Pregnancy outcome in women undergoing liver biopsy during pregnancy: A nationwide population-based cohort study.
Liver biopsy is an important procedure in the investigation of liver disease. We examined pregnancy outcomes in women who underwent liver biopsy during pregnancy. In a nationwide population-based cohort study we linked data from the Swedish Medical Birth Registry (for births between 1992 and 2011) with those from the Swedish Patient Registry. We identified 23 pregnancies exposed to liver biopsy. We calculated relative risks (RRs) for adverse pregnancy outcomes according to liver biopsy status using 1,953,887 unexposed pregnancies with and without a record of liver disease as reference. Our main outcome measures were stillbirth and preterm birth. There were no stillbirths in pregnancies exposed to liver biopsies compared with 0.3% stillbirths in unexposed pregnancies, and 3/23 (13%) exposed pregnancies were preterm (RR, 2.6; 95% confidence interval, 0.9-7.5). Compared to women with a record of liver disease, preterm birth was not increased in those exposed to liver biopsy (RR, 0.9; 95% confidence interval, 0.1-6.0). Except for an increased risk of small for gestational age birth in pregnancies exposed to liver biopsy (RR, 5.2; 95% confidence interval, 1.8-14.8), other adverse pregnancy outcomes were independent of liver biopsy status when the analysis was restricted to women with a diagnosis of liver disease. Compared with unexposed sibling pregnancies, pregnancies with a liver biopsy were 7 days shorter, but birth weights did not differ between the siblings (-67 g; P > 0.05). Apart from a moderately increased risk of preterm birth and small for gestational age, there was no association between liver biopsy during pregnancy and adverse pregnancy outcome; potential excess risks should be weighed against the advantages of having a liver biopsy that may influence clinical management of the patient and indirectly fetal health. (Hepatology 2017).


CircRNA_33702 Promotes Renal Fibrosis by Targeting the miR-29b-3p/WNT1-Inducible Signaling Pathway Protein 1 Pathway.
Growing evidence suggest that circular RNAs (circRNAs) are critical mediators in renal diseases. However, there have been very few reports about the role of circRNAs in renal fibrosis. In this study, circRNA_33702 was found to be upregulated, both in unilateral ureteral obstruction (UUO) mice and in TGF-β1-treated Boston University mouse proximal tubule cells. Furthermore, hsa_circ_0026331, homologous with mmu_circ_33702, was found to be upregulated in TGF-β1-treated HK-2 cells. Although knockdown of circRNA_33702 or hsa_circ_0026331 was shown to relieve the TGF-β1-induced expression of collagen I, collagen III, and fibronectin, overexpression of circRNA_33702 was found to exert an inhibitory effect on the expression of the same genes. Mechanistically, circRNA_33702 was demonstrated to bind directly with miR-29b-3p and inhibit its expression. MiR-29b-3p mimic was shown to inhibit the TGF-β1-induced expression of collagen I, collagen III, and fibronectin. Moreover, WNT1-inducible signaling pathway protein 1 (WISP1) was identified as a target of miR-29b-3p, and the expression of WISP1 was observed to be repressed by miR-29b-3p. Notably, knockdown of circRNA_33702 was found to attenuate the expression of collagen I, collagen III, and fibronectin by inhibiting the expression of WISP1, and the observed inhibitory effect can be reversed by miR-29b-3p inhibitor. Finally, inhibition of circRNA_33702 was shown to attenuate interstitial fibrosis in UUO mice via the miR-29b-3p/WISP1 axis. In general, our data show that circRNA_33702 may promote renal fibrosis via the miR-29b-3p/WISP1 axis, which may potentially be developed as a new therapeutic target. SIGNIFICANCE STATEMENT: This study's findings suggested that circRNA_33702 plays a profibrosis role and that circRNA_33702 with the homologous human hsa_circ_0026331 may be a novel therapeutic target of renal fibrosis.


Robust photoregulation of GABA(A) receptors by allosteric modulation with a propofol analogue.
Photochemical switches represent a powerful method for improving pharmacological therapies and controlling cellular physiology. Here we report the photoregulation of GABA(A) receptors (GABA(A)Rs) by a derivative of propofol (2,6-diisopropylphenol), a GABA(A)R allosteric modulator, which we have modified to contain photoisomerizable azobenzene. Using α(1)β(2)γ(2) GABA(A)Rs expressed in Xenopus laevis oocytes and native GABA(A)Rs of isolated retinal ganglion cells, we show that the trans-azobenzene isomer of the new compound (trans-MPC088), generated by visible light (wavelengths ~440 nm), potentiates the γ-aminobutyric acid-elicited response and, at higher concentrations, directly activates the receptors. cis-MPC088, generated from trans-MPC088 by ultraviolet light (~365 nm), produces little, if any, receptor potentiation/activation. In cerebellar slices, MPC088 co-applied with γ-aminobutyric acid affords bidirectional photomodulation of Purkinje cell membrane current and spike-firing rate. The findings demonstrate photocontrol of GABA(A)Rs by an allosteric ligand, and open new avenues for fundamental and clinically oriented research on GABA(A)Rs, a major class of neurotransmitter receptors in the central nervous system.


Genetics and phenotypes of RPE65 mutations in inherited retinal degeneration.
To characterize the spectrum of RPE65 mutations present in 453 patients with retinal dystrophy with an interest in understanding the range of functional deficits attributable to sequence variants in this gene. The 14 exons of RPE65 were amplified by polymerase chain reaction (PCR) from patients' DNA and analyzed for sequence changes by single-strand conformation polymorphism (SSCP) and direct sequencing. Haplotype analysis was performed using RPE65 intragenic polymorphisms. Patients were examined clinically and with visual function tests. Twenty-one different disease-associated DNA sequence changes predicting missense or nonsense point mutations, insertions, deletions, and splice site defects in RPE65 were identified in 20 patients in homozygous or compound heterozygous form. In one patient, paternal uniparental isodisomy (UPD) of chromosome 1 resulted in homozygosity for a probable functional null allele. Eight of the disease-associated mutations (Y79H, E95Q, E102X, D167Y, 669delCA, IVS7+4a-->g, G436V, and G528V) and one mutation likely to be associated with disease (IVS6+5g-->a) have not been reported previously. The most commonly occurring sequence variant identified in the patients studied was the IVS1+5g-->a mutation, accounting for 9 of 40 (22.5%) total disease alleles. This splice site mutation, as well as R91W, the most common missense mutation, exists on at least two different genetic backgrounds. The phenotype resulting from RPE65 mutations appears to be relatively uniform and independent of mutation class, suggesting that most missense mutations (15 of 40 disease alleles [37.5%]) result in loss of function. At young ages, this group of patients has somewhat better subjective visual capacity than is typically associated with Leber congenital amaurosis (LCA) type I, with a number of patients retaining some useful visual function beyond the second decade of life. RPE65 mutations account for a significant percentage (11.4%) of disease alleles in patients with early-onset retinal degeneration. The identification and characterization of patients with RPE65 mutations is likely to represent an important resource for future trials of rational therapies for retinal degeneration.


Activating transcription factor 1 directs Mhem atheroprotective macrophages through coordinated iron handling and foam cell protection.
Intraplaque hemorrhage (IPH) drives atherosclerosis through the dual metabolic stresses of cholesterol-enriched erythrocyte membranes and pro-oxidant heme/iron. When clearing tissue hemorrhage, macrophages are typically seen storing either iron or lipid. We have recently defined hemorrhage-associated macrophages (HA-mac) as a plaque macrophage population that responds adaptively to IPH. This study aimed to define the key transcription factor(s) involved in HO-1 induction by heme. To address this question, we used microarray analysis and transfection with siRNA and plasmids. To maintain physiological relevance, we focused on human blood-derived monocytes. We found that heme stimulates monocytes through induction of activating transcription factor 1 (ATF-1). ATF-1 coinduces heme oxygenase-1 (HO-1) and Liver X receptor beta (LXR-β). Heme-induced HO-1 and LXR-β were suppressed by knockdown of ATF-1, and HO-1 and LXR-β were induced by ATF-1 transfection. ATF-1 required phosphorylation for full functional activity. Expression of LXR-β in turn led to induction of other genes central to cholesterol efflux, such as LXR-α and ABCA1. This heme-directed state was distinct from known macrophage states (M1, M2, Mox) and, following the same format, we have designated them Mhem. These results show that ATF-1 mediates HO-1 induction by heme and drives macrophage adaptation to intraplaque hemorrhage. Our definition of an ATF-1-mediated pathway for linked protection from foam cell formation and oxidant stress may have therapeutic potential.


Allosteric Inhibition of the SARS-CoV-2 Main Protease: Insights from Mass Spectrometry Based Assays*.
The SARS-CoV-2 main protease (Mpro ) cleaves along the two viral polypeptides to release non-structural proteins required for viral replication. MPro is an attractive target for antiviral therapies to combat the coronavirus-2019 disease. Here, we used native mass spectrometry to characterize the functional unit of Mpro . Analysis of the monomer/dimer equilibria reveals a dissociation constant of Kd =0.14±0.03 μM, indicating MPro has a strong preference to dimerize in solution. We characterized substrate turnover rates by following temporal changes in the enzyme-substrate complexes, and screened small molecules, that bind distant from the active site, for their ability to modulate activity. These compounds, including one proposed to disrupt the dimer, slow the rate of substrate processing by ≈35 %. This information, together with analysis of the x-ray crystal structures, provides a starting point for the development of more potent molecules that allosterically regulate MPro activity.


Renewable energy powered membrane technology. 1. Development and characterization of a photovoltaic hybrid membrane system.
In isolated communities where potable water sources as well as energy grids are limited or nonexistent, treating brackish groundwater aquifers with small-scale desalination systems can be a viable alternative to existing water infrastructures. Given the unavailability of power in many such situations, renewable energy is an obvious solution to power such systems. However, renewable energy is an intermittent power supply and with regards to the performance of intermittently operated desalination systems, only very limited experience exists, both with regards to efficiency as well as water quality. In this paper, this lack of knowledge is addressed by evaluating a system operated with varying parameters (pressure and flow) with constant power as a step toward defining a safe operating window, and they provide a basis for interpreting future data obtained with a renewable energy source. Field trials were performed on a brackish (5300 mg/L TDS; 8290 microS/cm) bore in Central Australia with a photovoltaic-powered membrane filtration (PV-membrane) system. Four nanofiltration and reverse osmosis membranes (BW30, ESPA4, NF90, TFC-S) and a number of operation parameter combinations (transmembrane pressure, feed flow, TFC-S) and operating parameters transmembrane pressure and feed flow were investigated to find the best operating conditions for maximum drinking water production and minimum specific energy consumption (SEC). The ESPA4 membrane performed best for this brackish source, producing 250 L/h of excellent drinking water (257 mg/L TDS; 400 microS/ cm) at an SEC of 1.2 kWh/m3. The issue of brine disposal or reuse is also discussed and the article compares the salinity of the produced brine with livestock water. Since the feedwater is disinfected physically using ultrafiltration (UF), the brine is free from bacteria and most viruses and hence can be seen more as a reusable product stream than a waste stream with a disposal problem.


Bistability analyses of CD4+ T follicular helper and regulatory cells during Helicobacter pylori infection.
T follicular helper (Tfh) cells are a highly plastic subset of CD4+ T cells specialized in providing B cell help and promoting inflammatory and effector responses during infectious and immune-mediate diseases. Helicobacter pylori is the dominant member of the gastric microbiota and exerts both beneficial and harmful effects on the host. Chronic inflammation in the context of H. pylori has been linked to an upregulation in T helper (Th)1 and Th17 CD4+ T cell phenotypes, controlled in part by the cytokine, interleukin-21. This study investigates the differentiation and regulation of Tfh cells, major producers of IL-21, in the immune response to H. pylori challenge. To better understand the conditions influencing the promotion and inhibition of a chronically elevated Tfh population, we used top-down and bottom-up approaches to develop computational models of Tfh and T follicular regulatory (Tfr) cell differentiation. Stability analysis was used to characterize the presence of two bi-stable steady states in the calibrated Tfh/Tfr models. Stochastic simulation was used to illustrate the ability of the parameter set to dictate two distinct behavioral patterns. Furthermore, sensitivity analysis helped identify the importance of various parameters on the establishment of Tfh and Tfr cell populations. The core network model was expanded into a more comprehensive and predictive model by including cytokine production and signaling pathways. From the expanded network, the interaction between TGFB-Induced Factor Homeobox 1 (Tgif1) and the retinoid X receptor (RXR) was displayed to exert control over the determination of the Tfh response. Model simulations predict that Tgif1 and RXR respectively induce and curtail Tfh responses. This computational hypothesis was validated experimentally by assaying Tgif1, RXR and Tfh in stomachs of mice infected with H. pylori.


DNA intercalation optimized by two-step molecular lock mechanism.
The diverse properties of DNA intercalators, varying in affinity and kinetics over several orders of magnitude, provide a wide range of applications for DNA-ligand assemblies. Unconventional intercalation mechanisms may exhibit high affinity and slow kinetics, properties desired for potential therapeutics. We used single-molecule force spectroscopy to probe the free energy landscape for an unconventional intercalator that binds DNA through a novel two-step mechanism in which the intermediate and final states bind DNA through the same mono-intercalating moiety. During this process, DNA undergoes significant structural rearrangements, first lengthening before relaxing to a shorter DNA-ligand complex in the intermediate state to form a molecular lock. To reach the final bound state, the molecular length must increase again as the ligand threads between disrupted DNA base pairs. This unusual binding mechanism results in an unprecedented optimized combination of high DNA binding affinity and slow kinetics, suggesting a new paradigm for rational design of DNA intercalators.


Serial administration of a modified Richmond Agitation and Sedation Scale for delirium screening.
Because delirium is a common yet frequently unrecognized condition, this study sought to design a brief screening tool for a core feature of mental status and to validate the instrument as a serial assessment for delirium. Prospective cohort study. Tertiary VA Hospital in New England. A total of 95 veterans admitted to the medical service. A consensus panel developed a modified version of the Richmond Agitation and Sedation Scale (RASS) to capture alterations in consciousness. Upon admission, and daily thereafter, patients were screened with a modified RASS (mRASS) and independently underwent a comprehensive mental status interview by a geriatric expert, who determined whether the criteria for delirium were met. The sensitivity, specificity, and positive likelihood ratio (LR) of the mRASS for delirium are reported. As a single assessment, the mRASS had a sensitivity of 64% and a specificity of 93% for delirium (LR, 9.4). When used to detect change, serial mRASS assessments had a sensitivity of 74% and a specificity of 92% (LR, 8.9) in both prevalent and incident delirium. When prevalent cases were excluded, any change in the mRASS had a sensitivity of 85% and a specificity of 92% for incident delirium (LR, 10.2) When administered daily, the mRASS has good sensitivity and specificity for incident delirium. Given the brevity of the instrument (<30 seconds), consideration should be given to incorporating the modified RASS as a daily screening measure for consciousness and delirium.


Strategies of pain assessment used by nurses on surgical units.
The purpose of this study was to identify the criteria nurses actually used to assess postoperative pain and the kind of knowledge they drew on from past experience. A series of semistructured interviews were performed with 10 nurses while they conducted pain assessments with 30 postsurgical patients. Data were analyzed on the basis of Dahlgren and Fallsberg's (1991) analytic approach called phenomenography. Findings were compared with Sjostrom's 1995 study of nurses in Sweden. Nurses in this study, much like the group in Sweden, used three categories of criteria (i.e., how the patient looked, what the patient said, and experience with similar circumstances) and drew on their past experiences in four different ways (i.e., in terms of a typology of patients, a focus on listening to patients, what to look for, and what to do for patients) when assessing postoperative pain. In the most frequently reported strategy, nurses relied on criteria related to the patient's appearance and drew on their past experience in terms of what physical signs to look for (e.g., facial expressions, bodily movement, and heart rate). The variety of strategies used by nurses in this study opens an avenue for further investigating the linkage between underlying conceptions of pain (as essentially subjective vs. objective phenomenon) and the quality and effectiveness of nurses' assessments of pain and its management in postoperative and other contexts across nursing.


Removal of baboon and human antiporcine IgG and IgM natural antibodies by immunoadsorption. Results of in vitro and in vivo studies.
The safe and effective removal of xenoreactive antibodies in the peritransplant period is likely to be critical for the clinical application of xenotransplantation involving disparate donor species, such as the pig. In an effort to develop an improved method for antibody removal in xenotransplantation, we have studied reusable antihuman antibody (Ig) columns in vitro and in vivo. Two types of columns were tested: (1) an antihuman Ig column containing polyclonal sheep antihuman IgG (heavy- and light-chain-specific) conjugated to sepharose CL-4B (Ig-Therasorb), and (2) an antihuman Ig column using polyclonal antihuman IgM (mu-chain-specific) conjugated to sepharose. Passage of human or baboon plasma through the Ig-Therasorb column resulted in 97.5% and 78.4% mean reductions in total IgG and IgM, respectively. Reductions in total IgG and IgM correlated with lowering of antipig IgG (54-486 fold) and IgM (9-54 fold) antibody titers as assessed by pig endothelial cell ELISA. The ability of the Ig-Therasorb to significantly reduce IgM may be attributed to the light chain specificity of this column. With the anti-IgM column, marked reductions in total (82.6-83.9%) and antipig (27-54 fold) IgM in human and baboon plasma occurred, while levels of total and xenoreactive IgG were slightly affected. Other than a dilutional effect, neither column resulted in significant reduction in albumin, fibrinogen, factor 5, and factor 8. Repeated in vivo use of either column in baboons achieved reductions in IgG and IgM that closely followed the results of our in vitro studies. No subject morbidity or mortality occurred. Use of the Ig-Therasorb column with immunosuppression in two baboons receiving pig renal xenografts achieved sustained reductions in antipig antibodies and prevented hyperacute rejection. Subjects were sacrificed at 11 and 13 days posttransplant with functioning xenografts and were found to have no evidence of vascular xenograft rejection. We conclude that anti-Ig columns represent a safe and effective method for antibody removal, without several of the limitations of other antibody removal techniques. Also, columns appear to be safe for repeated antibody removal in the posttransplant period.


Aun (n = 1-16) clusters on the ZrO2(111) surface: a DFT+U investigation.
The growth patterns and electronic structures of Aun clusters (n = 1-16) supported on the monoclinic ZrO2(111) surface were investigated using a DFT+U approach. We found that the supported Aun clusters prefer quasi-planar structures and lay flat on the ZrO2 surface. This result agrees well with the experimental results. Both orbital overlap and dispersion interactions contribute to the interaction between the Aun cluster and the ZrO2 surface. Electrons were transferred from the ZrO2 surface to the Aun cluster. Small energy gaps between unoccupied states in the Aun clusters and occupied states in the ZrO2 surface were found, especially for the supported Aun clusters with odd n, which may indicate that more electrons are excited from the ZrO2 surface to the Aun cluster even under visible-light irradiation. In other words, the ZrO2 support may be involved in the photocatalytic process when Aun/ZrO2 is used as a photocatalyst.


Flagellar membrane fusion and protein exchange in trypanosomes; a new form of cell-cell communication?
Diverse structures facilitate direct exchange of proteins between cells, including plasmadesmata in plants and tunnelling nanotubes in bacteria and higher eukaryotes. Here we describe a new mechanism of protein transfer, flagellar membrane fusion, in the unicellular parasite Trypanosoma brucei. When fluorescently tagged trypanosomes were co-cultured, a small proportion of double-positive cells were observed. The formation of double-positive cells was dependent on the presence of extracellular calcium and was enhanced by placing cells in medium supplemented with fresh bovine serum. Time-lapse microscopy revealed that double-positive cells arose by bidirectional protein exchange in the absence of nuclear transfer. Furthermore, super-resolution microscopy showed that this process occurred in ≤1 minute, the limit of temporal resolution in these experiments. Both cytoplasmic and membrane proteins could be transferred provided they gained access to the flagellum. Intriguingly, a component of the RNAi machinery (Argonaute) was able to move between cells, raising the possibility that small interfering RNAs are transported as cargo. Transmission electron microscopy showed that shared flagella contained two axonemes and two paraflagellar rods bounded by a single membrane. In some cases flagellar fusion was partial and interactions between cells were transient. In other cases fusion occurred along the entire length of the flagellum, was stable for several hours and might be irreversible. Fusion did not appear to be deleterious for cell function: paired cells were motile and could give rise to progeny while fused. The motile flagella of unicellular organisms are related to the sensory cilia of higher eukaryotes, raising the possibility that protein transfer between cells via cilia or flagella occurs more widely in nature.


Risk factors for the development of NIDDM in Yonchon County, Korea.
To determine the risk factors for the development of NIDDM in Yonchon County of Korea. We studied 1,193 Korean nondiabetic subjects at baseline who participated in a 2-year follow-up study on diabetes in Yonchon County. A 75-g oral glucose tolerance test was performed 2 years after the baseline examination. Age, sex, and anthropometric and metabolic characteristics at baseline were analyzed simultaneously as potential predictors of conversion to NIDDM. We also designed a nested case-control study to determine the role of hyperinsulinemia and/or hyperproinsulinemia in the conversion to NIDDM in patients with newly developed diabetes and control subjects matched for age, sex, BMI, and waist-to-hip-ratio. At 2 years, 67 subjects developed diabetes, as defined by World Health Organization criteria. The age-adjusted incidence was significantly higher in men (6.4%) than in women (3.0%), and the incidence increased as age increased in both sexes. Multiple logistic regression analysis revealed age, male sex, and fasting and 2-h glucose levels to be significant risk factors for the development of NIDDM, whereas waist-to-hip ratio and BMI were not. In a nested case-control study, baseline proinsulin but not insulin levels were significantly higher in subjects who progressed to NIDDM than in those who did not. In the Korean population, beta-cell dysfunction, as measured by high proinsulin levels, seems to be associated with subsequent development of NIDDM, whereas regional and general obesity and fasting insulin levels, which may be a surrogate for insulin resistance, were not.


Influence of artisan bakery- or laboratory-propagated sourdoughs on the diversity of lactic acid bacterium and yeast microbiotas.
Seven mature type I sourdoughs were comparatively back-slopped (80 days) at artisan bakery and laboratory levels under constant technology parameters. The cell density of presumptive lactic acid bacteria and related biochemical features were not affected by the environment of propagation. On the contrary, the number of yeasts markedly decreased from artisan bakery to laboratory propagation. During late laboratory propagation, denaturing gradient gel electrophoresis (DGGE) showed that the DNA band corresponding to Saccharomyces cerevisiae was no longer detectable in several sourdoughs. Twelve species of lactic acid bacteria were variously identified through a culture-dependent approach. All sourdoughs harbored a certain number of species and strains, which were dominant throughout time and, in several cases, varied depending on the environment of propagation. As shown by statistical permutation analysis, the lactic acid bacterium populations differed among sourdoughs propagated at artisan bakery and laboratory levels. Lactobacillus plantarum, Lactobacillus sakei, and Weissella cibaria dominated in only some sourdoughs back-slopped at artisan bakeries, and Leuconostoc citreum seemed to be more persistent under laboratory conditions. Strains of Lactobacillus sanfranciscensis were indifferently found in some sourdoughs. Together with the other stable species and strains, other lactic acid bacteria temporarily contaminated the sourdoughs and largely differed between artisan bakery and laboratory levels. The environment of propagation has an undoubted influence on the composition of sourdough yeast and lactic acid bacterium microbiotas.


Robotic ureteroureterostomy in children with a duplex collecting system.
Duplex collecting system pathology can be handled using an ablative procedure or reconstructive procedure even in the light of a poorly functioning moiety. We propose that, when a reconstructive procedure is an option, a robotic ureteroureterostomy is safe and feasible. Fifteen children between the ages of 6 months and 10 years (mean 31.26 months) underwent transperitoneal robotic ureteroureterostomy for duplex collecting system pathology. The surgical procedure included transperitoneal robotic approach. Outcome measures included operative time, length of hospital stay, and resolution of symptoms. Mean operative time was 1.2 h (range 0.75-2.2 h) for the entire procedure, including the cystoscopic evaluation. Length of stay averaged 20.8 h (range 15-26 h). All postoperative imaging demonstrated intact, well-draining collecting systems. The presenting symptomatology resolved in all the patients in whom symptoms were present. Robotic ureteroureterostomy is feasible and safe in the pediatric population and should be considered part of the surgical armamentarium when upper tract preservation seems warranted.


Morphology and Phylogeny of a New Species of Anaerobic Ciliate, Trimyema finlayi n. sp., with Endosymbiotic Methanogens.
Many anaerobic ciliated protozoa contain organelles of mitochondrial ancestry called hydrogenosomes. These organelles generate molecular hydrogen that is consumed by methanogenic Archaea, living in endosymbiosis within many of these ciliates. Here we describe a new species of anaerobic ciliate, Trimyema finlayi n. sp., by using silver impregnation and microscopy to conduct a detailed morphometric analysis. Comparisons with previously published morphological data for this species, as well as the closely related species, Trimyema compressum, demonstrated that despite them being similar, both the mean cell size and the mean number of somatic kineties are lower for T. finlayi than for T. compressum, which suggests that they are distinct species. This was also supported by analysis of the 18S rRNA genes from these ciliates, the sequences of which are 97.5% identical (6 substitutions, 1479 compared bases), and in phylogenetic analyses these sequences grouped with other 18S rRNA genes sequenced from previous isolates of the same respective species. Together these data provide strong evidence that T. finlayi is a novel species of Trimyema, within the class Plagiopylea. Various microscopic techniques demonstrated that T. finlayi n. sp. contains polymorphic endosymbiotic methanogens, and analysis of the endosymbionts' 16S rRNA gene showed that they belong to the genus Methanocorpusculum, which was confirmed using fluorescence in situ hybridization with specific probes. Despite the degree of similarity and close relationship between these ciliates, T. compressum contains endosymbiotic methanogens from a different genus, Methanobrevibacter. In phylogenetic analyses of 16S rRNA genes, the Methanocorpusculum endosymbiont of T. finlayi n. sp. grouped with sequences from Methanomicrobia, including the endosymbiont of an earlier isolate of the same species, 'Trimyema sp.,' which was sampled approximately 22 years earlier, at a distant (∼400 km) geographical location. Identification of the same endosymbiont species in the two separate isolates of T. finlayi n. sp. provides evidence for spatial and temporal stability of the Methanocorpusculum-T. finlayi n. sp. endosymbiosis. T. finlayi n. sp. and T. compressum provide an example of two closely related anaerobic ciliates that have endosymbionts from different methanogen genera, suggesting that the endosymbionts have not co-speciated with their hosts.


Physiotherapists report improved understanding of and attitude toward the cognitive, psychological and social dimensions of chronic low back pain after Cognitive Functional Therapy training: a qualitative study.
What are physiotherapists' perspectives on managing the cognitive, psychological and social dimensions of chronic low back pain after intensive biopsychosocial training? Qualitative study design using semi-structured interviews to explore physiotherapists' perceptions of their identification and treatment of the biopsychosocial dimensions of chronic low back pain after intensive Cognitive Functional Therapy (CFT) training. Thirteen qualified physiotherapists from four countries who had received specific CFT training. The training involved supervised implementation of CFT in clinical practice with patients. Interviews were audio-recorded and transcribed verbatim. An interpretive descriptive analysis was performed using a qualitative software package. Four main themes emerged from the data: self-reported changes in understanding and attitudes; self-reported changes in professional practice; altered scope of practice; and increased confidence and satisfaction. Participants described increased understanding of the nature of pain, the role of patient beliefs, and a new appreciation of the therapeutic alliance. Changes in practice included use of new assessments, changes in communication, and adoption of a functional approach. Since undertaking CFT training, participants described a greater awareness of their role and scope of practice as clinicians in identifying and addressing these factors. Physiotherapists expressed confidence in their capacity and skill set to manage the biopsychosocial dimensions of chronic low back pain after CFT training, and identified a clear role for including these skills within the physiotherapy profession. Despite this, further clinical trials are needed to justify the time and cost of training, so that intensive CFT training may be made more readily accessible to clinicians, which to date has not been the case. [Synnott A, O'Keeffe M, Bunzli S, Dankaerts W, O'Sullivan P, Robinson K, O'Sullivan K (2016) Physiotherapists report improved understanding of and attitude toward the cognitive, psychological and social dimensions of chronic low back pain after Cognitive Functional Therapy training: a qualitative study.Journal of Physiotherapy62: 215-221].


Tidal breath analysis for infant pulmonary function testing. ERS/ATS Task Force on Standards for Infant Respiratory Function Testing. European Respiratory Society/American Thoracic Society.
The aim of this position paper is to provide recommendations pertaining to software and equipment requirements when analysing tidal breathing measurements in infants. These guidelines cover numerous aspects including terminology and definitions, equipment, data acquisition and analysis, and reporting of results, and highlight areas in which further research is needed before consensus can be reached. When collecting tidal breathing data in infants and children, equipment dead space and resistance must be minimized, all sources of leak eliminated, and a flowmeter with appropriate frequency response and linearity employed. Inspired gases should be corrected to body temperature, barometric pressure and saturated with water vapour conditions and efforts made to eliminate the various sources of drift in volume that can occur. In addition, the analogue-to-digital converter used to sample data must be capable of adequately resolving the highest and lowest flows required by the study. An adequate sampling rate must be used; 50-100 Hz may be sufficient for the determination of timing and volume parameters, especially in older infants, but rates of 200 Hz are recommended for analysis of the tidal breathing flow/volume loop and other sensitive parameters such as time to peak tidal expiratory flow/expiratory time. The potentially most troublesome aspect of tidal breath analysis from the computational point of view is the identification of the beginning and end of inspiration and expiration. Once methods and equipment for the measurement and analysis of tidal breathing in infants have been standardized, there is an urgent need to establish appropriate reference ranges for various key parameters so that they may be used more effectively in the clinical setting. Implementation of these recommendations should help to ensure that such measurements are as accurate as possible and that more meaningful comparisons can be made between data collected in different centres or with different equipment.


Ginsenosides attenuate methylglyoxal-induced impairment of insulin signaling and subsequent apoptosis in primary astrocytes.
Diabetes mellitus (DM), which is characterized by chronic hyperglycemia, is known to increase the risk of neurodegeneration. In type 2 diabetes, hyperglycemia could cause insulin resistance and neurodegeneration in various cells including neurons and astrocytes. Hyperglycemia is also known to result in the formation of advanced glycation end-products (AGE) Methylglyoxal (MG) is one of the most reactive AGE precursors in which its abnormal accumulation is usually found in diabetic patients and induces neuronal cell death in central nervous system. Ginseng is a herb that has been widely used to treat various diseases in traditional Chinese medicine. Ginsenosides, the pharmacologically active component isolated from ginseng, have been shown to have cryoprotective effects in different neural cells. In the present study we investigated the effects of MG in disturbing insulin signaling and leading to further cellular apoptosis in rat primary astrocytes. Furthermore, the protective effects of different subtypes of ginsenosides were studied. From the results, impairment of insulin signaling was found in astrocytes under MG treatment. Moreover, cleavage of caspase and Poly ADP ribose polymerase (PARP) was observed in line with insulin signaling disruption, showing the neurotoxic effects of MG towards astrocytes. The effects of ginsenosides in MG treated astrocytes were also investigated. After treatment, ginsenosides Rd and R-Rh2 were shown to ameliorate the cell viability of MG-treated astrocytes. In addition, Rd and R-Rh2 could improve insulin signaling and inhibit apoptosis, indicating that Rd, R-Rh2 and related compounds may have therapeutic potential in treating diabetes-induced neurodegeneration.


The behaviour and function of bottle cells during gastrulation of Xenopus laevis.
The behaviour of bottle cells in normal and microsurgically altered gastrulae and in cultured explants of Xenopus laevis was analysed, using time-lapse micrography, scanning electron microscopy (SEM) and cell tracing with fluorescein dextran amine (FDA). The results shed new light on the function of bottle cells. Bottle cells forming in vivo show a predominantly animal-vegetal apical contraction and a concurrent apical-basal elongation, whereas those forming in cultured explants show uniform apical contraction and remain rotund. Bottle cells forming in embryos with fewer subblastoporal cells contract more uniformly than those in normal embryos and release of normal bottle cells from supra- and subblastoporal cells results in immediate loss of the bottle shape. These results, and an analysis of the effects of bottle cell formation on the shapes and movements of surrounding tissues, show that unique shape of bottle cells and their probable function in development are not intrinsic properties but result from a modulation of the effect of a uniform and intrinsic apical contraction by the geometric and mechanical properties of the surrounding tissue. Mechanical simulations of bottle cell formation, using the finite element method, suggest how the site of bottle cell formation and the thickness and stiffness of adjacent tissues might change the effects of their formation. These results and FDA marking of prospective bottle cells and the adjacent deep mesodermal cells suggest that bottle cells function during their formation to initiate the involution of the prospective mesodermal mantle. Later they respread to deepen the archenteron and to form its peripheral wall.


Glycemic index and glycemic load are associated with some cardiovascular risk factors among the PREMIER study participants.
The clinical significance of glycemic index (GI) and glycemic load (GL) is inconclusive. This study was conducted to examine the association of GI and GL with clinical cardiovascular disease (CVD) risk factors including body weight, blood pressure (BP), serum lipids, fasting glucose, insulin and homocysteine over time among the PREMIER participants. PREMIER was an 18-month randomized lifestyle intervention trial, conducted from 2000 to 2002, designed to help participants reduce BP by following the Dietary Approaches to Stop Hypertension (DASH) dietary pattern, losing weight, reducing sodium and increasing physical activity. GI and GL were estimated from 24 h diet recall data at baseline, 6 and 18 months after intervention. PROC MIXED model was used to examine the association of changes in GI or GL with changes in CVD risk factors. A total of 756 randomized participants, 62% females and 34% African Americans and who averaged 50.0±0.3 years old and 95.3±0.7 kg, were included in this report. Neither GI nor GL changes was associated with changes in any risk factors at 6 months. At 18 months, however, the GI change was significantly and positively associated with total cholesterol (TC) change only (p<0.05, β=23.80±12.11 mg/dL or 0.62±0.31 mmol/L) with a significant age interaction. The GL change was significantly associated with TC (p=0.02, β=0.28±0.15 mg/dL or 0.01±0.00 mmol/L) positively and with low density lipoprotein cholesterol (LDL-C) changes negatively (p=0.03, β=-0.01±0.00 mg/dL or -0.00±0.00 mmol/L), and significant age interactions were observed for both. GI and GL was associated with TC and LDL-C after controlling for energy, fat and fiber intake and other potential confounders and the associations were modified by age. Further investigation into this relationship is important because of its potential clinical impact.


Fibroblast growth factor-2 in remodeling of the developing basement membrane zone in the trachea of infant rhesus monkeys sensitized and challenged with allergen.
Remodeling of the epithelial basement membrane zone (BMZ) involves increased deposition of collagen, resulting in thickening of the BMZ. The current study focuses on fibroblast growth factor-2 (FGF-2) in the tracheal BMZ in house dust mite allergen (HDMA)-sensitized infant rhesus monkeys, challenged with HDMA at a time when the BMZ is undergoing active postnatal development. To detect structural changes in the BMZ, we measured collagens I, III, and V. To detect changes in the function of the BMZ, we measured immunoreactivity of the heparan sulfate proteoglycan, perlecan, and FGF-2. We found significant thickening of the tracheal BMZ (p < 0.05) with each of these parameters. We also found that all HDMA tracheal samples expressed thin focal areas of the BMZ associated with leukocyte trafficking. These areas were depleted of perlecan and FGF-2; however, increased FGF-2 immunoreactivity was present in the adjacent basal cells. We conclude that basal cells and FGF-2 are involved with significant remodeling of the BMZ in the developing trachea of infant rhesus monkeys exposed to HDMA.


A role for mitochondrial dysfunction in perpetuating radiation-induced genomic instability.
Radiation-induced genomic instability (RIGI) manifests as a heritable increased rate of genetic alterations in the progeny of irradiated cells generations after the initial insult. The progeny can show an increased frequency of chromosomal translocations, deletions, mutations, micronuclei, and decreased plating efficiency. What perpetuates RIGI is unclear; however, persistently increased levels of reactive oxygen species (ROS) are frequently associated with genomically unstable clones. Furthermore, addition of free radical scavengers (e.g., DMSO, glycerol, and cationic thiol cysteamine) reduces the incidence of instability after irradiation, implicating a ROS-mediated role in RIGI induction. Because mitochondria are a major natural cellular source of ROS, we tested the hypothesis that mitochondrial dysfunction has a role in maintaining the elevated ROS levels in our irradiated, genetically unstable GM10115 Chinese hamster ovary cells. Amplex Red fluorometry measurements indicate that the relative contribution of uncoupler-sensitive mitochondrial hydrogen peroxide production to total cellular hydrogen peroxide generation is greater in unstable cells. Measurements of mitochondrial DNA levels and cell cytometric fluorescent measurements of Mitotracker Green FM indicate that differences in mitochondrial ROS production are not due to varying mitochondrial levels. However, mitochondrial respiration measured in digitonin-permeabilized cells is impaired in unstable clones. In addition, manganese superoxide dismutase, a major mitochondrial antioxidant enzyme, exhibits increased immunoreactivity but decreased enzyme activity in unstable clones, which along with decreased respiration rates may explain the increased levels of cellular ROS. These studies show that mitochondria from unstable cells are abnormal and likely contribute to the persistent oxidative stress in the unstable clones.


Chaotic frequency scaling in a coupled oscillator model for free rhythmic actions.
The question of how best to model rhythmic movements at self-selected amplitude-frequency combinations, and their variability, is a long-standing issue. This study presents a systematic analysis of a coupled oscillator system that has successfully accounted for the experimental result that humans' preferred oscillation frequencies closely correspond to the linear resonance frequencies of the biomechanical limb systems, a phenomenon known as resonance tuning or frequency scaling. The dynamics of the coupled oscillator model is explored by numerical integration in different areas of its parameter space, where a period doubling route to chaotic dynamics is discovered. It is shown that even in the regions of the parameter space with chaotic solutions, the model still effectively scales to the biomechanical oscillator's natural frequency. Hence, there is a solution providing for frequency scaling in the presence of chaotic variability. The implications of these results for interpreting variability as fundamentally stochastic or chaotic are discussed.


Development of a novel peptide aptamer-based immunoassay to detect Zika virus in serum and urine.
Zika virus (ZIKV) has been identified as a cause of adverse outcomes of pregnancy, including microcephaly and other congenital diseases. Most people infected with ZIKV do not show any symptoms. Development of a method to discriminate dengue virus (DENV) and ZIKV infections has been challenging, and efficient assays for patient management are limited, attributable to high levels of cross-reactivity among co-circulating Flaviviruses. Thus, there is an urgent need for a specific high-throughput diagnostic assay to discriminate ZIKV infections from other Flavivirus infections. Methods: A novel epitope peptide of the ZIKV envelope protein was predicted using three immune epitope database analysis tools and then further modified. A molecular docking study was conducted using three-dimensional structures of the ZIKV envelope and peptide. Experimentally, interactions between the selected peptides and virus were assessed via a fluorescence-linked sandwich immunosorbent assay (FLISA), and performance of peptide-linked sandwich FLISA was evaluated in virus-spiked human serum and urine. Results: The Z_10.8 peptide (KRAVVSCAEA) was predicted to be a suitable detector, with a higher binding affinity than other candidates based on four criteria (binding affinity, root mean square deviation, position of amine residue of lysine at the N-terminus, and interactive site) in a docking study. Z_10.8 was significantly more efficient at detecting ZIKV than the other two peptides, as shown in the direct FLISA (P < 0.001). Further, the equilibrium dissociation constant (Kd) for the Z_10.8 peptide was 706.0 ± 177.9 (mean ± SD, nM), with specificity to discriminate ZIKV from DENV. The limit of detection for the sandwich FLISA was calculated as 1×104 tissue culture infective dose (TCID)50/mL. The presence of serum or urine did not interfere with the performance of the Z_10.8-linked sandwich FLISA. Conclusion: Four criteria are suggested for the development of an in silico modeled peptide aptamer; this computerized peptide aptamer discriminated ZIKV from DENV via immunoassay.


Anatomical and clinical predictors of valve dysfunction and aortic dilation in bicuspid aortic valve disease.
Bicuspid aortic valve (BAV) is associated with early valvular dysfunction and proximal aorta dilation with high heterogeneity. This study aimed to assess the determinants of these complications. Eight hundred and fifty-two consecutive adults diagnosed of BAV referred from cardiac outpatient clinics to eight echocardiographic laboratories of tertiary hospitals were prospectively recruited. Exclusion criteria were aortic coarctation, other congenital disorders or intervention. BAV morphotype, significant valve dysfunction and aorta dilation (≥2 Z-score) at sinuses and ascending aorta were established. Three BAV morphotypes were identified: right-left coronary cusp fusion (RL) in 72.9%, right-non-coronary (RN) in 24.1% and left-non-coronary (LN) in 3.0%. BAV without raphe was observed in 18.3%. Multivariate analysis showed aortic regurgitation (23%) to be related to male sex (OR: 2.80, p<0.0001) and valve prolapse (OR: 5.16, p<0.0001), and aortic stenosis (22%) to BAV-RN (OR: 2.09, p<0.001), the presence of raphe (OR: 2.75, p<0.001), age (OR: 1.03; p<0.001), dyslipidaemia (OR: 1.77, p<0.01) and smoking (OR: 1.63, p<0.05). Ascending aorta was dilated in 76% without differences among morphotypes and associated with significant valvular dysfunction. By contrast, aortic root was dilated in 34% and related to male sex and aortic regurgitation but was less frequent in aortic stenosis and BAV-RN. Normofunctional valves are more prevalent in BAV without raphe. Aortic stenosis is more frequent in BAV-RN and associated with some cardiovascular risk factors, whereas aortic regurgitation (AR) is associated with male sex and sigmoid prolapse. Although ascending aorta is the most commonly dilated segment, aortic root dilation is present in one-third of patients and associated with AR. Remarkably, BAV-RL increases the risk for dilation of the proximal aorta, whereas BAV-RN spares this area.


Protein tyrosine phosphatase epsilon affects body weight by downregulating leptin signaling in a phosphorylation-dependent manner.
Molecular-level understanding of body weight control is essential for combating obesity. We show that female mice lacking tyrosine phosphatase epsilon (RPTPe) are protected from weight gain induced by high-fat food, ovariectomy, or old age and exhibit increased whole-body energy expenditure and decreased adiposity. RPTPe-deficient mice, in particular males, exhibit improved glucose homeostasis. Female nonobese RPTPe-deficient mice are leptin hypersensitive and exhibit reduced circulating leptin concentrations, suggesting that RPTPe inhibits hypothalamic leptin signaling in vivo. Leptin hypersensitivity persists in aged, ovariectomized, and high-fat-fed RPTPe-deficient mice, indicating that RPTPe helps establish obesity-associated leptin resistance. RPTPe associates with and dephosphorylates JAK2, thereby downregulating leptin receptor signaling. Leptin stimulation induces phosphorylation of hypothalamic RPTPe at its C-terminal Y695, which drives RPTPe to downregulate JAK2. RPTPe is therefore an inhibitor of hypothalamic leptin signaling in vivo, and provides controlled negative-feedback regulation of this pathway following its activation.


Comorbidity and early diagnosis of head and neck cancer in a Medicare population.
This study assessed potential opportunities for an early diagnosis of head and neck cancers (HNCA), and the role played by comorbidity, among contacts by Medicare patients with the healthcare system before their diagnosis. Comorbidity was hypothesized to affect the relationship between use of healthcare services and stage of disease. The study database (n =11,312) was constructed by linking files from the Surveillance, Epidemiology, and End Results Program 1991 through 1999 and those of the Center for Medicare and Medicaid Services Program. HNCA cases were comprised of the lip, oral cavity, pharynx, and larynx anatomic sites. Among persons with no alcohol and tobacco-related comorbidities (ATC), increasing numbers of physician visits were independently associated with a reduced risk of advanced stage at diagnosis for all anatomic sites, but especially the pharynx and larynx. Among persons with one or more ATC, physician visits displayed no association with stage at diagnosis with the exception of laryngeal tumors, where physician visits were strongly associated with reduced risk of late stage at diagnosis. Over 46% of cases with advanced stage tumors, most of whom also had ATC disease, had > or =11 physician visits in the year preceding their diagnosis of HNCA. Potential opportunities for an earlier diagnosis of HNCA were found among the numerous contacts with the healthcare system that patients had prior to their cancer diagnosis. Such opportunities were especially numerous and relevant for persons with ATC and form the basis for combining high-risk with opportunistic screening approaches.


Sensory reanimation of the hand by transfer of the superficial branch of the radial nerve to the median and ulnar nerve.
It remains a surgical challenge to treat high-grade nerve injuries of the upper extremity. Extra-anatomic reconstructions through the transfer of peripheral nerves have gained clinical importance over the past decades. This contribution outlines the anatomic and histomorphometric basis for the transfer of the superficial branch of the radial nerve (SBRN) to the median nerve (MN) and the superficial branch of the ulnar nerve (SBUN). The SBRN, MN, and SBUN were identified in 15 specimens and the nerve transfer performed. A favorable site for coaptation was chosen and its location described using relevant anatomical landmarks. Histomorphometric characteristics of donor and target were compared to evaluate the chances of a clinical success. A suitable location for dissecting the SBRN was identified prior to its first bifurcation. Coaptations were possible near the pronator quadratus muscle, approximately 22 cm distal to the lateral epicondyle of the humerus. The MN and SBUN had to be dissected interfasciculary over 82 ± 5.7 mm and 49 ± 5.5 mm, respectively. Histomorphometric analysis revealed sufficient donor-to-recipient axon ratios for both transfers and identified the SBRN as a suitable donor with high axon density. Our anatomic and histomorphometric results indicate that the SBRN is a suitable donor for the MN and SBUN at wrist level. The measurements show feasibility of this procedure and shall help in planning this sensory nerve transfer. High axon density in the SBRN identifies it or its branches an ideal candidate for sensory reanimation of fingers and thumbs.


Corneal collagen cross-linking in the stabilization of PRK, LASIK, thermal keratoplasty, and orthokeratology.
To describe the use of corneal collagen cross-linking (CXL) and its efficacy in the stabilization of keratorefractive procedures, including PRK, laser in-situ keratomileusis (LASIK), thermal keratoplasty, and orthokeratology. Since its introduction, CXL has quickly gained interest in the treatment of ectactic disorders due to its ability to increase the biomechanical stability of the cornea. In its earliest use, it has shown to be effective in the treatment of both keratoconus and post-LASIK ectasia. More recent studies of CXL in combination with keratorefractive procedures have shown varying degrees of success. CXL with PRK has shown to be effective in slowing or halting the progression of keratoconus, pellucid marginal degeneration, and post-LASIK ectasia, in addition to potentially decreasing or delaying the need for penetrating keratoplasty. Several small case series have also demonstrated improved stability and efficacy of PRK and LASIK when combined with CXL, as well as a potentially decreased risk of postprocedure ectasia. In conjunction with thermokeratoplasty and orthokeratology, CXL has shown improved but only temporary results in the treatment of keratoconus. Future studies are needed to determine the efficacy and long-term stability of CXL in combination with keratorefractive procedures, as well as to address possible complications.


New toothed flying reptile from Asia: close similarities between early Cretaceous pterosaur faunas from China and Brazil.
Despite the great increase in pterosaur diversity in the last decades, particularly due to discoveries made in western Liaoning (China), very little is known regarding pterosaur biogeography. Here, we present the description of a new pterosaur from the Jiufotang Formation that adds significantly to our knowledge of pterosaur distribution and enhances the diversity of cranial anatomy found in those volant creatures. Guidraco venator gen. et sp. nov. has an unusual upward-directed frontal crest and large rostral teeth, some of which surpass the margins of the skull and lower jaw when occluded. The new species is closely related to a rare taxon from the Brazilian Crato Formation, posing an interesting paleobiogeographic problem and supporting the hypothesis that at least some early Cretaceous pterosaur clades, such as the Tapejaridae and the Anhangueridae, might have originated in Asia. The association of the new specimen with coprolites and the cranial morphology suggest that G. venator preyed on fish.


Comparison of Long-term Complications in Patients on Haemodialysis and Peritoneal Dialysis Longer than 10 Years.
Depending on developments in dialysis techniques and new treatment strategies for comorbid diseases, life expectancy has increased. As a result, dialysis related long term complications could be seen more frequently. We investigated and compared long term complications of the Haemodialysis (HD) and Peritoneal Dialysis (PD) in patients with history if either mode at least 10years. A 13HD & 16PD patients were included to the study. Basic demographic parameters and prevalence of cardiovascular diseases (CVD), uraemic peripheral neuropathy (PNP), parathyroid adenoma, parathyroidectomy and acquired cystic disease (ACD) were assessed. HD patients were older than PD patients (p=0.035) and duration of dialysis was longer in HD patients (p=0.001). CVD was present in 18 patients (9 HD, 9 PD). There was no difference in presence of CVD between HD and PD patients (p=0.455). Valvular diseases (n=15), diastolic dysfunction (n=8), left ventricular hypertrophy (n=5), ischemic heart disease (n=3) and congestive heart failure (n=1) were investigated. Uraemic peripheral neuropathy was observed in 14 of the patients (8 HD, 6 PD patients). Eight patients had mixed type sensory motor neuropathy and 3 patients had mixed type sensorial neuropathy, 2 patients had demyelinating PNP, 1 patient had axonal PNP and 3 of them had CTS related to peripheral neuropathy. Parathyroid adenoma was detected in 4 patients (2 HD, 2 PD) and 3 patients (1 HD, 2 PD) had history of parathyroidectomy. Serum phosphate and iPTH levels were higher in HD patients (p=0.003, p=0.04, respectively). ACD was detected in 14 patients (7 HD, 7 PD). There was no difference between PD and HD patients (p=0.75). HD patients were older than PD patients and had longer duration of dialysis. The prevalence of long term complications was similar in HD and PD modalities. CVD especially valvular diseases were common complication in both modalities.


Kinematic Sunyaev-Zel'dovich Effect with Projected Fields: A Novel Probe of the Baryon Distribution with Planck, WMAP, and WISE Data.
The kinematic Sunyaev-Zel'dovich (KSZ) effect-the Doppler boosting of cosmic microwave background (CMB) photons due to Compton scattering off free electrons with nonzero bulk velocity-probes the abundance and the distribution of baryons in the Universe. All KSZ measurements to date have explicitly required spectroscopic redshifts. Here, we implement a novel estimator for the KSZ-large-scale structure cross-correlation based on projected fields: it does not require redshift estimates for individual objects, allowing KSZ measurements from large-scale imaging surveys. We apply this estimator to cleaned CMB temperature maps constructed from Planck and WMAP data and a galaxy sample from the Wide-field Infrared Survey Explorer (WISE). We measure the KSZ effect at 3.8σ-4.5σ significance, depending on the use of additional WISE galaxy bias constraints. We verify that our measurements are robust to possible dust emission from the WISE galaxies. Assuming the standard Λ cold dark matter cosmology, we directly constrain (f_{b}/0.158)(f_{free}/1.0)=1.48±0.19 (statistical error only) at redshift z≈0.4, where f_{b} is the fraction of matter in baryonic form and f_{free} is the free electron fraction. This is the tightest KSZ-derived constraint reported to date on these parameters. Astronomers have long known that baryons do not trace dark matter on ∼ kiloparsec scales and there has been strong evidence that galaxies are baryon poor. The consistency between the f_{b} value found here and the values inferred from analyses of the primordial CMB and big bang nucleosynthesis verifies that baryons approximately trace the dark matter distribution down to ∼ megaparsec scales. While our projected-field estimator is already competitive with other KSZ approaches when applied to current data sets (because we are able to use the full-sky WISE photometric survey), it will yield enormous signal-to-noise ratios when applied to upcoming high-resolution, multifrequency CMB surveys.


A complex containing p34cdc2 and cyclin B phosphorylates the nuclear lamin and disassembles nuclei of clam oocytes in vitro.
Cell-free extracts prepared from activated clam oocytes contain factors which induce phosphorylation of the single 67-kD lamin (L67), disassemble clam oocyte nuclei, and cause chromosome condensation in vitro (Dessev, G., R. Palazzo, L. Rebhun, and R. Goldman. 1989. Dev. Biol. 131:469-504). To identify these factors, we have fractionated the oocyte extracts. The nuclear lamina disassembly (NLD) activity, together with a protein kinase activity specific for L67, appear as a single peak throughout a number of purification steps. This peak also contains p34cdc2, cyclin B, and histone H1-kinase activity, which are components of the M-phase promoting factor (MPF). The NLD/L67-kinase activity is depleted by exposure of this purified material to Sepharose conjugated to p13suc1, and is restored upon addition of a p34cdc2/p62 complex from HeLa cells. The latter complex phosphorylates L67 and induces NLD in the absence of other clam oocyte proteins. Our results suggest that a single protein kinase activity (p34cdc2-H1 kinase, identical with MPF) phosphorylates the lamin and is involved in the meiotic breakdown of the nuclear envelope in clam oocytes.


Socioeconomic, lifestyle and dietary factors associated with dietary supplement use during pregnancy.
Information on dietary supplement (DS) use during pregnancy is largely lacking. Besides, little is known about the share of DS use as self-medication versus such use following a physician's advice/prescription. Our aim was to evaluate DS use and its socioeconomic, lifestyle and dietary correlates among pregnant women participating in the French NutriNet-Santé cohort study. Data were collected by self-administered web-based questionnaires. Food intake was assessed by repeated 24 h dietary records. 903 pregnant women provided data on their DS use (both "regular" DS and medication containing mainly vitamins/minerals). Supplement users were compared to non-users by unconditional logistic regression. DS use-in general and as regards folic acid in particular-was positively correlated with age, being primiparous, having higher income and belonging to a higher socioprofessional category. DS users had significantly higher dietary intakes of most vitamins and minerals. The proportion of DS users (e.g., those reporting use at least three days a week) increased significantly with the trimester of pregnancy (58.0%, 62.2% and 74.5%, respectively). 50.2% of women in their 1st trimester used folic acid. The proportion of iron users tripled from the 1st to the 3rd trimester (18.5 to 63.9%). DS use was prescribed or recommended by a physician in 86.7% of the cases. This study provided new and detailed information on DS use and its correlates during pregnancy. Even in this relatively well-educated population, folic acid supplementation at the beginning of pregnancy was inadequate and was associated with socioeconomic and demographic disparities.


Prevalence and evolutionary origins of the del(GJB6-D13S1830) mutation in the DFNB1 locus in hearing-impaired subjects: a multicenter study.
Mutations in GJB2, the gene encoding connexin-26 at the DFNB1 locus on 13q12, are found in as many as 50% of subjects with autosomal recessive, nonsyndromic prelingual hearing impairment. However, genetic diagnosis is complicated by the fact that 10%-50% of affected subjects with GJB2 mutations carry only one mutant allele. Recently, a deletion truncating the GJB6 gene (encoding connexin-30), near GJB2 on 13q12, was shown to be the accompanying mutation in approximately 50% of these deaf GJB2 heterozygotes in a cohort of Spanish patients, thus becoming second only to 35delG at GJB2 as the most frequent mutation causing prelingual hearing impairment in Spain. Here, we present data from a multicenter study in nine countries that shows that the deletion is present in most of the screened populations, with higher frequencies in France, Spain, and Israel, where the percentages of unexplained GJB2 heterozygotes fell to 16.0%-20.9% after screening for the del(GJB6-D13S1830) mutation. Our results also suggest that additional mutations remain to be identified, either in DFNB1 or in other unlinked genes involved in epistatic interactions with GJB2. Analysis of haplotypes associated with the deletion revealed a founder effect in Ashkenazi Jews and also suggested a common founder for countries in Western Europe. These results have important implications for the diagnosis and counseling of families with DFNB1 deafness.


Prognostic significance of the lymphocyte-to-monocyte ratio and the tumor-infiltrating lymphocyte to tumor-associated macrophage ratio in patients with stage T3N0M0 esophageal squamous cell carcinoma.
We assessed the prognostic significance of, and the relationship between, the pretreatment lymphocyte-to-monocyte ratio (LMR) and the TILs/tumor-associated macrophages (TAMs) ratio, in patients with esophageal squamous cell carcinoma (ESCC) of pathological stage T3N0M0 (pT3N0M0). A total of 220 newly diagnosed ESCC patients of stage pT3N0M0 who had not undergone neoadjuvant therapy were included. Densities of CD8+ TILs, CD4+ TILs, CD45RO+ TILs, and CD68+ TAMs were assessed by immunohistochemical staining of tissue microarray cores from all 220 pT3N0M0 ESCC patients (who underwent radical resection). Hematological biomarkers including lymphocyte and monocyte counts were obtained from routine preoperative blood test data, and the LMR and TILs/TAMs ratios calculated. Cutoff finder for survival prediction was plotted to find out the optimal cutoff point for each parameter. The LMR and TILs/TAMs ratios were interrelated. On univariate analyses of data from the entire cohort, the LMR, CD45RO/CD68 ratio, and CD8/CD68 ratio were significantly associated with both OS and disease-free survival. Only the CD45RO/CD68 ratio was independently prognostic of survival on multivariate analysis. The prognostic significance of the CD45RO/CD68 ratio was higher than that of the LMR. The CD45RO/CD68 ratio is a useful independent prognostic marker in patients with pT3N0M0 ESCC who have undergone complete resection without neoadjuvant therapy.


Altered Expression of the Transcription Factor Forkhead Box A1 (FOXA1) Is Associated With Poor Prognosis in Urothelial Carcinoma of the Upper Urinary Tract.
To determine the prognostic significance of Forkhead Box A1 (FOXA1) expression in patients with upper tract urothelial carcinoma (UTUC) undergoing radical nephroureterectomy (RNU). A retrospective analysis of 566 patients undergoing RNU at seven academic medical centers was performed. Tissue microarrays were subjected to immunohistochemistry using a commercially available polyclonal FOXA1 antibody. Logistic regression determined the association of FOXA1 expression with pathologic features and survival outcomes. Three hundred twenty-two men and 244 women were included. The pathologic distribution of specimens included 53% muscle-invasive or greater (≥pT2), 74% high-grade, 16% with flat architecture, 13% with necrosis, 21% with lymphovascular invasion, 18% with concomitant carcinoma in situ, and 8% with positive lymph nodes. The median FOXA1 score was 5.0 (range: 0-8). Lower FOXA1 expression was significantly correlated with advanced pathologic stage (≥pT3) (P = .02), concomitant carcinoma in situ (P = .006), and renal pelvis (vs ureter) location (P < .0001). At a median follow-up of 27.0 months (range: 3-196), 139 patients (25%) experienced disease recurrence and 121 (21%) died from the disease. In a multivariate model, lower FOXA1 expression was independently associated with disease recurrence (hazard ratio [HR]: 1.11, 95% confidence interval [CI]: 1.05-1.62, P = .04), cancer-specific mortality (HR: 1.17, 95% CI: 1.03-1.92, P = .04), and all-cause mortality (HR: 1.08, 95% CI: 1.02-1.18, P = .05). Lower FOXA1 expression is associated with adverse pathologic features and inferior survival outcomes for UTUC patients undergoing RNU. These data indicate lower FOXA1 expression may be a marker of aggressive disease in UTUC.


Correlation between histologic assessment and fluorescence in situ hybridization using MelanoSITE in evaluation of histologically ambiguous melanocytic lesions.
The 4-probe, multicolor, fluorescence in situ hybridization (FISH) panel targeting chromosomes 6 and 11 has shown promising sensitivity and specificity in distinguishing between benign nevi and malignant melanoma. Only a few studies have assessed the potential utility of FISH in classification of histologically ambiguous melanocytic lesions. In the United States, this assay is exclusively licensed to NeoGenomics Laboratories (Irvine, California), which provides the technical component and has developed an innovative service (MelanoSITE) allowing pathologists to interpret FISH results using a dedicated Web portal. Thus far, use of MelanoSITE as a diagnostic adjunct in the diagnosis of melanocytic lesions has not, to our knowledge, been reported in the literature. To analyze 1.5 years of experience with the MelanoSITE melanoma FISH assay in the evaluation of histologically ambiguous lesions in the context of second opinion and routine dermatopathology practice. A prospective histologic/FISH correlation study of 140 cases. Twenty-seven percent of abnormal FISH results were false-positive results because of tetraploidy. After correcting for known false-positive results, all lesions considered atypical nevi showed normal FISH signals. Abnormal FISH signals were reported in 30% of lesions considered histologically borderline and in 48% of lesions in which a diagnosis of melanoma was favored. Four-probe, multicolor FISH results for melanoma correlate with the microscopic assessments of histologically ambiguous lesions. Pathologists using MelanoSITE must be aware of the high rate of false-positive results from tetraploidy.


Predictors of acute kidney injury (AKI) in high-risk ST-elevation myocardial infarction (STEMI) patients: A single-center retrospective observational study.
Acute kidney injury (AKI) is a frequent complication in ST-elevation myocardial infarction (STEMI) patients. Factors other than contrast exposure have been suggested as major contributors to renal dysfunction in patients undergoing primary percutaneous coronary intervention (PPCI). Our aim was to assess the incidence and risk factors of AKI in high-risk STEMI patients, mostly treated by PPCI with implemented measures to prevent contrast-induced AKI. We retrospectively analyzed data of 245 STEMI patients (165 men, mean age 63.9 ± 11.9 years) admitted to the Department of Medical Intensive Care Unit. Demographic, clinical, and mortality data were compared between AKI and non-AKI group. AKI was defined as a 1.5-fold increase in serum creatinine from baseline level within 24-48 hours. AKI developed in 34/245 (13.9%) patients. PPCI was performed in 226/245 (92.2%) of all STEMI cases, with no difference between AKI and non-AKI group. There were significant differences between AKI and non-AKI group in diabetes mellitus (41.2% vs. 20.9%), prior MI (26.5% vs. 11.8%), prior resuscitation (38.2% vs. 12.4%), admission acute heart failure [AHF] (44.1% vs. 12.8%), in-hospital AHF (70.6% vs. 17.5%), and hospital-acquired infection [HAI] (79.4% vs. 18.0%). Significantly more AKI patients had increased admission CRP ≥25 mg/L (38.2% vs. 11.8%), peak CRP ≥50 mg/L (91.2% vs. 36%), admission troponin I ≥10 mg/L (44.1% vs. 24.6%), peak troponin I ≥50 mg/L (64.7% vs. 44.1%), peak NT-proBNP ≥400 pmol/L (82.4% vs. 27.5%), and ejection fraction <45% (76.5% vs. 33.6%). Mortality was significantly increased in AKI group, including in-hospital (52.9% vs. 7.1%), 30-day (64.7% vs. 10.7%) and 6-month mortality (70.6% vs. 13.7%). Significant independent predictors of AKI were prior resuscitation (OR 4.171, 95% CI 1.088-15.998), HAI (OR 7.974, 95% CI 1.992-31.912), and peak NT-proBNP (OR 21.261, 95% CI 2.357-191.795). To reduce the risk of AKI in STEMI patients, early diagnosis and treatment of AHF and HAIs are advisable.


Sex Differences in All-Cause Mortality in the Decade Following Complex Coronary Revascularization.
The poorer prognosis of coronary artery disease in females compared with males is related mainly to differences in baseline characteristics. In the SYNTAX (Synergy Between Percutaneous Coronary Intervention With Taxus and Cardiac Surgery) trial, the effect of treatment with percutaneous coronary intervention (PCI) versus coronary artery bypass grafting surgery (CABG) on mortality at 5 years differed significantly between females and males; however, the optimal revascularization beyond 5 years according to sex has not been evaluated. The aim of this study was to investigate the impact of sex on mortality and sex-treatment interaction at 10 years. The SYNTAXES (SYNTAX Extended Survival) study evaluated vital status up to 10 years in 1,800 patients with de novo 3-vessel and/or left main coronary artery disease randomized to treatment with PCI or CABG in the SYNTAX trial. All-cause death at 10 years was separately evaluated in female and male patients with complex coronary artery disease. Of 1,800 patients, 402 (22.3%) were female and 1,398 (77.7%) were males. Females had a higher 10-year mortality rate compared with males (32.8% vs. 24.7%; log-rank p = 0.002), but female sex was not an independent predictor of mortality (adjusted hazard ratio: 1.02; 95% confidence interval: 0.76 to 1.36). Mortality at 10 years tended to be lower after CABG than after PCI, with a similar treatment effect for female and male patients (adjusted hazard ratio for females: 0.90 [95% confidence interval: 0.54 to 1.51]; adjusted hazard ratio for males: 0.76 [95% confidence interval: 0.56 to 1.02]; p for interaction = 0.952). Female sex was not an independent predictor of mortality at 10 years in patients with complex coronary artery disease. The interaction between sex and treatment with PCI or CABG that was observed at 5 years was no longer present at 10 years. (Synergy Between PCI With TAXUS and Cardiac Surgery: SYNTAX Extended Survival [SYNTAXES], NCT03417050; SYNTAX Study: TAXUS Drug-Eluting Stent Versus Coronary Artery Bypass Surgery for the Treatment of Narrowed Arteries [SYNTAX], NCT00114972).


Virally delivered channelrhodopsin-2 safely and effectively restores visual function in multiple mouse models of blindness.
Previous work established retinal expression of channelrhodopsin-2 (ChR2), an algal cation channel gated by light, restored physiological and behavioral visual responses in otherwise blind rd1 mice. However, a viable ChR2-based human therapy must meet several key criteria: (i) ChR2 expression must be targeted, robust, and long-term, (ii) ChR2 must provide long-term and continuous therapeutic efficacy, and (iii) both viral vector delivery and ChR2 expression must be safe. Here, we demonstrate the development of a clinically relevant therapy for late stage retinal degeneration using ChR2. We achieved specific and stable expression of ChR2 in ON bipolar cells using a recombinant adeno-associated viral vector (rAAV) packaged in a tyrosine-mutated capsid. Targeted expression led to ChR2-driven electrophysiological ON responses in postsynaptic retinal ganglion cells and significant improvement in visually guided behavior for multiple models of blindness up to 10 months postinjection. Light levels to elicit visually guided behavioral responses were within the physiological range of cone photoreceptors. Finally, chronic ChR2 expression was nontoxic, with transgene biodistribution limited to the eye. No measurable immune or inflammatory response was observed following intraocular vector administration. Together, these data indicate that virally delivered ChR2 can provide a viable and efficacious clinical therapy for photoreceptor disease-related blindness.


Antiviral activity of Aloe hijazensis against some haemagglutinating viruses infection and its phytoconstituents.
Evaluation of the antiviral activities of flowers, flower-peduncles, leaves, and roots of Aloe hijazensis against haemagglutinating viruses of avian paramyxovirus type-1 (APMV-1), avian influenza virus type A (AI-H5N1), Newcastle disease virus (NDV), and egg-drop syndrome virus (EDSV) in specific pathogen free (SPF) chicken embryos were carried out. Extract of the flowers and leaves showed relatively higher activity than the extracts of other plant parts. Thirteen compounds were isolated from both the flowers and flower-peduncles of A. hijazensis. The isolated compounds were classified into: five anthraquinones; ziganein, ziganein-5-methyl ether, aloesaponarin I, chrysophanol, aloe-emodin, one dihydroisocoumarin; feralolide, four flavonoids; homoplantaginin, isoorientin, luteolin 7-glucuronopyranoside, isovitexin, one phenolic acid; p-coumaric acid, the anthrone; barbaloin together with aloenin. Eleven compounds were attributed to the flowers and seven to the flower-peduncles. Homoplantaginin and luteolin 7-glucuronopyranoside are reported here for the first time from Aloe spp. To the best of our knowledge, this is the first report on the chemical composition and biological activity of those plant parts.


Hydrodissection of conjunctival flap during trabeculectomy in eyes with conjunctival scarring caused by trachoma.
Conjunctival scarring caused by trachoma represents a substantial challenge even to the most experienced cataract/glaucoma surgeon. We describe a simple technique of hydrodissection of conjunctival flap during combined trabeculectomy and phacoemulsification in eyes with conjunctival scarring caused by trachoma, and discuss its impact on surgical outcome. A prospective study was conducted in 10 Omani patients, aged 48 to 83 who underwent combined trabeculectomy and phacoemulsification with intraocular lens (IOL) implantation. The conjunctival flap was prepared after hydrodissection using a mixture of 1 mL of balanced salt solution (BSS) and 1 mL of lignocaine 2% with 1:200,000 adrenaline. Outcome measures that were evaluated were visual function, intraocular pressure (IOP) and bleb characteristics. Patients were followed-up postoperatively for a minimum period of 6 months. The mean age of patients was 60.4 years (48-83) and a functioning bleb after 6 months was seen in 8 (80%) cases. IOP < 20 mm of Hg (applanation tonometry) without antiglaucoma medications was found in 8 cases; 1 case required one antiglaucoma medication and 1 case required more than one antiglaucoma medication for control of IOP. The latter was reoperated for control of IOP. Nonprogression of visual field was found in 8 (80%) cases. Preparation of conjunctival flap by hydrodissection was found to be a simple modification of the conventional technique during combined trabeculectomy and phacoemulsification in trachomatous eyes. It reduced conjunctival dissection, use of cautery and possibility of button-holing of the conjunctiva during dissection in scarred eyes. The minimal tissue dissection involved in this procedure reduced wound healing and thereby increased chances of a functioning bleb, control of IOP and nonprogression of visual field compared to combined surgery employing a conventional method (nonhydrodissection method) of conjunctival flap dissection.


Photoconductivity and magnetoconductance effects on vacuum vapor deposition films of weak charge-transfer complexes.
Thin films of weak charge-transfer (CT) complexes (pyrene/dimethylpyromellitdiimide or pyrene/pyromellitic dianhydride) were prepared on an interdigitated platinum electrode by vacuum vapor deposition. Their photoconductivity and magnetoconductance (MC) effects were investigated, and mobile triplet excitons (probably CT excitons) were detected by time-resolved ESR (TRESR) at room temperature. The MC effect on the photocurrent was observed and analyzed by quantum-mechanical simulation assuming two types of collision mechanisms between the electron and hole carriers and between the trapped triplet excitons and mobile carriers. A successful simulation was achieved when the parameters (g, D, E, and polarization) determined by TRESR and the effective hyperfine splitting estimated from an ab initio molecular-orbital calculation were used.


Durability of ChAdOx1 nCoV-19 (AZD1222) vaccine and hybrid humoral immunity against variants including omicron BA.1 and BA.4 6 months after vaccination (COV005): a post-hoc analysis of a randomised, phase 1b-2a trial.
COVID-19 vaccine rollout is lagging in Africa, where there has been a high rate of SARS-CoV-2 infection. We aimed to evaluate the effect of SARS-CoV-2 infection before vaccination with the ChAdOx-nCoV19 (AZD1222) vaccine on antibody responses through to 180 days. We did an unmasked post-hoc immunogenicity analysis after the first and second doses of AZD1222 in a randomised, placebo-controlled, phase 1b-2a study done in seven locations in South Africa. AZD1222 recipients who were HIV-uninfected, were stratified into baseline seropositive or seronegative groups using the serum anti-nucleocapsid (anti-N) immunoglobulin G (IgG) electroluminescence immunoassay to establish SARS-CoV-2 infection before the first dose of AZD1222. Binding IgG to spike (anti-S) and receptor binding domain (anti-RBD) were measured before the first dose (day 0), second dose (day 28), day 42, and day 180. Neutralising antibody (NAb) against SARS-CoV-2 variants D614G, beta, delta, gamma, and A.VOI.V2, and omicron BA1 and BA.4 variants, were measured by pseudovirus assay (day 28, day 42, and day 180). This trial is registered with ClinicalTrials.gov, NCT04444674, and the Pan African Clinicals Trials Registry, PACTR202006922165132. Of 185 individuals who were randomly assigned to AZD1222, we included 91 individuals who were baseline seropositive and 58 who were baseline seronegative, in the final analysis. In the seropositive group, there was little change of anti-S IgG (and anti-RBD IgG) or neutralising antibody (NAb) titres at day 42 compared with at day 28. Anti-S (and anti-RBD) IgG geometric mean concentrations (GMCs) were higher throughout in the seropositive compared with the seronegative group, including at day 180 (GMCs 517·8 [95% CI 411·3-651·9] vs 82·1 [55·2-122·3] BAU/mL). Also D614G NAb geometric mean titres (GMTs) were higher in the seropositive group than the seronegative group, as was the percentage with titres of at least 185 (80% putative risk reduction threshold [PRRT] against wild-type-alpha COVID-19), including at day 180 (92·0% [74·0-99·0] vs 18·2% [2·3-51·8). Similar findings were observed for beta, A.VOI.V2, and gamma. For delta, BA.1, and BA.4, NAb GMTs and the proportion with titres above the PRRT were substantially higher in the seropositive compared with seronegative group at day 28 and day 42, but no longer differed between the groups by day 180. A single dose of AZD1222 in the general African population, where COVID-19 vaccine coverage is low and SARS-CoV-2 seropositivity is 90%, could enhance the magnitude and quality of antibody responses to SARS-CoV-2. The Bill & Melinda Gates Foundation, the South African Medical Research Council, the UK Research and Innovation, the UK National Institute for Health Research, and the South African Medical Research Council. For the Zulu translation of the abstract see Supplementary Materials section.


Streptococcus bovis-infected total hip arthroplasty.
The case of a 51-year-old man who underwent a total hip arthroplasty following failed AO screw fixation of a subcapital femoral neck fracture is reported. Infection of the prosthesis with Streptococcus bovis type 1 followed a febrile illness. Further investigation revealed an occult premalignant polyp in the proximal colon. Colonic neoplasia and S. bovis bacteremia are associated with endocarditis; however, S. bovis is a rare pathogen infecting joint prostheses and should raise the possibility of a gastrointestinal lesion.


Industrial health services in large plants.
An industrial area or a large industry can be most usefully served by setting up an occupational health centre in close co-operation with other health services. This centre should strive to achieve not only the adaptation of the worker to the machine and to the work, but also-and even more important-the adaptation of the machine and the working methods to the physical and mental capacities of men.The industrial medical service has to work in close co-operation with the social services of the plant, but should not be considered as a part of these services, because the programme of the medical department is based on different principles and has different aims. The medical department should be directly under the supervision of the top management. But an industrial health service should not be the exponent of either the management or the worker; it should be based on general medical principles and knowledge. It is very important that the workers should fully support the activities of the medical department, and this can best be achieved by encouraging representatives of the workers to take a special interest in the work and keeping them fully informed about it.The preventive work of an industrial health centre includes pre-employment and periodical medical examinations, job analysis and placement, supervision of environmental conditions, health guidance, health statistics, and co-operation with other health agencies. The therapeutic work includes first aid and accident treatment and, under certain conditions, full medical care of the workers and their families. Experience has shown that the activity of such a centre is of great value to the health of the workers; it is also of great importance to the industry itself, even from the financial point of view.


Insight into the modulation of Shaw2 Kv channels by general anesthetics: structural and functional studies of S4-S5 linker and S6 C-terminal peptides in micelles by NMR.
The modulation of the Drosophila Shaw2 Kv channel by 1-alkanols and inhaled anesthetics is correlated with the involvement of the S4-S5 linker and C-terminus of S6, and consistent with stabilization of the channel's closed state. Structural analysis of peptides from S4-S5 (L45) and S6 (S6c), by nuclear magnetic resonance and circular dichroism spectroscopy supports that an α-helical conformation was adopted by L45, while S6c was only in an unstable/dynamic partially folded α-helix in dodecylphosphocholine micelles. Solvent accessibility and paramagnetic probing of L45 revealed that L45 lies parallel to the surface of micelles with charged and polar residues pointing towards the solution while hydrophobic residues are buried inside the micelles. Chemical shift perturbation introduced by 1-butanol on residues Gln320, Thr321, Phe322 and Arg323 of L45, as well as Thr423 and Gln424 of S6c indicates possible anesthetic binding sites on these two important components in the channel activation apparatus. Diffusion measurements confirmed the association of L45, S6c and 1-butanol with micelles which suggests the capability of 1-butanol to influence a possible interaction of L45 and S6c in the micelle environment.


Host genetic factors influence disease progression in chronic hepatitis C.
Progressive hepatic fibrosis and cirrhosis develops in 20% to 30% of patients with chronic hepatitis C virus (HCV). We propose that host genetic factors influencing fibrogenesis may account for some of the variability in progression of this disease. In progressive fibrosis of other organs, particularly heart and kidney, production of the profibrogenic cytokine, transforming growth factor beta1 (TGF-beta1), may be enhanced by angiotensin II, the principal effector molecule of the renin-angiotensin system. The inheritance of polymorphisms in TGF-beta1, interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-alpha), and genes of the renin-angiotensin system was examined in 128 patients with chronic HCV. The influence of genotypes on the stage of hepatic fibrosis was tested after adjustment for potential confounders (age, gender, alcohol consumption, portal inflammation, and steatosis), which may have independent effects on histological severity. The stage of fibrosis was 0 in 30 (23.4%), 1 in 44 (34.4%), 2 in 27 (21.1%), and 3 or 4 in 27 (21.1%). A statistically significant relationship was seen between inheritance of high TGF-beta1- and angiotensinogen (AT)-producing genotypes and the development of progressive hepatic fibrosis. This association persisted after correcting for potential confounders. Patients who inherited neither of the profibrogenic genotypes had no or only minimal fibrosis. Knowledge of these polymorphisms may have prognostic significance in patients with chronic HCV and may direct more aggressive therapy towards those patients with an increased risk of disease progression. The documentation of a significant relationship between AT genotype and fibrosis raises the novel suggestion that angiotensin II may be another mediator of extracellular matrix production in the liver.


Association of specific biotypes in patients with Parkinson disease and disease progression.
To identify biotypes in patients with newly diagnosed Parkinson disease (PD) and to test whether these biotypes could explain interindividual differences in longitudinal progression. In this longitudinal analysis, we use a data-driven approach clustering PD patients from the Parkinson's Progression Markers Initiative (n = 314, age 61.0 ± 9.5, years 34.1% female, 5 years of follow-up). Voxel-level neuroanatomic features were estimated with deformation-based morphometry (DBM) of T1-weighted MRI. Voxels with deformation values that were significantly correlated (p < 0.01) with clinical scores (Movement Disorder Society-sponsored revision of the Unified Parkinson's Disease Rating Scale Parts I-III and total score, tremor score, and postural instability and gait difficulty score) at baseline were selected. Then, these neuroanatomic features were subjected to hierarchical cluster analysis. Changes in the longitudinal progression and neuroanatomic pattern were compared between different biotypes. Two neuroanatomic biotypes were identified: biotype 1 (n = 114) with subcortical brain volumes smaller than heathy controls and biotype 2 (n = 200) with subcortical brain volumes larger than heathy controls. Biotype 1 had more severe motor impairment, autonomic dysfunction, and much worse REM sleep behavior disorder than biotype 2 at baseline. Although disease durations at the initial visit and follow-up were similar between biotypes, patients with PD with smaller subcortical brain volume had poorer prognosis, with more rapid decline in several clinical domains and in dopamine functional neuroimaging over an average of 5 years. Robust neuroanatomic biotypes exist in PD with distinct clinical and neuroanatomic patterns. These biotypes can be detected at diagnosis and predict the course of longitudinal progression, which should benefit trial design and evaluation.


Analysis of the expression of toll-like receptors 2 and 4 and cytokine production during experimental Leishmania chagasi infection.
Toll-like receptors (TLRs) recognise pathogen-derived molecules and influence immunity to control parasite infections. This study aimed to evaluate the mRNA expression of TLRs 2 and 4, the expression and production of the cytokines interleukin (IL)-12, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-17, IL-10 and transforming growth factor (TGF)-β and the production of nitric oxide (NO) in the spleen of mice infected with Leishmania chagasi. It also aimed to evaluate any correlations between mRNA expression TLR2 and 4 and cytokines and NO production. Infection resulted in increased TLR2-4, IL-17, TNF-α and TGF-β mRNA expression during early infection, with decreased expression during late infection correlating with parasite load. IFN-γ and IL-12 mRNA expression decreased at the peak of parasitism. IL-10 mRNA expression increased throughout the entire time period analysed. Although TGF-β, TNF-α and IL-17 were highly produced during the initial phase of infection, IFN-γ and IL-12 exhibited high production during the final phase of infection. IL-10 and NO showed increased production throughout the evaluated time period. In the acute phase of infection, there was a positive correlation between TLR2-4, TNF-α, IL-17, NO, IL-10 and TGF-β expression and parasite load. During the chronic phase of infection, there was a positive correlation between TLR2-4, TNF-α, IL-17 and TGF-β expression and parasite load. Our data suggest that infection by L. chagasi resulted in modulation of TLRs 2 and 4 and cytokines.


Alternate splicing of mRNAs encoding human mast cell growth factor and localization of the gene to chromosome 12q22-q24.
Human mast cell growth factor (MGF) complementary DNAs (cDNAs) were cloned from HeLa cells using the polymerase chain reaction with oligonucleotides corresponding to murine and human MGF sequences. Sequencing of the cloned human MGF polymerase chain reaction products revealed two types of cDNA: a full length form corresponding in size to the murine cDNA, and an alternately spliced clone with a deletion of the sixth exon of the gene. Since membrane-bound MGF is predicted to be proteolytically cleaved within the sequences encoded by exon 6 to generate a soluble protein, this alternately spliced cDNA would likely encode a noncleavable, membrane-bound form of MGF. No difference in biological activity on human bone marrow cells was observed with recombinant, soluble forms of both types of human MGF protein. Our previous localization of the murine MGF gene to the Sl locus on chromosome 10 suggested (via conserved linkage groups) that the human MGF gene would be located on human chromosome 12. Therefore, rodent-human somatic cell hybrids with or without an entire human chromosome 12 and hybrids retaining partial 12 were tested by Southern blot analysis and used to show the presence of the human Mgf locus at chromosome region 12q. Chromosomal in situ hybridization localized the gene to 12q22-q24 in the region predicted by the comparative mapping of the murine Mgf/Sl locus.


B-vitamin status and concentrations of homocysteine in Austrian omnivores, vegetarians and vegans.
A vegetarian diet is considered to promote health and longevity and reduce the risk of cardiovascular diseases and cancer. However, a vegetarian diet may be deficient in some nutrients. Exclusion of animal products in vegetarian diets may affect the status of certain B-vitamins, and further cause the rise of plasma homocysteine concentration. The nutritional status of various B-vitamins (B(1), B(2), B(6), B(12), folic acid) and the concentration of homocysteine in blood plasma of omnivores (n = 40), vegetarians (n = 36) and vegans (n = 42) in Austria was evaluated. The evaluation was done using the functional parameters erythrocyte transketolase (ETK), glutathione reductase (EGR) and glutamic oxaloacetic transaminase (EGOT) activation coefficients. Enzyme activity was measured photometrically. The quantity of vitamins B(1), B(2) and B(6) in urine and the concentrations of vitamin B(6) and homocysteine in plasma were determined by HPLC methods with fluorescence detection. Plasma concentration of vitamin B(12) and folic acid were measured with radioimmunoassay. Most of the subjects showed a satisfying vitamin B(1) status. Vegans presented a significantly lower mean plasma vitamin B(12) concentration than omnivores and vegetarians and deficiency in 2.4% of the volunteers but the highest mean value of plasma folate among the investigated groups. A deficient status of folate was found in 18% of omnivores and in approximately 10% of vegans and vegetarians. The status of riboflavin is considered to be deficient in about 10% of omnivores and vegetarians and in over 30% of vegans. According to the activation coefficient of GOT, approximately one third of all subjects showed vitamin B(6) deficiency. Elevated homocysteine concentration in plasma was observed in 66% of the vegans and about 45-50% of the omnivores and vegetarians. Vegan subjects had significantly higher mean plasma homocysteine levels than omnivores. Thiamin and folate need not be a problem in a well-planned vegan diet. Vitamins B(12) and B(2) may need attention in the strict vegan diet, especially regarding elevated homocysteine levels in plasma. Pyridoxine status appeared to be independent of the diet.


Eosinophil-Cryptococcus neoformans interactions in vivo and in vitro.
Eosinophils are components of inflammatory responses to a variety of pathogens. Although a variety of beneficial and harmful functions have been ascribed to these cells, their role in protection against infectious agents remains uncertain. Previous studies have reported eosinophilic pneumonia in mice infected intratracheally with Cryptococcus neoformans. We confirmed this observation and studied the inflammatory response in the lung at day 14 by light and electron microscopy. Immunostaining for glucuronoxylomannan showed isolated cryptococci inside the eosinophilic cuffs. Eosinophils were found to be in close association with C. neoformans in vivo. Cryptococci were associated with eosinophils within eosinophilic perivascular cuffs, within granulomas, and lining the alveolar space. To further investigate this phenomenon in vitro, we isolated rat peritoneal eosinophils and studied cryptococcus-eosinophil interactions in the presence and absence of anti-capsular immunoglobulin G1 (IgG1) and IgE monoclonal antibody (MAb). Eosinophils phagocytosed C. neoformans only in the presence of specific antibody. Phagocytosis was rapid, and dense rings that appeared to consist of granule contents were formed around the organisms. Mast cells were observed to occasionally phagocytose C. neoformans in vitro in the presence of IgE MAb. Our observations suggest that eosinophils may be effector cells against C. neoformans.