A comparison of psychosocial factors related to dental anxiety among Turkish and Finnish pre-adolescents.
To assess psychological, individual and maternal factors related to dental anxiety among pre-adolescents in the different cultural settings of Turkey and Finland. A cross-sectional study of Finnish (n = 338) and Turkish (n = 611) pre-adolescents aged 10-12 years old by paired matching of questionnaires for the subjects and their mothers. These questionnaires, along with modified dental anxiety scales, were designed to collect data on the pre-adolescents' and their mothers' oral and dietary behaviours in relation to psychosocial factors. Multiple linear regression and factor analyses as well as descriptive and correlation statistics were applied. The Turkish pre-adolescents (TP) reported poorer dental health (50%) and more frequent gingival bleeding (26%) than did the Finnish pre-adolescents (FP) (26%, 0%), (p < 0.05). A similar trend occurred for reported toothache (70% and 17% for TP and FP respectively [p = 0.00]). Mean dental anxiety (DA) for TP and their mothers, and Finnish counterparts respectively were 9.64 +/- 6.01, 8.70 +/- 4.53 and 7.39 +/- 4.51, 6.02 +/- 5.50 (p < 0.05). Among TP, those with low DA reported high levels of toothbrushing and dietary self-efficacy (DSES) and self-esteem (p < 0.05); FP showed similar trends for DSES and self-esteem. Principal component analysis revealed that maternal DA and regular dental visits largely accounted for primary associations with DA among TP. Appreciating the impact of various psychosocial factors may provide a better understanding of DA among preadolescents, and thus may reduce dental avoidance and behavioural management problems for the overall improvement of young patients' oral health.


Distribution of genital Lactobacillus strains shared by female sex partners.
The prevalence of bacterial vaginosis among lesbians is high. We assessed whether unique Lactobacillus strains were shared by female sex partners. Cultures of vaginal and rectal specimens for detection of Lactobacillus organisms were performed for women who reported having had sex with women. Lactobacilli were identified on the basis of DNA homology and were typed and fingerprinted by repetitive element sequence-based polymerase chain reaction (rep-PCR). Of 237 women, Lactobacillus crispatus was detected in 98% and Lactobacillus gasseri in 21%. Detection of L. gasseri was associated with recent receptive digital-vaginal sex (P = .04) and increased bacterial vaginosis risk (odds ratio, 4.2; 95% confidence interval, 1.4-13.4). Of 31 couples monogamous for >3 months, rep-PCR fingerprints were identical in both members in 23 (77%).


The tree-dimensional structure of aspergillus niger pectin lyase B at 1.7-A resolution.
The three-dimensional structure of Aspergillus niger pectin lyase B (PLB) has been determined by crystallographic techniques at a resolution of 1.7 A. The model, with all 359 amino acids and 339 water molecules, refines to a final crystallographic R factor of 16.5%. The polypeptide backbone folds into a large right-handed cylinder, termed a parallel beta helix. Loops of various sizes and conformations protrude from the central helix and probably confer function. The largest loop of 53 residues folds into a small domain consisting of three antiparallel beta strands, one turn of an alpha helix, and one turn of a 3(10) helix. By comparison with the structure of Erwinia chrysanthemi pectate lyase C (PelC), the primary sequence alignment between the pectate and pectin lyase subfamilies has been corrected and the active site region for the pectin lyases deduced. The substrate-binding site in PLB is considerably less hydrophilic than the comparable PelC region and consists of an extensive network of highly conserved Trp and His residues. The PLB structure provides an atomic explanation for the lack of a catalytic requirement for Ca2+ in the pectin lyase family, in contrast to that found in the pectate lyase enzymes. Surprisingly, however, the PLB site analogous to the Ca2+ site in PelC is filled with a positive charge provided by a conserved Arg in the pectin lyases. The significance of the finding with regard to the enzymatic mechanism is discussed.


A fat-derived metabolite regulates a peptidergic feeding circuit in Drosophila.
Here, we show that the enzymatic cofactor tetrahydrobiopterin (BH4) inhibits feeding in Drosophila. BH4 biosynthesis requires the sequential action of the conserved enzymes Punch, Purple, and Sepiapterin Reductase (Sptr). Although we observe increased feeding upon loss of Punch and Purple in the adult fat body, loss of Sptr must occur in the brain. We found Sptr expression is required in four adult neurons that express neuropeptide F (NPF), the fly homologue of the vertebrate appetite regulator neuropeptide Y (NPY). As expected, feeding flies BH4 rescues the loss of Punch and Purple in the fat body and the loss of Sptr in NPF neurons. Mechanistically, we found BH4 deficiency reduces NPF staining, likely by promoting its release, while excess BH4 increases NPF accumulation without altering its expression. We thus show that, because of its physically distributed biosynthesis, BH4 acts as a fat-derived signal that induces satiety by inhibiting the activity of the NPF neurons.


Distribution and diversity of the enterococcal surface protein (esp) gene in animal hosts and the Pacific coast environment.
This study sought to evaluate the distribution of the enterococcal surface protein (esp) gene in Enterococcus faecium in the Pacific coast environment as well as the distribution and diversity of the gene in Northern California animal hosts. Over 150 environmental samples from the Pacific coast environment (sand, surf zone, fresh/estuarine, groundwater, and storm drain) were screened for the esp gene marker in E. faecium, and the marker was found in 37% of the environmental samples. We examined the host specificity of the gene by screening various avian and mammalian faecal samples, and found the esp gene to be widespread in nonhuman animal faeces. DNA sequence analysis performed on esp polymerase chain reaction amplicons revealed that esp gene sequences were not divergent between hosts. Our data confirm recent findings that the E. faecium variant of the esp gene is not human-specific. Our results suggest that the use of the esp gene for microbial source tracking applications may not be appropriate at all recreational beaches.


Moving droplets between closed and open microfluidic systems.
In electric-field-mediated droplet microfluidics, there are two distinct architectures - closed systems using parallel-plate electrodes and open systems using coplanar electrodes fabricated on an open substrate. An architecture combining both closed and open systems on a chip would facilitate many of the chemical and biological processes now envisioned for the laboratory on a chip. To accomplish such an integration requires a means to move droplets back and forth between the two. This paper presents an investigation of the requirements for such manipulation of both water and oil droplets. The required wetting conditions for a droplet to cross the open/closed boundary is revealed by a force balance analysis and predictions of this model are compared to experimental results. Water droplets can be moved between closed and open systems by electrowetting actuation; droplet detachment from the upper plate is facilitated by the use of beveled edge. The force model predicts that driving an oil droplet from a closed to an open structure requires an oleophobic surface. This prediction has been tested and confirmed using <100> silicon wafers made oleophobic by re-entrant microstructures etched into the surface.


Identification of nontraditional dental hygienists and their educational degrees. A strategy for baccalaureate program recruitment.
A pilot project was conducted to increase student applicant interest in a degree-completion program. Since applicants reported a preference for employment in nontraditional settings, and questioned whether further education was needed, a strategy was developed to identify dental hygienists in Michigan who were so employed, and to establish their educational levels. Through telephone interviews, nontraditional dental hygienists were asked to identify additional professionals that were known within the nontraditional network. Nontraditional employment was defined as any setting other than a private dental office, with job responsibilities not confined to patient care; dental hygiene educators were excluded. The interviews were conducted by one graduate student, one day a week, for 15 weeks. The interviewer generated a list of 44 Michigan dental hygienists employed in nontraditional job settings as defined in this study. In terms of educational levels, 23% had certificate/associate degrees (n = 10) as their highest degrees, 41% had bachelor's degrees (n = 18), and 36% had master's degrees (n = 16). Non-patient care was the job description for 93% of the nontraditional settings (n = 41). The implied need for baccalaureate and/or graduate education for nontraditional employment has positively affected recruitment for the degree-completion program. The relationship that has been established with the network of nontraditional dental hygienists has also improved recruitment efforts and enhanced the curriculum. This project suggests that the definition of nontraditional dental hygiene employment should not be limited to settings which involve patient care. Similar projects should be initiated in other states to improve interest in baccalaureate education.


Quantification of Fusarium solani f. sp. glycines isolates in soybean roots by colony-forming unit assays and real-time quantitative PCR.
Fusarium solani f. sp. glycines (FSG; syn. F. virguliforme Akoi, O'Donnell, Homma & Lattanzi) is a soil-borne fungus that infects soybean roots and causes sudden death syndrome (SDS), a widespread and destructive soybean disease. The goal of this study was to develop and use a real-time quantitative polymerase chain reaction (QPCR) assay to compare the accumulation of genomic DNA among 30 FSG isolates in inoculated soybean roots. Isolates differed significantly (P < or = 0.05) in their DNA accumulation on a susceptible soybean cultivar when detected and quantified using a FSG-specific probe/primers set derived from the sequences of the nuclear-encoded, mitochondrial small subunit ribosomal RNA gene. QPCR results that were normalized as the fold change over the sample collection times after inoculation were significantly (P < or = 0.001) correlated with the log(10) transformed colony-forming unit (CFU) values of FSG obtained from plating of inoculated ground roots on FSG semi-selective agar medium. Several isolates were identified that accumulated more FSG DNA and had higher CFU values than the reference isolate FSG1 (Mont-1). Compared to other isolates, FSG5 was the most aggressive root colonizer based on DNA accumulation and CFU values in infested roots. The described QPCR assay should provide more specificity, greater sensitivity, and less variability than alternatives to the culturing-dependent and time-consuming plating assays. Evaluation of isolate relative DNA differences on host plants using the QPCR approach provides useful information for evaluating isolates based on the extent and/or degree of colonization on soybean roots and for selecting isolates for breeding SDS-resistant soybean lines.


Prevalence of osteopenia and osteoporosis in Thai women.
To determine the age-specific and age-adjusted prevalence of osteopenia and osteoporosis in pre- and postmenopausal Thai women. This was a descriptive study of 1,935 Thai women ranging in age from 40 to 80 years, with randomly selected strata using multistage sampling and stratifying from six representative provinces of the country. After recruiting, all the women were interviewed by a well-trained interviewer using structured questionnaires. Bone mineral density (BMD) of lumbar spine 1-4 and nondominant hip was measured by a dual energy photon absorptiometer. The investigators were trained and standardized; inter- and intraobserver variations were measured periodically. Every BMD outcome was re-examined by the specialist. Age-specific prevalence of osteoporosis and osteopenia were determined using both Thai and Western standard BMD values. Age-adjusted prevalence of osteopenia and osteoporosis was calculated using the age-adjusted direct method. Using the Thai BMD reference, the age-specific prevalence of osteoporosis among Thai women rose progressively with increasing age to more than 50% after the age of 70. The age-adjusted prevalence of osteoporosis also rose progressively. It was 19.8%, 13.6%, and 10% for lumbar spine, femoral neck, and intertrochanteric. The age-adjusted prevalence of osteoporosis indicates the overall magnitude of that condition in the population or country. In our study, using a Western BMD reference resulted in a misleadingly high prevalence of osteoporosis in the population of Asian countries. It is important to calculate the age-adjusted prevalence of osteopenia and osteoporosis to address the overall magnitude of the problem in Thai women. This will allow us to predict the socioeconomic impact of preventable chronic conditions such as osteoporosis. The results obtained from this study are important data for public health policy: maximizing bone mass throughout life as well as detection of important risk factors is essential.


Expression of Nitric Oxide-Transporting Aquaporin-1 Is Controlled by KLF2 and Marks Non-Activated Endothelium In Vivo.
The flow-responsive transcription factor Krüppel-like factor 2 (KLF2) maintains an anti-coagulant, anti-inflammatory endothelium with sufficient nitric oxide (NO)-bioavailability. In this study, we aimed to explore, both in vitro and in human vascular tissue, expression of the NO-transporting transmembrane pore aquaporin-1 (AQP1) and its regulation by atheroprotective KLF2 and atherogenic inflammatory stimuli. In silico analysis of gene expression profiles from studies that assessed the effects of KLF2 overexpression in vitro and atherosclerosis in vivo on endothelial cells, identifies AQP1 as KLF2 downstream gene with elevated expression in the plaque-free vessel wall. Biomechanical and pharmaceutical induction of KLF2 in vitro is accompanied by induction of AQP1. Chromosome immunoprecipitation (CHIP) confirms binding of KLF2 to the AQP1 promoter. Inflammatory stimulation of endothelial cells leads to repression of AQP1 transcription, which is restrained by KLF2 overexpression. Immunohistochemistry reveals expression of aquaporin-1 in non-activated endothelium overlying macrophage-poor intimae, irrespective whether these intimae are characterized as being plaque-free or as containing advanced plaque. We conclude that AQP1 expression is subject to KLF2-mediated positive regulation by atheroprotective shear stress and is downregulated under inflammatory conditions both in vitro and in vivo. Thus, endothelial expression of AQP1 characterizes the atheroprotected, non-inflamed vessel wall. Our data provide support for a continuous role of KLF2 in stabilizing the vessel wall via co-temporal expression of eNOS and AQP1 both preceding and during the pathogenesis of atherosclerosis.


Effects of limit feeding a high-grain diet on puberty and reproductive performance of ewes.
In Exp. 1, 39 ewe lambs (initial BW 29 +/- .1 kg) were used in a 2 x 2 factorial arrangement with two breeds (Polypay and Targhee) and two diets (high-forage and high-grain) to examine the effects of restricting intake of a high-grain diet on onset of puberty and lamb performance. The experimental diets were fed from 13 to 40 wk of age. The high-forage diet was composed of 80% alfalfa cubes and 20% supplement. The high-grain diet was 90% concentrate and 10% alfalfa cubes. Feed intake of the diets was controlled so that lambs fed the high-grain diet had the same growth rate as those fed the high-forage diet. Plasma insulin concentration was higher (P < .001) for lambs fed the high-grain diet than for those fed the high-forage diet. However, no diet effect on age at puberty was observed. In Exp. 2, 88 mature Targhee ewes were used to determine the effects of limiting intake of a high-grain diet on pregnancy and number of lambs born per ewe. Intake of the high-grain diet was restricted to achieve similar ME intake between diets. There were no diet effects on number of lambs born per ewe or subsequent lamb performance. Ewes fed the high-forage diet had a lower (P < .001) weight gain than those fed the high-grain diet. It was concluded that restricting intake of high-grain diets is an effective alternative to feeding high-forage diets for meeting the nutrient requirements of ewe lambs and gestating ewes; however, no improvements on reproductive performance were observed.


Integrin β1-enriched extracellular vesicles mediate monocyte adhesion and promote liver inflammation in murine NASH.
Hepatic recruitment of monocyte-derived macrophages (MoMFs) contributes to the inflammatory response in non-alcoholic steatohepatitis (NASH). However, how hepatocyte lipotoxicity promotes MoMF inflammation is unclear. Here we demonstrate that lipotoxic hepatocyte-derived extracellular vesicles (LPC-EVs) are enriched with active integrin β1 (ITGβ1), which promotes monocyte adhesion and liver inflammation in murine NASH. Hepatocytes were treated with either vehicle or the toxic lipid mediator lysophosphatidylcholine (LPC); EVs were isolated from the conditioned media and subjected to proteomic analysis. C57BL/6J mice were fed a diet rich in fat, fructose, and cholesterol (FFC) to induce NASH. Mice were treated with anti-ITGβ1 neutralizing antibody (ITGβ1Ab) or control IgG isotype. Ingenuity® Pathway Analysis of the LPC-EV proteome indicated that ITG signaling is an overrepresented canonical pathway. Immunogold electron microscopy and nanoscale flow cytometry confirmed that LPC-EVs were enriched with activated ITGβ1. Furthermore, we showed that LPC treatment in hepatocytes activates ITGβ1 and mediates its endocytic trafficking and sorting into EVs. LPC-EVs enhanced monocyte adhesion to liver sinusoidal cells, as observed by shear stress adhesion assay. This adhesion was attenuated in the presence of ITGβ1Ab. FFC-fed, ITGβ1Ab-treated mice displayed reduced inflammation, defined by decreased hepatic infiltration and activation of proinflammatory MoMFs, as assessed by immunohistochemistry, mRNA expression, and flow cytometry. Likewise, mass cytometry by time-of-flight on intrahepatic leukocytes showed that ITGβ1Ab reduced levels of infiltrating proinflammatory monocytes. Furthermore, ITGβ1Ab treatment significantly ameliorated liver injury and fibrosis. Lipotoxic EVs mediate monocyte adhesion to LSECs mainly through an ITGβ1-dependent mechanism. ITGβ1Ab ameliorates diet-induced NASH in mice by reducing MoMF-driven inflammation, suggesting that blocking ITGβ1 is a potential anti-inflammatory therapeutic strategy in human NASH. Herein, we report that a cell adhesion molecule termed integrin β1 (ITGβ1) plays a key role in the progression of non-alcoholic steatohepatitis (NASH). ITGβ1 is released from hepatocytes under lipotoxic stress as a cargo of extracellular vesicles, and mediates monocyte adhesion to liver sinusoidal endothelial cells, which is an essential step in hepatic inflammation. In a mouse model of NASH, blocking ITGβ1 reduces liver inflammation, injury and fibrosis. Hence, ITGβ1 inhibition may serve as a new therapeutic strategy for NASH.


Prevalence, Correlates and Outcomes of Smoking in Pregnant Women with HIV: A National Observational Study in Italy.
Background: Few studies have evaluated in pregnant women with HIV the prevalence of smoking and its associations with maternal and neonatal outcomes. Objectives: to assess the prevalence of smoking among women with HIV in early pregnancy and the association between smoking and pregnancy outcomes in this particular population. Methods: We used data from a multicenter observational study to define the prevalence of smoking in women with HIV in early pregnancy, and the role of smoking status and intensity as risk factors for adverse maternal and neonatal outcomes. Main outcome measures were fetal growth restriction [FGR], preterm delivery [PD] and low birthweight [LB], evaluated in univariate and multivariate analyses. Results: The overall (2001-2018) prevalence of reported smoking (at least one cigarette/day) was 25.6% (792/3097), with a significant decrease in recent years (19.0% in 2013-2018). Women who smoked were less commonly African, had lower body mass index, older age, a longer history of HIV infection and higher CD4 counts. In univariate analyses, smokers were significantly more likely to have PD, LB, FGR and detectable HIV viral load at third trimester. Multivariable analyses confirmed for smokers a significantly higher risk of LB (adjusted odds ratio [AOR]: 1.69, 95%CI 1.22-2.34) and FGR (AOR 1.88, 95%CI 1.27-2.80), while the associations with detectable HIV and PD were not maintained. Conclusions: The common prevalence of smoking among pregnant women with HIV and its association with adverse outcomes indicates that smoking cessation programs in this population may have a significant impact on neonatal and maternal health.


Industrial requirements in food safety.
The principles of establishing industrial requirements in food safety are described, taking risk potentials all along the food chain into their respective account. Regulations will, in the future, lead to increased autocontrol in production. The rapid changes in food technology require constant adaptation to new problems, to keep the global quality of food at a high level. Regulatory authorities will, in the new European market, concentrate on enforcement of 'essential requirements' while industrialists will follow good manufacturing practices. Open dialogue between the latter, the former and the scientific community is highly desirable since mutual knowledge of the problem will help maintain a high level of food safety, for the benefit of everybody.


Prevalence and factors associated with family planning during COVID-19 pandemic in Bangladesh: A cross-sectional study.
The COVID-19 pandemic has negatively impacted health systems worldwide, including in Bangladesh, limiting access to family planning information (FP) and services. Unfortunately, the evidence on the factors linked to such disruption is limited, and no study has addressed the link among Bangladeshis. This study aimed to examine the socioeconomic, demographic, and other critical factors linked to the use of FP in the studied areas during the COVID-19 pandemic. The characteristics of the respondents were assessed using a cross-sectional questionnaire survey and descriptive statistics. The variables that were substantially linked with FP usage were identified using a Chi-square test. In addition, a multivariate logistic regression model was used to identify the parameters linked to FP in the study areas during the COVID-19 pandemic. The prevalence of FP use among currently married 15-49 years aged women was 36.03% suggesting a 23% (approximately) decrease compared to before pandemic data. Results also showed that 24.42% of the respondents were using oral contraceptive pills (OCP) which is lower than before pandemic data (61.7%). Multivariate regression analysis provided broader insight into the factors affecting FP use. Results showed that woman's age, education level of the respondents, working status of the household head, locality, reading a newspaper, FP workers' advice, currently using OCP, ever used OCP, husbands' supportive attitude towards OCP use, duration of the marriage, ever pregnant, the number of children and dead child were significantly associated with FP use in the study areas during COVID-19 pandemic. This study discusses unobserved factors that contributed to a reduction in FP use and identifies impediments to FP use in Bangladesh during the COVID-19 epidemic. This research further adds to our understanding of FP usage by revealing the scope of the COVID-19 pandemic's impact on FP use in Bangladesh's rural and urban areas.


Evaluation of the Performance of Helmet Prototypes Fabricated from Acrylonitrile Butadiene Styrene Composites Filled with Natural Resource.
The performance of helmet prototypes fabricated from acrylonitrile butadiene styrene composites filled with oil palm empty fruit bunch fibers was evaluated. The fibers were produced using a milling procedure, while the composites were fabricated using a single-screw extrusion. The physical characteristics of the produced fibers, which are water content, size, and density, were investigated. In addition, the mechanical properties of the produced helmets, including shock absorption, yield stress, frequency, and head injury criterion (HIC), were examined. The impact strength of the produced helmets increases with the rise of filler content. In addition, the helmets were also able to withstand a considerable pressure such that the transmitted pressure was far under the maximum value acceptable by the human skull. The present work also found that HICs exhibited by the investigated helmet prototypes fulfill all the practical guidelines as permitted by the Indonesian government. In terms of novelty, such innovation can be considered the first invention in Indonesia since the endorsement of the use of motorcycle helmets.


Nephrotic syndrome.
On the basis of observational studies, the most common cause of nephrotic syndrome in school-aged children is minimal change disease. On the basis of research evidence and consensus, corticosteroids are considered first-line therapy for treatment of nephrotic syndrome. On the basis of consensus, prednisone therapy should be initiated at doses of 60 mg/m2 per day (2 mg/kg per day) administered for 4 to 6 weeks, followed by 40 mg/m2 per dose (1.5 mg/kg) every other day for at least 6 to 8 weeks. On the basis of consensus and expert opinion, it is important to recognize and manage the complications that can arise in patients with nephrotic syndrome, such as dyslipidemia, infection, and thrombosis. On the basis of research evidence, consensus, and expert opinion, several alternative therapies have been observed to have variable efficacy in children with both corticosteroid-dependent and corticosteroid-resistant nephrotic syndrome, although caution must be exercised in the administration of these corticosteroid-sparing medications secondary to toxic adverse effects. On the basis of observational studies, the course of nephrotic syndrome in most patients is that of relapse and remission.


RAFT-Polymerization-Induced Self-Assembly and Reorganizations: Ultrahigh-Molecular-Weight Polymer and Morphology-Tunable Micro-/Nanoparticles in One Pot.
A one-pot method is introduced for the successful synthesis of narrow-distributed (Đ = 1.22) vinyl polymer with both ultrahigh molecular weight (UHMW) (M w = 1.31 × 106 g mol-1 ) and micro-/nanomorphology under mild conditions. The method involves the following four stages: homogeneous polymerization, polymerization-induced self-assembly (PISA), PISA and reorganization, and PISA and multiple reorganizations. The key points to the production of UHMW polystyrene are to minimize radical termination by segregating radicals in different nanoreactors and to ensure sufficient chain propagation by promoting further reorganizations of these reactors in situ. This method therefore endows polymeric materials with the outstanding properties of both UHMW and tunable micro-/nanoparticles under mild conditions in one pot.


SUMOylation of the Hyperpolarization-Activated Cyclic Nucleotide-Gated Channel 2 Increases Surface Expression and the Maximal Conductance of the Hyperpolarization-Activated Current.
Small Ubiquitin-like Modifier (SUMO) is a ∼10 kDa peptide that can be post-translationally added to a lysine (K) on a target protein to facilitate protein-protein interactions. Recent studies have found that SUMOylation can be regulated in an activity-dependent manner and that ion channel SUMOylation can alter the biophysical properties and surface expression of the channel. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channel surface expression can be regulated in an activity-dependent manner through unknown processes. We hypothesized that SUMOylation might influence the surface expression of HCN2 channels. In this manuscript, we show that HCN2 channels are SUMOylated in the mouse brain. Baseline levels of SUMOylation were also observed for a GFP-tagged HCN2 channel stably expressed in Human embryonic kidney (Hek) cells. Elevating GFP-HCN2 channel SUMOylation above baseline in Hek cells led to an increase in surface expression that augmented the hyperpolarization-activated current (Ih) mediated by these channels. Increased SUMOylation did not alter Ih voltage-dependence or kinetics of activation. There are five predicted intracellular SUMOylation sites on HCN2. Site-directed mutagenesis indicated that more than one K on the GFP-HCN2 channel was SUMOylated. Enhancing SUMOylation at one of the five predicted sites, K669, led to the increase in surface expression and IhGmax. The role of SUMOylation at additional sites is currently unknown. The SUMOylation site at K669 is also conserved in HCN1 channels. Aberrant SUMOylation has been linked to neurological diseases that also display alterations in HCN1 and HCN2 channel expression, such as seizures and Parkinson's disease. This work is the first report that HCN channels can be SUMOylated and that this can regulate surface expression and Ih.


First insights into the entry process of hyperthermophilic archaeal viruses.
A decisive step in a virus infection cycle is the recognition of a specific receptor present on the host cell surface, subsequently leading to the delivery of the viral genome into the cell interior. Until now, the early stages of infection have not been thoroughly investigated for any virus infecting hyperthermophilic archaea. Here, we present the first study focusing on the primary interactions between the archaeal rod-shaped virus Sulfolobus islandicus rod-shaped virus 2 (SIRV2) (family Rudiviridae) and its hyperthermoacidophilic host, S. islandicus. We show that SIRV2 adsorption is very rapid, with the majority of virions being irreversibly bound to the host cell within 1 min. We utilized transmission electron microscopy and whole-cell electron cryotomography to demonstrate that SIRV2 virions specifically recognize the tips of pilus-like filaments, which are highly abundant on the host cell surface. Following the initial binding, the viral particles are found attached to the sides of the filaments, suggesting a movement along these appendages toward the cell surface. Finally, we also show that SIRV2 establishes superinfection exclusion, a phenomenon not previously described for archaeal viruses.


[A comparative analysis of proteins from persistent and antigen-defective strains of tick-borne encephalitis virus].
Structural (E) and three nonstructural (NS1, NS3, and NS5) proteins of persistent and antigen-defective strains of tick-borne encephalitis virus are compared by immunoblotting with monoclonal antibodies to the corresponding proteins of strain Sofyin. Appreciable phenotypical differences were revealed between antigen-defective strains, but no immunological modifications as concerns the studied antigenic structures. The size of the reference NS3 protein differed from that of the persistent virus strains. Similar proteins detected in the antigenic preparations of strain Zausaev may have notable modifications.


Arrays of 3D double-network hydrogels for the high-throughput discovery of materials with enhanced physical and biological properties.
Synthetic hydrogels are attractive biomaterials due to their similarity to natural tissues and their chemical tunability, which can impart abilities to respond to environmental cues, e.g. temperature, pH and light. The mechanical properties of hydrogels can be enhanced by the generation of a double-network. Here, we report the development of an array platform that allows the macroscopic synthesis of up to 80 single- and double-network hydrogels on a single microscope slide. This new platform allows for the screening of hydrogels as 3D features in a high-throughput format with the added dimension of significant control over the compressive and tensile properties of the materials, thus widening their potential application. The platform is adaptable to allow different hydrogels to be generated, with the potential ability to tune and alter the first and second network, and represents an exciting tool in material and biomaterial discovery.


Is gastroesophageal reflux contribute to the development chronic cough by triggering pulmonary fibrosis.
Previous studies have shown that the prevalence of abnormal acid reflux in fibrotic lung disease patients is high, and in particular, patients with secondary pulmonary fibrosis show higher esophageal acid exposure than normal controls. There are also some findings that, in patients with pathological reflux, pulmonary fibrosis may develop. The aim of this study is to investigate if pulmonary fibrosis is involved in the pathogenesis of chronic cough due to Gastroesophageal Reflux. A prospective study was performed in twenty-one patients with chronic cough due to gastroesophageal reflux who was diagnosed as reflux esophagitis by upper gastrointestinal endoscopy, histology, and in ten healthy controls without GER or any lung disease. All participitants underwent laryngoscopic examination and gastroesophageal scintigraphy with late lung imaging. Bronchoalveolar lavage fluid total and differential cell counts, T and B cell subsets, and the concentrations of IL- 1β and TNF-α were measured. Reflux extending into the proximal esophagus was noted in 52.5%, and posterior laryngitis was present in 90.5% of the patients. No evidence of pulmonary aspiration was noted in the patients with reflux on scintigraphic examination. No significant difference was found between the GER and control groups in terms of cellular content, IL-1β and TNF-α levels or mean T cell subsets and B cell counts in bronchoalveolar lavage fluid. Forced expiratory volume in one second, forced vital capacity FEV1/FVC, total lung capacity, and carbon monoxide diffusion capacity values were within normal limits in the gastroesophageal reflux group. Our findings do not support the hypothesis that gastroesophageal reflux leads to chronic cough by triggering alveolar epithelial injury and subsequent pulmonary fibrosis.


Deployment experiences and mental health problems as predictors of post-deployment unemployment length: a prospective, register-based study among Danish soldiers.
To test responses of formerly deployed soldiers (FDS) to a questionnaire on deployment experiences in combination with screening levels of post-traumatic stress disorder (PTSD) and depression approximately 6 months after homecoming as predictors of the subsequent probability of gaining employment when unemployed within 5 years post-deployment. Danish FDS responders (n=3935) and non-responders (n=3046) to a 6-month post-deployment screening questionnaire after returning from a first-ever deployment to Kosovo, Iraq or Afghanistan (2002 to 2012) were included in the study and followed in public registers from 6 months to 5 years post-deployment. We tested Cox regression models including deployment experiences (1a), screening levels of PTSD and depressive symptoms (1b), and their combination (2) for FDS responders. For all FDS, a secondary model included a measure on whether they responded to the questionnaire (3). Neither the deployment experiences (1a) of exposure to danger and combat (HR=1.00, 95% CI=0.97 to 1.03) and witnessing consequences of war (HR=1.01, 95% CI=0.96 to 1.06), or the screening levels (1b) of PTSD (HR=1.06, 95% CI=0.84 to 1.33) and depressive symptoms (HR=0.82, 95% CI=0.64 to 1.06) were significant predictors of transitioning from unemployment to employment. Similar results were found for the combined model (2). A tendency among non-respondents (3) to have a lower probability of transitioning from unemployment to employment was found (HR=0.90, 95% CI=0.81 to 1.00). Deployment experiences, PTSD and depressive symptoms, as measured at 6-month screening questionnaire, did not predict differences in the probability of gaining employment when unemployed within 5 years post-deployment. However, the findings suggest that those with the least probability of transitioning from unemployment to employment can be found among the non-responders to the post-deployment screening questionnaire.


Combined Resistance and Plyometric Training Is More Effective Than Plyometric Training Alone for Improving Physical Fitness of Pubertal Soccer Players.
The purpose of this study was to compare the effects of combined resistance and plyometric/sprint training with plyometric/sprint training or typical soccer training alone on muscle strength and power, speed, change-of-direction ability in young soccer players. Thirty-one young (14.5 ± 0.52 years; tanner stage 3-4) soccer players were randomly assigned to either a combined- (COMB, n = 14), plyometric-training (PLYO, n = 9) or an active control group (CONT, n = 8). Two training sessions were added to the regular soccer training consisting of one session of light-load high-velocity resistance exercises combined with one session of plyometric/sprint training (COMB), two sessions of plyometric/sprint training (PLYO) or two soccer training sessions (CONT). Training volume was similar between the experimental groups. Before and after 7-weeks of training, peak torque, as well as absolute and relative (normalized to torque; RTD r ) rate of torque development (RTD) during maximal voluntary isometric contraction of the knee extensors (KE) were monitored at time intervals from the onset of contraction to 200 ms. Jump height, sprinting speed at 5, 10, 20-m and change-of-direction ability performances were also assessed. There were no significant between-group baseline differences. Both COMB and PLYO significantly increased their jump height (Δ14.3%; ES = 0.94; Δ12.1%; ES = 0.54, respectively) and RTD at mid to late phases but with greater within effect sizes in COMB in comparison with PLYO. However, significant increases in peak torque (Δ16.9%; p < 0.001; ES = 0.58), RTD (Δ44.3%; ES = 0.71), RTD r (Δ27.3%; ES = 0.62) and sprint performance at 5-m (Δ-4.7%; p < 0.001; ES = 0.73) were found in COMB without any significant pre-to-post change in PLYO and CONT groups. Our results suggest that COMB is more effective than PLYO or CONT for enhancing strength, sprint and jump performances.


Lead impact on nutrition, energy reserves, respiration and stress protein (hsp 70) level in Porcellio scaber (Isopoda) populations differently preconditioned in their habitats.
The impact of lead on food consumption, energy metabolism and the stress protein (hsp 70) level was investigated in the woodlouse Porcellio scaber (Isopoda), a common representative of the saprophagous soil macrofauna. To examine possible acclimation or tolerance to lead in woodlice from a contaminated habitat, animals of two populations, one deriving from a lead-contaminated artillery range and one from an uncontaminated control stand, were exposed to a series of lead concentrations under otherwise constant laboratory conditions for a maximum of 80 days. The applied lead concentrations (at a maximum 7945 mg/kg food dry wt) did not have any significant quantitative effect on the food consumption of the isopods, although the population pre-exposed in the artillery range showed a tendency toward a higher food uptake than the control population. After 80 days of exposure, both populations showed an equal trend toward increasing their respiration as lead concentrations, that they had been fed on, were increased. Accordingly, the glycogen content of the body, in both populations, was elevated with increasing lead concentrations in the food. This effect was more pronounced in the pre-exposed isopod population than in the one from the control stand. The non-pre-exposed isopods showed a general tendency toward a lower protein content of their bodies than the pre-exposed ones, although no effect of the lead on this parameter could be statistically proven. The ability of the artillery range isopods to synthesise stress proteins (hsp 70) in response to lead contamination decreased at much lower lead concentrations in their food than in the non-pre-exposed control population, even though the artillery range isopods seemed to be equally or even slightly better equipped with energy storage products. Even though the better nutrient status of these animals might refer to some lead tolerance of the pre-exposed population, the stress protein data suggest that a metal-resistant Porcellio population did not evolve in this lead-contaminated site.


"You Don't Feel": The Experience of Youth Benzodiazepine Misuse in Ireland.
There are negative effects to inappropriate use of benzodiazepines, yet the percentage of young people in Ireland experimenting with benzodiazepines has increased. There is a paucity of research about why Irish young people misuse benzodiazepines. In this study, people between 18 and 25 years attending substance misuse services in the south of Ireland (N = 13) were interviewed in a semi-structured style between June 2012 and April 2013. Content analysis was performed. The main motivations for benzodiazepine misuse were to self-regulate negative emotions and to induce dissociation from their environment. Interviewees also described the consequences of benzodiazepine misuse, such as disengagement from family relationships and other protective environments such as school and sports clubs. The consequences of chronic misuse were discussed, such as the compulsion to take more benzodiazepines despite experiencing severe side-effects. The incidence of paradoxical aggression on benzodiazepines is also explored. Education about benzodiazepines and their risks to young people, families, and the public may reduce benzodiazepine misuse. Future research on the role of trauma and mental health in young people's substance misuse is needed.


Investigating the influence of intermittent and continuous mechanical loading on skin through non-invasive sampling of IL-1α.
Pressure ulcers (PUs) are a major burden to both patients, carers and the healthcare system. It is therefore important to identify patients at risk and detect pressure ulcers at an early stage of their development. The pro-inflammatory cytokine IL-1α is a promising indicator of tissue damage. The aim of this study was to compare the temporal skin response, by means of IL-1α expression, to different loading regimens and to investigate the presence of individual variability. The sacrum of eleven healthy volunteers was subjected to two different loading protocols. After a baseline measurement, the left and right side of the sacrum were subjected to continuous and intermittent loading regimen, respectively, at a pressure of 100 mmHg. Data was collected every 20 min, allowing for a total experimental time of 140 min. Sebum, collected at ambient conditions using Sebutape, was analyzed for the pro-inflammatory cytokine IL-1α. Most robust results were obtained using a baseline normalization approach on individual data. The IL-1α level significantly changed upon load application and removal (p<0.05) for both loading regimens. Highest IL-1α ratio increase, 3.7-fold, was observed for 1 h continuous loading. During the refractory periods for both loading regimen the IL-1α levels were still found to be up-regulated compared to baseline (p<0.05). The IL-1α level increased significantly for the two initial loading periods (p<0.05), but stabilized during the final loading period for both loading regimens. Large individual variability in IL-1α ratio was observed in the responses, with median values of 1.91 (range 1.49-3.08), and 2.52 (range 1.96-4.29), for intermittent and continuous loading, respectively, although the differences were not statistically significant. Cluster analysis revealed the presence of two distinct sub-populations, with either a low or high response to the applied loading regimen. The measurement after the first loading period proved to be representative for the subsequent measurements on each site. This study revealed that trends in normalized IL-1α provided an early indicator for tissue status following periods of mechanical loading and refractory unloaded conditions. Additionally, the observed individual variability in the response potentially identifies patients at risk of developing PUs.


Perceptual-Motor and Perceptual-Cognitive Skill Acquisition in Soccer: A Systematic Review on the Influence of Practice Design and Coaching Behavior.
Facilitating players' skill acquisition is a major challenge within sport coaches' work which should be supported by evidence-based recommendations outlining the most effective practice and coaching methods. This systematic review aimed at accumulating empirical knowledge on the influence of practice design and coaching behavior on perceptual-motor and perceptual-cognitive skill acquisition in soccer. A systematic search was carried out according to the PRISMA guidelines across the databases SPORTDiscus, PsycInfo, MEDLINE, and Web of Science to identify soccer-specific intervention studies conducted in applied experimental settings (search date: 22nd November 2020). The systematic search yielded 8,295 distinct hits which underwent an independent screening process. Finally, 34 eligible articles, comprising of 35 individual studies, were identified and reviewed regarding their theoretical frameworks, methodological approaches and quality, as well as the interventions' effectiveness. These studies were classified into the following two groups: Eighteen studies investigated the theory-driven instructional approaches Differential Learning, Teaching Games for Understanding, and Non-linear Pedagogy. Another seventeen studies, most of them not grounded within a theoretical framework, examined specific aspects of practice task design or coaches' instructions. The Downs and Black checklist and the Template for Intervention Description and Replication were applied to assess the quality in reporting, risk of bias, and the quality of interventions' description. Based on these assessments, the included research was of moderate quality, however, with large differences across individual studies. The quantitative synthesis of results revealed empirical support for the effectiveness of coaching methodologies aiming at encouraging players' self-exploration within representative scenarios to promote technical and tactical skills. Nevertheless, "traditional" repetition-based approaches also achieved improvements with respect to players' technical outcomes, yet, their impact on match-play performance remains widely unexplored. In the light of the large methodological heterogeneity of the included studies (e.g., outcomes or control groups' practice activities), the presented results need to be interpreted by taking the respective intervention characteristics into account. Overall, the current evidence needs to be extended by theory-driven, high-quality studies within controlled experimental designs to allow more consolidated and evidence-based recommendations for coaches' work.


The relative timing of mutations in a breast cancer genome.
Many tumors have highly rearranged genomes, but a major unknown is the relative importance and timing of genome rearrangements compared to sequence-level mutation. Chromosome instability might arise early, be a late event contributing little to cancer development, or happen as a single catastrophic event. Another unknown is which of the point mutations and rearrangements are selected. To address these questions we show, using the breast cancer cell line HCC1187 as a model, that we can reconstruct the likely history of a breast cancer genome. We assembled probably the most complete map to date of a cancer genome, by combining molecular cytogenetic analysis with sequence data. In particular, we assigned most sequence-level mutations to individual chromosomes by sequencing of flow sorted chromosomes. The parent of origin of each chromosome was assigned from SNP arrays. We were then able to classify most of the mutations as earlier or later according to whether they occurred before or after a landmark event in the evolution of the genome, endoreduplication (duplication of its entire genome). Genome rearrangements and sequence-level mutations were fairly evenly divided earlier and later, suggesting that genetic instability was relatively constant throughout the life of this tumor, and chromosome instability was not a late event. Mutations that caused chromosome instability would be in the earlier set. Strikingly, the great majority of inactivating mutations and in-frame gene fusions happened earlier. The non-random timing of some of the mutations may be evidence that they were selected.


[Nosological profile of sudden cardiac death in athletes].
The review of the literature on sudden cardiac death in sport considers its main causes. The clinical and morphological characteristics are represented by the cardiac structure and electrophysiological pathology that causes sudden death in athletes.


The pathway of pepsin-catalysed transpeptidation. Evidence for the reactive species being the anion of the acceptor molecule.
1. The inhibition of pepsin-catalysed hydrolysis of N-acetyl-l-phenylalanyl-l-phenylalanylglycine by the acyl product and product analogues was studied at pH4.3. 2. The acyl product, N-acetyl-l-phenylalanine, gives rise to linear competitive inhibition at pH4.3, whereas at pH2.1 it shows linear non-competitive behaviour. 3. The extent of transpeptidation to N-acetyl-l-[(3)H]phenylalanine during the pepsin-catalysed hydrolysis of N-acetyl-l-phenylalanyl-l-phenylalanyl-glycine is significant at pH4.7, but is undetectable at pH1.3. 4. Both the inhibition and transpeptidation experiments are consistent with the anion of the acceptor molecule being the substrate in pepsin-catalysed transpeptidation. This conclusion supports the formulation of pepsin-catalysed reactions put forward by Knowles et al. (1970).


Indwelling pleural catheters: evidence for and management.
The incidence of pleural disease is increasing, and pleural medicine is increasingly recognised as a subspecialty within respiratory medicine. This often requires additional training time. Once underresearched, the last decade has seen an explosion in evidence related to the management of pleural disease. One of the cornerstones of pleural effusion management is the insertion of an indwelling pleural catheter. This allows patient-centred outpatient management and now has a robust evidence base. This article summarises evidence as well serves as a practical guide to the management of any complications related to an indwelling pleural catheter that might present on an acute take.


Highly diversified coronaviruses in neotropical bats.
Bats host a broad diversity of coronaviruses (CoVs), including close relatives of human pathogens. There is only limited data on neotropical bat CoVs. We analysed faecal, blood and intestine specimens from 1562 bats sampled in Costa Rica, Panama, Ecuador and Brazil for CoVs by broad-range PCR. CoV RNA was detected in 50 bats representing nine different species, both frugivorous and insectivorous. These bat CoVs were unrelated to known human or animal pathogens, indicating an absence of recent zoonotic spill-over events. Based on RNA-dependent RNA polymerase (RdRp)-based grouping units (RGUs) as a surrogate for CoV species identification, the 50 viruses represented five different alphacoronavirus RGUs and two betacoronavirus RGUs. Closely related alphacoronaviruses were detected in Carollia perspicillata and C. brevicauda across a geographical distance exceeding 5600 km. Our study expands the knowledge on CoV diversity in neotropical bats and emphasizes the association of distinct CoVs and bat host genera.


[Interaction of hypoxia and haemostasis--hypoxia as a prothrombotic factor at high altitude?].
For an extended period of time various research projects have been conducted on the relationship of hypoxia and haemostasis. The enclosed article contains the conclusion to which extent lack of oxygen can activate the coagulation system and induce a prothrombotic state. The majority of studies proved a shortening of coagulation times during acute exposure to hypoxia, whereas activated parameters of coagulation and fibrinolysis like prothrombin fragment F1+2 as well as thrombin-antithrombin III complexes and D-dimer remained mostly unmodified. It is suggested that a prolonged sojourn at high altitudes could lead to activation of the coagulation system through an increase of haematocrit and blood viscosity. Recently it was proven that people living at high altitudes show an enhanced risk of stroke incidents. The significance of the change in haemostasis on that outcome has not yet been part of the research. However, it has been proven that the activity of the coagulation system does not play a pathophysiological part in the development of acute mountain sickness and high altitude pulmonary edema. Recent studies also demonstrated that moderate hypoxia during long haul flights may not be the main trigger in inducing deep vein thrombosis in passengers.


Treatment pattern changes in high-risk patients newly initiated on statin monotherapy in a managed care setting.
For high-risk patients who do not achieve guideline-recommended LDL-C levels, more intensive treatment including statin-uptitration to higher doses or potency, as well as combination therapy may be considered. A better understanding of statin treatment patterns in real-world clinical practice may contribute to improved lipid-lowering management in these patients. We determined treatment pattern changes among patients with high risk of cardiovascular disease who were not at low-density lipoprotein cholesterol (LDL-C) goal on statin monotherapy. Treatment pattern changes were evaluated among patients newly initiated on statins between January 1, 2006, and August 31, 2009, in the HealthCore Integrated Research Database. Rates and mean time to first and second treatment changes were examined in patients with claims for coronary heart disease (CHD), atherosclerotic vascular disease (AVD), and diabetes mellitus during 12 months before index, who were not at LDL-C <70 mg/dL at their first-eligible LDL-C test (≥ 4 weeks after index). Therapy change was assessed for 12 months after the LDL-C result. Of 11,473 eligible subjects, 61.3% had diabetes, 26.6% had CHD and AVD, and 12.1% had CHD and AVD and diabetes. At index, patients were prescribed medium-potency levels of statins, including simvastatin (44.7%), atorvastatin (31.5%), and other statins (23.8%). Mean ± SD LDL-C before statin initiation was 138 ± 34 mg/dL, and at the first-eligible LDL-C result after index, it was 101 ± 25 mg/dL. During follow-up, 7444 subjects (64.9%) experienced a first treatment change, with mean time to change of 93.8 ± 92 days, whereas 4029 (36.1%) had no treatment change. Discontinuation of index therapy occurred in 46.9% of subjects and medication switches or titration in 18.0% (index statin titration, switch to other statins, other lipid-lowering therapies [LLT], including ezetimibe). Of the discontinuers, 27.4% restarted LLT. Of subjects with a first treatment change who did not discontinue, 48.9% experienced a second therapy change. Results were similar between the 3 high-risk groups. In this managed-care setting, among patients with high risk of cardiovascular disease who were not at LDL-C goal, statins were usually started at medium-potency doses without being titrated up, whereas nearly one-half had a discontinuation of LLT within 12 months. These treatment patterns indicate the need for better patient and provider education as well as other system-wide modifications to improve medication adherence.


A UK national audit of hereditary and acquired angioedema.
Hereditary angioedema (HAE) and acquired angioedema (AAE) are rare life-threatening conditions caused by deficiency of C1 inhibitor (C1INH). Both are characterized by recurrent unpredictable episodes of mucosal swelling involving three main areas: the skin, gastrointestinal tract and larynx. Swelling in the gastrointestinal tract results in abdominal pain and vomiting, while swelling in the larynx may be fatal. There are limited UK data on these patients to help improve practice and understand more clearly the burden of disease. An audit tool was designed, informed by the published UK consensus document and clinical practice, and sent to clinicians involved in the care of HAE patients through a number of national organizations. Data sets on 376 patients were received from 14 centres in England, Scotland and Wales. There were 55 deaths from HAE in 33 families, emphasizing the potentially lethal nature of this disease. These data also show that there is a significant diagnostic delay of on average 10 years for type I HAE, 18 years for type II HAE and 5 years for AAE. For HAE the average annual frequency of swellings per patient affecting the periphery was eight, abdomen 5 and airway 0·5, with wide individual variation. The impact on quality of life was rated as moderate or severe by 37% of adult patients. The audit has helped to define the burden of disease in the UK and has aided planning new treatments for UK patients.


Induction of RANTES/CCL5 by herpes simplex virus is regulated by nuclear factor kappa B and interferon regulatory factor 3.
Chemokines regulate migration of leukocytes to sites of infection. In this work, we have shown that the chemokine RANTES/CCL5 is produced after herpes simplex virus (HSV) infection of macrophages and fibroblasts and provide data on the underlying molecular mechanism. Reporter gene assays showed HSV-induced RANTES production to be regulated at the transcriptional level. Expression of RANTES was blocked by N-tosyl-L-phenylalanine, an inhibitor of the nuclear factor kappa B (NF-kappa B) pathway and also in cell lines stably expressing a dominant-negative mutant of I kappa B kinase beta. Cell lines stably expressing a dominant-negative mutant of interferon regulatory factor 3 (IRF-3) also produced dramatically decreased levels of RANTES after infection compared with the control cell line. In contrast, overexpression of dominant-negative p38 and also treatment with SB203580, an inhibitor of p38, did not significantly affect expression of RANTES. Taken together, these data suggest that HSV induces transcriptional activation of the RANTES gene through the transcription factors NF-kappa B and IRF-3.


Psammomatous Squamous Cell Carcinoma of the Skin.
Psammoma bodies (PBs) are concentric, lamellated calcifications commonly observed in malignancies such as papillary thyroid carcinoma and serous carcinoma of the ovary in which they may serve prognostic value. PBs are rare in cutaneous squamous cell carcinoma (cSCC), with only 1 previously reported case. Here, we present 3 cases of cSCC displaying PBs. One case occurred in the setting of end-stage renal disease, whereas the other 2 cases were in patients who did not have comorbid conditions that might predispose to hypercalcemia and dystrophic calcification. All 3 tumors demonstrated classic immunophenotypic findings of cSCC. Our findings indicate that PBs are a rare but recurrent phenomenon in cSCC, with unknown prognostic significance. The potential for PB formation in cSCC should be kept in mind, as this may represent a diagnostic pitfall in tumors with limited sampling or unusual morphologies.


Evaluation of resting state networks in patients with gliomas: connectivity changes in the unaffected side and its relation to cognitive function.
In this study, we investigated changes in resting state networks (RSNs) in patients with gliomas located in the left hemisphere and its relation to cognitive function. We hypothesized that long distance connection, especially between hemispheres, would be affected by the presence of the tumor. We further hypothesized that these changes would correlate with, or reflect cognitive changes observed in patients with gliomas. Resting state functional MRI datasets from 12 patients and 12 healthy controls were used in the analysis. The tumor's effect on three well-known RSNs including the default mode network (DMN), executive control network (ECN), and salience network (SN) identified using independent component analysis were investigated using dual regression analysis. Scores of neuropsychometric testing (WAIS-III and WMS-R) were also compared. Compared to the healthy control group, the patient group showed significant decrease in functional connectivity in the right angular gyrus/inferior parietal lobe of the ventral DMN and in the dorsolateral prefrontal cortex of the left ECN, whereas a significant increase in connectivity in the right ECN was observed in the right parietal lobe. Changes in connectivity in the right ECN correlated with spatial memory, while that on the left ECN correlated with attention. Connectivity changes in the ventral DMN correlated with attention, working memory, full IQ, and verbal IQ measures. Although the tumors were localized in the left side of the brain, changes in connectivity were observed in the contralateral side. Moreover, these changes correlated with some aspects of cognitive function indicating that patients with gliomas may undergo cognitive changes even in the absence of or before the onset of major symptoms. Evaluation of resting state networks could be helpful in advancing our hodological understanding of brain function in glioma cases.


Cystatin, a protein inhibitor of cysteine proteinases. Improved purification from egg white, characterization, and detection in chicken serum.
The protein from chicken egg white that inhibits cysteine proteinases, and has been named 'cystatin', was purified by ovomucin precipitation, affinity chromatography on carboxymethylpapain-Sepharose and chromatofocusing. The final purification step separated two major forms of the protein (pI 6.5 and 5.6), with a total recovery of about 20% from egg white. By use of affinity chromatography and immunodiffusion it was shown that the inhibitor is also present at low concentrations in the serum of male and female chickens. Tryptic peptide maps of the separated forms 1 and 2 of egg-white cystatin were closely similar, and each form had the N-terminal sequence Ser-Glx-Asx. The two forms showed complete immunological identity, and neither contained carbohydrate. Ki values for the inhibition of cysteine proteinases were as follows: papain (less than 1 X 10(-11)M), cathepsin B (8 X 10(-10)M), cathepsin H (about 2 X 10(-8)M) and cathepsin L (about 3 X 10(-12)M). Some other cysteine proteinases, and several non-cysteine proteinases, were found not to be significantly inhibited by cystatin. The inhibition of the exopeptidase dipeptidyl peptidase I by cystatin was confirmed and the Ki found to be 2 X 10(-10)M. Inhibitor complexes with active cysteine proteinases and the inactive derivatives formed by treatment with iodoacetate, E-64 [L-trans-epoxysuccinylleucylamido(4-guanidino)butane] and benzyloxycarbonylphenylalanylalanyldiazomethane were demonstrated by isoelectric focusing and cation-exchange chromatography. The complexes dissociated in sodium dodecyl sulphate/polyacrylamide-gel electrophoresis (with or without reduction) with no sign of fragmentation of the inhibitor. Cystatin was found not to contain a free thiol group, and there was no indication that disulphide exchange plays any part in the mechanism of inhibition.


[Adhesion pili in Yersinia pestis].
Y. pestis cells cultivated at 37 degrees C are capable of agglutinating red blood cells of some animals, which is due to the appearance of pili. The adhesion pili consist of protein subunits with a molecular weight of the order of 12000 daltons, their isoionic point being at pH 4.7. The reaction of hemagglutination was inhibited by the mixture of ganglyosides, while the preliminary treatment of red blood cells with neuraminidases increased its effectiveness. The pili are supposed to take part in the expression of virulence.


Total elbow arthroplasty following traumas: mid-term results.
Total elbow arthroplasty (TEA) has been normally indicated for chronic inflammatory arthropathy. In the last decades this surgery has also evolved as an acceptable procedure following traumatic lesions of this joint. The aim of this study was to analyze the mid-term outcomes of TEA for the treatment of complex fractures and non-unions of the elbow in selected patients. Between May 2002 and December 2011, 34 patients with these indications were surgically treated with TEA and divided in two groups. Group 1 included 16 cases of fractures and Group 2 18 of non-union. All patients were clinically assessed using the Mayo Elbow Performance Score (MEPS). A statistical analysis was performed in order to investigate the outcomes of these patients and compare the two groups. Similar results were observed in both groups between operated and non-operated arms. No differences in clinical results were documented between groups. TEA following traumas can be considered as a valid treatment in old patients in complex fractures of the elbow region with low functional demands and in non-unions. Outcomes are influenced by the collaborative capacities of the patients.


On predicting multi-route and multimedia residential exposure to chlorpyrifos and diazinon.
This paper formulates regression models and examines their ability to associate exposures to chlorpyrifos and diazinon in residences with information obtained from questionnaires and environmental sampling of the National Human Exposure Assessment Survey Arizona (NHEXAS-AZ) database. A knowledge-based list of 29 potential exposure determinants was assembled from information obtained from six questionnaires administered in the course of the study. This list was used to select the independent variables of each model statistically and electronically. Depending on the data type of dependent and independent variables, four classes of regression models were developed to determine desired associations. Route-specific exposures were estimated using the indirect method of exposure estimation and measurements from the NHEXAS-AZ field study. The stepwise procedure was used to construct regression models. Significance level at P=0.10 was used for entry and retention of independent variables in a model. Twelve significant regression models were formulated to quantify associations among exposures and other variables in the NHEXAS-AZ database. Route-specific exposures to pesticides associate significantly with questionnaire-based variables such as preparation of pesticides, use of pesticide inside the house, and income level; and with concentration variables in three media: dermal wipe, sill wipe, and indoor air. Models formulated in this study may be used to estimate exposures to each of the pesticides. Yet, the use of these models must incorporate clear statements of the assumptions made in the formulation as well as the coefficient of determination and the confidence and prediction intervals of the dependent variable.


Thermodynamics of wheat germ agglutinin-sialyloligosaccharide interactions by proton nuclear magnetic resonance.
The thermodynamic parameters that characterize the binding of wheat germ agglutinin isolectin I (WGA I) to the alpha 2-3 isomer of (N-acetylneuraminyl)lactose have been determined by 360-MHz proton nuclear magnetic resonance spectroscopy. The chemical exchange of the ligand between the free and bound sites resulted in a broadening and upfield shifting of the N-acetyl methyl resonance [Kronis, K.A., & Carver, J.P. (1985) Biochemistry (preceding paper in this issue)] which has allowed the determination of the equilibrium constant, KD, and the dissociation rate constant, kD. In this paper, the analysis of the temperature dependence of the KD values between 25.4 and 51.6 degrees C yielded equilibrium parameters indicative of a large entropy barrier to binding: delta H degree = -13.3 +/- 1.0 kcal mol-1 and delta S degree = -31.9 +/- 2.4 cal mol-1 K-1. The Arrhenius plot of the effect of temperature on the dissociation rate (kD) and the plot of 1n (kD/T) vs. 1/T indicated that the transition complex represented an unfavorable energy state compared to the dissociated molecules with an activation energy (EA) of +18.0 kcal mol-1 and enthalpy and entropy of dissociation (delta HD not equal to and delta SD not equal to) values of +17.4 +/- 0.3 kcal mol-1 and +13.4 +/- 1.2 cal mol-1 K-1, respectively. The driving force for this binding reaction is the large negative delta H degree with a small enthalpic barrier to association (delta HA = +4.1 kcal mol-1).(ABSTRACT TRUNCATED AT 250 WORDS)


Chitotriosidase Levels in the Colostrum from Mothers of Term and Preterm Infants.
Background: Breast milk Chitotriosidase (Chit 1) shows antifungal effect and has an active role in the natural immune response against certain pathogens. The aim of this study was to compare colostrum Chit 1 levels from mothers of term and preterm infants. Materials and Methods: The study included 72 mothers of 32 preterm and 40 term infants (gestational age; 33.7 ± 1.8 vs. 39.1 ± 1.1 weeks, birth weight; 1931.7 ± 539.8 vs. 3350.9 ± 419.7 g). Breast milk samples were taken at postnatal 24-48 hours. Chit 1 level was evaluated with the quantitative calorimetric method. Results: No significant difference was determined between the term and preterm groups in terms of maternal age, education level, weight gain in pregnancy, and body mass index (BMI). The median colostrum Chit 1 level was higher in the preterm group, but the difference was not statistically significant between two groups (p = 0.43). There is no association between colostrum Chit 1 level, maternal age, gravida, BMI, infant gender, income level, and pre-eclampsia. The colostrum Chit 1 level of mothers who had weight gain exceeding the recommended limits was significantly lower than mothers with weight gain within the recommended limits in the term group (4346.2 vs. 4914.2, p = 0.021). A negative correlation was determined between the birthweight of term infants and the colostrum Chit 1 levels (p = 0.045, r = -0.319). Conclusion: Although the data need to be validated by further investigation, the observations made in this study seem to indicate that colostrum Chit-1 may have role in the protection of preterm infants.


An Integrated HOCl-Producing E-Scaffold Is Active against Monomicrobial and Polymicrobial Biofilms.
Oxidizing agents like hypochlorous acid (HOCl) have antimicrobial activity. We developed an integrated electrochemical scaffold, or e-scaffold, that delivers a continuous low dose of HOCl aimed at targeting microbial biofilms without exceeding concentrations toxic to humans as a prototype of a device being developed to treat wound infections in humans. In this work, we tested the device against 33 isolates of bacteria (including isolates with acquired antibiotic resistance) grown as in vitro biofilms alongside 12 combinations of dual-species in vitro biofilms. Biofilms were grown on the bottoms of 12-well plates for 24 h. An integrated e-scaffold was placed atop each biofilm and polarized at 1.5 V for 1, 2, or 4 h. HOCl was produced electrochemically by oxidizing chloride ions (Cl-) in solution to chlorine (Cl2); dissolved Cl2 spontaneously dissociates in water to produce HOCl. The cumulative concentration of HOCl produced at the working electrode in each well was estimated to be 7.89, 13.46, and 29.50 mM after 1, 2, and 4 h of polarization, respectively. Four hours of polarization caused an average reduction of 6.13 log10 CFU/cm2 (±1.99 log10 CFU/cm2) of viable cell counts of monospecies biofilms and 5.53 log10 CFU/cm2 (±2.31 log10 CFU/cm2) for the 12 dual-species biofilms studied. The described integrated e-scaffold reduces viable bacterial cell counts in biofilms formed by an array of antibiotic-susceptible and -resistant bacteria alone and in combination.


Abductor muscle performance in total hip arthroplasty with and without trochanteric osteotomy. Radiographic and mechanical analyses.
Total hip arthroplasty was performed for unilateral hip disease in 26 patients with trochanteric osteotomy (TS) and 27 patients without osteotomy (NS). Early postoperative recovery of function, pain, mobility, radiographic changes, and abduction torques were recorded and evaluated. Theoretic analysis of hip mechanics based on radiographic measurements was compared with measured abduction torques for each patient. Function, pain, and mobility ratings were similar for both groups pre- and postoperation. Radiographic measurements demonstrated better mechanical reconstruction of hips in both groups. There was a trend, but no statistically significant correlation, of mechanical parameters and measured isokinetic or isometric abduction torques at the hip joint in the TS group. In the NS group, no correlation was found between mechanical parameters and measured torques. A significant improvement in recovery of torque was found in the NS group when compared to the TS patients.


V-ATPase as an effective therapeutic target for sarcomas.
Malignant tumors show intense glycolysis and, as a consequence, high lactate production and proton efflux activity. We investigated proton dynamics in osteosarcoma, rhabdomyosarcoma, and chondrosarcoma, and evaluated the effects of esomeprazole as a therapeutic agent interfering with tumor acidic microenvironment. All sarcomas were able to survive in an acidic microenvironment (up to 5.9–6.0 pH) and abundant acidic lysosomes were found in all sarcoma subtypes. V-ATPase, a proton pump that acidifies intracellular compartments and transports protons across the plasma membrane, was detected in all cell types with a histotype-specific expression pattern. Esomeprazole administration interfered with proton compartmentalization in acidic organelles and induced a significant dose-dependent toxicity. Among the different histotypes, rhabdomyosarcoma, expressing the highest levels of V-ATPase and whose lysosomes are most acidic, was mostly susceptible to ESOM treatment.


[Supraconservative breast surgery].
Currently, 30% of the breast biopsies are performed for patients with nonpalpable lesions. The surgical management of these lesions had to evolve to a better three-dimensional targeting and a reduction of the tissue traumatism. The ABBI procedure allows the percutaneous one bloc excision of suspicious mammographically detected lesions with a diameter of less than 2cm. We prospectively evaluated this procedure as a therapeutic tool. Of the 10 malignant lesions with a pathologic size <10mm, 9 (90%) were completely resected with the ABBI device (no residual disease at re-excision of the biopsy site). The results of this preliminary study suggest a potential therapeutic role of the ABBI procedure in the therapeutic arsenal against mammary lesions.


Coxiella burnetii: turning hostility into a home.
Coxiella burnetii, the causative agent of the human disease Q fever, is a unique intracellular bacterial pathogen. Coxiella replicates to high numbers within a pathogen-derived lysosome-like vacuole, thriving within a low pH, highly proteolytic and oxidative environment. In 2009, researchers developed means to axenically culture Coxiella paving the way for the development of tools to genetically manipulate the organism. These advances have revolutionized our capacity to examine the pathogenesis of Coxiella. In recent years, targeted and random mutant strains have been used to demonstrate that the Dot/Icm type IV secretion system is essential for intracellular replication of Coxiella. Current research is focused towards understanding the unique cohort of over 130 effector proteins that are translocated into the host cell. Mutagenesis screens have been employed to identify effectors that play important roles for the biogenesis of the Coxiella-containing vacuole and intracellular replication of Coxiella. A surprisingly high number of effector mutants demonstrate significant intracellular growth defects, and future studies on the molecular function of these effectors will provide great insight into the pathogenesis of Coxiella. Already, this expanse of new data implicates many eukaryotic processes that are targeted by the arsenal of Coxiella effectors including autophagy, apoptosis and vesicular trafficking.


Isotope metallomics approaches for medical research.
Metallomics is a rapidly evolving field of bio-metal research that integrates techniques and perspectives from other "-omics" sciences (e.g. genomics, proteomics) and from research vocations further afield. Perhaps the most esoteric of this latter category has been the recent coupling of biomedicine with element and isotope geochemistry, commonly referred to as isotope metallomics. Over the course of less than two decades, isotope metallomics has produced numerous benchmark studies highlighting the use of stable metal isotope distribution in developing disease diagnostics-e.g. cancer, neurodegeneration, osteoporosis-as well as their utility in deciphering the underlying mechanisms of such diseases. These pioneering works indicate an enormous wealth of potential and provide a call to action for researchers to combine and leverage expertise and resources to create a clear and meaningful path forward. Doing so with efficacy and impact will require not only building on existing research, but also broadening collaborative networks, bolstering and deepening cross-disciplinary channels, and establishing unified and realizable objectives. The aim of this review is to briefly summarize the field and its underpinnings, provide a directory of the state of the art, outline the most encouraging paths forward, including their limitations, outlook and speculative upcoming breakthroughs, and finally to offer a vision of how to cultivate isotope metallomics for an impactful future.


Urine bisphenol-A level in relation to obesity and overweight in school-age children.
Bisphenol-A (BPA) is a potential endocrine disruptor impacting metabolic processes and increasing the risk of obesity. To determine whether urine BPA level is associated with overweight/obesity in school-age children, we examined 1,326 students in grades 4-12 from three schools (one elementary, one middle, and one high school) in Shanghai. More than 98% of eligible students participated. Total urine BPA concentration was measured and anthropometric measures were taken by trained research staff. Information on risk factors for childhood obesity was collected for potential confounders. Age- and gender-specific weight greater than 90(th) percentile of the underlying population was the outcome measure. After adjustment for potential confounders, a higher urine BPA level (≥2 µg/L), at the level corresponding to the median urine BPA level in the U.S. population, was associated with more than two-fold increased risk of having weight >90(th) percentile among girls aged 9-12 (adjusted odds ratio (aOR) = 2.32, 95% confidence interval: 1.15-4.65). The association showed a dose-response relationship with increasing urine BPA level associated with further increased risk of overweight (p = 0.006 for trend test). Other anthropometric measures of obesity showed similar results. The same association was not observed among boys. This gender difference of BPA effect was consistent with findings from experimental studies and previous epidemiological studies. Our study suggests that BPA could be a potential new environmental obesogen. Widespread exposure to BPA in the human population may also be contributing to the worldwide obesity epidemic.


The manoeuvrability hypothesis to explain the maintenance of bilateral symmetry in animal evolution.
The overwhelming majority of animal species exhibit bilateral symmetry. However, the precise evolutionary importance of bilateral symmetry is unknown, although elements of the understanding of the phenomenon have been present within the scientific community for decades. Here we show, with very simple physical laws, that locomotion in three-dimensional macro-world space is itself sufficient to explain the maintenance of bilateral symmetry in animal evolution. The ability to change direction, a key element of locomotion, requires the generation of instantaneous "pushing" surfaces, from which the animal can obtain the necessary force to depart in the new direction. We show that bilateral is the only type of symmetry that can maximize this force; thus, an actively locomoting bilateral body can have the maximal manoeuvrability as compared to other symmetry types. This confers an obvious selective advantage on the bilateral animal. These considerations imply the view that animal evolution is a highly channelled process, in which bilateral and radial body symmetries seem to be inevitable.


High temperature ferroelectric behaviour in α-MnO2 nanorods realised through enriched oxygen vacancy induced non-stoichiometry.
Nanostructuring followed by incorporation of defect induced non-stoichiometry is an emerging field of prominence due to its capacity to introduce unprecedented properties in materials with potential applications. In this work, crystalline α-MnO2 nanorods are synthesised using a facile co-precipitation method to exhibit ferroelectric behaviour for the first time. The evolution mechanism of the nanorods is investigated using XRD, HRTEM and FTIR spectra, while their thermal stability is probed using TGA/DTA. The novel properties observed are the result of structural rearrangements sparked by electrons in mixed valence cations (Mn3+/Mn4+). The high density of Jahn-Teller active Mn3+ cations breaks the inversion symmetry in α-MnO2, thereby altering the atomic environment inducing distortion in the basic MnO6 octahedra. Since variable temperature XRD analysis confirms the phase stability of the crystal structure up to very high temperatures, the ferroelectric phase exhibited by the material below Tc is an outcome of the combined effects of orbital ordering (OO) of the eg electron in Mn3+ and charge ordering (CO) of Mn3+ and Mn4+ cations. This is confirmed by DSC analysis. The breakdown of the ferroelectric nature is identified to originate as a result of octahedral tilting as suggested by temperature-dependent Raman studies. Magnetic and electrical transport studies provide additional evidence of a CO ferroelectric phase as they predict the existence of double-exchange hopping conduction and surface ferromagnetism in the sample.


Holocene eruptive activity of el chichon volcano, chiapas, Mexico.
Geologic and radiometric-age data indicate that El Chichón was frequently and violently active during the Holocene, including eruptive episodes about 600, 1250, and 1700 years ago and several undated, older eruptions. These episodes, involving explosive eruptions of sulfur-rich magma and associated dome-growth processes, were apparently separated by intervals of approximately 350 to 650 years. Some of El Chichón's eruptions may correlate with unusual atmospheric phenomena around A.D. 1300 and possibly A.D. 623.


Catecholamine-synthesizing enzymes and chromogranin proteins in drug-induced proliferative lesions of the rat adrenal medulla.
Both epinephrine (E) and norepinephrine (NE) cells in the rat adrenal medulla are able to proliferate in response to pharmacologic stimulation. However, previous biochemical studies have suggested that drug-induced or spontaneous pheochromocytomas in rats are almost invariably NE-producing. To resolve these apparently conflicting data, immunocytochemical techniques were utilized to establish functional profiles of adrenal medullary lesions classified as pheochromocytoma or nodular hyperplasia in rats treated chronically with a phosphodiesterase inhibitor which induced pheochromocytomas. Sixteen of 17 pheochromocytomas and all hyperplastic nodules stained positively for tyrosine hydroxylase and dopamine beta-hydroxylase, consistent with an ability to produce NE. No lesion of either type stained for phenylethanolamine N-methyltransferase, consistent with an inability to produce epinephrine. Lesions of both types showed variable staining for chromogranin proteins. The findings indicate that qualitative functional differences cannot be used to discriminate hyperplastic nodules from small pheochromocytomas in rats. Some lesions currently classified as hyperplastic nodules might in fact be small pheochromocytomas. Others might represent diffuse hyperplasia within pre-existing islands of NE-cells in a background of hyperplastic epinephrine-cells.


Intracellular signaling pathways and the regulation of cell adhesion.
Adhesion molecules play an essential role in the host immune response by mediating the adhesive interactions that are essential for immune cell trafficking and activation. Integrins are one family of adhesion receptors that leukocytes utilize to interact with other cells and with components of the extracellular matrix. Since leukocytes rapidly alternate between adhesive and nonadhesive states, the functional activity of integrins expressed on leukocytes is carefully and precisely regulated. Resting T lymphocytes express integrin receptors, but they mediate minimal cell adhesion. However, activation of the T cell results within minutes in increased integrin functional activity that occurs without a change in the level of integrin expression on the cell surface. Increased integrin-mediated adhesion appears to be a general response of T cells to activation, since a diverse array of activation stimuli are capable of inducing this rapid increase in integrin functional activity. We have used DNA-mediated gene transfer and site-directed mutagenesis to elucidate the intracellular signaling pathways that regulate integrin-mediated cell adhesion. Our studies have revealed two important general themes. First, the lipid kinase phosphatidylinositol 3-kinase (PI 3-K) plays a role in integrin regulation mediated by many regulators of integrin function. Second, there are cell-specific differences in the signaling pathways that regulate integrin function. These studies illustrate the complex nature of the signaling pathways that regulate lymphocyte adhesion.


Learning flexible features for conditional random fields.
Extending traditional models for discriminative labeling of structured data to include higher-order structure in the labels results in an undesirable exponential increase in model complexity. In this paper, we present a model that is capable of learning such structures using a random field of parameterized features. These features can be functions of arbitrary combinations of observations, labels and auxiliary hidden variables. We also present a simple induction scheme to learn these features, which can automatically determine the complexity needed for a given data set. We apply the model to two real-world tasks, information extraction and image labeling, and compare our results to several other methods for discriminative labeling.


Association of MEFV gene mutations with rheumatoid factor levels in patients with rheumatoid arthritis.
Rheumatoid arthritis (RA) is a systemic autoimmune disease primarily affecting the joints. Arthritis disorders are associated with mutations of the Mediterranean fever (MEFV) gene. This gene has already been identified as being responsible for familial Mediterranean fever. The aim of this study was to explore the frequency and clinical significance of MEFV gene mutations in a cohort of Turkish patients with RA. The study included 101 patients with RA and 110 healthy controls. Genomic DNA was isolated and genotyped using polymerase chain reaction and restriction fragment length polymorphism for the 5 MEFV gene mutations (M694V, M680I, V726A, E148Q, and P369S). Carrier rates of MEFV gene mutations were 31 (30.7%) of 101 and 26 (23.6%) of 110 in the RA and healthy control groups, respectively (P > 0.05; odds ratio, 1.4; 95% CI, 0.77-2.65). Whereas deformed joint count was relatively higher in the mutation carrier group than those of the noncarrier group, the rheumatoid factor levels were significantly higher in the carrier group of patients with RA (P = 0.001). The results of this study suggest that MEFV gene mutations are not positively associated with a predisposition to develop RA but might increase the severity of RA. Further research is needed to determine the actual pathogenic role of MEFV mutations in this disease.


Nuclear Factor κB-COX2 Pathway Activation in Non-myelinating Schwann Cells Is Necessary for the Maintenance of Neuropathic Pain in vivo.
Chronic neuropathic pain leads to long-term changes in the sensitivity of both peripheral and central nociceptive neurons. Glial fibrillary acidic protein (GFAP)-positive glial cells are closely associated with the nociceptive neurons including astrocytes in the central nervous system (CNS), satellite glial cells (SGCs) in the sensory ganglia, and non-myelinating Schwann cells (NMSCs) in the peripheral nerves. Central and peripheral GFAP-positive cells are involved in the maintenance of chronic pain through a host of inflammatory cytokines, many of which are under control of the transcription factor nuclear factor κB (NFκB) and the enzyme cyclooxygenase 2 (COX2). To test the hypothesis that inhibiting GFAP-positive glial signaling alleviates chronic pain, we used (1) a conditional knockout (cKO) mouse expressing Cre recombinase under the hGFAP promoter and a floxed COX2 gene to inactivate the COX2 gene specifically in GFAP-positive cells; and (2) a tet-Off tetracycline transactivator system to suppress NFκB activation in GFAP-positive cells. We found that neuropathic pain behavior following spared nerve injury (SNI) significantly decreased in COX2 cKO mice as well as in mice with decreased glial NFκB signaling. Additionally, experiments were performed to determine whether central or peripheral glial NFκB signaling contributes to the maintenance of chronic pain behavior following nerve injury. Oxytetracycline (Oxy), a blood-brain barrier impermeable analog of doxycycline was employed to restrict transgene expression to CNS glia only, leaving peripheral glial signaling intact. Signaling inactivation in central GFAP-positive glia alone failed to exhibit the same analgesic effects as previously observed in animals with both central and peripheral glial signaling inhibition. These data suggest that the NFκB-COX2 signaling pathway in NMSCs is necessary for the maintenance of neuropathic pain in vivo.


Proportional changes in body fluid with hemodialysis evaluated by dual-energy X-ray absorptiometry and transthoracic bioimpedance with particular emphasis on the thoracic region.
Alterations in body composition during extracorporeal hemodialysis (HD) were investigated in 12 hemodialysis patients (9 males and 3 females, mean age 50 +/- 15 years) with a mean ultrafiltration of 2.6 +/- 1.0 L. Analysis was performed using a dual-energy x-ray absorptiometry technique (DXA), which measures 3 principal components of the body: fat mass (FM): lean body mass (LBM), i.e., all soft tissues excluding fat; and bone mineral content (BMC). These 3 components were calculated for the whole body and for different body regions (namely, the thorax, trunk, lower limbs, and upper limbs). The thoracic cage region could be defined manually, separately from the trunk, and its tissue composition was calculated. DXA analysis was performed concomitant with a measurement of the basal thoracic impedance (TFI) by bioimpedance cardiography prior to and 1 h after dialysis. We found a significant decrease in the total LBM, from 55.8 +/- 8.8 to 53.3 +/- 9.3 kg (p < 0.05), but no change in either the FM or BMC. Moreover, there was a disproportional reduction in the LBM in different regions, being significantly greater in the thorax (7.47 +/- 3.7%) than in the other body regions (trunk 4.3 +/- 2.0%, lower limbs 5.4 +/- 2.1%, and upper limbs 4.7 +/- 1.5%). Regarding bioimpedance, a stronger significant correlation was detected between the percentage changes in the TFI and the changes in thoracic fluid (r = 0.80, p < 0.01) than between the changes in the TFI and the changes in the total body fluid (r = 0.63, p < 0.01). The absolute values of the TFI were also significantly and negatively correlated to the thoracic lean mass to fat mass ratio, both before and after HD (r = 0.82, p < 0.001 and r = 0.86, p < 0.001, respectively). In conclusion, DXA is a very sensitive technique to detect fluid changes during HD in the thorax when the thoracic cavity is defined as a region of interest as well as for the whole body. The data also indicate an extracellular compartmental imbalance between different regions with a significantly greater change in the thoracic region. Transthoracic bioimpedance is a useful technique for evaluating the HD induced changes in the thoracic fluid, rather than total body fluid.


Neurosurgery training in India during the COVID-19 pandemic: straight from the horse's mouth.
The COVID-19 pandemic has forced medical professionals throughout the world to adapt to the changing medical scenario. The objective of this survey was to assess the change in neurosurgical training in India following the COVID-19 pandemic. Between May 7, 2020, and May 16, 2020, a validated questionnaire was circulated among neurosurgical residents across India by social media, regarding changes in the department's functioning, patient interaction, surgical exposure, changes in academics, and fears and apprehensions associated with the pandemic. The responses were kept anonymous and were analyzed for changes during the COVID-19 pandemic compared to before the pandemic. A total of 118 residents from 29 neurosurgical training programs across 17 states/union territories of the country gave their responses to the survey questionnaire. The survey revealed that the surgical exposure of neurosurgical residents has drastically reduced since the onset of the COVID-19 pandemic, from an average of 39.86 surgeries performed/assisted per month (median 30) to 12.31 per month (median 10), representing a decrease of 67.50%. The number of academic sessions has fallen from a median of 5 per week to 2 per week. The survey uncovered the lack of universal guidelines and homogeneity regarding preoperative COVID-19 testing. The survey also reveals reluctance toward detailed patient examinations since the COVID-19 outbreak. The majority of respondents felt that the COVID-19 pandemic will hamper their operative and clinical skills. Fear of rescheduling or deferring of licensing examinations was significantly higher among those closest to the examination (p = 0.002). The adverse impact of the pandemic on neurosurgical training needs to be addressed. While ensuring the safety of the residents, institutes and neurosurgical societies/bodies must take it upon themselves to ensure that their residents continue to learn and develop neurosurgical skills during these difficult times.


Digital parallelism, perpendicularity, and rectangles.
The slope of digital line segments is defined and an algorithm to evaluate it is presented. Parallelism and perpendicularity of two digital line segments are also defined. Finally, rectangular digital regions are defined and characterized, and an algorithm that determines whether or not a given digital region is a digital rectangle is presented.


Associations with retinal nerve fiber layer measures in the EPIC-Norfolk Eye Study.
To describe GDxVCC retinal nerve fiber layer (RNFL) measures and associations in a predominantly white British population. The EPIC-Norfolk Eye Study is nested within a large multicenter cohort study, the European Prospective Investigation of Cancer. RNFL measurements were taken using the GDxVCC. Generalized estimating equation models were used to assess associations of RNFL measures with age, sex, body mass index (BMI), height, blood pressure, social class, education level, alcohol intake, smoking status, axial length, intraocular pressure, and lens status. Models were linearly adjusted for typical scan score to handle scans with atypical retardation. There were complete data from 11,030 eyes of 6309 participants with mean age 68 years (48-90 years). Older age (-1.53 μm/decade [95% confidence interval {CI} -1.73, -1.33], P < 0.001), male sex (-0.44 μm [95% CI -0.04, -0.84], P = 0.031), shorter axial length (-0.15 μm/mm [95% CI -0.02, -0.28], P = 0.024), and pseudophakia (-0.49 μm [95% CI -0.94, -0.04], P = 0.033) were associated with thinner RNFL after adjustment for possible confounders. Higher BMI was associated with a thinner RNFL in men only (-0.30 μm/5 kg/m(2) [95% CI -0.58, -0.02], P = 0.039). This analysis of associations with RNFL thickness in a largely healthy population may provide insight into the determinants of glaucoma, suggesting higher risk in those who are older, in men, and in men with a higher BMI.


[Risks and side effects of diagnostic bronchoalveolar lavage in ventilated patients].
Bronchoalveolar lavage is used increasingly often as a diagnostic tool in critically ill patients undergoing mechanical ventilation. Most patients, even those with haemodynamic instability or respiratory failure, tolerate the procedure without marked adverse effects. A relatively small proportion of patients, however, will experience potentially deleterious side effects like prolonged deterioration of pulmonary mechanics, oxygen desaturation or sepsis-like symptoms. Despite all precautions, it is not possible to date to identify increased-risk patients beforehand. The potential benefits, namely, early diagnosis of an aetiological agent who is responsible for infection, must therefore be carefully weighted against the risks.