. Trait and social anxiety are definite problems for many adults who stutter. Clinical implications of these findings for the diagnosis and treatment of adult who stutter are discussed. The reader will be able to: (a) describe the process of conducting a systematic review and meta-analysis; (b) describe the possible impact of publication bias on meta-analysis results; (c) explain the impact of a chronic disorder like stuttering on levels of trait anxiety; (d) explain the impact of stuttering on levels of social anxiety; (e) interpret the results of meta-analysis when applied to differences in anxiety between adult people who stutter and those who do not stutter; and (f) describe implications for fluency enhancing treatments.


Impact of Venetoclax Exposure on Clinical Efficacy and Safety in Patients with Relapsed or Refractory Chronic Lymphocytic Leukemia.
Venetoclax is a selective, potent, first-in-class B-cell lymphoma-2 inhibitor that restores apoptosis in cancer cells and has demonstrated efficacy in a variety of hematological malignancies. The objective of this research was to characterize the relationship between venetoclax exposures and efficacy and safety in patients with relapsed or refractory (R/R) chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL). A total of 272 and 338 patients from four clinical studies were pooled for the exposure-efficacy and exposure-safety analyses, respectively. Demographics, baseline disease characteristics, and select co-medications were evaluated for their impact on efficacy (lymphocytes, tumor size, objective response [OR]) and safety (neutropenia and infection). Higher venetoclax concentrations led to a more rapid decrease in lymphocyte counts and tumor size, which translated into patients more rapidly achieving OR. The 17p deletion somatic mutation was not identified, in any of the analyses, to affect the responsiveness of patients to venetoclax. Model-based simulations of lymphocyte counts and tumor size estimated an OR rate (ORR) of 84.8 % (95 % confidence interval 81.5-88.0 %) at a venetoclax dosage of 400 mg daily, with minimal increase in ORR at higher doses. The safety analyses of the adverse events (grade 3 or higher) of neutropenia and infection indicated that higher average venetoclax concentrations were not associated with an increase in adverse events. The exposure-response analyses indicated that a venetoclax dosage regimen of 400 mg daily results in a high (>80 %) probability of achieving OR in R/R CLL/SLL patients, with minimal probability of increasing neutropenia or infection with higher exposures.


Effects of a Mindfulness Meditation App on Subjective Well-Being: Active Randomized Controlled Trial and Experience Sampling Study.
Mindfulness training (MT) includes a variety of contemplative practices aimed at promoting intentional awareness of experience, coupled with attitudes of nonjudgment and curiosity. Following the success of 8-week, manualized group interventions, MT has been implemented in a variety of modalities, including smartphone apps that seek to replicate the success of group interventions. However, although smartphone apps are scalable and accessible to a wider swath of population, their benefits remain largely untested. This study aimed to investigate a newly developed MT app called Wildflowers, which was codeveloped with the laboratory for use in mindfulness research. It was hypothesized that 3 weeks of MT through this app would improve subjective well-being, attentional control, and interoceptive integration, albeit with weaker effects than those published in the 8 week, manualized group intervention literature. Undergraduate students completed 3 weeks of MT with Wildflowers (n=45) or 3 weeks of cognitive training with a game called 2048 (n=41). State training effects were assessed through pre- and postsession ratings of current mood, stress level, and heart rate. Trait training effects were assessed through pre- and postintervention questionnaires canvassing subjective well-being and behavioral task measures of attentional control and interoceptive integration. State and trait training data were analyzed in a multilevel model using emergent latent factors (acceptance, awareness, and openness) to summarize the trait questionnaire battery. Analyses revealed both state and trait effects specific to MT; participants engaging in MT demonstrated improved mood (r=.14) and a reduction of stress (r=-.13) immediately after each training session compared with before the training session and decreased postsession stress over 3 weeks (r=-.08). In addition, MT relative to cognitive training resulted in greater improvements in attentional control (r=-.24). Interestingly, both groups demonstrated increased subjective ratings of awareness (r=.28) and acceptance (r=.23) from pre- to postintervention, with greater changes in acceptance for the MT group trending (r=.21). MT, using a smartphone app, may provide immediate effects on mood and stress while also providing long-term benefits for attentional control. Although further investigation is warranted, there is evidence that with continued usage, MT via a smartphone app may provide long-term benefits in changing how one relates to their inner and outer experiences. ClinicalTrials.gov NCT03783793; https://clinicaltrials.gov/ct2/show/NCT03783793 (Archived by WebCite at http://www.webcitation.org/75EF2ehst).


Enzymatic efficiency of erythrocyte antioxidant barrier and lipid peroxidation in children from families with high risk of early atherosclerosis.
The most important risk factors contributing to the development of atherosclerosis include lipid disorders and the predisposition to early ischaemic heart disease in the family. Atherosclerotic process proceeds with age and it develops as a result of oxide LDL modification at the level of vascular wall. Oxygen-free radicals take part in this process, which may probably be opposed by the antioxidant system of the body. The aim of this study was to compare the intensity of lipid peroxidation and the activity of antioxidant enzymes in children from the families with the risk of early atherosclerosis and in children without such predisposition. The activity of katalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were determined and the concentration of malonic dialdehyde--a lipid peroxidation marker was established. The study was conducted on 76 children aged 4-17 years, mean age 12 +/- 0.6 years. The risk group consisted of 56 patients with the history of hypercholesterolaemia and early atherosclerosis in the members of their families up to 45 years of age. Control group was formed of 20 subjects without such history. MDA concentration as well as the activity of antioxidant enzymes were determined with the use of adequate methods of spectrophotometry. The results obtained were subject to statistical analysis. The activity of antioxidant enzymes displayed considerable fluctuations in both groups of children, but these differences remained statistically insignificant in all the cases. Higher MDA concentrations in serum and in erythrocytes were observed in the risk group. These differences proved statistically significant (alpha < 0.05). On the basis of the present study and the analysis performed, it was found that the activity of antioxidant enzymes (CAT, SOD, GSH-Px) cannot serve as a parameter differentiating between children from the families with the risk of early atherosclerosis and children without such predisposition. Children with positive family history of hypercholesterolaemia and early atherosclerosis may demonstrate intensive lipid peroxidation, but this hypothesis requires further investigations.


Liver regeneration and alpha-fetoprotein messenger RNA expression in the retrorsine model for hepatocyte transplantation.
Recently, we described a new model for hepatocyte transplantation with nearly total replacement of the liver by exogenous hepatocytes (E. Laconi et al., Am. J. Pathol., 153: 319-329, 1998). The model is based on the mitoinhibitory effect of the pyrrolizidine alkaloid retrorsine on hepatocytes in the resident liver while transplanted hepatocytes proliferate. In this study, we exploit this novel approach to address the important and controversial issue of whether hepatocytes, when proliferating extensively, undergo dedifferentiation and give rise to foci of undifferentiated hepatocytes. Genetically marked hepatocytes (isolated from normal Dipeptidyl peptidase IV+ Fischer 344 rats) were delivered intraportally (2 x 10(6) cells) into the liver of retrorsine-treated Dipeptidyl peptidase IV- mutant Fischer 344 rats in conjunction with partial hepatectomy. Transplanted hepatocytes were detected histochemically or immunohistochemically, and cell proliferation was studied by in situ hybridization for histone-3 mRNA. Expression of alpha-fetoprotein (AFP) mRNA, a marker of hepatocyte dedifferentiation, was also revealed by in situ hybridization. One day after partial hepatectomy and hepatocyte transplantation, endogenous hepatocytes and oval cells expanding in the liver expressed histone-3 mRNA (cells had entered S phase); 2 days later, transplanted hepatocytes and nonparenchymal cells also expressed histone-3 mRNA. Although the majority of endogenous hepatocytes did not divide and became arrested as quiescent megalocytes, the exogenous hepatocytes, as well as newly formed small hepatocytes, most probably derived from liver progenitor cells, underwent extensive proliferation. After 7-14 days, the nonparenchymal cells stopped proliferating, but transplanted hepatocytes and small endogenous hepatocytes continued to proliferate for 1 month, forming foci of dividing parenchymal cells. Although many of the hepatocytes in clusters were in S phase (histone-3 mRNA positive), none expressed AFP mRNA. In contrast, high expression of AFP mRNA was observed in proliferating oval and transitional cells, forming duct-like structures of cytokeratin-19-positive cells. From these studies, we conclude that hepatocyte proliferation in the adult liver is not associated with dedifferentiation.


Studies of surface of metal nanoparticles in a flowing liquid with XPS.
Studying surface of catalyst nanoparticles in a flowing liquid is important for understanding the underlying mechanism of a reaction performed in liquid. We report the design of a reaction cell system of Si3N4 window covering the flowing liquid with an electron-transmissible membrane. By using metal nanoparticles as a catalyst dispersed in a solvent, examination of the surface of catalyst nanoparticles in a flowing liquid was demonstrated by observation of Ag 3d photoemission feature when a liquid containing Ag nanoparticles was flowing through this system.


Use of the scapular spine in lumbar fusion procedures: cadaveric feasibility study. Laboratory investigation.
Graft sources for lumbar fusion include synthetic materials, donor grafts, and autologous bone such as the iliac crest. Considering the data indicating that autologous bone grafts generate the best results for fusion, the next logical step is to seek alternative donor sites in an attempt to reduce the complications associated with these procedures. To the authors' knowledge, autologous scapula has not been explored as a potential source for posterior lumbar fusion graft material. Therefore, the following study was performed to verify the utility of this bone in these procedures. Six adult cadavers (mean age 71 years), four formalin-fixed and two fresh specimens, were used in this study. With the cadaver in the prone position, an incision was made over the spine of the scapula. Soft tissues were stripped from the middle of this region of the scapula, and bone segments were removed with a bone saw and used for a posterior lumbar fusion procedure. A mean length of 11.5 cm was measured for the spine of the scapula and the mean thicknesses of this bone at its medial part, segment just medial to the spinoglenoid notch, and acromion were 1 cm, 2.2 cm, and 2.5 cm, respectively. No obvious injury to surrounding vessels or nerves was found using this procedure, and adequate fusion was achieved with it. Following clinical testing, such a bone substitute as autologous scapular spine might be a reasonable alternative to iliac crest grafts for use in posterior lumbar fusion procedures.


Assessment of quality of life in outpatients with advanced cancer: the accuracy of clinician estimations and the relevance of spiritual well-being--a Hoosier Oncology Group Study.
To evaluate the association between quality-of-life (QOL) impairment as reported by patients and QOL impairment as judged by nurses or physicians, with and without consideration of spiritual well-being (SWB). A total of 163 patients with advanced cancer were enrolled onto a therapeutic trial, and cross-sectional data were derived from clinical and demographic questionnaires obtained at baseline, including assessment of patient QOL and SWB. Clinicians rated the QOL impairment of their patients as mild, moderate, or severe. Clinician-estimated QOL impairment and patient-derived QOL categories were compared. Correlation coefficients were estimated to associate QOL scores using different instruments. The analysis of variance method was used to compare Functional Assessment of Cancer Therapy-General scores on categorical variables. There was no significant association between self-assessment scores and marital status, education level, performance status, or predicted life expectancy. However, a strong relationship between SWB and QOL was noted (P <.0001). Clinician-estimated QOL impairment matched the level of patient-derived QOL correctly in approximately 60% of cases, with only slight variation depending on the method of categorizing patient-derived QOL scores. The accuracy of clinician estimates was not associated with the level of SWB. Interestingly, a subset analysis of the inaccurate estimates revealed an association between lower SWB and clinician underestimation of QOL impairment (P =.0025). Clinician estimates of QOL impairment were accurate in more than 60% of patients. SWB is strongly associated with QOL, but it is not associated with the overall accuracy of clinicians' judgments about QOL impairment.


Abnormal motor function and dopamine neurotransmission in DYT1 DeltaGAG transgenic mice.
A single GAG deletion in Exon 5 of the TOR1A gene is associated with a form of early-onset primary dystonia showing less than 40% penetrance. To provide a framework for cellular and systems study of DYT1 dystonia, we characterized the genetic, behavioral, morphological and neurochemical features of transgenic mice expressing either human wild-type torsinA (hWT) or mutant torsinA (hMT1 and hMT2) and their wild-type (WT) littermates. Relative to human brain, hMT1 mice showed robust neural expression of human torsinA transcript (3.90x). In comparison with WT littermates, hMT1 mice had prolonged traversal times on both square and round raised-beam tasks and more slips on the round raised-beam task. Although there were no effects of genotype on rotarod performance and rope climbing, hMT1 mice exhibited increased hind-base widths in comparison to WT and hWT mice. In contrast to several other mouse models of DYT1 dystonia, we were unable to identify either torsinA- and ubiquitin-positive cytoplasmic inclusion bodies or nuclear bleb formation in hMT1 mice. High-performance liquid chromatography with electrochemical detection was used to determine cerebral cortical, striatal, and cerebellar levels of dopamine (DA), norepinephrine, epinephrine, serotonin, 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindoleacetic acid. Although there were no differences in striatal DA levels between WT and hMT1 mice, DOPAC and HVA concentrations and DA turnover (DOPAC/DA and HVA/DA) were significantly higher in the mutants. Our findings in DYT1 transgenic mice are compatible with previous neuroimaging and postmortem neurochemical studies of human DYT1 dystonia. Increased striatal dopamine turnover in hMT1 mice suggests that the nigrostriatal pathway may be a site of functional neuropathology in DYT1 dystonia.


Transdetermination of plant cells.
Tissues derived from the leaf lamina of Nicotiana tabacum L. cv. Havana 425 plants require cytokinin for continuous proliferation in culture, whereas tissues derived from the cortex of the stem are cytokinin autotrophic. Both phenotypes persist when the two types of cells are cloned, indicating that leaf- and cortex-derived cells are determined to express different cytokinin requirements in culture. We showed that cultures derived from leaf and cortex tissues of plants regenerated from cloned leaf and cortex cells exhibit the cytokinin requirement of comparable tissues of seed-grown plants. This provides direct evidence that plant cells can undergo transdetermination and that this process has an epigenetic basis.


Patterns of care for patients with cancer before and after the establishment of a hospice ward.
A majority of people die in acute hospital care, in a culture of quickness primary aimed at curing. There are indications that hospice care in various forms will spread during the present decade in Sweden. The hospice philosophy contributes to a culture of slowness, where the patient is not only allowed but is encouraged to live at his own pace. As a first step in establishing for whom hospice inpatient care is the best alternative, patient flow to a newly started hospice ward was studied. All patients (n = 1464) who died at the Medical Centre Hospital with a primary diagnosis of cancer during the period of study were compared regarding place of death, diagnosis, gender, age and length of the last period of care. The year before the ward was established, 82% of patients died in acute hospital care compared with 59% during the hospice ward's third year of operation. A total of 315 patients (22%) died in the hospice ward during the study period. The percentage of these patients over the age of 80 was significantly higher (p = 0.0001), and they also had a longer continuous final period of care. More women (57%) than men (43%) were cared for at the hospice ward. The hospice ward has in this way influenced the pattern of care in the hospital.


Colony stimulating factors for chemotherapy induced febrile neutropenia.
Febrile neutropenia is a frequent event for cancer patients undergoing chemotherapy and it is potentially a life threatening situation. The current treatment is supportive care plus antibiotics. Colony stimulating factors (CSF) are cytokines that stimulate and accelerate the production of one or more cellular lines in bone marrow. Some clinical trials addressed the question of whether the addition of CSF to antibiotics (ATB) could improve the outcomes of patients with febrile neutropenia. The results of these trials are conflicting and no definitive conclusion could be reached. To evaluate the safety and effectiveness of adding colony stimulating factors to ATB when treating febrile neutropenia caused by cancer chemotherapy. The search covered the major electronic databases: CANCERLIT, EMBASE, LILACS, MEDLINE, SCI and The Cochrane Controlled Trials Register. Experts were consulted and references from the relevant articles scanned. We looked for all randomized controlled trials (RCTs) that compare CSF plus antibiotics versus antibiotics alone for the treatment of established febrile neutropenia in adults and children. Two of the reviewers independently selected, critically appraised and extracted data from the studies. A meta-analysis of the select studies was performed, using Review Manager. More than 8000 references were screened. Thirteen studies were included. The overall mortality was not influenced by the use of CSF [Odds Ratio (OR) = 0.68; 95% Confidence Interval (CI) = 0.43 to 1.08; p=0.1]. A marginally significant result was obtained for the use of CSF in reducing infection related mortality [OR= 0.51; 95% CI = 0.26 to 1.00; p=0.05], but this result was highly influenced by one study. When this study is excluded from our analysis, this possible benefit disappears [OR= 0.85; 95% CI = 0.33 to 2.20; p= 0.7]. The group of patients treated with CSF had a shorter length of hospitalization [Hazard Ratio (HR) = 0.63; 95% CI = 0.49 to 0.82; p=0.0006] and a shorter time to neutrophil recovery [HR= 0.32; 95% CI = 0.23 to 0.46; p < 0.00001]. The use of CSF in patients with febrile neutropenia due to cancer chemotherapy does not affect overall mortality, but reduces the amount of time spent in hospital and the neutrophil recovery period. It was not clear whether CSF has an effect on infection-related mortality.


Dietary taste patterns and diet quality of female nurses around the night shift.
Night shift workers are at risk of making poor food choices: e.g. sleep deprivation may lead to higher food intake with innate preferred tastes, such as sweet, savoury and fatty foods. Therefore, better insight in dietary taste patterns of night shift workers may improve the understanding of their food choices. This observational study assessed dietary taste patterns of 120 female night shift working nurses and compared them to 307 women of a reference population. Dietary intake, assessed with 24-h dietary recalls, was combined with a taste intensity database, including taste profiles of 557 foods. The contribution to the daily intake of 6 taste clusters was assessed: fat, neutral, sweet/fat, sweet/sour, salt/umami/fat and bitter. During night shifts, nurses consumed a significantly higher energy percentage (en%) of 'neutral' (5.9 en%), 'sweet/sour' (8.1 en%) and 'sweet/fat' (6.5 en%) tasting foods and a lower en% of 'fat' (- 17.1 en%) and 'bitter' (- 2.1 en%) tasting foods than outside the night shift. They consumed a larger en% from foods with a 'sweet/sour' (1.9 en%) taste and a lower en% from foods with a 'bitter' (- 2.1 en%) taste than the reference population, irrespective of age, BMI and smoking status. A higher en% and gram% of 'fat' tasting foods and a higher gram% 'fat/salt/umami' tasting foods were associated with lower diet quality. Our results only partly support our hypothesis that nurses would select foods with more innate taste preferences. In addition, fat and savoury tasting foods were negatively associated with their diet quality.


Transcriptome-Wide Studies of RNA-Targeted Small Molecules Provide a Simple and Selective r(CUG)exp Degrader in Myotonic Dystrophy.
Myotonic dystrophy type 1 (DM1) is caused by a highly structured RNA repeat expansion, r(CUG)exp, harbored in the 3' untranslated region (3' UTR) of dystrophia myotonica protein kinase (DMPK) mRNA and drives disease through a gain-of-function mechanism. A panel of low-molecular-weight fragments capable of reacting with RNA upon UV irradiation was studied for cross-linking to r(CUG)expin vitro, affording perimidin-2-amine diazirine (1) that bound to r(CUG)exp. The interactions between the small molecule and RNA were further studied by nuclear magnetic resonance (NMR) spectroscopy and molecular modeling. Binding of 1 in DM1 myotubes was profiled transcriptome-wide, identifying 12 transcripts including DMPK that were bound by 1. Augmenting the functionality of 1 with cleaving capability created a chimeric degrader that specifically targets r(CUG)exp for elimination. The degrader broadly improved DM1-associated defects as assessed by RNA-seq, while having limited effects on healthy myotubes. This study (i) provides a platform to investigate molecular recognition of ligands directly in disease-affected cells; (ii) illustrates that RNA degraders can be more specific than the binders from which they are derived; and (iii) suggests that repeating transcripts can be selectively degraded due to the presence of multiple ligand binding sites.


Simultaneous quantification of five biflavonoids in rat plasma by LC-ESI-MS/MS and its application to a comparatively pharmacokinetic study of Selaginella doederleinii Hieron extract in rats.
Selaginella doederleinii Hieron is a widely used as folk Chinese medicine for treatment of different cancers. Our previous investigations have confirmed that the total biflavonoids in ethyl acetate extract from S. doederleinii (SDEA) have favorable anticancer potentials. However, the in vivo process of its bioactive ingredients remains unknown. In this paper, a sensitive and reliable method was developed for simultaneous quantification of main five biflavonoids, including amentoflavone, robustaflavone, 2″,3″-dihydro-3',3″-biapigenin, 3',3″-binaringenin and delicaflavone in the ethyl acetate extract of S. doederleinii (SDEA extract) in rat plasma by high-performance liquid chromatography with electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS). Chromatographic separation was performed using an Ultimate® XB-C18 (100×2.1mm, 3.5μm) with gradient elution of water (0.5% acetic acid) and acetonitrile at 0.2mL/min. All analytes with internal standard (chrysin) were detected using selective reaction monitoring (SRM) in negative ionization mode. The method showed a good linearity over a wide concentration range (r2>0.99). The limits of quantification for the biflavonoids were less than 10ng/mL. The developed method was applied to the comparatively pharmacokinetic study of the five biflavonoids after oral or intravenous administration of SDEA extract in rats. In addition, in silico assessments of permeability and solubility of these biflavonoids were also performed to understand their poor bioavailability. It is the first time to report the in vivo process profiles of the biflavonoids of SDEA extract in rats.


Providers perspectives on self-regulation impact their use of responsive feeding practices in child care.
Supporting children's self-regulation in eating through caregivers' practice of responsive feeding is paramount to obesity prevention, and while much attention has been given to supporting children's self-regulation in eating through parents' responsive feeding practices in the home setting, little attention has been given to this issue in childcare settings. This qualitative study examines childcare providers' perspectives on using responsive feeding practices with young children (2-5years). Individual semi-structured interviews were conducted with providers until saturation was reached. Data was analyzed using thematic analysis. The final sample included 18 providers who were employed full-time in Head Start or state-licensed center-based childcare programs, cared for children (2-5y), and were directly responsible for serving meals and snacks. Providers were primarily (67%) employed in childcare programs that served children from low-income families and received reimbursement for meals and snacks from the US Department of Agriculture's Child and Adult Care Food Program. Three factors emerged that shaped childcare providers' experiences using responsive feeding practices: the providers' perspectives about whether or not young children can self-regulate food intake, their understanding of Child and Adult Care Food Program (CACFP) portion size regulations, and the availability of food at the center where they worked. Future research should examine how childcare providers' understanding of children's ability to self-regulate their food intake, the appropriate use of the CACFP regulations in relationship to serving sizes, and having food available to offer seconds promotes providers' use of responsive feeding practices in center-based childcare programs and children's dietary behaviors.


Macroglial plasticity and the origins of reactive astroglia in experimental autoimmune encephalomyelitis.
Accumulations of hypertrophic, intensely glial fibrillary acidic protein-positive (GFAP(+)) astroglia, which also express immunoreactive nestin and vimentin, are prominent features of multiple sclerosis lesions. The issues of the cellular origin of hypertrophic GFAP(+)/vimentin(+)/nestin(+) "reactive" astroglia and also the plasticities and lineage relationships among three macroglial progenitor populations-oligodendrocyte progenitor cells (OPCs), astrocytes and ependymal cells-during multiple sclerosis and other CNS diseases remain controversial. We used genetic fate-mappings with a battery of inducible Cre drivers (Olig2-Cre-ER(T2), GFAP-Cre-ER(T2), FoxJ1-Cre-ER(T2) and Nestin-Cre-ER(T2)) to explore these issues in adult mice with myelin oligodendrocyte glycoprotein peptide-induced experimental autoimmune encephalomyelitis (EAE). The proliferative rate of spinal cord OPCs rose fivefold above control levels during EAE, and numbers of oligodendroglia increased as well, but astrogenesis from OPCs was rare. Spinal cord ependymal cells, previously reported to be multipotent, did not augment their low proliferative rate, nor give rise to astroglia or OPCs. Instead, the hypertrophic, vimentin(+)/nestin(+), reactive astroglia that accumulated in spinal cord in this multiple sclerosis model were derived by proliferation and phenotypic transformation of fibrous astroglia in white matter, and solely by phenotypic transformation of protoplasmic astroglia in gray matter. This comprehensive analysis of macroglial plasticity in EAE helps to clarify the origins of astrogliosis in CNS inflammatory demyelinative disorders.


Erythrocyte lysis and Xenopus laevis oocyte rupture by recombinant Plasmodium falciparum hemolysin III.
Malaria kills more than 1 million people per year worldwide, with severe malaria anemia accounting for the majority of the deaths. Malaria anemia is multifactorial in etiology, including infected erythrocyte destruction and decrease in erythrocyte production, as well as destruction or clearance of noninfected erythrocytes. We identified a panspecies Plasmodium hemolysin type III related to bacterial hemolysins. The identification of a hemolysin III homologue in Plasmodium suggests a potential role in host erythrocyte lysis. Here, we report the first characterization of Plasmodium falciparum hemolysin III, showing that the soluble recombinant P. falciparum hemolysin III is a pore-forming protein capable of lysing human erythrocytes in a dose-, time-, and temperature-dependent fashion. The recombinant P. falciparum hemolysin III-induced hemolysis was partially inhibited by glibenclamide, a known channel antagonist. Studies with polyethylene glycol molecules of different molecular weights indicated a pore size of approximately 3.2 nm. Heterologous expression of recombinant P. falciparum hemolysin III in Xenopus oocytes demonstrated early hypotonic lysis similar to that of the pore-forming aquaporin control. Live fluorescence microscopy localized transfected recombinant green fluorescent protein (GFP)-tagged P. falciparum hemolysin III to the essential digestive vacuole of the P. falciparum parasite. These transfected trophozoites also possessed a swollen digestive vacuole phenotype. Native Plasmodium hemolysin III in the digestive vacuole may contribute to lysis of the parasitophorous vacuole membrane derived from the host erythrocyte. After merozoite egress from infected erythrocytes, remnant P. falciparum hemolysin III released from digestive vacuoles could potentially contribute to lysis of uninfected erythrocytes to contribute to severe life-threatening anemia.


Glucocorticoids inhibit glucose transport and glutamate uptake in hippocampal astrocytes: implications for glucocorticoid neurotoxicity.
Glucocorticoids (GCs), the adrenal steroid hormones secreted during stress, can damage the hippocampus and impair its capacity to survive coincident neurological insults. This GC endangerment of the hippocampus is energetic in nature, as it can be prevented when neurons are supplemented with additional energy substrates. This energetic endangerment might arise from the ability of GCs to inhibit glucose transport into both hippocampal neurons and astrocytes. The present study explores the GC inhibition in astrocytes. (1) GCs inhibited glucose transport approximately 15-30% in both primary and secondary hippocampal astrocyte cultures. (2) The parameters of inhibition agreed with the mechanisms of GC inhibition of glucose transport in peripheral tissues: A minimum of 4 h of GC exposure were required, and the effect was steroid specific (i.e., it was not triggered by estrogen, progesterone, or testosterone) and tissue specific (i.e., it was not triggered by GCs in cerebellar or cortical cultures). (3) Similar GC treatment caused a decrease in astrocyte survival during hypoglycemia and a decrease in the affinity of glutamate uptake. This latter observation suggests that GCs might impair the ability of astrocytes to aid neurons during times of neurologic crisis (i.e., by impairing their ability to remove damaging glutamate from the synapse).


Multi-Species Phylogeography of Arid-Zone Sminthopsinae (Marsupialia: Dasyuridae) Reveals Evidence of Refugia and Population Expansion in Response to Quaternary Change.
Historical population contraction and expansion events associated with Pleistocene climate change are important drivers of intraspecific population structure in Australian arid-zone species. We compared phylogeographic patterns among arid-adapted Dasyuridae (Sminthopsis and Planigale) with close phylogenetic relationships and similar ecological roles to investigate the drivers of phylogeographic structuring and the importance of historical refugia. We generated haplotype networks for two mitochondrial (control region and cytochrome b) and one nuclear (omega-globin) gene from samples distributed across each species range. We used ΦST to test for a genetic population structure associated with the four Pilbara subregions, and we used expansion statistics and Bayesian coalescent skyline analysis to test for signals of historical population expansion and the timing of such events. Significant population structure associated with the Pilbara and subregions was detected in the mitochondrial data for most species, but not with the nuclear data. Evidence of population expansion was detected for all species, and it likely began during the mid-late Pleistocene. The timing of population expansion suggests that these species responded favorably to the increased availability of arid habitats during the mid-late Pleistocene, which is when previously patchy habitats became more widespread. We interpret our results to indicate that the Pilbara region could have acted as a refugium for small dasyurids.


[Antimicrobial activity of piperacillin in vitro].
There was an increase in the resistance of clinical isolates to piperacillin as compared to that in 1987-1988. It was shown on the kinetic models that there was difference in the effect of the inhibitory and subinhibitory concentrations of the drug on microorganisms in various growth phases. No post-antibiotic action against Staphylococcus aureus and Pseudomonas aeruginosa and a slightly pronounced dose-dependent effect with respect to Escherichia coli were noted.


Serious fatal and nonfatal non-AIDS-defining illnesses in Europe.
Little is known about the incidence and risk factors for serious non-AIDS-defining events. The incidence of non-AIDS events (malignancies, end-stage renal disease, liver failure, pancreatitis, cardiovascular disease), and AIDS after January 1, 2001, was calculated; Poisson regression was used to investigate factors associated with non-AIDS and AIDS. Among 12,844 patients, 1058 were diagnosed with a non-AIDS event [incidence 1.77 per 100 person-years of follow-up; 95% confidence interval (CI): 1.66 to 1.87]; 462 patients (43.7%) died. The incidence of AIDS (1025 diagnoses; 339 deaths, 33.1%) was 1.72 per 100 person-years of follow-up (1.61 to 1.83). After adjustment, older age [incidence rate ratio (IRR): 1.71 per 10 years older, 95% CI: 1.60 to 1.83], diabetes (IRR: 1.49, 95% CI: 1.22 to 1.82) and hypertension (IRR: 1.63, 95% CI: 1.43 to 1.87) were associated with non-AIDS events. Compared with patients without an event, there was a 4-fold increased risk of death after an AIDS event (relative hazard: 4.14; 95% CI 3.47 to 4.94) and almost a 7-fold increased risk of death after a non-AIDS event (relative hazard: 6.72; 95% CI: 5.61 to 8.05). Non-AIDS events were common in the combination antiretroviral therapy era and associated with considerably mortality. Evidence on the impact of modifying immunodeficiency and lifestyle-related factors on the risk of non-AIDS events in HIV-infected persons is an important but unmet research need.


Glucose repression of STA1 expression is mediated by the Nrg1 and Sfl1 repressors and the Srb8-11 complex.
In the yeast Saccharomyces diastaticus, expression of the STA1 gene, which encodes an extracellular glucoamylase, is negatively regulated by glucose. Here we demonstrate that glucose-dependent repression of STA1 expression is imposed by both Sfl1 and Nrg1, which serve as direct transcriptional repressors. We show that Nrg1 acts only on UAS1, and Sfl1 acts only on UAS2, in the STA1 promoter. When bound to its specific site, Sfl1 (but not Nrg1) prevents the binding to UAS2 of two transcriptional activators, Ste12 and Tec1, required for STA1 expression. We also found that Sfl1 contributes to STA1 repression by binding to the promoter and inhibiting the expression of FLO8, a gene that encodes a third transcriptional activator involved in STA1 expression. In addition, we show that the levels of Nrg1 and Sfl1 increase in glucose-grown cells, suggesting that the effects of glucose are mediated, at least in part, through an increase in the abundance of these repressors. NRG1 and SFL1 expression requires the Srb8-11 complex, and correspondingly, the Srb8-11 complex is also necessary for STA1 repression. However, our evidence indicates that the Srb8-11 complex does not associate with either the SFL1 or the NRG1 promoter and thus plays an indirect role in activating NRG1 and SFL1 expression.


The Neural Bases of Egocentric Spatial Representation for Extracorporeal and Corporeal Tasks: An fMRI Study.
(1) Background: Humans use reference frames to elaborate the spatial representations needed for all space-oriented behaviors such as postural control, walking, or grasping. We investigated the neural bases of two egocentric tasks: the extracorporeal subjective straight-ahead task (SSA) and the corporeal subjective longitudinal body plane task (SLB) in healthy participants using functional magnetic resonance imaging (fMRI). This work was an ancillary part of a study involving stroke patients. (2) Methods: Seventeen healthy participants underwent a 3T fMRI examination. During the SSA, participants had to divide the extracorporeal space into two equal parts. During the SLB, they had to divide their body along the midsagittal plane. (3) Results: Both tasks elicited a parieto-occipital network encompassing the superior and inferior parietal lobules and lateral occipital cortex, with a right hemispheric dominance. Additionally, the SLB > SSA contrast revealed activations of the left angular and premotor cortices. These areas, involved in attention and motor imagery suggest a greater complexity of corporeal processes engaging body representation. (4) Conclusions: This was the first fMRI study to explore the SLB-related activity and its complementarity with the SSA. Our results pave the way for the exploration of spatial cognitive impairment in patients.