Impact of regulatory light chain mutation K104E on the ATPase and motor properties of cardiac myosin.
Mutations in the cardiac myosin regulatory light chain (RLC, MYL2 gene) are known to cause inherited cardiomyopathies with variable phenotypes. In this study, we investigated the impact of a mutation in the RLC (K104E) that is associated with hypertrophic cardiomyopathy (HCM). Previously in a mouse model of K104E, older animals were found to develop cardiac hypertrophy, fibrosis, and diastolic dysfunction, suggesting a slow development of HCM. However, variable penetrance of the mutation in human populations suggests that the impact of K104E may be subtle. Therefore, we generated human cardiac myosin subfragment-1 (M2β-S1) and exchanged on either the wild type (WT) or K104E human ventricular RLC in order to assess the impact of the mutation on the mechanochemical properties of cardiac myosin. The maximum actin-activated ATPase activity and actin sliding velocities in the in vitro motility assay were similar in M2β-S1 WT and K104E, as were the detachment kinetic parameters, including the rate of ATP-induced dissociation and the ADP release rate constant. We also examined the mechanical performance of α-cardiac myosin extracted from transgenic (Tg) mice expressing human wild type RLC (Tg WT) or mutant RLC (Tg K104E). We found that α-cardiac myosin from Tg K104E animals demonstrated enhanced actin sliding velocities in the motility assay compared with its Tg WT counterpart. Furthermore, the degree of incorporation of the mutant RLC into α-cardiac myosin in the transgenic animals was significantly reduced compared with wild type. Therefore, we conclude that the impact of the K104E mutation depends on either the length or the isoform of the myosin heavy chain backbone and that the mutation may disrupt RLC interactions with the myosin lever arm domain.


Coping with COVID: risk and resilience factors for mental health in a German representative panel study.
The coronavirus disease 2019 (COVID-19) pandemic might affect mental health. Data from population-representative panel surveys with multiple waves including pre-COVID data investigating risk and protective factors are still rare. In a stratified random sample of the German household population (n = 6684), we conducted survey-weighted multiple linear regressions to determine the association of various psychological risk and protective factors assessed between 2015 and 2020 with changes in psychological distress [(PD; measured via Patient Health Questionnaire for Depression and Anxiety (PHQ-4)] from pre-pandemic (average of 2016 and 2019) to peri-pandemic (both 2020 and 2021) time points. Control analyses on PD change between two pre-pandemic time points (2016 and 2019) were conducted. Regularized regressions were computed to inform on which factors were statistically most influential in the multicollinear setting. PHQ-4 scores in 2020 (M = 2.45) and 2021 (M = 2.21) were elevated compared to 2019 (M = 1.79). Several risk factors (catastrophizing, neuroticism, and asking for instrumental support) and protective factors (perceived stress recovery, positive reappraisal, and optimism) were identified for the peri-pandemic outcomes. Control analyses revealed that in pre-pandemic times, neuroticism and optimism were predominantly related to PD changes. Regularized regression mostly confirmed the results and highlighted perceived stress recovery as most consistent influential protective factor across peri-pandemic outcomes. We identified several psychological risk and protective factors related to PD outcomes during the COVID-19 pandemic. A comparison of pre-pandemic data stresses the relevance of longitudinal assessments to potentially reconcile contradictory findings. Implications and suggestions for targeted prevention and intervention programs during highly stressful times such as pandemics are discussed.


Laparoscopic repair of vesicoureteral reflux utilizing the Lich-Gregoir technique in the pig model.
To date, laparoscopic urological surgery has largely been limited to diagnostic or ablative procedures. Herein we report our experience with laparoscopic reconstructive surgery to perform an extravesical ureteral reimplantation. Seven anesthetized pigs with iatrogenic ureteral reflux underwent a laparoscopic extravesical ureteral reimplantation. The newly created ureteral tunnel varied from 2 to 4 cm. In 3 pigs, the tunnel was created with tacking staples, while in the other 4 pigs, the tunnel was created with intracorporeal suturing techniques using a 3-zero polyglyconate running suture. The procedure required an average of 132 minutes. There was one anesthetic death. There were no urinary tract infections. At 3 to 8 weeks after reimplantation, the cystograms were repeated on 5 pigs. One of 2 stapled reimplant pigs still had reflux; 1 of 3 sewn reimplant pigs had reflux. At 6 months following the reimplantation, only 1 pig had residual grade I reflux and this was a sutured reimplantation. None of the stapled reimplantations exhibited any residual reflux on the surgical side; however, in 1 animal a submucosal staple was noted at the time of harvest.


Thioredoxin reductase is essential for thiol/disulfide redox control and oxidative stress survival of the anaerobe Bacteroides fragilis.
Results of this study showed that the anaerobic, opportunistic pathogen Bacteroides fragilis lacks the glutathione/glutaredoxin redox system and possesses an extensive number of putative thioredoxin (Trx) orthologs. Analysis of the genome sequence revealed six Trx orthologs and an absence of genes required for synthesis of glutathione and glutaredoxins. In addition, it was shown that the thioredoxin reductase (TrxB)/Trx system is the major or sole redox system for thiol/disulfide cellular homeostasis in this anaerobic bacterium. Expression of the B. fragilis trxB gene was induced following treatment with diamide or H(2)O(2) or exposure to oxygen. This inducible trxB expression was OxyR independent. Northern blot hybridization analysis showed that the trxB mRNA was cotranscribed with lolA as a bicistronic transcript or was present as a monocistronic transcript that was also highly induced under the same conditions. The role of LolA, a prokaryotic periplasmic lipoprotein-specific molecular chaperone in the thiol/disulfide redox system, is unknown. A trxB deletion mutant was more sensitive to the effects of diamide and oxygen than the parent strain. In addition, the trxB mutant was unable to grow in culture media without addition of a reductant. Furthermore, the trxB mutant was not able to induce intraabdominal abscess formation in a mouse model, whereas the parent strain was. Taken together, these data strongly suggest that TrxB/Trx is the major, if not the sole, thiol/disulfide redox system in this anaerobe required for survival and abscess formation in a peritoneal cavity infection model.


Evidence for Bidirectional Noninnocent Behavior of a Formazanate Ligand in Ruthenium Complexes.
Redox series of the complexes [Ru(L)(L')2](n), L = 1,5-diphenyl-3-(4-tolyl)-formazanate and L' = 2,4-pentanedionate (acac(-)), 2,2'-bipyridine (bpy), or 2-phenylazopyridine (pap), were studied by cyclic and differential pulse voltammetry and by TD-DFT-supported spectroelectrochemistry (UV-vis-NIR, EPR). The precursors [Ru(III)(L(-))(acac(-))2], [Ru(II)(L(-))(bpy)2]ClO4, and [Ru(II)(L(-))(pap)2]ClO4 were identified in their indicated oxidation states by X-ray crystal structure determination. The six-membered formazanato-ruthenium chelate rings have an envelope conformation with puckering of the metal. DFT calculations indicate a pronounced sensitivity of the N-N bond lengths toward the ligand oxidation state. Several electrochemically accessible charge states were analyzed, and the derived oxidation numbers Ru(II), Ru(III), or Ru(IV), L' or (L')(•-), and L(-), L(•2-), or the new formazanyl ligand L(•) for the two-way noninnocent formazanate reflect the increasing acceptor effect of the ancillary ligands L' in the series acac(-) < bpy < pap.


An acidic pH/reduction dual-stimuli responsive nanoprobe for enhanced CT imaging of tumours in vivo.
Computed tomography (CT) is one of the most frequently used diagnostic imaging modalities in clinics. However, the fast clearance of CT contrast agents through the kidney and short circulation time severely restrict their in vivo applications. Herein, taking advantage of the biocompatible CBT condensation reaction, we rationally designed and synthesized a new smart acidic pH/glutathione (GSH) dual-stimuli responsive nanoprobe (1) which can intermolecularly undergo condensation and form a nanoparticle assembly (I-NPs) in the tumour microenvironment. In vivo CT imaging results indicated that probe 1 could be successfully applied for enhanced CT imaging of tumours in nude mice with a low dose of 21.79 mg I per kg body weight, which may offer a promising tool for precise tumor diagnosis.


A pilot study of the effect of sodium thiosulfate on urinary lithogenicity and associated metabolic acid load in non-stone formers and stone formers with hypercalciuria.
Sodium thiosulfate (STS) reduced calcium stone formation in both humans and genetic hypercalciuric stone forming (GHS) rats. We sought to measure urine chemistry changes resulting from STS administration in people. DESIGN SETTING PARTICIPANTS MEASUREMENTS: STS was given to healthy and hypercalciuric stone forming adults. Five normal non-stone forming adults (mean age 33 years), and 5 people with idiopathic hypercalciuria and calcium kidney stones (mean age 66 years) participated. Two baseline 24-hour urine collections were performed on days 2 and 3 of 3 days of self-selected diets. Subjects then drank STS 10 mmol twice a day for 7 days and did urine collections while repeating the self-selected diet. Results were compared by non-parametric Wilcoxon signed rank test. The primary outcome was the resulting change in urine chemistry. STS administration did not cause a significant change in urinary calcium excretion in either group. In both groups, 24 hour urinary ammonium (P = 0.005) and sulfate excretion (P = 0.007) increased, and urinary pH fell (P = 0.005); citrate excretion fell (P<0.05) in hypercalciuric participants but not in non-stone formers. Among stone formers with hypercalciuria, 3 of 5 patients had measurement of serum HCO3 concentration after the STS period: it did not change. The net effect was an increase in supersaturation of uric acid, and no change in supersaturation of calcium oxalate or calcium phosphate. The basis for studies demonstrating that STS prevented stones in rats and people was not reflected by the changes in urine chemistry reported here. Although serum HCO3 did not change, urine tests suggested an acid load in both non-stone forming and hypercalciuric stone-forming participants. The long term safety of STS needs to be determined before the drug can be tested in humans for long-term prevention of stone recurrence.


Multisite performance evaluation of an enzyme-linked immunosorbent assay for detection of Giardia, Cryptosporidium, and Entamoeba histolytica antigens in human stool.
A novel fecal antigen detection assay for fresh and frozen human samples that detects but does not differentiate Giardia spp, Cryptosporidium spp, and Entamoeba histolytica, the Tri-Combo parasite screen, was compared to three established enzyme-linked immunosorbent assays (ELISAs) at three international sites. It exhibited 97.9% sensitivity and 97.0% specificity, with positive and negative predictive values of 93.4% and 99.1%, respectively. The Tri-Combo test proved a reliable means to limit the use of individual parasite ELISAs to positive samples.


Adjusting for collider bias in genetic association studies using instrumental variable methods.
Genome-wide association studies have provided many genetic markers that can be used as instrumental variables to adjust for confounding in epidemiological studies. Recently, the principle has been applied to other forms of bias in observational studies, especially collider bias that arises when conditioning or stratifying on a variable that is associated with the outcome of interest. An important case is in studies of disease progression and survival. Here, we clarify the links between the genetic instrumental variable methods proposed for this problem and the established methods of Mendelian randomisation developed to account for confounding. We highlight the critical importance of weak instrument bias in this context and describe a corrected weighted least-squares procedure as a simple approach to reduce this bias. We illustrate the range of available methods on two data examples. The first, waist-hip ratio adjusted for body-mass index, entails statistical adjustment for a quantitative trait. The second, smoking cessation, is a stratified analysis conditional on having initiated smoking. In both cases, we find little effect of collider bias on the primary association results, but this may propagate into more substantial effects on further analyses such as polygenic risk scoring and Mendelian randomisation.


Effect of Tinospora crispa on thioacetamide-induced liver cirrhosis in rats.
This study was conducted to determine the effect of ethanolic extract of the dried stems of Tinospora crispa in a male rat model of hepatic fibrosis caused by the hepatotoxin, thioacetamide. The extract was gavaged daily to the rats, at doses of 100 and 200 mg/kg along with thioacetamide at a dose of 200 mg/kg twice weekly. To assess the effectivity of extract, against thioacetamide, the activity of aminotransferases (alanine aminotransferase, aspartate aminotransferase), alkaline phosphatase (AP); and bilirubin were measured, together with morphological and histopathological indices in the liver of healthy and thioacetamide-treated rats. A significant increase in the activity of liver enzymes, bilirubin and G-glutamyl transferase and gross and histopathological changes were determined. Although previous in vitro study established that this extract had strong antioxidant activity, this in vivo study establishes that this extract contains hepatotoxins whose identity may be quite different from those compounds with antioxidant properties. The study confirms that complete reliance on data obtained using in vitro methodologies may lead to erroneous conclusions pertaining to the safety of phytopharmaceuticals.


The Mobility of the Cap Domain Is Essential for the Substrate Promiscuity of a Family IV Esterase from Sorghum Rhizosphere Microbiome.
Metagenomics offers the possibility to screen for versatile biocatalysts. In this study, the microbial community of the Sorghum bicolor rhizosphere was spiked with technical cashew nut shell liquid, and after incubation, the environmental DNA (eDNA) was extracted and subsequently used to build a metagenomic library. We report the biochemical features and crystal structure of a novel esterase from the family IV, EH0, retrieved from an uncultured sphingomonad after a functional screen in tributyrin agar plates. EH0 (optimum temperature [Topt], 50°C; melting temperature [Tm], 55.7°C; optimum pH [pHopt], 9.5) was stable in the presence of 10 to 20% (vol/vol) organic solvents and exhibited hydrolytic activity against p-nitrophenyl esters from acetate to palmitate, preferably butyrate (496 U mg-1), and a large battery of 69 structurally different esters (up to 30.2 U mg-1), including bis(2-hydroxyethyl)-terephthalate (0.16 ± 0.06 U mg-1). This broad substrate specificity contrasts with the fact that EH0 showed a long and narrow catalytic tunnel, whose access appears to be hindered by a tight folding of its cap domain. We propose that this cap domain is a highly flexible structure whose opening is mediated by unique structural elements, one of which is the presence of two contiguous proline residues likely acting as possible hinges, which together allow for the entrance of the substrates. Therefore, this work provides a new role for the cap domain, which until now was thought to be an immobile element that contained hydrophobic patches involved in substrate prerecognition and in turn substrate specificity within family IV esterases. IMPORTANCE A better understanding of structure-function relationships of enzymes allows revelation of key structural motifs or elements. Here, we studied the structural basis of the substrate promiscuity of EH0, a family IV esterase, isolated from a sample of the Sorghum bicolor rhizosphere microbiome exposed to technical cashew nut shell liquid. The analysis of EH0 revealed the potential of the sorghum rhizosphere microbiome as a source of enzymes with interesting properties, such as pH and solvent tolerance and remarkably broad substrate promiscuity. Its structure resembled those of homologous proteins from mesophilic Parvibaculum and Erythrobacter spp. and hyperthermophilic Pyrobaculum and Sulfolobus spp. and had a very narrow, single-entry access tunnel to the active site, with access controlled by a capping domain that includes a number of nonconserved proline residues. These structural markers, distinct from those of other substrate-promiscuous esterases, can help in tuning substrate profiles beyond tunnel and active site engineering.


[Features of hospital-acquired infections in newborns with culture-positive].
To determine the prevalence of nosocomial infection (NI) in newborns (NB) as well as the etiology, frequency mortality related to these infections in a tertiary-care hospital. A retrospective epidemiological study was carried out with NB admitted to a tertiary-care neonatology hospital service from January 2006 to December 2008 who complied with selection criteria. All NB between the gestational ages of 25 to 44 weeks, NI supported by positive culture and either local or systemic infection were included. Descriptive statistic was used. NI prevalence in the Neonatal Service was an average of 30.4%. The most common microorganisms isolated in first event cultures (n = 100) were Gram-positive bacteria such as coagulase-negative staphylococci found in 55 patients (55%), followed by Gram-negative bacteria present in 44 patients (44%) and fungi such as Candida albicans and Candida sp. from 8 patients (8%); as to the second event (n = 32), Gram-negative bacteria were isolated from 20 patients (62.5%), Gram-positive bacteria such as coagulase-negative staphylococci were present in 6 patients (18.7%) and fungi such as Candida albicans were found in 5 patients (15.6%). Finally, regarding the third event (n = 18), Gram-negative bacteria were present in 16 patients (88.9%), Gram-positive bacteria were found in 15 patients (83.3%) such as coagulase-negative staphylococci in 10 patients (55.5%), and fungi such as Candida sp. in two (11.1%). Twelve (12%) who died secondary to IN, two suffered three NI events. Awareness of the various characteristics of NI in the work area (etiology, prevalence, and final outcome) is of great importance to design new interventions.


Choice of conjunctival incisions for horizontal rectus muscle surgery--a survey of American Association for Pediatric Ophthalmology and Strabismus members.
To assess the use of limbal versus fornix conjunctival incisions in primary and repeat horizontal rectus muscle surgeries among pediatric ophthalmologists and adult strabismus surgeons. A 2-page questionnaire was mailed to members of the American Association for Pediatric Ophthalmology and Adult Strabismus. Participants were asked about their use of limbal, fornix, or other incision types in their pediatric and adult strabismus patient population. A total of 1,022 questionnaires were circulated. The net overall response rate was 27.8%. For first-time strabismus surgeries in pediatric patients, 40.8% of respondents used limbal incisions in the majority of their cases, 58.1% used fornix incisions, and 1.1% used other incision types. For reoperations, 58.1% used limbal-based incisions, 39.1% used fornix incisions, and 2.1% used other incision types. In the majority of their primary surgeries on adults, 40.1% used limbal incisions, 53.5% preferred fornix incisions, 1.4% used other incision types, and 4.9% answered "not applicable" (available for respondents who operated exclusively on either children or adults). On reoperations, limbal incisions were used by 63.4% of respondents, 29.9% used fornix incisions, 1.4% used other incision types, and 5.3% answered "not applicable." Among those surveyed, limbal incisions were preferred for greater intraoperative exposure and better teaching of junior surgeons; fornix incisions were thought to cause less postoperative pain and inflammation and lead to more rapid soft-tissue healing.


Interfacial adhesion between functionalized polyethylene surface and graphene via molecular dynamic simulation.
In this study, interfacial adhesion between functionalized polyethylene (PE) surfaces and graphene were examined using molecular simulation. Various functional groups including amino, carboxy, hydroxy, cyano, isocyanato, oxo, and ethylamino were used to cover the PE surface with surface densities of 0.48, 1.30, and 4.84 groups per nm(2). The interfacial adhesion between the modified PE surfaces and the graphene was quantified via calculation of work of separation (Wsep), the amount of the required work to separate two surfaces without occurring any relaxation and diffusion phenomena. Insertion of the functional groups on the PE surface decreased the amount of Wsep, except for the oxo, amino, and higher densities of the carboxy groups. Increasing the surface group density enhanced the adhesion due to decreasing the surface atomic roughness and increasing the atomic density at the interface. In addition, the effect of surface group rearrangement was investigated via calculation of the work of adhesion (Wadh) while sufficient time had been devoted to relax the interface. The surface reorganization during the relaxation process significantly enhanced adhesion due to eliminating the surface roughness and increasing the surface atomic density.


Adherence to Secondary Prevention and Influential Factors in Individuals with Coronary Angioplasty.
To explore the relation between adherence to secondary prevention and factors that influence on said adherence in people with acute coronary syndrome, who underwent percutaneous coronary angioplasty in a clinic in Medellín. Cross-sectional study on a random sample of 128 volunteer patients. A questionnaire was used for sociodemographic variables, the "Scale to measure therapeutic adherence for patients with chronic diseases, based on explicit behaviors" by Trujano, Vega, and Nava and the "Instrument to evaluate adherence by patients according to influential cardiovascular risk factors" validated by Consuelo Ortiz. Socioeconomic factors influenced in very low manner on the adherence to secondary prevention; factors related with the therapy did so moderately and patient factors influenced in low manner. No relation was found between the health provider factor and said adherence. Factors exist that influence in a lesser or higher measure on adherence to secondary prevention and which must be recognized in people with coronary angioplasty to design strategies to improve this aspect of self-care.


An examination of gender imbalance in Scottish adolescents' vocational interests.
This paper documents Scottish adolescents' vocational interest types. Our research is based on the responses of 1,306 pupils from 18 secondary schools to an empirically verified online interest inventory test. Our results are threefold. First, the structural validity of the test with the Scottish sample is confirmed by evaluating the underlying circumplex structure of Holland's RIASEC vocational interests. Second, gender distribution along the six primary vocational interest dimensions is consistent with the research literature: young men scoring higher on the Realistic vocational interest and young women scoring higher on the Social dimension. Finally, we observe that across dimensions, vocational interests of young women are less diverse than those of young men. We discuss how these dissimilarities could lead to differences in education choice and career decision-making.


Saliva-based methods for SARS-CoV-2 testing in low- and middle-income countries.
As the coronavirus disease 2019 (COVID-19) continues to disproportionately affect low- and middle-income countries, the need for simple, accessible and frequent diagnostic testing grows. In lower-resource settings, case detection is often limited by a lack of available testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To address global inequities in testing, alternative sample types could be used to increase access to testing by reducing the associated costs. Saliva is a sensitive, minimally invasive and inexpensive diagnostic sample for SARS-CoV-2 detection that is appropriate for asymptomatic surveillance, symptomatic testing and at-home collection. Saliva testing can lessen two major challenges faced by lower- and middle-income countries: constrained resources and overburdened health workers. Saliva sampling enables convenient self-collection and requires fewer resources than swab-based methods. However, saliva testing for SARS-CoV-2 diagnostics has not been implemented on a large scale in low- and middle-income countries. While numerous studies based in these settings have demonstrated the usefulness of saliva sampling, there has been insufficient attention on optimizing its implementation in practice. We argue that implementation science research is needed to bridge this gap between evidence and practice. Low- and middle-income countries face many barriers as they continue their efforts to provide mass COVID-19 testing in the face of substantial inequities in global access to vaccines. Laboratories should look to replicate successful approaches for sensitive detection of SARS-CoV-2 in saliva, while governments should act to facilitate mass testing by lifting restrictions that limit implementation of saliva-based methods.


Redox Modulation and Induction of Ferroptosis as a New Therapeutic Strategy in Hepatocellular Carcinoma.
Ferroptosis, a newly discovered form of cell death mediated by reactive oxygen species (ROS) and lipid peroxidation, has recently been shown to have an impact on various cancer types; however, so far there are only few studies about its role in hepatocellular carcinoma (HCC). The delicate equilibrium of ROS in cancer cells has found to be crucial for cell survival, thus increased levels may trigger ferroptosis in HCC. In our study, we investigated the effect of different ROS modulators and ferroptosis inducers on a human HCC cell line and a human hepatoblastoma cell line. We identified a novel synergistic cell death induction by the combination of Auranofin and buthionine sulfoxime (BSO) or by Erastin and BSO at subtoxic concentrations. We found a caspase-independent, redox-regulated cell death, which could be rescued by different inhibitors of ferroptosis. Both cotreatments stimulated lipid peroxidation. All these findings indicated ferroptotic cell death. Both cotreatments affected the canonical ferroptosis pathway through GPX4 downregulation. We also found an accumulation of Nrf2 and HO-1, indicating an additional effect on the non-canonical pathway. Our results implicate that targeting these two main ferroptotic pathways simultaneously can overcome chemotherapy resistance in HCC.


Application of Digital Image Based on Machine Learning in Media Art Design.
In digital media art, expressive force is an important art form of media. This paper studies digital images that have the same effect when applied to media art. The research object is media art images, and the application effect of the proposed algorithm is related to the media art images. The development of digital image technology has brought revolutionary changes to traditional media art expression techniques. In this paper, a partial-pixel interpolation technique based on convolutional neural network is proposed. Supervised training of convolutional neural networks requires predetermining the input and target output of the network, namely, integer image and fractional image in this paper. To solve the problem that the subpixel sample cannot be obtained, this paper first analyzes the imaging principle of digital image and proposes a subpixel sample generation algorithm based on Gaussian low-pass filter and polyphase sampling. From the perspective of rate distortion optimization, the purpose of pixel motion compensation is to improve the accuracy of interframe prediction. Therefore, this paper defines pixel motion compensation as an interframe regression problem, that is, the mapping process of the reference image integral pixel sample to the current image sample to be encoded. In this paper, a generalized partial-pixel interpolation model is proposed for bidirectional prediction. The partial-pixel interpolation of bidirectional prediction is regarded as a binary regression model; that is, the integral pixel reference block in two directions is mapped to the current block to be coded. It further studies how to apply the trained digital images to media art design more flexibly and efficiently.


Interferon-gamma-induced membrane PAF-receptor expression confers tumor cell susceptibility to NK perforin-dependent lysis.
Perforin is known to display a membranolytic activity on tumor cells. Nevertheless, perforin release during natural killer (NK)-cell activation is not sufficient to induce membrane target-cell damage. On the basis of the ability of perforin to interact with phospholipids containing a choline phosphate headgroup, we identify the platelet-activating factor (PAF) and its membrane receptor as crucial components in tumor cell killing activity of human resting NK cells. We demonstrate for the first time that upon activation, naive NK cells release the choline phosphate-containing lysolipid PAF, which binds to perforin and acts as an agonist on perforin-induced membrane damage. PAF is known to incorporate cell membranes using a specific receptor. Here we show that interferon-gamma (IFN-gamma) secreted from activated NK cells ends in PAF-receptor expression on perforin-sensitive K562 cells but not on perforin-resistant Daudi cells. In order to prove the capacity of PAF to interact simultaneously with its membrane PAF receptor and with perforin, we successfully co-purified the 3 components in the presence of bridging PAF molecules. The functional activity of this complex was further examined. The aim was to determine whether membrane PAF-receptor expression on tumor cells, driven to express this receptor, could render them sensitive to the perforin lytic pathway. The results confirmed that transfection of the PAF-receptor complementary DNA into major histocompatibility complex class I and Fas-receptor negative tumor cells restored susceptibility to naive NK cells and perforin attack. Failure of IFN-gamma to induce membrane PAF receptor constitutes the first described mechanism for tumor cells to resist the perforin lytic pathway.


Motor neuron disease and polio in Scotland.
An analysis of mortality and morbidity rates for motor neuron disease (MND) in Scotland has confirmed earlier observations that the disease is more common in men and older age groups. The geographical distribution is non-uniform and related to discharge rates for all neurological diseases. Discharge and mortality rates are increasing but there has been no decline in populations who would have been vaccinated against polio.


CT Scan Differences of Pulmonary TB According to Presence of Pleural Effusion.
Subpleural micronodules and interlobular septal thickening are common CT scan findings in TB pleural effusion. These CT scan features could help us differentiate between TB pleural effusion and nonTB empyema. Does the frequency of subpleural micronodules and interlobular septal thickening correlate with the presence of pleural effusion in patients with pulmonary TB? CT scan findings of pulmonary TB, micronodules and their distribution (peribronchovascular, septal, subpleural, centrilobular, and random), large opacity (consolidation/macronodule), cavitation, tree-in-buds, bronchovascular bundle thickening, interlobular septal thickening, lymphadenopathy, and pleural effusion were retrospectively analyzed. Patients were divided into two groups according to the presence of pleural effusion. Clinicoradiologic findings of the two groups were then analyzed. We presented Benjamini-Hochberg critical value for multiple testing correction of CT scan findings, with a false discovery rate of 0.05. Of a total of 338 consecutive patients diagnosed with pulmonary TB who underwent CT scans, 60 were excluded because of coexisting pulmonary diseases. The frequency of subpleural nodules (47/68, 69% in pulmonary TB with pleural effusion vs 30/210, 14% in pulmonary TB without effusion, P < .001, Benjamini-Hochberg [B-H] critical value = 0.0036) and interlobular septal thickening (55/68, 81% vs 134/210, 64%, P = .009, B-H critical value = 0.0107) was significantly higher in the group of patients with pulmonary TB with pleural effusion than in the group without pleural effusion. In contrast, tree-in-buds (20/68, 29% vs 101/210, 48%, P = .007, B-H critical value = 0.0071) were less frequently seen in patients with pulmonary TB with pleural effusion. Subpleural nodules and septal thickening were more common in pulmonary TB patients with pleural effusion than in those without pleural effusion. TB involvement of the lymphatics in the peripheral interstitium could be associated with the development of pleural effusion.


Influence of γ-Lactones on Monolinolein/Water Bulk and Emulsified Mesophases.
The emulsification of two flavor compounds of the γ-lactone type with monolinolein liquid crystalline submicron particles is reported. The stabilization is ensured by the copolymer F127. γ-Nonalactone can be loaded in bicontinuous cubic monolinolein particles at a larger level (up to 20 wt %) than γ-decalactone (less than 15 wt %). The phase behavior of the ternary monolinolein/water/γ-nonalactone system was studied. The large γ-nonalactone content solubilized into cubosomes was corroborated by the observation of a wide cubic V2 range in the ternary phase diagram. Surprisingly, no inverted hexagonal phase was found in the system. On the contrary, the incorporation of γ-decalactone in the lipid particles gave rise to a dispersion of inverted hexagonal phase, which corresponds to a classical behavior of an oily additive. We finally determined the internal phase of particles including 10 wt % of γ-nonalactone upon increasing the F127 content. We thus found that γ-nonalactone restricts very significantly the interaction of the emulsifier with the cubosomes' interior.


Early positive end-expiratory pressure in the adult respiratory distress syndrome.
This prospective study was designed to determine the effect of positive end-expiratory pressure (PEEP) instituted early in the course of adult respiratory distress syndrome (ARDS). Seventy-nine (7%) of 1,200 patients admitted to the surgical intensive care unit were selected because of a high probability that ARDS would develop, and were randomized into two treatment groups. Of the 79 patients, 45 were immediately treated with 5 cm H2O of end-expiratory pressure (early PEEP group), and 34 received PEEP only when severe hypoxemia developed (late PEEP group). The incidence of ARDS was significantly lower in the early PEEP group than in the late PEEP group (20% vs 53%; P less than .002). Fewer pulmonary deaths occurred in this group (11% vs 29%; P = .02), and there was less pulmonary morbidity. This study supports the efficacy of early low-level PEEP in the treatment of patients for whom there is a high probability that ARDS will develop.


Therapeutic news in ALS.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by death of motor neurons in the cortex and the spinal cord. This loss of motor neurons causes progressive weakness and amyotrophy. To date, the median duration of survival in patients with ALS, from first symptoms to death, is estimated to be 36 months. Currently the treatment is limited to two options: riluzole which prolongs survival for a few months and edaravone which is available in only a few countries and also has a small impact on disease progression. There is an urgent need for more effective drugs in this disease to significantly improve progression. Over the last 30 years, all trials have failed to find a curative drug for ALS. This is due, partially, to the heterogeneity of the clinical features and the pathophysiology of motor neuron death. We present in this review the various treatment options currently being developed for ALS, with an emphasis on the range of therapeutic approaches being explored, from old drugs tested in a new indication to innovative drugs obtained via biotechnology or gene therapy.


Multimorbidity and Genetic Characteristics of DICER1 Syndrome Based on Systematic Review.
It has been reported that germline DICER1 mutations correlate with a distinctive human disease syndrome. Many published studies within this field have been conducted based on rare cases. We systematically searched bibliographic databases, including PubMed, Embase, and COSMIC for articles which are related to diseases covered by DICER1 syndrome. The weighted summary of mutation frequencies among patients with pleuropulmonary blastoma (PPB), cystic nephroma (CN), and Sertoli-Leydig cell tumor (SLCT) were calculated. Forty-nine eligible articles were included. In total, 72 cases with multimorbidity of DICER1 syndrome were identified. More females (n=46, 64%) presented with multimorbidity than males (n=18, 25%) and the remaining 8 patients' sex were unknown. Nineteen of 72 patients with multimorbidity suffered from another disease that was not yet included in DICER1 syndrome, which would provide potential phenotypes of DICER1 syndrome. The germline DICER1 mutation frequencies in PPB, CN, and SLCT were 66.9%, 73.2%, and 57.1%, respectively. The somatic DICER1 mutation frequencies of PPB, CN, and SLCT were 92.4%, 87.9%, and 43.3%, respectively. Majority of patients with multimorbidity of DICER1 syndrome were mutation positive individuals so that multimorbidity may suggest the possible germline mutation of these patients and their relatives.


[The afferent and efferent connections of field 7 in the parietal cortex with field 18a of the visual cortex in the rat].
By means of administration of horseradish peroxidase to the field 7 in the parietal cortex of rat, studies have been made on antero- and retrograde axonal transport of the enzyme to reveal bilateral connections of this field with the field 18a of the visual cortex. It was shown that the field 18a is a source of afferent fibers which reach the field 7, more evident projections being observed at the ipsilateral side. Labeled neurones in the field 18a were found mostly in layer III, and to a lesser extent-in layer V. Single labeled neurones were also found in layers II and VI. Labeled terminals of efferent fibers originating from cells in the field 7 were located in layers I, II-III and V of the field 18a. Maximum density of the terminals was registered in layer II-III, less one in layer V, and very negligible-in layer I. It should be noted that labeled terminals were revealed in those cortical regions, which contained labeled neurones which gave origin to afferent fibers to the field 7.


A Disseminated Cryptococcus gattii VGIIa Infection in a Citron-Crested Cockatoo (Cacatua sulphurea citrinocristata) in Québec, Canada.
Cryptococcus gattii infection in mammals and birds has been confined historically to tropical and subtropical regions in Australia, Southeast Asia, Africa, and South America. Since the early 2000s, numerous reports describe the emergence of C. gattii on the Pacific Coast of North America. We report on a C. gattii infection in an 8-year-old male citron-crested cockatoo (Cacatua sulphurea citrinocristata) hatched on the Canadian Pacific Coast and raised in the province of Québec, Canada. The bird developed a slow growing ulcerated, fleshy, crusty, and hemorrhagic mass infiltrating the left lower rhamphotheca. Cryptococcus gattii infection was confirmed by cytologic examination of a fine needle aspirate of the mass, and results of fungal culture and sequencing. The genotype of the strain was determined to be VGIIa sequence type 20, the strongly overrepresented subgroup found on the Canadian Pacific coast. Minimum inhibitory concentrations for multiple antifungal drugs were determined. The bird received fluconazole but died acutely 55 days after initial presentation. Postmortem examination revealed a disseminated infection, with involvement of the beak, lungs, spleen, and brain.


Responses to the primary health care needs of Aboriginal and Torres Strait Islander women experiencing violence: A scoping review of policy and practice guidelines.
It is demonstrated that primary health care (PHC) providers are sought out by women who experience violence. Given the disproportionate burden of violence experienced by Aboriginal and Torres Strait Islander women, it is essential there is equitable access to appropriate PHC services. This review aimed to analyse whether Australian PHC policy accounts for the complex needs of Aboriginal and Torres Strait Islander women experiencing violence and the importance of PHC providers responding to violence in culturally safe ways. Using the Arskey and O'Malley framework, an iterative scoping review determined the policies for analysis. The selected policies were analysed against concepts identified as key components in responding to the needs of Aboriginal and Torres Strait Islander women experiencing violence. The key components are Family Violence, Violence against Aboriginal and Torres Strait Islander Women, Social Determinants of Aboriginal and Torres Strait Islander Health and Wellbeing, Cultural Safety, Holistic Health, Trauma, Patient-Centred Care and Trauma-and-Violence-Informed Care. Following a search of Australian government websites, seven policies were selected for analysis. Principally, no policy embedded or described best practice across all key components. The review demonstrates the need for a specific National framework supporting Aboriginal and Torres Strait Islander women who seek support from PHC services, as well as further policy analysis and review. SO WHAT?: Aboriginal and Torres Strait Islander women disproportionately experience more severe violence, with complex impact, than other Australian women. PHC policy and practice frameworks must account for this, together with the intersection of contemporary manifestations of colonialism and historical and intergenerational trauma.


FGF2 regulates proliferation of neural crest cells, with subsequent neuronal differentiation regulated by LIF or related factors.
Two of the key early events in the development of the peripheral nervous system are the proliferation of neural crest precursor cells and their subsequent differentiation into different neural cell types. We present evidence that members of the fibroblast growth factor family, (FGF1 or FGF2) act directly on the neural crest cells in vitro to stimulate proliferation in the presence of serum. These findings correlate with in situ hybridisation analysis, which shows FGF2 mRNA is expressed in cells both in the neural tube and within newly formed sensory ganglia (dorsal root ganglia, DRG) at embryonic day 10 in the mouse, when neural crest precursors are proliferating within the DRG. This data infers an autocrine/paracrine loop for FGF regulation of proliferation. Evidence supporting this notion is provided by the finding that part of the endogenous proliferative activity in the NC cultures is related to FGF. It was also found, in early neural crest cultures, that exogenous FGF completely inhibited neuronal differentiation, probably as a direct consequence of its mitogenic activity. In order to stimulate neuronal differentiation significantly, it was necessary to remove the FGF and replace it with leukemia inhibitory factor (LIF) or related factors. Under these conditions, 50% of the cells differentiated into neurons, which developed a sensory neuron morphology and were immunoreactive for the sensory markers CGRP and substance P. These data support a model of neural crest development, whereby multipotential neural crest precursor cells are stimulated to divide by FGF and subsequent development into sensory neurons is regulated by LIF or other cytokines with a similar signalling mechanism.


Decreased stress responsivity of central and peripheral catecholaminergic systems in aged 344/N Fischer rats.
We investigated the effects of stress on central and peripheral sympatho-adrenal and sympatho-neural functions in healthy, intact young (3-4 mo) and aged (24 mo) male Fischer 344/N rats. Extracellular fluid (ECF) levels of the catecholamines norepinephrine (NE), dihydroxyphenylglycol (DHPG), methoxyhydroxyphenylglycol (MHPG), and dihydroxyphenylacetic acid (DOPAC) were obtained by microdialysis in the paraventricular nucleus (PVN) of the hypothalamus at baseline and during immobilization (IMMO). The baseline levels of these substances were similar in both age groups, and their concentrations increased significantly in response to IMMO. The IMMO-induced increases of NE and MHPG, however, were significantly smaller in old than in young rats. Plasma levels of the catecholamines NE, DHPG, MHPG, DOPAC, dihydroxyphenylalanine (DOPA), epinephrine (EPI), dopamine (DA), and HVA were also determined in young and old rats during IMMO. Basal levels of these substances were significantly higher in old than in young rats. The magnitude of the IMMO-induced increases in the majority of these compounds however, was significantly smaller in old than in young rats. We conclude that, at the basal state, aging in the Fischer rat is associated with normal PVN ECF, but high plasma catecholamine levels; at stress state, however, old rats have substantially lesser activation of their central and peripheral catecholaminergic systems than young rats.


Rates of adherence and persistence with allopurinol therapy among gout patients in Israel.
To assess the adherence and persistence with allopurinol therapy among gout patients and to identify risk factors for therapy discontinuation. The study population included adults in Maccabi Healthcare Services, a 2-million member health maintenance organization in Israel, who were diagnosed with gout between 2002 and 2008. Adherence with allopurinol was retrospectively assessed by calculating the proportion of days covered of dispensed prescriptions. Persistence was assessed by calculating the mean proportion of follow-up days covered with allopurinol for every study participant. A total of 7644 patients were identified. Among men, the incidence of gout was strongly associated with age, ranging from 0.5 per 1000 among adults younger than 45 years to more than 36 per 1000 among elderly men aged 85 or older). A total of 1331 gout patients (17% of the study population) were adherent to allopurinol therapy, 36% and 47% had partial and poor adherence, respectively. Persistence analysis indicated that the average duration until therapy was discontinued was similar among men (358 days) and women (379 days). Women aged 45-64 years, non-married individuals, those of low socioeconomic status and those with lower body weight were more likely to discontinue therapy. Logistic regression (n = 2471, 32% of the study sample) showed a 4.5 risk of non-compliance among 45- to 65-year-old women. Better compliance was achieved among those with comorbidities, particularly among patients with concomitant cardiovascular disease. Only one out of six gout patients is adherent with allopurinol. Intervention programmes to increase adherence with treatment should focus on high-risk populations.


TRAIP is a regulator of the spindle assembly checkpoint.
Accurate chromosome segregation during mitosis is temporally and spatially coordinated by fidelity-monitoring checkpoint systems. Deficiencies in these checkpoint systems can lead to chromosome segregation errors and aneuploidy, and promote tumorigenesis. Here, we report that the TRAF-interacting protein (TRAIP), a ubiquitously expressed nucleolar E3 ubiquitin ligase important for cellular proliferation, is localized close to mitotic chromosomes. Its knockdown in HeLa cells by RNA interference (RNAi) decreased the time of early mitosis progression from nuclear envelope breakdown (NEB) to anaphase onset and increased the percentages of chromosome alignment defects in metaphase and lagging chromosomes in anaphase compared with those of control cells. The decrease in progression time was corrected by the expression of wild-type but not a ubiquitin-ligase-deficient form of TRAIP. TRAIP-depleted cells bypassed taxol-induced mitotic arrest and displayed significantly reduced kinetochore levels of MAD2 (also known as MAD2L1) but not of other spindle checkpoint proteins in the presence of nocodazole. These results imply that TRAIP regulates the spindle assembly checkpoint, MAD2 abundance at kinetochores and the accurate cellular distribution of chromosomes. The TRAIP ubiquitin ligase activity is functionally required for the spindle assembly checkpoint control.


Pretrained transformer framework on pediatric claims data for population specific tasks.
The adoption of electronic health records (EHR) has become universal during the past decade, which has afforded in-depth data-based research. By learning from the large amount of healthcare data, various data-driven models have been built to predict future events for different medical tasks, such as auto diagnosis and heart-attack prediction. Although EHR is abundant, the population that satisfies specific criteria for learning population-specific tasks is scarce, making it challenging to train data-hungry deep learning models. This study presents the Claim Pre-Training (Claim-PT) framework, a generic pre-training model that first trains on the entire pediatric claims dataset, followed by a discriminative fine-tuning on each population-specific task. The semantic meaning of medical events can be captured in the pre-training stage, and the effective knowledge transfer is completed through the task-aware fine-tuning stage. The fine-tuning process requires minimal parameter modification without changing the model architecture, which mitigates the data scarcity issue and helps train the deep learning model adequately on small patient cohorts. We conducted experiments on a real-world pediatric dataset with more than one million patient records. Experimental results on two downstream tasks demonstrated the effectiveness of our method: our general task-agnostic pre-training framework outperformed tailored task-specific models, achieving more than 10% higher in model performance as compared to baselines. In addition, our framework showed a potential to transfer learned knowledge from one institution to another, which may pave the way for future healthcare model pre-training across institutions.


Obstructive Sleep Apnea in Pregnancy: Assessment of Current Screening Tools.
To estimate the predictive ability of current obstructive sleep apnea (OSA) screening tools and the individual questions in these tools at identifying pregnant women who have OSA. A total of 293 third-trimester patients were enrolled between 2010 and 2012, and 248 patients had sleep monitor results. Recruited participants completed a questionnaire consisting of six OSA screening tools and overnight portable sleep monitoring. Predictive ability of the screening tools for OSA compared with results of the diagnosis from overnight sleep monitoring was estimated using the area under receiver operating characteristic curves. Two hundred eighteen (88%) of the patients were OSA-negative, and 30 patients (12%) were OSA-positive based on sleep monitoring results. Compared with OSA-negative patients, OSA-positive patients had greater body mass indices and neck circumferences as well as significantly higher rates of hypertension (chronic and gestational), pregestational diabetes mellitus, asthma, and preeclampsia. The overall predictive ability of the screening tools for OSA was modest (area under receiver operating characteristic curves 0.62-0.695). However, individual components of the questionnaire were strongly associated with OSA. We found that none of the studied OSA screening tools accurately detected OSA in pregnant women in the third trimester. We have identified proposed elements of a new screening tool based on promising components from several tools that may more accurately detect patients with OSA. II.


Programmed synthesis of triarylnitroimidazoles via sequential cross-coupling reactions.
Transition-metal-catalyzed programmed sequential arylation reactions of 2-chloro-4-nitro-1H-imidazoles were achieved. The methods are general and were applied in a chemoselective manner for the synthesis of different multiarylated 4-nitroimidazoles bearing three different aryl groups. A salient feature is Pd-catalyzed hetero-hetero coupling at the C5 position through a NO2 directed cross-dehydrogenative coupling (CDC) approach.


Fertilization of cultured Xenopus oocytes and use in studies of maternally inherited molecules.
The methods described here of fertilizing stage VI oocytes are lengthy and quite difficult techniques. They would become more attractive if the success rate (i.e., the number of fertilizations compared to the numbers of matured oocytes) could be improved. An important step toward this for the host transfer technique would be to monitor carefully the status of mature Xenopus females ovaries in relation to cyclical HCG stimulation, so that we could predict more accurately whether stage VI oocytes are fertilizable. The in vitro technique would obviously be improved if oocytes could be fertilized without removing their membranes, perhaps by using oviduct extracts. So far, this approach has had only limited success. It seems that the rewards of using these techniques could be great, in terms of understanding the maternal contribution to development. Although our experiments have not yet shown that oocyte injection of DNA has any advantage over egg injection, it is clear that it is possible to make "mRNA-minus mutants" by this approach. In the message depletion experiments mentioned here, we targeted the cleavage of an mRNA which is of low abundance in the full grown oocyte, but preliminary experiments have shown that we can deplete more abundant messages and produce specific phenotypes. Of course such experiments need to be controlled to show that the effect is specific, and the best proof that this is the case is to rescue the effect with injection of the appropriate mRNA. Finally, it seems likely that the method can be used to study the function of both localized molecules, such as the putative primordial germ cell (PGC) or dorsal determinants, and more ubiquitous molecules such as cytoskeletal elements.


Assesssment of Health Facilities for Implementation of Non-communicable Disease Package.
Non-communicable Diseases are an alarming public health emergency in Nepal. Owing to the risk of NCD's in Nepal, Government of Nepal has developed a Multisectoral Action Plan for Non-communicable Disease 2014-2020 and has adopted the World Health Organization Package of Essential Non-communicable Disease protocol. Prior for its implementation in Nepal, baseline study has been carried out to assess the status of health facilities in Nepal. A descriptive cross-sectional study was carried out in Kailali and Ilam district encompassing a total of 92 health facilities. A set of structured questionnaire and interview guideline was used to obtain the data. Collected data was transferred to Microsoft Excel, cleaned and analyzed in SPSS 16.0. Descriptive analysis was performed to express the frequencies and relative frequencies Results: Of the total health facilities, 49 and 43 health facilities of Ilam and Kailali were interviewed. The hospital of Ilam consisted all the procedure, equipment and medicine for the management of NCDs whilst, health posts lacked Oxygen services. Only 592 posts were fulfilled out of 704 sanctioned post in both the districts of which only 161 were trained in management of NCDs. Atotal of 231 patients were diagnosed with NCDs before the day of study in all the health facilities of both districts. Study reveals the gaps in capacity of health institution and system in terms of training, supply, equipments, and diagnostics. However, training of health workers, supply of essential medicines and improvising the service delivery would supplement the effective implementation of PEN in Nepal.


Transpupillary Two-photon In vivo Imaging of the Mouse Retina.
The retina transforms light signals from the environment into electrical signals that are propagated to the brain. Diseases of the retina are prevalent and cause visual impairment and blindness. Understanding how such diseases progress is critical to formulating new treatments. In vivo microscopy in animal models of disease is a powerful tool for understanding neurodegeneration and has led to important progress towards treatments of conditions ranging from Alzheimer's disease to stroke. Given that the retina is the only central nervous system structure inherently accessible by optical approaches, it naturally lends itself towards in vivo imaging. However, the native optics of the lens and cornea present some challenges for effective imaging access. This protocol outlines methods for in vivo two-photon imaging of cellular cohorts and structures in the mouse retina at cellular resolution, applicable for both acute- and chronic-duration imaging experiments. It presents examples of retinal ganglion cell (RGC), amacrine cell, microglial, and vascular imaging using a suite of labeling techniques including adeno-associated virus (AAV) vectors, transgenic mice, and inorganic dyes. Importantly, these techniques extend to all cell types of the retina, and suggested methods for accessing other cellular populations of interest are described. Also detailed are example strategies for manual image postprocessing for display and quantification. These techniques are directly applicable to studies of retinal function in health and disease.


The structure of the tetanus toxin reveals pH-mediated domain dynamics.
The tetanus neurotoxin (TeNT) is a highly potent toxin produced by Clostridium tetani that inhibits neurotransmission of inhibitory interneurons, causing spastic paralysis in the tetanus disease. TeNT differs from the other clostridial neurotoxins by its unique ability to target the central nervous system by retrograde axonal transport. The crystal structure of the tetanus toxin reveals a "closed" domain arrangement stabilised by two disulphide bridges, and the molecular details of the toxin's interaction with its polysaccharide receptor. An integrative analysis combining X-ray crystallography, solution scattering and single particle electron cryo-microscopy reveals pH-mediated domain rearrangements that may give TeNT the ability to adapt to the multiple environments encountered during intoxication, and facilitate binding to distinct receptors.


High-throughput Identification of Gene Regulatory Sequences Using Next-generation Sequencing of Circular Chromosome Conformation Capture (4C-seq).
The identification of regulatory elements for a given target gene poses a significant technical challenge owing to the variability in the positioning and effect sizes of regulatory elements to a target gene. Some progress has been made with the bioinformatic prediction of the existence and function of proximal epigenetic modifications associated with activated gene expression using conserved transcription factor binding sites. Chromatin conformation capture studies have revolutionized our ability to discover physical chromatin contacts between sequences and even within an entire genome. Circular chromatin conformation capture coupled with next-generation sequencing (4C-seq), in particular, is designed to discover all possible physical chromatin interactions for a given sequence of interest (viewpoint), such as a target gene or a regulatory enhancer. Current 4C-seq strategies directly sequence from within the viewpoint but require numerous and diverse viewpoints to be simultaneously sequenced to avoid the technical challenges of uniform base calling (imaging) with next generation sequencing platforms. This volume of experiments may not be practical for many laboratories. Here, we report a modified approach to the 4C-seq protocol that incorporates both an additional restriction enzyme digest and qPCR-based amplification steps that are designed to facilitate a greater capture of diverse sequence reads and mitigate the potential for PCR bias, respectively. Our modified 4C method is amenable to the standard molecular biology lab for assessing chromatin architecture.


Cardioprotection with beta-blockers: myths, facts and Pascal's wager.
Beta-blockers were documented to reduce reinfarction rate more than 3 decades ago and subsequently touted as being cardioprotective for a broad spectrum of cardiovascular indications such as hypertension, diabetes, angina, atrial fibrillation as well as perioperatively in patients undergoing surgery. However, despite lowering blood pressure, beta-blockers have never shown to reduce morbidity and mortality in uncomplicated hypertension. Also, beta-blockers do not prevent heart failure in hypertension any better than any other antihypertensive drug class. Beta-blockers have been shown to increase the risk on new onset diabetes. When compared with nondiuretic antihypertensive drugs, beta-blockers increase all-cause mortality by 8% and stroke by 30% in patients with new onset diabetes. Beta-blockers are useful for rate control in patients with chronic atrial fibrillation but do not help restore sinus rhythm or have antifibrillatory effects in the atria. Beta-blockers provide symptomatic relief in patients with chronic stable angina but do not reduce the risk of myocardial infarction. Adverse effects of beta-blockers are common including fatigue, dizziness, depression and sexual dysfunction. However, beta-blockers remain a cornerstone in the management of patients having suffered a myocardial infarction and for patients with heart failure. Thus, recent evidence argues against universal cardioprotective properties of beta-blockers but attest to their usefulness for specific cardiovascular indications.


Laboratory diagnosis and genetic analysis of an echovirus 30-associated outbreak of aseptic meningitis in Taiwan in 2001.
A large outbreak of aseptic meningitis occurred from April to November 2001 in Taiwan. Of the 1,130 enterovirus-infected patients, echovirus 30 (E30) infection was diagnosed in 188 (16.6%), with the patients having various clinical manifestations including aseptic meningitis (73.9%), young infant fever (6.9%), respiratory symptoms or herpangina (13.3%), or others (5.9%). The majority of the E30-infected patients were between 3 and 10 years old. Of the 264 E30 strains identified, 94.3, 71, and 67.4% were isolated from RD, MRC-5, and A549 cells, respectively. Primary isolation of E30 required mean times of 3.7 days for RD cells and 4.1 days for MRC-5 and A549 cells. Among all E30-positive patients, virus was most frequently isolated from throat swab specimens (85.2%) and, to a lesser extent, stool (76.4%) or cerebrospinal fluid (70.1%) specimens. The virus isolates were initially identified as echovirus 4 (E4) on the basis of immunofluorescence staining with anti-E4 and anti-E30 (Bastianni prototype) monoclonal antibodies. However, upon performance of the neutralization test, E30-specific reverse transcription-PCR, and sequencing of the VP1 gene, the results identified these isolates as E30, not E4, indicating that the reagent used to type E30, which is produced with the Bastianni strain as the immunogen, is inadequate for the identification of recent E30 isolates in Taiwan. Phylogenetic analyses of the VP1 genes of these isolates showed that their sequences differed from those of E30 isolates from the GenBank database by 9.1 to 25.2%, suggesting that this outbreak was caused by a new variant strain of E30 introduced into Taiwan in 2000 that resulted in the widespread aseptic meningitis epidemic in 2001.


Person centred care and shared decision making: implications for ethics, public health and research.
This paper presents a systematic account of ethical issues actualised in different areas, as well as at different levels and stages of health care, by introducing organisational and other procedures that embody a shift towards person centred care and shared decision-making (PCC/SDM). The analysis builds on general ethical theory and earlier work on aspects of PCC/SDM relevant from an ethics perspective. This account leads up to a number of theoretical as well as empirical and practice oriented issues that, in view of broad advancements towards PCC/SDM, need to be considered by health care ethics researchers. Given a PCC/SDM-based reorientation of health care practice, such ethics research is essential from a quality assurance perspective.


Radio- and nano-chemistry of aqueous Ga(iii) ions anchored onto graphene oxide-modified complexes.
The gallium-68 radiolabelling of new functional graphene oxide composites is reported herein along with kinetic stability investigations of the radio-nanohybrids under different environments and insights into their surface characteristics by SEM and XPS. The present work highlights the potential of graphene oxides as nanocarriers for small molecules such as bis(thiosemicarbazonato) complexes to act as multifunctional platforms for rapid and effective radioimaging agent incorporation.


Rosehip extract and wound healing: A review.
Each year, over 100 million patients are afflicted with new scars from medical procedures worldwide. Natural compounds have shown promise in the treatment of scars and skin disorders. Rosehip oil (RO), produced from the pressed fruit of the rosehip (Rosa canina L.) plant, is used in the pharmaceutical, cosmetic, and food industries. The use of this plant in the treatment of scars has yet to be reviewed. This review aims to analyze the current findings on the use of RO in the treatment of postsurgical scars. This literature search considered published journal articles (clinical trials or literature reviews). Studies were identified by searching electronic databases (PubMed and MEDLINE) and reference lists of respective articles. Additional articles were identified through Google Scholar. Only articles available in English were included in this review. There is a scarcity of high-quality studies assessing the therapeutic potential of RO. From the two human clinical trials using RO, there is some evidence to suggest its potential as an active ingredient in topical formulations for the treatment of wounds. Topical treatments containing RO extract may reduce the size and erythema of postsurgical scars through the polarization of macrophages and the inhibition of inflammatory cytokines. Some evidence suggests that RO may improve postsurgical scars. At present, there is insufficient evidence to recommend the use of RO for the treatment of wounds. Further investigation is required to establish its therapeutic effects on human skin and its potential use as an ingredient in topical formulations.


The Structural Biology of Bcl-xL.
Interactions between the pro-survival and pro-apoptotic members of the Bcl-2 family of proteins dictate whether a cell lives or dies. Much of our knowledge of the molecular details of these interactions has come from biochemical and structural studies on the pro-survival protein Bcl-xL. The first high-resolution structure of any Bcl-2 family member was of Bcl-xL, which revealed the conserved topology amongst all family members. Subsequent structures of Bcl-xL complexes with pro-apoptotic ligands demonstrated the general features of all pro-survival:pro-apoptotic complexes. Structural studies involving Bcl-xL were also the basis for the discovery of the first small-molecule pro-survival protein inhibitors, leading ultimately to the development of a new class of drugs now successfully used for cancer treatment in the clinic. This article will review our current knowledge of the structural biology of Bcl-xL and how this has impacted our understanding of the molecular details of the intrinsic apoptotic pathway.


Evaluation of Gentian cystatin C reagent on Abbott Ci8200 and calculation of glomerular filtration rate expressed in mL/min/1.73 m(2) from the cystatin C values in mg/L.
Estimation of the glomerular filtration rate (GFR) is essential when evaluating patients with kidney disease and treating patients with drugs eliminated from the circulation by the kidneys. Cystatin C has been shown in several studies to be superior to creatinine in the estimation of GFR. At our hospitals, there is an increasing demand for cystatin C and at present we perform approximately 1500 cystatin C analyses a month. We thus need the assay available 24 h/day and to have it on our routine chemistry instrument to minimize handling time per test and time to reported test results. We have evaluated a new cystatin C immunoassay from Gentian (Gentian, Moss, Norway) on Architect ci8200 (Abbott Laboratories, Abbott Park, Ill., USA). A prerequisite at our hospital is that cystatin C results are reported as a calculated GFR in mL/min/1.73 m(2), so we also made a comparison with iohexol clearance. The Gentian cystatin C assay showed good agreement with the corresponding assay from Dade Behring (Deerfield, Ill., USA) and good inter-laboratory concordance. The assay has very low total imprecision, good linearity and strong correlation with iohexol clearance (R (2) = 0.956). The equation for the correlation curve is: y = 79.901x(-1.4389). There was low inter-laboratory variation between the three laboratories involved in the cystatin C evaluation, and thus all three laboratories can use the same equation for calculating the estimated GFR.


Association of Cumulative Colorectal Surgery Hospital Costs, Readmissions, and Emergency Department/Observation Stays with Insurance Type.
Medicare's Hospital Readmission Reduction Program disproportionately penalizes safety-net hospitals (SNH) caring for vulnerable populations. This study assessed the association of insurance type with 30-day emergency department visits/observation stays (EDOS), readmissions, and cumulative costs in colorectal surgery patients. Retrospective inpatient cohort study using the National Surgical Quality Improvement Program (2013-2019) with cost data in a SNH. The odds of EDOS and readmissions and cumulative variable (index hospitalization and all 30-day EDOS and readmissions) costs were modeled adjusting for frailty, case status, presence of a stoma, and open versus laparoscopic surgery. The cohort had 245 private, 195 Medicare, and 590 Medicaid/uninsured cases, with a mean age 55.0 years (SD = 13.3) and 52.9% of the cases were performed on male patients. Most cases were open surgeries (58.7%). Complication rates were 41.8%, EDOS 12.0%, and readmissions 20.1%. Medicaid/uninsured had increased odds of urgent/emergent surgeries (aOR = 2.15, CI = 1.56-2.98, p < 0.001) and complications (aOR = 1.43, CI = 1.02-2.03, p = 0.042) versus private patients. Medicaid/uninsured versus private patients had higher EDOS (16.6% versus 4.1%) and readmissions (22.9% versus 14.3%) rates and higher odds of EDOS (aOR = 4.81, CI = 2.57-10.06, p < 0.001), and readmissions (aOR = 1.62, CI = 1.07-2.50, p = 0.025), while Medicare patients had similar odds versus private. Cumulative variable cost %change was increased for Medicare and Medicaid/uninsured, but Medicaid/uninsured was similar to private after adjusting for urgent/emergent cases. Increased urgent/emergent cases in Medicaid/uninsured populations drive increased complications odds and higher costs compared to private patients, suggesting lack of access to outpatient care. SNH care for higher cost populations, receive lower reimbursements, and are penalized by value-based programs. Increasing healthcare access for Medicaid/uninsured patients could reduce urgent/emergent surgeries, resulting in fewer complications, EDOS/readmissions, and costs.


Microsatellites for ecologists: a practical guide to using and evaluating microsatellite markers.
Recent improvements in genetic analysis and genotyping methods have resulted in a rapid expansion of the power of molecular markers to address ecological questions. Microsatellites have emerged as the most popular and versatile marker type for ecological applications. The rise of commercial services that can isolate microsatellites for new study species and genotype samples at reasonable prices presents ecologists with the unprecedented ability to employ genetic approaches without heavy investment in specialized equipment. Nevertheless, the lack of accessible, synthesized information on the practicalities and pitfalls of using genetic tools impedes ecologists' ability to make informed decisions on using molecular approaches and creates the risk that some will use microsatellites without understanding the steps needed to evaluate the quality of a genetic data set. The first goal of this synthesis is to provide an overview of the strengths and limitations of microsatellite markers and the risks, cost and time requirements of isolating and using microsatellites with the aid of commercial services. The second goal is to encourage the use and consistent reporting of thorough marker screening to ensure high quality data. To that end, we present a multistep screening process to evaluate candidate loci for inclusion in a genetic study that is broadly targeted to both novice and experienced geneticists alike.


Highly Controlled Diffusion Drug Release from Ureasil-Poly(ethylene oxide)-Na+-Montmorillonite Hybrid Hydrogel Nanocomposites.
In this work, we report the effects of incorporation of variable amounts (1-20 wt %) of sodium montmorillonite (MMT) into a siloxane-poly(ethylene oxide) hybrid hydrogel prepared by the sol-gel route. The aim was to control the nanostructural features of the nanocomposite, improve the release profile of the sodium diclofenac (SDCF) drug, and optimize the swelling behavior of the hydrophilic matrix. The nanoscopic characteristics of the siloxane-cross-linked poly(ethylene oxide) network, the semicrystallinity of the hybrid, and the intercalated or exfoliated structure of the clay were investigated by X-ray diffraction, small-angle X-ray scattering, and differential scanning calorimetry. The correlation between the nanoscopic features of nanocomposites containing different amounts of MMT and the swelling behavior revealed the key role of exfoliated silicate in controlling the water uptake by means of a flow barrier effect. The release of the drug from the nanocomposite displayed a stepped pattern kinetically controlled by the diffusion of SDCF molecules through the mass transport barrier created by the exfoliated silicate. The sustained SDCF release provided by the hybrid hydrogel nanocomposite could be useful for the prolonged treatment of painful conditions, such as arthritis, sprains and strains, gout, migraine, and pain after surgical procedures.


Genotoxic and biochemical effects of dimethylamine.
The genotoxicity of dimethylamine (DMA) was investigated in the D7 strain of Saccharomyces cerevisiae. DMA was able to induce mitotic gene conversion and point reverse mutation in the presence of metabolic activation (S9 fraction). To study the co-mutagenicity/co-carcinogenicity or toxicity of DMA, changes in xenobiotic metabolizing enzymes were studied in purified microsomes from DMA-induced mice or rats receiving a single or three consecutive doses (25 or 50 mg/kg body wt). Pentoxyresorufin O-dealkylase (class IIB1 P450, PROD), ethoxyresorufin O-deethylase (IA1, EROD) and p-nitrophenol hydroxylase (IIE1, pNPH) were all affected by DMA treatment with a loss of activity of 62, 55 and 54% in the mouse, or 64, 25 and 56% in the rat for PROD, EROD and pNPH activities, respectively, at the highest tested dose. No significant alteration of P450 IIIA-like activity was seen. Results indicated that the metabolites of DMA induced genetic activity in yeast. In addition, a clear non-specific hepatotoxic effect was recorded as a significant reduction in activity of the selected monooxygenases.


White Matter Hyperintensities Contribute to Language Deficits in Primary Progressive Aphasia.
To determine the contribution of white matter hyperintensities (WMH) to language deficits while accounting for cortical atrophy in individuals with primary progressive aphasia (PPA). Forty-three individuals with PPA completed neuropsychological assessments of nonverbal semantics, naming, and sentence repetition plus T2-weighted and fluid-attenuated inversion recovery scans. Using three visual scales, we rated WMH and cerebral ventricle size for both scan types. We used Spearman correlations to evaluate associations between the scales and scans. To test whether visual ratings-particularly of WMH-are associated with language, we compared a base model (including gray matter component scores obtained via principal component analysis, age, and days between assessment and MRI as independent variables) with full models (ie, the base model plus visual ratings) for each language variable. Visual ratings were significantly associated within and between scans and were significantly correlated with age but not with other vascular risk factors. Only the T2 scan ratings were associated with language abilities. Specifically, controlling for other variables, poorer naming was significantly related to larger ventricles (P = 0.033) and greater global (P = 0.033) and periventricular (P = 0.049) WMH. High global WMH (P = 0.034) were also correlated with worse sentence repetition skills. Visual ratings of global brain health were associated with language deficits in PPA independent of cortical atrophy and age. While WMH are not unique to PPA, measuring WMH in conjunction with cortical atrophy may elucidate more accurate brain structure-behavior relationships in PPA than cortical atrophy measures alone.


Tumor response and survival predicted by post-therapy FDG-PET/CT in anal cancer.
To evaluate the response to therapy for anal carcinoma using post-therapy imaging with positron emission tomography (PET)/computed tomography and F-18 fluorodeoxyglucose (FDG) and to compare the metabolic response with patient outcome. This was a prospective cohort study of 53 consecutive patients with anal cancer. All patients underwent pre- and post-treatment whole-body FDG-PET/computed tomography. Patients had been treated with external beam radiotherapy and concurrent chemotherapy. Whole-body FDG-PET was performed 0.9-5.4 months (mean, 2.1) after therapy completion. The post-therapy PET scan did not show any abnormal FDG uptake (complete metabolic response) in 44 patients. Persistent abnormal FDG uptake (partial metabolic response) was found in the anal tumor in 9 patients. The 2-year cause-specific survival rate was 94% for patients with a complete vs. 39% for patients with a partial metabolic response in the anal tumor (p = 0.0008). The 2-year progression-free survival rate was 95% for patients with a complete vs. 22% for patients with a partial metabolic response in the anal tumor (p < 0.0001). A Cox proportional hazards model of survival outcome indicated that a complete metabolic response was the most significant predictor of progression-free survival in our patient population (p = 0.0003). A partial metabolic response in the anal tumor as determined by post-therapy FDG-PET is predictive of significantly decreased progression-free and cause-specific survival after chemoradiotherapy for anal cancer.


Analysis of the multi-item dimensionality of patients' perceived value in hospital services.
The purpose of this research is to analyse the multi-item dimensionality of patients' perceived value in hospital service. A total of 564 patients administered to Gulhane Military Medical Academy were surveyed. Sample population was specified through convenience sampling procedure. A face to face survey was conducted by three interviewers in August 2008. Confirmatory Data Analysis was carried out to reach the final data analysis. A scale of overall perceived value in health service was developed, which was composed of three dimensions and represented by 21 items. These dimensions were: Functional value, emotional value, and social value. The results indicated that perceived value was a multidimensional construct that affects the whole process of service purchasing. If the hospitals managers give importance to the sub-dimensions of the patient value inclined to their own hospitals and they regularly examine the situation of these dimensions, they can move one step ahead in achieving patient satisfaction and loyalty.


The relationship between the structural and orthogonal compressive properties of trabecular bone.
In this study, cubes of trabecular bone with a wide range of structural properties were scanned on a micro-computed tomography system to produce complete three-dimensional digitizations from which morphological and architectural parameters could be measured in a nondestructive manner. The cubes were then mechanically tested in uniaxial compression in three orthogonal directions and to failure in one direction to find the orthogonal tangent elastic moduli and ultimate strengths. After testing, the cubes were weighed and ashed to determine the apparent and ash densities. A high correlation between the basic stereologic measurements was found, indicating that there is a relationship between the amount of bone and number of trabeculae in cancellous bone. Regression analysis was used to estimate the modulus and ultimate strength; these regressions accounted for 68-90% of the variance in these measures. These relationships were dependent on the metaphyseal type and donor, with the modulus also dependent on the direction of testing. This indicates that the properties of the individual trabeculae, as well as their amount and organization, may be important in predicting the mechanical properties of cancellous bone.


Molecular diversity of arbuscular mycorrhizal fungi in onion roots from organic and conventional farming systems in the Netherlands.
Diversity and colonization levels of naturally occurring arbuscular mycorrhizal fungi (AMF) in onion roots were studied to compare organic and conventional farming systems in the Netherlands. In 2004, 20 onion fields were sampled in a balanced survey between farming systems and between two regions, namely, Zeeland and Flevoland. In 2005, nine conventional and ten organic fields were additionally surveyed in Flevoland. AMF phylotypes were identified by rDNA sequencing. All plants were colonized, with 60% for arbuscular colonization and 84% for hyphal colonization as grand means. In Zeeland, onion roots from organic fields had higher fractional colonization levels than those from conventional fields. Onion yields in conventional farming were positively correlated with colonization level. Overall, 14 AMF phylotypes were identified. The number of phylotypes per field ranged from one to six. Two phylotypes associated with the Glomus mosseae-coronatum and the G. caledonium-geosporum species complexes were the most abundant, whereas other phylotypes were infrequently found. Organic and conventional farming systems had similar number of phylotypes per field and Shannon diversity indices. A few organic and conventional fields had larger number of phylotypes, including phylotypes associated with the genera Glomus-B, Archaeospora, and Paraglomus. This suggests that farming systems as such did not influence AMF diversity, but rather specific environmental conditions or agricultural practices.


Letter regarding the article: Trends, and Outcomes of Transcatheter Aortic Valve Implantation in Aortic Insufficiency; A Nationwide Readmission Database Analysis.
With great interest, we have read the article "Trends, and Outcomes of Transcatheter Aortic Valve Implantation In Aortic Insufficiency; A Nationwide Readmission Database Analysis" by Ullah et al. For its clarity and conciseness, we applaud the author for his extensive research on this delicate topic. The authors have concisely written several scenarios including the current understanding of trends, and potential outcome of aortic insufficiency following transcatheter aortic valve implantation (TAVI). Nonetheless, we thought it would be helpful to add a few more things to the article's conclusion to strengthen it. The inclusion of the pathophysiology of coronary microcirculation disorder, and endothelial dysfunction in patients with aortic stenosis and then examine the impact of TAVI on these conditions can improve the results. Second, echocardiographic data regarding right and left ventricular dimensions and function or tricuspid regurgitation presence and its severity could lead to a better understanding of the mechanism.


The mouse NKR-P1B:Clr-b recognition system is a negative regulator of innate immune responses.
NKR-P1B is a homodimeric type II transmembrane C-type lectinlike receptor that inhibits natural killer (NK) cell function upon interaction with its cognate C-type lectin-related ligand, Clr-b. The NKR-P1B:Clr-b interaction represents a major histocompatibility complex class I (MHC-I)-independent missing-self recognition system that monitors cellular Clr-b levels. We have generated NKR-P1B(B6)-deficient (Nkrp1b(-/-)) mice to study the role of NKR-P1B in NK cell development and function in vivo. NK cell inhibition by Clr-b is abolished in Nkrp1b(-/-) mice, confirming the inhibitory nature of NKR-P1B(B6). Inhibitory receptors also promote NK cell tolerance and responsiveness to stimulation; hence, NK cells expressing NKR-P1B(B6) and Ly49C/I display augmented responsiveness to activating signals vs NK cells expressing either or none of the receptors. In addition, Nkrp1b(-/-) mice are defective in rejecting cells lacking Clr-b, supporting a role for NKR-P1B(B6) in MHC-I-independent missing-self recognition of Clr-b in vivo. In contrast, MHC-I-dependent missing-self recognition is preserved in Nkrp1b(-/-) mice. Interestingly, spontaneous myc-induced B lymphoma cells may selectively use NKR-P1B:Clr-b interactions to escape immune surveillance by wild-type, but not Nkrp1b(-/-), NK cells. These data provide direct genetic evidence of a role for NKR-P1B in NK cell tolerance and MHC-I-independent missing-self recognition.


MetfOrmin BenefIts Lower Extremities with Intermittent Claudication (MOBILE IC): randomized clinical trial protocol.
Peripheral artery disease (PAD) affects over 230 million people worldwide and is due to systemic atherosclerosis with etiology linked to chronic inflammation, hypertension, and smoking status. PAD is associated with walking impairment and mobility loss as well as a high prevalence of coronary and cerebrovascular disease. Intermittent claudication (IC) is the classic presenting symptom for PAD, although many patients are asymptomatic or have atypical presentations. Few effective medical therapies are available, while surgical and exercise therapies lack durability. Metformin, the most frequently prescribed oral medication for Type 2 diabetes, has salient anti-inflammatory and promitochondrial properties. We hypothesize that metformin will improve function, retard the progression of PAD, and improve systemic inflammation and mitochondrial function in non-diabetic patients with IC. 200 non-diabetic Veterans with IC will be randomized 1:1 to 180-day treatment with metformin extended release (1000 mg/day) or placebo to evaluate the effect of metformin on functional status, PAD progression, cardiovascular disease events, and systemic inflammation. The primary outcome is 180-day maximum walking distance on the 6-min walk test (6MWT). Secondary outcomes include additional assessments of functional status (cardiopulmonary exercise testing, grip strength, Walking Impairment Questionnaires), health related quality of life (SF-36, VascuQoL), macro- and micro-vascular assessment of lower extremity blood flow (ankle brachial indices, pulse volume recording, EndoPAT), cardiovascular events (amputations, interventions, major adverse cardiac events, all-cause mortality), and measures of systemic inflammation. All outcomes will be assessed at baseline, 90 and 180 days of study drug exposure, and 180 days following cessation of study drug. We will evaluate the primary outcome with linear mixed-effects model analysis with covariate adjustment for baseline 6MWT, age, baseline ankle brachial indices, and smoking status following an intention to treat protocol. MOBILE IC is uniquely suited to evaluate the use of metformin to improve both systematic inflammatory responses, cellular energetics, and functional outcomes in patients with PAD and IC. The prospective MOBILE IC trial was publicly registered (NCT05132439) November 24, 2021.


Stochastic segmentation models for array-based comparative genomic hybridization data analysis.
Array-based comparative genomic hybridization (array-CGH) is a high throughput, high resolution technique for studying the genetics of cancer. Analysis of array-CGH data typically involves estimation of the underlying chromosome copy numbers from the log fluorescence ratios and segmenting the chromosome into regions with the same copy number at each location. We propose for the analysis of array-CGH data, a new stochastic segmentation model and an associated estimation procedure that has attractive statistical and computational properties. An important benefit of this Bayesian segmentation model is that it yields explicit formulas for posterior means, which can be used to estimate the signal directly without performing segmentation. Other quantities relating to the posterior distribution that are useful for providing confidence assessments of any given segmentation can also be estimated by using our method. We propose an approximation method whose computation time is linear in sequence length which makes our method practically applicable to the new higher density arrays. Simulation studies and applications to real array-CGH data illustrate the advantages of the proposed approach.


Characterization 0.1 wt.% Nanomaterial/Photopolymer Composites with Poor Nanomaterial Dispersion: Viscosity, Cure Depth and Dielectric Properties.
Notably, 3D printing techniques such as digital light processing (DLP) have the potential for the cost-effective and flexible production of polymer-based piezoelectric composites. To improve their properties, conductive nanomaterials can be added to the photopolymer to increase their dielectric properties. In this study, the microstructure, viscosity, cure depth, and dielectric properties of ultraviolet (UV) light curable 0.1 wt.% nanomaterial/photopolymer composites are investigated. The composites with multi-walled carbon nanotubes (MWCNTs), graphene nanoplatelets (GNPs), and carbon black (CB) are pre-dispersed in different solvents (acetone, isopropyl alcohol, and ethanol) before adding photopolymer and continuing dispersion. For all prepared suspensions, a reduction in viscosity is observed, which is favorable for 3D printing. In contrast, the addition of 0.1 wt.% nanomaterials, even with poor dispersion, leads to curing depth reduction up to 90% compared to pristine photopolymer, where the nanomaterial dispersion is identified as a contributing factor. The formulation of MWCNTs dispersed in ethanol is found to be the most promising for increasing the dielectric properties. The post-curing of all composites leads to charge immobility, resulting in decreased relative permittivity.


Nurse managers' leadership roles in mining primary healthcare settings in Gauteng.
Nurse managers carry the light of leadership in mining primary healthcare settings (mPHCs). They must ensure that this light shines for their followers to improve the quality of service rendered in primary healthcare. However, the nurse managers in mPHCs are confusing their leadership roles with management roles. The existing policies such as operational management of mPHCs are also not clarifying the leadership roles of nurse managers. The purpose of this study was to understand the nurse managers' leadership roles in mPHCs. This study was conducted in mPHCs, situated in Gauteng province, in the West Rand in 2017. A qualitative, exploratory, descriptive and contextual research design was employed in this study. The participants were selected using a non-probability purposive sampling method. Ten participants formed part of the study. To collect data, individual in-depth interviews were conducted. Giorgi's descriptive thematic data analysis was used to analyse data. Measures to ensure trustworthiness and ethical principles were adhered to. Three themes emanated: confusion of leadership roles with management roles, confusion of leadership roles with clinical roles and confusion of leadership roles with resources management roles. The importance of describing nurse managers' leadership roles surfaced from this study to eradicate confusion. The nurse managers in mPHCs are now understanding their leadership roles, and that leadership roles, management roles, clinical roles and resources management roles are distinguishable.


Binding and state of aggregation of spin-labeled cecropin AD in phospholipid bilayers: effects of surface charge and fatty acyl chain length.
The binding and state of aggregation of cecropin in large unilamellar vesicles of different surface potential and varying acyl chain length were examined using a Cys-33 spin-labeled derivative of cecropin AD (CAD). Association isotherms of the peptide were measured for vesicles of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) containing 5, 15, and 30 mol % 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (POPG). The isotherms display a concentration-dependent positive cooperativity indicating the possible formation of cecropin aggregates in the lipid phase. The critical aqueous concentration for aggregation was dependent on the fraction of POPG, suggesting the involvement of acidic lipids in the formation and stabilization of the putative aggregate. Our data also indicate that cooperativity depends on the state of side-chain ionization of an acidic residue that titrates between pH 7 and 4.4. The binding of spin-labeled Cys-33 CAD was found to be influenced by the acyl chain length of the host lipid. The association isotherm of the peptide for dilaureoyl-sn-glycero-3-phosphatidylcholine vesicles containing 30 mol % dilauroyl-sn-glycero-3-phosphatidylglycerol (DLPG) differed significantly from that in POPC/POPG and could be interpreted in terms of a monomer-monomer partitioning between the aqueous and lipid phases. ESR line-shape analysis was consistent with peptide aggregation in dioleoyl-sn-glycero-3-phosphatidylglycerol vesicles but not in DLPG vesicles.(ABSTRACT TRUNCATED AT 250 WORDS)


Two isoforms of HOXA9 function differently but work synergistically in human MLL-rearranged leukemia.
HOXA9 plays a critical role in both normal hematopoiesis and leukemogenesis, particularly in the development and maintenance of mixed lineage leukemia (MLL)-rearranged leukemia. Through reverse transcription-polymerase chain reaction (RT-PCR) analysis of HOXA9 transcripts in human leukemia and normal bone marrow samples, we identified a truncated isoform of HOXA9, namely HOXA9T, and found that both HOXA9T and canonical HOXA9 were highly expressed in leukemia cell lines bearing MLL rearrangements, relative to human normal bone marrow cells or other subtypes of leukemia cells. A frameshift in HOXA9T in exon I causes a premature stop codon upstream of the PBX-binding domain and the homeodomain, which leads to the generation of a non-homeodomain-containing protein. Unlike the canonical HOXA9, HOXA9T alone cannot transform normal bone marrow progenitor cells. Moreover, HOXA9T cannot cooperate with MEIS1 to transform cells, despite the presence of a MEIS1-binding domain. Remarkably, although the truncated isoforms of many proteins function as dominant-negative competitors or inhibitors of their full-length counterparts, this is not the case for HOXA9T; instead, HOXA9T synergized with HOXA9 in transforming mouse normal bone marrow progenitor cells through promoting self-renewal and proliferation of the cells. Collectively, our data indicate that both truncated and full-length forms of HOXA9 are highly expressed in human MLL-rearranged leukemia, and the truncated isoform of HOXA9 might also play an oncogenic role by cooperating with canonical HOXA9 in cell transformation and leukemogenesis.


Isolation of the microtubule-vesicle motor kinesin from rat liver: selective inhibition by cholestatic bile acids.
Vesicular transport is supported by microtubule-based, force-transducing adenosine triphosphatases (ATPases), such as kinesin, a ubiquitous motor enzyme that has been well studied in neuronal tissues. Although vesicular transport is important for hepatocellular secretory and clearance activities, the role of kinesin in liver function is poorly understood. Furthermore, the effects of bile acids on kinesin are unknown. Kinesin was purified from rat liver cytosol by conventional chromatography and microtubule affinity binding and was characterized by immunoblotting with domain-specific kinesin antibodies and amino acid sequencing of tryptic fragments. Kinesin activity was measured with and without bile acids using an in vitro motility assay and ATPase assays. Immunoblot analysis and partial amino acid sequencing of purified kinesin showed that the sequence at the heavy chain of hepatic kinesin is nearly identical to that of brain kinesin. Purified kinesin transported microtubules in vitro with a velocity of approximately 0.5 microns/s; this activity was significantly inhibited by 0.5-1 mmol/L taurochenodeoxycholate but not by tauroursodeoxycholate. At a dose of 1 mmol/L, chenodeoxycholate conjugates, but not ursodeoxycholate or cholate conjugates, directly inhibited the ATPase activities of kinesin and another microtubule motor, cytoplasmic dynein. Cholestatic concentrations of chenodeoxycholate conjugates directly inhibit the activity of microtubule motors, suggesting a possible mechanism for impairment of vesicular transport in cholestasis.