Noncanonical compensation of zygotic X transcription in early Drosophila melanogaster development revealed through single-embryo RNA-seq.
When Drosophila melanogaster embryos initiate zygotic transcription around mitotic cycle 10, the dose-sensitive expression of specialized genes on the X chromosome triggers a sex-determination cascade that, among other things, compensates for differences in sex chromosome dose by hypertranscribing the single X chromosome in males. However, there is an approximately 1 hour delay between the onset of zygotic transcription and the establishment of canonical dosage compensation near the end of mitotic cycle 14. During this time, zygotic transcription drives segmentation, cellularization, and other important developmental events. Since many of the genes involved in these processes are on the X chromosome, we wondered whether they are transcribed at higher levels in females and whether this might lead to sex-specific early embryonic patterning. To investigate this possibility, we developed methods to precisely stage, sex, and characterize the transcriptomes of individual embryos. We measured genome-wide mRNA abundance in male and female embryos at eight timepoints, spanning mitotic cycle 10 through late cycle 14, using polymorphisms between parental lines to distinguish maternal and zygotic transcription. We found limited sex-specific zygotic transcription, with a weak tendency for genes on the X to be expressed at higher levels in females. However, transcripts derived from the single X chromosome in males were more abundant that those derived from either X chromosome in females, demonstrating that there is widespread dosage compensation prior to the activation of the canonical MSL-mediated dosage compensation system. Crucially, this new system of early zygotic dosage compensation results in nearly identical transcript levels for key X-linked developmental regulators, including giant (gt), brinker (brk), buttonhead (btd), and short gastrulation (sog), in male and female embryos.


The ocular morphology of the southern hemisphere lamprey geotria australis gray, with special reference to optical specialisations and the characterisation and phylogeny of photoreceptor types.
This paper describes the ocular morphology of young adults of the southern hemisphere lamprey Geotria australis, the sole representative of the Geotriidae, and makes comparisons with those of holarctic lampreys (Petromyzontidae). As previously reported for the holarctic lamprey Ichthyomyzon unicuspis [Collin and Fritzsch, 1993], the lens of G. australis is non-spherical and possesses a cone-shaped posterior that may be capable of mediating variable focus. The avascular retina of G. australis is well differentiated, containing three retinal ganglion cell populations, three layers of horizontal cells and three photoreceptor types. In contrast to petromyzontids that contain only two photoreceptor types (short and long), G. australis possesses one rod-like (R1) and two cone-like (C1 and C2) photoreceptors. Although the rod-like receptor in G. australis may be homologous with the short receptors of holarctic lampreys, the two cone-like receptors have morphological characteristics that differ markedly from those of the long receptors of their holarctic counterparts. The features which distinguish the two cone-like receptors from those of the long receptor type in holarctic lampreys are the characteristics of the mitochondria and the presence of large amounts of two different types of stored secretory material in the endoplasmic reticulum of the myoid (refractile bodies). The endoplasmic reticulum of each receptor type has a different shape and staining profile and is polymorphic, each showing a continuum of distension. It is proposed that the presence of two cone-like photoreceptors with different characteristics would increase the spectral range of G. australis and thus be of value during the parasitic phase, when this lamprey lives in the surface marine waters. The irideal flap, present in G. australis but not petromyzontids, would assist in reducing intraocular flare during life in surface waters. The results of this study, which are discussed in the context of the proposed evolution of lampreys, emphasise that it is important to take into account the characteristics of the eyes of southern hemisphere lampreys when making generalizations about the eyes of lampreys as a whole.


Experiences of virus, retrovirus and retrovirus-like particles in Chinese hamster ovary (CHO) and hybridoma cells used for production of protein therapeutics.
Garnick and coworkers indicated that they experienced two independent MVM outbreaks in a period where approximately 2000 fermentations were performed, hypothesizing that such events were rare but inevitable consequences of very large scale operations. In GIs experience over the last 12 years we have seen no incidence of MVM (or any other virus) in close to 3000 fermentations, albeit at lower volumes than produced at Genentech; GI has used 250-2500L bioreactors for manufacturing whereas Genentech have reported using 100-10,000L bioreactors. Nonetheless, volumes of complex media in the same range as used at Genentech have been used at GI with no observations of viral contamination events. The reason for this is not clear. However, GI's experience in combination with experience from sub-contract testing agencies who service the majority of the biotechnology industry may call the inevitability of an MVM outbreak into question. It would appear that very few adventitious viral contaminations of cell cultures have occurred in industry in the last decade. Interestingly, the frequency of contamination events appear to be lower in CHO cells than in hybridoma cells. It should be noted, however, that these conclusions are not statistically based and the scope of the above survey was somewhat limited. RVLPs are present in both CHO and hybridoma cells. The characteristics of both are compared in Table 4. C-type particles from hybridoma cells are more abundant as a rule than those from CHO cells. Although the majority of C-type particles produced by hybridoma cells appear to be non-infective (in S+L- assays), approximately one in a million particles are competent to replicate in S+L- cells. The evidence that C-type particles can replicate in human cells has proved difficult to reproduce consistently. It is likely that replication of xenotropic hybridoma C-type particles in human cells is inefficient or restricted to only a small number of specific cell lines. C-type RVLPs from CHO cells are produced less abundantly than those from hybridoma cells and are not competent to replicate due to a defective endonuclease gene. However, over the last two decades the use of hybridoma cells and products derived from these cells has not provided any evidence of transmission of these viruses to humans; in addition they can be readily removed or inactivated. Thus, neither agent would appear to constitute a significant risk to pharmaceutical products made from their respective host cells. Nonetheless, given the difference in relative safety profiles between RVLPs from CHO and hybridoma cells it is not unreasonable to propose that safety factors (clearance factors in removal/inactivation studies in excess of the reduction of virus loads to zero) required should be less for a CHO process than for a hybridoma process.


Prevalence and correlates of suicidal ideation among the general population in China during the COVID-19 pandemic.
The coronavirus disease 2019 (COVID-19) pandemic is a major threat to the public. However, the comprehensive profile of suicidal ideation among the general population has not been systematically investigated in a large sample in the age of COVID-19. A national online cross-sectional survey was conducted between February 28, 2020 and March 11, 2020 in a representative sample of Chinese adults aged 18 years and older. Suicidal ideation was assessed using item 9 of the Patient Health Questionnaire-9. The prevalence of suicidal ideation and its risk factors was evaluated. A total of 56,679 participants (27,149 males and 29,530 females) were included. The overall prevalence of suicidal ideation was 16.4%, including 10.9% seldom, 4.1% often, and 1.4% always suicidal ideation. The prevalence of suicidal ideation was higher in males (19.1%) and individuals aged 18-24 years (24.7%) than in females (14.0%) and those aged 45 years and older (11.9%). Suicidal ideation was more prevalent in individuals with suspected or confirmed infection (63.0%), frontline workers (19.2%), and people with pre-existing mental disorders (41.6%). Experience of quarantine, unemployed, and increased psychological stress during the pandemic were associated with an increased risk of suicidal ideation and its severity. However, paying more attention to and gaining a better understanding of COVID-19-related knowledge, especially information about psychological interventions, could reduce the risk. The estimated prevalence of suicidal ideation among the general population in China during COVID-19 was significant. The findings will be important for improving suicide prevention strategies during COVID-19.


Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film.
Sensitive quantitation of multiple cytokines can provide important diagnostic information during infection, inflammation and immunopathology. In this study sensitive immunoassay detection of human cytokines IL-1β, IL-6, IL-12p70 and TNFα is shown for singleplex and multiplex formats using a novel miniaturized ELISA platform. The platform uses a disposable plastic multi-syringe aspirator (MSA) integrating 8 disposable fluoropolymer microfluidic test strips, each containing an array of ten 200 μm mean i.d. microcapillaries coated with a set of monoclonal antibodies. Each MSA device thus performs 10 tests on 8 samples, delivering 80 measurements. Unprecedented levels of sensitivity were obtained with the novel fluoropolymer microfluidic material and simple colorimetric detection in a flatbed scanner. The limit of detection for singleplex detection ranged from 2.0 to 15.0 pg ml(-1), i.e. 35 and 713 femtomolar for singleplex cytokine detection, and the intra- and inter-assay coefficient of variation (CV) remained within 10%. In addition, a triplex immunoassay was developed for measuring IL-1β, IL-12p70 and TNFα simultaneously from a given sample in the pg ml(-1) range. These assays permit high sensitivity measurement with rapid <15 min assay or detection from undiluted blood serum. The portability, speed and low-cost of this system are highly suited to point-of-care testing and field diagnostics applications.


T-cells play the classics with a different spin.
The immune system uses much of the classic machinery of cell biology, but in ways that put a different spin on organization and function. Striking recent examples include the demonstration of intraflagellar transport protein and hedgehog contributions to the immune synapse, even though immune cells lack a primary cilium that would be the typical setting for this machinery. In a second example, lymphocytes have their own subfamily of integrins, the β2 subfamily, and only integrins in this family form a stable adhesion ring using freely mobile ligands, a key feature of the immunological synapse. Finally, we showed recently that T-cells use endosomal sorting complexes required for transport (ESCRTs) at the plasma membrane to generate T-cell antigen receptor-enriched microvesicles. It is unusual for the ESCRT pathway to operate at the plasma membrane, but this may allow a novel form of cell-cell communication by providing a multivalent ligand for major histocompatibility complex-peptide complexes and perhaps other receptors on the partnering B-cell. Immune cells are thus an exciting system for novel cell biology even with classical pathways that have been studied extensively in other cell types.


Intralesional cytokine therapy in cancer: a pilot study of GM-CSF infusion in mesothelioma.
Systemic cytokine therapy in cancer has major side effects, and we reasoned that the local infusion of cytokines into tumors could induce local immunologic responses with minimal toxicity and potentially strong systemic anticancer effects. This study investigated the toxicity and effectiveness of intralesional granulocyte/macrophage-colony-stimulating factor (GM-CSF) infusion in solid-tumor masses. We studied 14 patients (12 men, two women) with malignant mesothelioma (MM), aged 60 years (range, 46-70 years), with stage 2 disease, in whom the tumor was of sufficient size and accessibility for an intralesional catheter to be inserted. Recombinant human GM-CSF (Molgramostim; Schering Plough) was infused intralesionally for 8 weeks, by using a portable pump, at a dose of 2.5-10 micrograms/kg/day. One patient using GM-CSF developed histologically confirmed necrosis of tumor surrounding the distal catheter, one developed a marked lymphocytic infiltrate in the tumor and had a partial response measured by chest computed tomography (CT) scan, 10 progressed, and three had no response. Neutrophilia with morphologic evidence of neutrophil activation and clinical features suggestive of neutrophil plugging of blood vessels occurred at doses > 5 micrograms/kg/day. In vitro, GM-CSF doubled human neutrophil/CD11b/CD18 expression, suggesting that neutrophil clumping as seen in vivo might be due to integrin upregulation. Intralesional infusion of cytokines is feasible but can be associated with systemic toxicity and has considerable technical problems. It produces a localized immune reaction with tumor regression in a minority of patients.


Impact of prenatal diagnosis on the birth prevalence of neural tube defects, Atlanta, 1990-1991.
To determine the impact of prenatal diagnosis on the birth prevalence of neural tube defects (NTDs) in Atlanta during 1990 through 1991. Live-born and stillborn infants with NTDs who were at least 20 weeks' gestation were ascertained by the Metropolitan Atlanta Congenital Defects Program (MACDP), a population-based birth defects registry. Prenatally diagnosed NTD-affected pregnancies were ascertained from the four perinatal centers and the three genetic laboratories operating in Atlanta during 1990 through 1991. Fetal death certificates were also reviewed for potential cases. During 1990 through 1991, MACDP ascertained 59 NTD cases, for a birth prevalence of 0.77/1000 live births. During this period, an additional 28 NTD-affected pregnancies were detected prenatally and terminated before 20 weeks' gestation. The adjusted NTD rate during 1990 through 1991, which includes prenatally diagnosed cases, was 1.13/1000 live births. Prenatal diagnosis is making a substantial impact on the birth prevalence of NTDs in Atlanta. However, since NTD rates in Atlanta were 2 to 2.5 per 1000 live births in 1970, prenatal diagnosis and termination of pregnancy does not completely account for the declining rate of NTDs.


Essential components of schizophrenia care: a Delphi approach.
Other than medication and access to in-patient care, there is little published consensus on the essential components of schizophrenia care. This paper reports a Delphi approach to the identification of these components. This approach identifies and measures the degree of consensus, using a structured iterative series of questionnaires by a group of experienced UK and Irish psychiatrists (n = 15) on the essential components. Pharmacotherapy, in-patient care and community mental health team (CMHT) staffing were explicitly excluded from the study. In total, 106 components were identified and their importance was rated twice. There was consensus on 92% of the components and strong consensus on 64%. The Delphi results were refined by the participants to produce 10 essential components of care.


Optimal Self-Assembly of Linked Constructs and Catenanes via Spatial Confinement.
How to direct the self-assembly of simple templates toward constructs with complex shape and topology is still an open problem. Recent advancements have made it possible to self-assemble various types of knotted constructs, but targeting general multicomponent topologies, that is, links and catenanes, has proved much harder. Here, we study how the yield and complexity of self-assembled links depends on both intrinsic and extrinsic system properties and, particularly, template shape and spatial confinement. We show that slit confinement does not necessarily suppress linking but can rather enhance it significantly thanks to entropic effects. We also found that only a limited set of binary links are recurrent for different template shapes. These privileged topologies include all those experimentally realized so far plus a few additional ones, such as the 772 and 782 links that, hence, ought to be ideal candidates for broadening the current class of constructs with addressable topology.


Understanding causal pathways within health systems policy evaluation through mediation analysis: an application to payment for performance (P4P) in Tanzania.
The evaluation of payment for performance (P4P) programmes has focused mainly on understanding contributions to health service coverage, without unpacking causal mechanisms. The overall aim of the paper is to test the causal pathways through which P4P schemes may (or may not) influence maternal care outcomes. We used data from an evaluation of a P4P programme in Tanzania. Data were collected from a sample of 3000 women who delivered in the 12 months prior to interview and 200 health workers at 150 health facilities from seven intervention and four comparison districts in Tanzania in January 2012 and in February 2013. We applied causal mediation analysis using a linear structural equation model to identify direct and indirect effects of P4P on institutional delivery rates and on the uptake of two doses of an antimalarial drug during pregnancy. We first ran a series of linear difference-in-difference regression models to test the effect of P4P on potential mediators, which we then included in a linear difference-in-difference model evaluating the impact of P4P on the outcome. We tested the robustness of our results to unmeasured confounding using semi-parametric methods. P4P reduced the probability of women paying for delivery care (-4.5 percentage points) which mediates the total effect of P4P on institutional deliveries (by 48%) and on deliveries in a public health facility (by 78%). P4P reduced the stock-out rate for some essential drugs, specifically oxytocin (-36 percentage points), which mediated the total effect of P4P on institutional deliveries (by 22%) and deliveries in a public health facility (by 30%). P4P increased kindness at delivery (5 percentage points), which mediated the effect of P4P on institutional deliveries (by 48%) and on deliveries in a public health facility (by 49%). P4P increased the likelihood of supervision visits taking place within the last 90 days (18 percentage points), which mediated 15% of the total P4P effect on the uptake of two antimalarial doses during antenatal care (IPT2). Kindness during deliveries and the probability of paying out of pocket for delivery care were the mediators most robust to unmeasured confounding. The effect of P4P on institutional deliveries is mediated by financing and human resources factors, while uptake of antimalarials in pregnancy is mediated by governance factors. Further research is required to explore additional and more complex causal pathways.


How to Count Our Microbes? The Effect of Different Quantitative Microbiome Profiling Approaches.
Next-generation sequencing (NGS) has instigated the research on the role of the microbiome in health and disease. The compositional nature of such microbiome datasets makes it however challenging to identify those microbial taxa that are truly associated with an intervention or health outcome. Quantitative microbiome profiling overcomes the compositional structure of microbiome sequencing data by integrating absolute quantification of microbial abundances into the NGS data. Both cell-based methods (e.g., flow cytometry) and molecular methods (qPCR) have been used to determine the absolute microbial abundances, but to what extent different quantification methods generate similar quantitative microbiome profiles has so far not been explored. Here we compared relative microbiome profiling (without incorporation of microbial quantification) to three variations of quantitative microbiome profiling: (1) microbial cell counting using flow cytometry (QMP), (2) counting of microbial cells using flow cytometry combined with Propidium Monoazide pre-treatment of fecal samples before metagenomics DNA isolation in order to only profile the microbial composition of intact cells (QMP-PMA), and (3) molecular based quantification of the microbial load using qPCR targeting the 16S rRNA gene. Although qPCR and flow cytometry both resulted in accurate and strongly correlated results when quantifying the bacterial abundance of a mock community of bacterial cells, the two methods resulted in highly divergent quantitative microbial profiles when analyzing the microbial composition of fecal samples from 16 healthy volunteers. These differences could not be attributed to the presence of free extracellular prokaryotic DNA in the fecal samples as sample pre-treatment with Propidium Monoazide did not improve the concordance between qPCR-based and flow cytometry-based QMP. Also lack of precision of qPCR was ruled out as a major cause of the disconcordant findings, since quantification of the fecal microbial load by the highly sensitive digital droplet PCR correlated strongly with qPCR. In conclusion, quantitative microbiome profiling is an elegant approach to bypass the compositional nature of microbiome NGS data, however it is important to realize that technical sources of variability may introduce substantial additional bias depending on the quantification method being used.


Targeting Multicopy Prophage Genes for the Increased Detection of Borrelia burgdorferi Sensu Lato (s.l.), the Causative Agents of Lyme Disease, in Blood.
The successful treatment of Lyme disease (LD) is contingent on accurate diagnosis. However, current laboratory detection assays lack sensitivity in the early stages of the disease. Because delayed diagnosis of LD incurs high healthcare costs and great suffering, new highly sensitive tests are in need. To overcome these challenges, we developed an internally controlled quantitative PCR (Ter-qPCR) that targets the multicopy terminase large subunit (terL) gene encoded by prophages that are only found in LD-causing bacteria. The terL protein helps phages pack their DNA. Strikingly, the detection limit of the Ter-qPCR was analytically estimated to be 22 copies and one bacterial cell in bacteria spiked blood. Furthermore, significant quantitative differences was observed in terms of the amount of terL detected in healthy individuals and patients with either early or late disease. Together, the data suggests that the prophage-targeting PCR has significant power to improve success detection for LD. After rigorous clinical validation, this new test could deliver a step-change in the detection of LD. Prophage encoded markers are prevalent in many other pathogenic bacteria rendering this approach highly applicable to bacterial identification in general.


dally, a Drosophila glypican, controls cellular responses to the TGF-beta-related morphogen, Dpp.
Decapentaplegic (Dpp) is a Drosophila member of the Transforming Growth Factor-beta (TGF-beta)/Bone Morphogenetic Protein (BMP) superfamily of growth factors. Dpp serves as a classical morphogen, where concentration gradients of this secreted factor control patterning over many cell dimensions. Regulating the level of Dpp signaling is therefore critical to its function during development. One type of molecule proposed to modulate growth factor signaling at the cell surface are integral membrane proteoglycans. We show here that division abnormally delayed (dally), a Drosophila member of the glypican family of integral membrane proteoglycans is required for normal Dpp signaling during development, affecting cellular responses to this morphogen. Ectopic expression of dally+ can alter the patterning activity of Dpp, suggesting a role for dally+ in modulating Dpp signaling strength. These findings support a role for members of the glypican family in controlling TGF-beta/BMP activity in vivo by affecting signaling at the cell surface.


Evaluation of Molecular Methods for the Detection and Quantification of Pathogen-Derived Nucleic Acids in Sediment.
The accurate detection of pathogens in environmental matrices, such as sediment, is critical in understanding pathogen fate and behavior in the environment. In this study, we assessed the usefulness of methods for the detection and quantification of Vibrio spp. and norovirus (NoV) nucleic acids in sediment. For bacteria, a commonly used direct method using hexadecyltrimethylammonium bromide (CTAB) and phenol-chloroform-isoamyl alcohol (PCI) extraction was optimized, whereas for NoV, direct and indirect (virus elution-concentration) methods were evaluated. For quantification, commercially available quantitative PCR (qPCR) and reverse transcription qPCR (RT-qPCR) kits were tested alongside a digital PCR (dPCR) approach. CTAB-based extraction combined with 16 h polyethylene glycol 6000 (PEG6000) precipitation was found to be suitable for the direct extraction of high abundance bacterial and viral nucleic acids. For the indirect extraction of viral RNA, beef extract-based elution followed by PEG6000 precipitation and extraction using the NucliSENS® MiniMag® Nucleic Acid Purification System and the PowerViral® Environmental RNA/DNA Isolation Kit and qRT-PCR resulted in 83-112 and 63-69% recoveries of NoV, respectively. dPCR resulted in lower viral recoveries (47 and 9%) and ~4 orders of magnitude lower Vibrio concentrations (3.6-4.6 log10 gc/100 g sediment) than was observed using qPCR. The use of internal controls during viral quantification revealed that the RT step was more affected by inhibitors than the amplification. The methods described here are suitable for the enumeration of viral and/or bacterial pathogens in sediment, however the use of internal controls to assess efficiency is recommended.


A multispecies overkill simulation of the end-Pleistocene megafaunal mass extinction.
A computer simulation of North American end-Pleistocene human and large herbivore population dynamics correctly predicts the extinction or survival of 32 out of 41 prey species. Slow human population growth rates, random hunting, and low maximum hunting effort are assumed; additional parameters are based on published values. Predictions are close to observed values for overall extinction rates, human population densities, game consumption rates, and the temporal overlap of humans and extinct species. Results are robust to variation in unconstrained parameters. This fully mechanistic model accounts for megafaunal extinction without invoking climate change and secondary ecological effects.


β-ionone induces cell cycle arrest and apoptosis in human prostate tumor cells.
3-Hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase is the rate-limiting activity in the mevalonate pathway that provides essential intermediates for posttranslational modification of growth-associated proteins. Assorted dietary isoprenoids found in plant foods suppress HMG CoA reductase and have cancer chemopreventive activity. β-Ionone, a cyclic sesquiterpene and an end-ring analog of β-carotene, induced concentration-dependent inhibition of the proliferation of human DU145 (IC50 = 210 μmol/L) and LNCaP (IC50 = 130 μmol/L) prostate carcinoma cells and PC-3 prostate adenocarcinoma cells (IC50 = 130 μmol/L). Concomitantly, β-ionone-induced apoptosis and cell cycle arrest at the G1 phase in DU145 and PC-3 cells were shown by fluorescence microscopy, flow cytometry, and TUNEL reaction, and downregulation of cyclin-dependent kinase 4 (Cdk4) and cyclin D1 proteins. Growth suppression was accompanied by β-ionone-induced downregulation of reductase protein. A blend of β-ionone (150 μmol/L) and trans, trans-farnesol (25 μmol/L), an acyclic sesquiterpene that putatively initiates the degradation of reductase, suppressed the net growth of DU145 cells by 73%, an impact exceeding the sum of those of β-ionone (36%) and farnesol (22%), suggesting a synergistic effect. β-ionone, individually or in combination with other HMG CoA reductase suppressors, may have potential in prostate cancer chemoprevention and/or therapy.


Selective mutism: a consensus based care pathway of good practice.
Selective mutism (SM) now acknowledged as an anxiety condition, tends to be a poorly understood, highly complex and vastly under-recognised clinical entity. Children with SM are a vulnerable group as the condition is not the remit of any one professional group. This inevitably leads to delay in formal diagnosis and management. There is a lack of systematic research on which to base guidelines for management. To develop, agree and validate key principles underlying the management of SM through a consensus process involving international experts, in order to create a local care pathway. A local multi-agency consultation process developed 11 statements, which were felt to be the key principles underpinning a potential care pathway for managing SM. Thirteen recognised experts from North America, Europe and Australia participated in a modified Delphi process involving two rounds using a Likert-scale and free commentary. Both quantitative and qualitative analyses were used in the validation or revision of the statements at each stage. Response rates were 100% for Round 1 and 84.6% for Round 2. Despite the differing professional backgrounds and service contexts, by successive revision and/or revalidation of statements, it was possible to arrive at a consensus about key principles relating to early recognition, assessment and intervention. The agreed key principles are presented together with the resulting local care pathway. Through a Delphi process, agreement was reached by a multidisciplinary group of professionals, on key principles that underpin the timely identification, assessment and management of children with SM. These include the potential for staff in school/preschool settings to identify SM and that intervention programmes should generally be based in these settings. Children with SM should receive assessment for possible coexisting disorders, whether developmental, emotional or behavioural and additional specific intervention given for these. Agreement was reached as to what constitutes clinical progress, intervals for monitoring progress, criteria for referral onwards for multidisciplinary specialist assessment and the role of selective serotonin reuptake inhibitor (SSRI) medication. A consensus methodology has been successfully used to compensate for the lack of evidence base and harness the expertise of a relatively small number of experienced professionals in order to provide a basis for the future development of services.


Dissection of the FCGR3A association with RA: increased association in men and with autoantibody positive disease.
Genome-wide association studies in rheumatoid arthritis (RA) have failed to examine the FCGR gene cluster because of the confounding effects of segmental duplication. This study aimed to replicate previous candidate gene studies that had identified a significant association between the FCGR3A-158V allele and RA and then sought to estimate specific subgroup effects. FCGR3A-158F/V genotyping was undertaken in a UK Caucasian replication cohort comprising 2049 patients with RA and 1156 controls. Subgroup analyses assessing the magnitude of association according to gender and autoantibody (rheumatoid factor (RF) and cyclic citrullinated peptide (CCP)) status were undertaken in a pooled cohort of 2963 patients with RA and 1731 controls. Logistic regression was used to test for interaction between FCGR3A and HLA-DRB1 shared epitope (SE) alleles. In the combined RA cohort, borderline association with homozygosity was found for the FCGR3A-158V allele (OR 1.2, p=0.05), which was stronger in men (OR 1.7, p=0.01). Stratification by autoantibody status showed an increased risk in RF and CCP positive RA. Analysis of the FCGR3A-158V and HLA-DRB1 SE interaction revealed roles for both genes in susceptibility to autoantibody positive RA, with no evidence of interaction. FCGR3A is a risk factor for the development of autoantibody positive RA, particularly in men, with evidence of a multiplicative effect with HLA-DRB1 SE.


Neuropeptide Y in rat sympathetic neurons is altered by genetic hypertension and by age.
We have used immunocytochemistry to quantitate neuronal neuropeptide Y in superior cervical ganglia of a strain of normotensive Wistar-Otago rats and a related genetically hypertensive strain over the age range 1-60 weeks. The numbers of neuropeptide Y-immunoreactive cells and total ganglionic cell numbers were both greater in ganglia of young normotensive than in those of hypertensive rats. Between 10 and 60 weeks of age, peptide immunoreactivity and total cell numbers both fell in normotensive rat ganglia but remained constant in ganglia from hypertensive rats. Densitometric analysis showed that the concentrations of neuropeptide Y were similar in neurons of age-matched individuals of both strains, but during aging there was a substantial decline in neuronal peptide content that was similar in both strains and that was not accompanied by any decline in neuronal immunoreactivity for tyrosine hydroxylase. Our results suggest that there is a developmental abnormality of neuropeptide Y in sympathetic neurons of this strain of genetically hypertensive rat and that, furthermore, the aging process is accompanied by a selective loss of neuronal neuropeptide Y that is independent of blood pressure status.


Antipsychotic-like effect of GLP-1 agonist liraglutide but not DPP-IV inhibitor sitagliptin in mouse model for psychosis.
Recent studies indicate a high comorbidity between type-2 diabetes mellitus (T2DM) and neurological disorders. Many are associated with abnormalities in dopamine neurotransmission such as schizophrenia. Because most of the antipsychotic drugs aggravate pre-existing insulin resistance in type-2 diabetics, there is a need to search for alternative antipsychotics. Glucagon like peptide-1 (GLP-1) is a gut hormone primarily involved in glucose homeostasis. GLP-1 agonist (liraglutide) and dipeptidyl peptidase-IV (DPP-IV) inhibitor (sitagliptin) are the US-FDA approved medications for the management of T2DM. However, little is known about their role in dopamine mediated neurological disorders like schizophrenia. To address this, we used apomorphine-induced cage climbing behavior as a murine model for psychosis and examined for potential antipsychotic-like effect of liraglutide and sitagliptin. While acute liraglutide treatment (50 μg/kg; i.p.) significantly attenuated apomorphine (3 mg/kg, s.c.) induced cage climbing, sitagliptin (50mg/kg; i.p.) failed to elicit such effect. This is the first preclinical evidence for antipsychotic-like effect of GLP-1 receptor agonist. These results open an opportunity to explore GLP-1 analogs for their potential to modulate spectrum of dopamine-mediated neurological disorders.


Temperature effect on the SARS-CoV-2: A molecular dynamics study of the spike homotrimeric glycoprotein.
Rapid spread of SARS-CoV-2 virus have boosted the need of knowledge about inactivation mechanisms to minimize the impact of COVID-19 pandemic. Recent studies have shown that SARS-CoV-2 virus can be disabled by heating, the exposure time for total inactivation depending on the reached temperature (e.g. more than 45 min at 329 K or less than 5 min at 373 K. In spite of recent crystallographic structures, little is known about the molecular changes induced by the temperature. Here, we unravel the molecular basis of the effect of the temperature over the SARS-CoV-2 spike glycoprotein, which is a homotrimer with three identical monomers, by executing atomistic molecular dynamics (MD) simulations at 298, 310, 324, 338, 358 and 373 K. Furthermore, both the closed down and open up conformational states, which affect the accessibility of receptor binding domain, have been considered. Our results suggest that the spike homotrimer undergoes drastic changes in the topology of the hydrogen bonding interactions and important changes on the secondary structure of the receptor binding domain (RBD), while electrostatic interactions (i.e. salt bridges) are mainly preserved. The proposed inactivation mechanism has important implications for engineering new approaches to fight the SARS-CoV-2 coronavirus, as for example, cleaving or reorganizing the hydrogen bonds through chaotropic agents or nanoparticles with local surface resonant plasmon effect.


Detection of Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia akari in skin biopsy specimens using a multiplex real-time polymerase chain reaction assay.
Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia akari are the most common causes of spotted fever group rickettsioses indigenous to the United States. Infected patients characteristically present with a maculopapular rash, often accompanied by an inoculation eschar. Skin biopsy specimens are often obtained from these lesions for diagnostic evaluation. However, a species-specific diagnosis is achieved infrequently from pathologic specimens because immunohistochemical stains do not differentiate among the causative agents of spotted fever group rickettsiae, and existing polymerase chain reaction (PCR) assays generally target large gene segments that may be difficult or impossible to obtain from formalin-fixed tissues. This work describes the development and evaluation of a multiplex real-time PCR assay for the detection of these 3 Rickettsia species from formalin-fixed, paraffin-embedded (FFPE) skin biopsy specimens. The multiplex PCR assay was specific at discriminating each species from FFPE controls of unrelated bacterial, viral, protozoan, and fungal pathogens that cause skin lesions, as well as other closely related spotted fever group Rickettsia species. This multiplex real-time PCR demonstrates greater sensitivity than nested PCR assays in FFPE tissues and provides an effective method to specifically identify cases of Rocky Mountain spotted fever, rickettsialpox, and R. parkeri rickettsiosis by using skin biopsy specimens.


Reciprocity constraints on the matrix of reflection from optically anisotropic surfaces.
We derive certain constraints on the reflection matrix for reflection from a plane, nonmagnetic, optically anisotropic surface using a reciprocity theorem stated long ago by Van de Hulst [Light Scattering by Small Particles (Wiley, 1957)] in the context of scattering of polarized light. The constraints are valid for absorbing and chiral media and can be used as tools to check the consistency of derived expressions for such matrices in terms of the intrinsic parameters of the reflecting medium as illustrated by several examples.


Effect of overexpressing rsmA from Pseudomonas aeruginosa on virulence of select phytotoxin-producing strains of P. syringae.
The GacS/GacA two-component system functions mechanistically in conjunction with global post-transcriptional regulators of the RsmA family to allow pseudomonads and other bacteria to adapt to changing environmental stimuli. Analysis of this Gac/Rsm signal transduction pathway in phytotoxin-producing pathovars of Pseudmonas syringae is incomplete, particularly with regard to rsmA. Our approach in studying it was to overexpress rsmA in P. syringae strains through introduction of pSK61, a plasmid constitutively expressing this gene. Disease and colonization of plant leaf tissue were consistently diminished in all P. syringae strains tested (pv. phaseolicola NPS3121, pv. syringae B728a, and BR2R) when harboring pSK61 relative to these isolates harboring the empty vector pME6031. Phaseolotoxin, syringomycin, and tabtoxin were not produced in any of these strains when transformed with pSK61. Production of protease and pyoverdin as well as swarming were also diminished in all of these strains when harboring pSK61. In contrast, alginate production, biofilm formation, and the hypersensitive response were diminished in some but not all of these isolates under the same growth conditions. These results indicate that rsmA is consistently important in the overarching phenotypes disease and endophtyic colonization but that its role varies with pathovar in certain underpinning phenotypes in the phytotoxin-producing strains of P. syringae.


Poly(L-lysine)-graft-dextran copolymer: amazing effects on triplex stabilization under physiological pH and ionic conditions (in vitro).
Triplex DNA formation involving unmodified triplex-forming oligonucleotides (TFOs) is very unstable under physiological conditions. Here, we report a novel strategy to stabilize both purine and pyrimidine motif triplex DNA within the rat alpha1 (I) collagen gene promoter under physiologically relevant conditions by a poly(L-lysine)- graft -dextran copolymer. Using an in vitro electrophoretic mobility shift assay, we show that the copolymer almost completely abrogates the inhibitory effects of physiological concentrations of monovalent cations, particularly potassium ion (K+), on purine motif triplex formation involving very low concentrations of an unmodified guanine-rich TFO. Of importance, pH dependency in pyrimidine motif triplex formation involving an unmodified cytosine-rich TFO is also significantly overcome by the copolymer. Finally, the triplex-stabilizing efficiency of the copolymer is remarkably higher than that of other oligocations, like spermine and spermidine. We suggest that the ability of the graft copolymer to stabilize triplex DNA under physiologically relevant pH and salt concentrations will be a cue for further progress in the antigene strategy.


Involvement of Astrocytes in Alzheimer's Disease from a Neuroinflammatory and Oxidative Stress Perspective.
Alzheimer disease (AD) is a frequent and devastating neurodegenerative disease in humans, but still no curative treatment has been developed. Although many explicative theories have been proposed, precise pathophysiological mechanisms are unknown. Due to the importance of astrocytes in brain homeostasis they have become interesting targets for the study of AD. Changes in astrocyte function have been observed in brains from individuals with AD, as well as in AD in vitro and in vivo animal models. The presence of amyloid beta (Aβ) has been shown to disrupt gliotransmission, neurotransmitter uptake, and alter calcium signaling in astrocytes. Furthermore, astrocytes express apolipoprotein E and are involved in the production, degradation and removal of Aβ. As well, changes in astrocytes that precede other pathological characteristics observed in AD, point to an early contribution of astroglia in this disease. Astrocytes participate in the inflammatory/immune responses of the central nervous system. The presence of Aβ activates different cell receptors and intracellular signaling pathways, mainly the advanced glycation end products receptor/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway, responsible for the transcription of pro-inflammatory cytokines and chemokines in astrocytes. The release of these pro-inflammatory agents may induce cellular damage or even stimulate the production of Aβ in astrocytes. Additionally, Aβ induces the appearance of oxidative stress (OS) and production of reactive oxygen species and reactive nitrogen species in astrocytes, affecting among others, intracellular calcium levels, NADPH oxidase (NOX), NF-κB signaling, glutamate uptake (increasing the risk of excitotoxicity) and mitochondrial function. Excessive neuroinflammation and OS are observed in AD, and astrocytes seem to be involved in both. The Aβ/NF-κB interaction in astrocytes may play a central role in these inflammatory and OS changes present in AD. In this paper, we also discuss therapeutic measures highlighting the importance of astrocytes in AD pathology. Several new therapeutic approaches involving phenols (curcumin), phytoestrogens (genistein), neuroesteroids and other natural phytochemicals have been explored in astrocytes, obtaining some promising results regarding cognitive improvements and attenuation of neuroinflammation. Novel strategies comprising astrocytes and aimed to reduce OS in AD have also been proposed. These include estrogen receptor agonists (pelargonidin), Bambusae concretio Salicea, Monascin, and various antioxidatives such as resveratrol, tocotrienol, anthocyanins, and epicatechin, showing beneficial effects in AD models.


Isolation of Salmonella from urban wild brown rats (Rattus norvegicus) in the West Midlands, UK.
A 6-month study was undertaken to investigate the prevalence of Salmonella enterica in wild urban brown rats (Rattus norvegicus) in the West Midlands. Samples were obtained of faecal droppings (n = 100) and from rectal swabs (n = 50) of rat carcases collected from active infestation sites. A subset of the rats (n = 25) had additional swab samples taken of the fur, paws and tail. Five (10%) of the rectal swabs were positive for Salmonella by direct plating onto XLD media. No further samples were positive following pre-enrichment and selective culture. A total of eight (8%) faecal samples were positive for Salmonella; two by direct plating and a further six following enrichment. All positive faecal samples were fresh or moist upon collection. None of the samples obtained from the outer surfaces of the rat were positive. Additionally, rat faeces were spiked with Salmonella and sampled periodically to determine survival in drying faeces exposed to a typical indoor environment. Salmonella could be recovered by direct culture up to 86 days. These results demonstrate a regional variability in the carriage of Salmonella in urban rats compared to other studies and that Salmonella longevity in faecal pellets is sufficient to present a potential contamination risk in the absence of an active infestation.


Screening for lung cancer with CT: a preliminary cost-effectiveness analysis.
To appraise the potential cost-effectiveness of lung cancer screening with CT. Incremental cost-effectiveness ratios are estimated for two hypothetical cohorts followed up over time. One cohort was screened over the first 5 years of the study period; the other cohort received usual care. Cost streams are projected for each cohort under alternative sets of parameters/ assumptions and from the perspective of national payer groups. Cohort cost differentials arise as a result of screening and variations in stage-specific treatment. Cohort life expectancies are also projected, and they too differ as a consequence of variations in the stage distribution at diagnosis. The ratios of these cost and life-expectancy differences are used to judge the expected economic value of screening. Results are analyzed for a "worst-case" scenario, ie, with the highest cost and lowest yield assumptions. Under these conditions, screening with CT costs approximately $48,000 per life-year gained, if screening results in 50% of lung cancers detected at localized stage. Smaller proportions of cancer detected at a localized stage result in higher cost-effectiveness ratios, and vice versa. If screening for lung cancer is effective, it is likely to be cost-effective if the screening process can detect > 50% of cancers at localized stage.


Intestinal Inflammation Induced by Soybean Meal Ingestion Increases Intestinal Permeability and Neutrophil Turnover Independently of Microbiota in Zebrafish.
Intestinal inflammation is a condition shared by several intestinal chronic diseases, such as Crohn's disease and ulcerative colitis, with severely detrimental consequences in the long run. Current mammalian models have considerably increased understanding of this pathological condition, highlighting the fact that, in most of the cases, it is a highly complex and multifactorial problem and difficult to deal with. Thus, there is an increasingly evident need for alternative animal models that could offer complementary approaches that have not been exploited in rodents, thereby contributing to a different view on the disease. Here, we report the effects of a soybean meal-induced intestinal inflammation model on intestinal integrity and function as well as on neutrophil recruitment and microbiota composition in zebrafish. We find that the induced intestinal inflammation process is accompanied by an increase in epithelial permeability in addition to changes in the mRNA levels of different tight junction proteins. Conversely, there was no evidence of damage of epithelial cells nor an increase in their proliferation. Of note, our results show that this intestinal inflammatory model is induced independently of the presence of microbiota. On the other hand, this inflammatory process affects intestinal physiology by decreasing protein absorption, increasing neutrophil replacement, and altering microbiota composition with a decrease in the diversity of cultivable bacteria.


The effect of old-age pensions on health care utilization patterns and insurance uptake in Mexico.
As old-age pensions continue to expand around the world in response to population ageing, policymakers increasingly wish to understand their impact on healthcare demand. In this paper, we examine the effects of supplemental income to older adults on healthcare use patterns, expenditures and insurance uptake in Yucatan, Mexico. We use a longitudinal survey for individuals aged 70 or older and an individual fixed-effects difference-in-difference approach to understand the effect of an income supplement on healthcare use patterns, out-of-pocket expenditures and health insurance uptake patterns. The implementation of the old-age pension was associated with increased use of healthcare with nuanced effects on the type of care. Old-age pensions increase the use of formal healthcare by 15 percentage points (95% CI 6.1 to 23.9) for those with healthcare use at baseline and by 7.5 percentage points (95% CI 3.7 to 11.3) for those without healthcare use at baseline. We find no evidence of greater out-of-pocket expenditures, likely because old-age pensions were associated with a 4.2 percentage point (95% CI 1.5 to 6.9) increase in use of public health insurance. Old-age pensions can shift healthcare demand towards formal services and eliminate financial barriers to basic care. Pension benefits can also increase the uptake of insurance programmes. These results demonstrate how social programmes can complement each other This highlights the potential role of old-age pensions in achieving universal health coverage for individuals at older ages.


Sodium thiosulfate improves renal function and oxygenation in L-NNA-induced hypertension in rats.
Sodium thiosulfate, a reversible oxidation product of hydrogen sulfide, has vasodilating and anti-oxidative properties, making it an attractive agent to alleviate damaging effects of hypertension. In experimental settings, inhibition of nitric oxide synthase causes hypertension, renal dysfunction and damage. We hypothesized that thiosulfate would attenuate renal injury and improve renal function, hemodynamics and the efficiency of oxygen utilization for sodium reabsorption in hypertensive renal disease. Additionally, thiosulfate co-administration would further improve these variables when compared to inhibiting the renin-angiotensin system alone. Nitric oxide synthase was inhibited in Sprague Dawley rats by administering N-ω-nitro-L-arginine (L-NNA) in the food for three weeks. After one week, rats were split into two groups; without and with thiosulfate in the drinking water. In a parallel study, rats given N-ω-nitro-L-arginine and the angiotensin converting enzyme inhibitor lisinopril at a relatively low dose in their food were divided into two groups; without and with thiosulfate in the drinking water. Treatment with thiosulfate alleviated hypertension (mean 190 vs. 229 mmHg), lowered plasma urea (mean 11.3 vs. 20.0 mmol/L) and improved the terminal glomerular filtration rate (mean 503 vs. 260 μl/min/100 gbw), effective renal plasma flow (mean 919 vs. 514 μl/min/100 gbw) and oxygen utilization for sodium reabsorption (mean 14.3 vs. 8.6 μmol/μmol). Combining thiosulfate with lisinopril further lowered renal vascular resistance (mean 43 vs. 63 mmHg/ml/min/100 gbw) and prevented glomerulosclerosis. Thus, our results suggest that thiosulfate has therapeutic potential in hypertensive renal disease and might be of value when added to standard antihypertensive therapies.


Comparison of Quality of Life Before and After Septoplasty With Short Form-36.
The authors aimed to evaluate quality of life after septal surgery with Short Form-36 survey, and the effectiveness of the survey. Nasal symptoms (nasal obstruction, facial pain, catarrh), and general quality of life (using the Turkish version of the Short Form-36 questionnaire) were assessed preoperatively and at 1st and 6th postoperative months. Acoustic rhinometry and rhinomanometry were assessed preoperatively and at 6th postoperative month. Data from 78 patients were analyzed. This prospective clinical study was conducted on patients complaining of nasal obstruction with nasal septal deviation. Seventy-eight patients were included in the study. Fourty-two patients (53.8%) were male and 36 patients (46.2%) were female. At 1st postoperative month, nasal obstruction, facial pain and catarrh scores significantly improved in all, 46, 18 patients, respectively (P < 0.0001). At 6th postoperative month, scores nonsignificantly worsened by 1 point in 12, 6, 12 patients, respectively. Nasal volume significantly increased and total resistance significantly decreased at 6th month (P < 0.0001). Compared to preoperative values, all items except social function and bodily pain significantly increased at 1st postoperative month. Compared to values at 1st postoperative month, only bodily pain score decreased at 6th postoperative month. One patient presented with septal perforation. Septoplasty is a well-established technique to reduce nasal obstruction and improve quality of life. SF-36 may be used as a reliable measure of changes in quality of life after septal surgery.


Evidence-based practice in a multiprofessional context.
Healthcare today is a complex system with increasing needs of specific knowledge of evaluation of research and implementation into clinical practice. A critical issue is that we all apply evidence-based practice (EBP) with standardised methods and continuing and systematic improvements. EBP includes both scientific and critical assessed experience-based knowledge. For the individual, this means applying evidence-based knowledge to a specific situation, and for the organisation, it means catering for a systematic critical review and evaluation and compiling research into guidelines and programmes. In 2009, the County Council of Jönköping had approximately 335,000 inhabitants and the healthcare organisation had more than 10,000 employees. As the County Council actively promotes clinical improvement, it is interesting to explore how healthcare employees think about and act upon EBP. The aim of this survey was therefore to describe factors that facilitate or hinder the application of EBP in the clinical context. A quantitative study was performed with a questionnaire to healthcare staff employed in the County Council of Jönköping in 2009. The questionnaire consisted of questions concerning which factors are experienced to affect the development of evidence-based healthcare. There were 59 open and closed questions, divided into the following areas: • Sources of knowledge used in practice • Barriers to finding and evaluating research reports and guidelines • Barriers to changing practice on the basis of best evidence • Facilitating factors for changing practice on the basis of best evidence • Experience in finding, evaluating and using different sources of evidence. The participants were selected using the county council's staff database and included medical, caring and rehabilitative staff within hospitals, primary care, dentistry and laboratory medicine. The inclusion criteria were permanent employment and clinical work. Invitations were sent to 5787 persons to participate in the study and 1445 persons answered the questionnaire. Knowledge used in daily clinical practice was mainly based on information about the patient, personal experience and local guidelines. Twenty per cent answered that they worked 'in the way they always had', and 11% responded that they used evidence from research as a basis for change. The participants experienced that EBP was not used enough in clinical healthcare and explained this with practical and structural barriers, which they thought should be better monitored by the organisation and directors. Overall, the results indicate that the scientific evidence for healthcare is not used sufficiently as a base for decisions in daily practice as well as for changing practice. This is more prominent among assistant staff. As a consequence, this might affect the care of the patients in a negative way. Increased awareness of EBP and a stronger evidence-based approach are keys in the ongoing improvement work in the county. Local guidelines seem to be a way to implement knowledge. But, as the arena of activities is complex and the employees have diverse education levels, different strategies to facilitate and promote EBP are necessary.


Older people's experience of proactive welfare rights advice: qualitative study of a South Asian community.
Many older people in the UK require means-tested and health-related benefits to supplement low incomes in retirement and pay for additional resources required to cope with ill-health. Ethnic minority older people have lower uptake of welfare services than white older people. This study investigated routes to the service, barriers to claiming and explored the impact of additional financial resources among ethnic minority elders by evaluating a novel welfare rights advice service which facilitated access to state benefit entitlements. Qualitative study using data from one-to-one interviews with ethnic minority elders from Newcastle upon Tyne, UK analysed using the Framework method. Participants were recruited to this study from among ethnic minority elders attending a full benefits assessment offered by Newcastle Welfare Rights Service. Twenty-two South Asian participants aged between 50 and 81 were interviewed. Nineteen participants were above state retirement age, 15 of whom were on means-tested state benefits. Knowledge of state entitlements was extremely low. Sixteen qualified for non-means-tested health benefits; six qualified for further means-tested state benefits. Additional resources had a considerable impact on participants and their families. Participants could better afford essential items such as food, bills, shoes, clothes and 'one off' payments. Less stress, increased independence and better quality of life were reported. Welfare rights advice also had a positive impact on carers, none of whom knew what they or their relatives were entitled to. As with older people of all backgrounds, facilitating access to state benefit entitlements with appropriate services is an important way of increasing the resources of ethnic minority older people on low incomes and/or in poor health. Such services can also significantly improve quality of life for carers. As the numbers of ethnic minority older people will rise over the next few decades, it is necessary to meet this need with linguistically and culturally appropriate welfare rights services. To do otherwise will exacerbate existing income and health inequalities.


The second myocardial infarction: Higher risk factor burden and earlier second myocardial infarction in women compared with men. The Northern Sweden MONICA study.
Several studies have examined various parameters and experiences when patients suffer their first myocardial infarction (MI), but knowledge about when they suffer their second MI is limited. To compare risk factors for MI, that is, diabetes, hypertension and smoking, for the first and second MI events in men and women affected by two MIs and to analyse the time intervals between the first and second MIs. A retrospective cohort study of 1017 patients aged 25-74 years with first and second MIs from 1990 through 2009 registered in the Northern Sweden MONICA registry. More women than men have diabetes and hypertension and are smokers at the first MI. Similar differences between the genders remain at the time of the second MI for diabetes and hypertension, although both risk factors have increased. Smoking decreased at the second MI without any remaining difference between genders. Women suffer their second MI within a shorter time interval than men do. Within 16 months of their first MI, 50% of women had a second MI. The corresponding time interval for men was 33 months. Patients affected by an MI should be made aware of their risk of recurrent MI and that the risk of recurrence is highest during the first few years after an MI. In patients affected by two MIs, women have a higher risk factor burden and suffer their second MI earlier than men do and thus may need more aggressive and more prompt secondary prevention.


S-Nitrosoglutathione Reductase Dysfunction Contributes to Obesity-Associated Hepatic Insulin Resistance via Regulating Autophagy.
Obesity is associated with elevated intracellular nitric oxide (NO) production, which promotes nitrosative stress in metabolic tissues such as liver and skeletal muscle, contributing to insulin resistance. The onset of obesity-associated insulin resistance is due, in part, to the compromise of hepatic autophagy, a process that leads to lysosomal degradation of cellular components. However, it is not known how NO bioactivity might impact autophagy in obesity. Here, we establish that S-nitrosoglutathione reductase (GSNOR), a major protein denitrosylase, provides a key regulatory link between inflammation and autophagy, which is disrupted in obesity and diabetes. We demonstrate that obesity promotes S-nitrosylation of lysosomal proteins in the liver, thereby impairing lysosomal enzyme activities. Moreover, in mice and humans, obesity and diabetes are accompanied by decreases in GSNOR activity, engendering nitrosative stress. In mice with a GSNOR deletion, diet-induced obesity increases lysosomal nitrosative stress and impairs autophagy in the liver, leading to hepatic insulin resistance. Conversely, liver-specific overexpression of GSNOR in obese mice markedly enhances lysosomal function and autophagy and, remarkably, improves insulin action and glucose homeostasis. Furthermore, overexpression of S-nitrosylation-resistant variants of lysosomal enzymes enhances autophagy, and pharmacologically and genetically enhancing autophagy improves hepatic insulin sensitivity in GSNOR-deficient hepatocytes. Taken together, our data indicate that obesity-induced protein S-nitrosylation is a key mechanism compromising the hepatic autophagy, contributing to hepatic insulin resistance.


Spin-state-correlated optical properties of copper(ii)-nitroxide based molecular magnets.
Molecular magnets based on copper(ii) ions and stable nitroxide radicals exhibit promising switchable behavior triggered by a number of external stimuli; however, their spin-state-correlated optical properties vital for photoinduced switching have not been profoundly investigated to date. Herein, the electronic absorption spectra of single crystals of three representatives of this unique family are studied experimentally and theoretically in the visible and near-IR regions. We established that the color of the complexes is mainly determined by optical properties of the nitroxide radicals, whereas the Cu(hfac)2 fragment contributes to the near-IR range with the intensity smaller by an order of magnitude. The thermochromism of these complexes evident upon thermal spin state switching is mainly caused by a spectral shift of the absorption bands of the nitroxides. The vibrational progression observed in the visible range for single crystals as well as for solutions of pure nitroxides is well reproduced by DFT calculations, where the C-C stretching mode governs the observed progression. The analysis of the spectra of single crystals in the near-IR region reveals changes in the energy and in the intensity of the copper(ii) d-d transitions, which are well reproduced by SOC-NEVPT2 calculations and owe to the flip of the Jahn-Teller axis in the coordination environment of copper. Further strategies for designing bidirectional magnetic photoswitches using these appealing compounds are discussed.


Fructus phyllanthi tannin fraction induces apoptosis and inhibits migration and invasion of human lung squamous carcinoma cells in vitro via MAPK/MMP pathways.
Fructus phyllanthi tannin fraction (PTF) from the traditional Tibetan medicine Fructus phyllanthi has been found to inhibit lung and liver carcinoma in mice. In this study we investigated the anticancer mechanisms of PTF in human lung squamous carcinoma cells in vitro. Human lung squamous carcinoma cell line (NCI-H1703), human large-cell lung cancer cell line (NCI-H460), human lung adenocarcinoma cell line (A549) and human fibrosarcoma cell line (HT1080) were tested. Cell viability was detected with MTT assay. Cell migration and invasion were assessed using a wound healing assay and a transwell chemotaxis chambers assay, respectively. Cell apoptosis was analyzed with flow cytometric analysis. The levels of apoptosis-related and metastasis-related proteins were detected by Western blot and immunofluorescence. PTF dose-dependently inhibited the viability of the 3 human lung cancer cells. The IC50 values of PTF in inhibition of NCI-H1703, NCI-H460, and A549 cells were 33, 203, and 94 mg/L, respectively. PTF (15, 30, and 60 mg/L) dose-dependently induced apoptosis of NCI-H1703 cells. Treatment of NCI-H1703 and HT1080 cells with PTF significantly inhibited cell migration, and reduced the number of invasive cells through Matrigel. Furthermore, PTF dose-dependently down-regulated the expression of phosphor-ERK1/2, MMP-2 and MMP-9, up-regulated the expression of phosphor-JNK, but had no significant effect on the expression of ERK1/2 or JNK. PTF induces cell apoptosis and inhibits the migration and invasion of NCI-H1703 cells by decreasing MPPs expression through regulation of the MAPK pathway.


Longer hospital stay, more complications, and increased mortality but substantially improved function after knee replacement in older patients.
Total knee replacement (TKR) is being increasingly performed in elderly patients, yet there is little information on specific requirements and complication rates encountered by this group. We assessed whether elderly patients undergoing TKR had different length of stay, requirements, complication rates, and functional outcomes compared to younger counterparts. We analyzed prospectively gathered data on 3,144 consecutive primary TKRs (in 2,092 patients aged less than 75 years, 694 patients aged between 75 and 80 years, and 358 patients aged over 80 years at the time of surgery). Incidence of blood transfusion, urinary catheterization, postoperative confusion, cardiac arrhythmia, and 1-year mortality increased with age, even after adjusting for confounding factors, whereas the incidences of chest infection and mortality at 1 month were highest in those aged 75-80. Rates of thromboembolism, prosthetic infection, and revision were similar in the 3 age groups. All groups showed similar substantial improvements in American Knee Society (AKS) knee scores, which were maintained at 5 years. Older patients had smaller improvements in AKS function score, which deteriorated between 3 and 5 years postoperatively, in contrast to the younger group. Elderly people stand to gain considerably from TKR, particularly in terms of pain relief, and they should not be denied surgery based solely on age. However, they should be warned that they can expect a longer length of stay, a higher requirement for blood transfusion and/or urinary catheterization, and more medical complications postoperatively. Mortality was also higher in the older age groups. The risks have been quantified to assist in perioperative counselling, informed consent, and healthcare planning.


Abnormal desmoglein expression by squamous cell carcinoma cells.
Abnormal expression of cell adhesion molecules and related proteins has been observed in various carcinoma cells. We compared expression patterns of desmosomal cadherins, E-cadherin, and cytoplasmic plaque proteins of four different human squamous cell carcinoma cell lines and in vivo squamous cell carcinoma cells with those of normal human keratinocytes. Unlike normal human keratinocytes, the squamous cell carcinoma cells, both in culture and in vivo, exhibited diminished or unusual expression of desmoglein 3 and desmoglein 1, which bear pemphigus vulgaris and pemphigus foliaceus antigens, respectively. Abnormal expression of E-cadherin and cytoplasmic plaque proteins such as desmoplakin and plakoglobin was also observed. Western blotting study demonstrated that three squamous cell carcinoma cell lines expressed two desmogleins with a predominant 150 kDa molecule, and a minor 130 kDa one. Although these molecular sizes were similar to those of cultured normal human keratinocytes, the 130 kDa desmoglein, which usually carries pemphigus antigenic epitopes, was weakly or negatively reactive with pemphigus vulgaris serum. One squamous cell carcinoma cell line showed a doublet of 140 and 145 kDa bands in addition to the 130 kDa band. All the carcinoma cell lines constantly expressed desmoglein 2 and desmoglein 3 mRNA, whereas cultured normal human keratinocytes always expressed desmoglein 1 and desmoglein 3 mRNA, with or without desmoglein 2 mRNA. These findings indicate that the squamous cell carcinoma cells revealed abnormal expression of desmoglein isoforms, which may be related to tumor cell kinetics such as cell invasion and metastasis.


Ancient DNA reveals elephant birds and kiwi are sister taxa and clarifies ratite bird evolution.
The evolution of the ratite birds has been widely attributed to vicariant speciation, driven by the Cretaceous breakup of the supercontinent Gondwana. The early isolation of Africa and Madagascar implies that the ostrich and extinct Madagascan elephant birds (Aepyornithidae) should be the oldest ratite lineages. We sequenced the mitochondrial genomes of two elephant birds and performed phylogenetic analyses, which revealed that these birds are the closest relatives of the New Zealand kiwi and are distant from the basal ratite lineage of ostriches. This unexpected result strongly contradicts continental vicariance and instead supports flighted dispersal in all major ratite lineages. We suggest that convergence toward gigantism and flightlessness was facilitated by early Tertiary expansion into the diurnal herbivory niche after the extinction of the dinosaurs.


Bioengineeredin vitro3D model of myotonic dystrophy type 1 human skeletal muscle.
Myotonic dystrophy type 1 (DM1) is the most common hereditary myopathy in the adult population. The disease is characterized by progressive skeletal muscle degeneration that produces severe disability. At present, there is still no effective treatment for DM1 patients, but the breakthroughs in understanding the molecular pathogenic mechanisms in DM1 have allowed the testing of new therapeutic strategies. Animal models andin vitrotwo-dimensional cell cultures have been essential for these advances. However, serious concerns exist regarding how faithfully these models reproduce the biological complexity of the disease. Biofabrication tools can be applied to engineer human three-dimensional (3D) culture systems that complement current preclinical research models. Here, we describe the development of the firstin vitro3D model of DM1 human skeletal muscle. Transdifferentiated myoblasts from patient-derived fibroblasts were encapsulated in micromolded gelatin methacryloyl-carboxymethyl cellulose methacrylate hydrogels through photomold patterning on functionalized glass coverslips. These hydrogels present a microstructured topography that promotes myoblasts alignment and differentiation resulting in highly aligned myotubes from both healthy and DM1 cells in a long-lasting cell culture. The DM1 3D microtissues recapitulate the molecular alterations detected in patient biopsies. Importantly, fusion index analyses demonstrate that 3D micropatterning significantly improved DM1 cell differentiation into multinucleated myotubes compared to standard cell cultures. Moreover, the characterization of the 3D cultures of DM1 myotubes detects phenotypes as the reduced thickness of myotubes that can be used for drug testing. Finally, we evaluated the therapeutic effect of antagomiR-23b administration on bioengineered DM1 skeletal muscle microtissues. AntagomiR-23b treatment rescues both molecular DM1 hallmarks and structural phenotype, restoring myotube diameter to healthy control sizes. Overall, these new microtissues represent an improvement over conventional cell culture models and can be used as biomimetic platforms to establish preclinical studies for myotonic dystrophy.


Co-occurrence among three divergent plant-castrating fungi in the same Silene host species.
The competitive exclusion principle postulates that different species can only coexist in sympatry if they occupy distinct ecological niches. The goal of this study was to understand the geographical distribution of three species of Microbotryum anther-smut fungi that are distantly related but infect the same host plants, the sister species Silene vulgaris and S. uniflora, in Western Europe. We used microsatellite markers to investigate pathogen distribution in relation to host specialization and ecological factors. Microbotryum violaceo-irregulare was only found on S. vulgaris at high elevations in the Alps. Microbotryum lagerheimii could be subdivided into two genetically differentiated clusters, one on S. uniflora in the UK and the second on S. vulgaris in the Alps and Pyrenees. The most abundant pathogen species, M. silenes-inflatae, could be subdivided into four genetic clusters, co-occurring in the Alps, the UK and the Pyrenees, and was found on both S. vulgaris and S. uniflora. All three fungal species had high levels of homozygosity, in agreement with the selfing mating system generally observed in anther-smut fungi. The three pathogen species and genetic clusters had large range overlaps, but occurred at sites with different elevations, temperatures and precipitation levels. The three Microbotryum species thus do not appear to be maintained by host specialization or geographic allopatry, but instead may occupy different ecological niches in terms of environmental conditions.


Centrosomes and microtubules: wedded with a ring.
How centrosomes nucleate microtubule growth is a question that has puzzled cell biologists for decades. It has been suspected for some time that a centrosome contains multiple copies of a basic microtubule-nucleating structure, each of which is responsible for nucleating a single microtubule. This suspicion has now been confirmed. A ring of gamma-tubulin molecules, associated with a large protein complex, apparently serves as the long-sought-after microtubule-nucleating structure.


Upholding an ideal image of palliative work in the face of obstacles.
Homecare workers play a pivotal role in palliative care, but research on their experiences is limited. The aim of this study was to describe how homecare workers in Sweden experience their work in palliative homecare. This qualitative study used open-ended interviews with a purposive sample of 15 homecare workers. The data was analysed by means of qualitative content analysis. One main theme emerged from the analysis: upholding an ideal image of palliative work. This had four subthemes: doing good; doing their best work; feeling work satisfaction; and facing difficulties. Palliative care performed by homecare workers is demanding. Homecare workers need more practical support and would benefit from a care model that involves working with registered nurses, as a team, to provide humanistic care to dying patients and their relatives.


Costs, benefits and the evolution of inducible defences: a case study with Daphnia pulex.
Phenotypic plasticity is one major source of variation in natural populations. Inducible defences, which can be considered threshold traits, are a form of plasticity that generates ecological and evolutionary consequences. A simple cost-benefit model underpins the maintenance and evolution of these threshold, inducible traits. In this model, a rank-order switch in expected fitness, defined by costs and benefits of induction between defended and undefended morphs, predicts the risk level at which individuals should induce defences. Here, taking predator-induced morphological defences in Daphnia pulex as a threshold trait, we provide the first comprehensive investigation into the costs and benefits of a threshold trait, and how they combine to reflect fitness and predict the switchpoint at which induction should occur. We develop reaction norms that show genetic variation in switchpoints. Further experiments show that induction can confer a survival benefit and a cost in terms of lifetime reproductive success. Together, these two traits combine to estimate expected fitness and can predict the switchpoint between an undefended and a defended strategy. The predictions match the reaction norm data for clones that experience these costs and benefits, and correspond well to independent field data on induction. However, predictions do not, and cannot, match for clones that do not gain a benefit from induction. This study confirms that a simple theory, based on life history costs and benefits, is a sufficient framework for understanding the ecology and evolution of inducible, threshold traits.


The elasticity of the epidemic growth rate to observed weather patterns with an application to yellow rust.
ABSTRACT We extend a previously developed method that quantifies the sensitivity of the exponential epidemic growth rate, r, to weather changes, through a pathogen's life cycle components (basic reproduction number, latent period, and mean and standard deviation of the spore production curve). Here a method is developed to study the elasticities of the system and subsequently the model is linked to observed weather patterns. This enables a direct comparison between the effects of different weather variables (temperature, surface wetness duration, and light quantity) under realistic weather scenarios. The three sites studied represent areas within the United Kingdom with contrasting climates. Yellow rust, caused by Puccinia striiformis, on winter wheat is studied as a key application. Our results show that temperature and more importantly changes in temperature through their effect on pathogen reproduction have the largest effect on r. The long latent period at low winter temperatures is not a key component in the epidemic development, which is contrary to general beliefs. The results combined with long term average yellow rust severity patterns show that it is winter survival and not summer survival that controls the eventual disease severity. The results also show that within the current United Kingdom spraying regime on wheat crops against yellow rust, the first spray should mainly affect the basic reproduction number, i.e., should be a protectant spray, whereas the second spray should also affect the latent period, i.e., should also have curative action.


Housing conditions of home-based care patients in Botswana and the implications for HIV care.
People living with HIV or with a terminal disease face frequent health risks due to opportunistic infections which affect their way of life and dramatically increase their risk of death. Increasingly, terminally ill patients in Botswana are cared for at home, hence the need to understand their housing conditions. Despite a high volume of HIV-related studies undertaken in Botswana, little research has examined the relationship between patients' housing conditions and quality of care at home. Using existing government data from the Botswana 2001 Population and Housing Census, we investigated the housing situation for the households in the national home-based care (HBC) programme. Data were available for 8 872 households having residents enrolled in the programme. The findings indicate that the majority of patients under HBC are faced with housing-related features that can affect their quality of care and chances of recovery. The largest proportion of households had six or more individuals sharing one or two rooms, which raises health concerns for the spread of communicable diseases. In rural areas, few of the HBC households had indoor piped water, proper waste disposal or flush toilets. The lack of basic amenities such as these creates problems for patients as well as for caregivers. We advocate for policy that will address and cater to housing as part of a comprehensive and effective HBC programme. Housing determines where, how and when homecare is provided for sick individuals; therefore, achieving healthy living environments and housing assistance for HBC patients must be addressed by the government. It is recommended that an evaluation of a patient's home should be a first step in their enrolment in the HBC programme. Thereafter, HIV-infected or affected household members should be advised and assisted to make the necessary housing changes to maintain quality of life or accelerate the homecare patient's recovery.


A systematic review of animal models for experimental neuroma.
Peripheral neuromas can result in an unbearable neuropathic pain and functional impairment. Their treatment is still challenging, and their optimal management is to be defined. Experimental research still plays a major role, but - although numerous neuroma models have been proposed on different animals - there is still no single model recognised as being the reference. Several models show advantages over the others in specific aspects of neuroma physiopathology, prevention or treatment, making it unlikely that a single model could be of reference. A reproducible and standardised model of peripheral neuroma would allow better comparison of results from different studies. We present a systematic review of the literature on experimental in vivo models, analysing advantages and disadvantages, specific features and indications, with the goal of providing suggestions to help their standardisation. Published models greatly differ in the animal and the nerve employed, the mechanisms of nerve injury and the evaluation methods. Specific experimental models exist for terminal neuromas and neuromas in continuity (NIC). The rat is the most widely employed animal, the rabbit being the second most popular model. NIC models are more actively researched, but it is more difficult to generate such studies in a reproducible manner. Nerve transection is considered the best method to cause terminal neuromas, whereas partial transection is the best method to cause NIC. Traditional histomorphology is the historical gold-standard evaluation method, but immunolabelling, reverse transcriptase-polymerase chain reaction (RT-PCR) and proteomics are gaining increasing popularity. Computerised gait analysis is the gold standard for motor-recovery evaluation, whereas mechanical testing of allodynia and hyperalgesia reproducibly assesses sensory recovery. This review summarises current knowledge on experimental neuroma models, and it provides a useful tool for defining experimental protocols. Furthermore, it could help future research to define standard experimental model(s) of peripheral neuromas, allowing better comparison of results and improvement of our understanding of such a complex disease.


Molecular phylogeny and evolution of morphology in the social amoebas.
The social amoebas (Dictyostelia) display conditional multicellularity in a wide variety of forms. Despite widespread interest in Dictyostelium discoideum as a model system, almost no molecular data exist from the rest of the group. We constructed the first molecular phylogeny of the Dictyostelia with parallel small subunit ribosomal RNA and a-tubulin data sets, and we found that dictyostelid taxonomy requires complete revision. A mapping of characters onto the phylogeny shows that the dominant trend in dictyostelid evolution is increased size and cell type specialization of fruiting structures, with some complex morphologies evolving several times independently. Thus, the latter may be controlled by only a few genes, making their underlying mechanisms relatively easy to unravel.


Is there a place for sentinel technique in treatment of vaginal cancer?: feasibility, clinical experience, and results.
To evaluate the clinical feasibility of sentinel lymph node (SLN) technique and the role of single-photon emission computed tomography with CT (SPECT/CT) compared to lymphoscintigraphy for detection of SLN in vaginal cancer. The study was performed in a prospective, unicentric setting. Patients with vaginal carcinoma were scheduled for surgery and SLN labeling by peritumoral injection of 10-MBq technetium Tc 99m nanocolloid and patent blue. After 30 minutes, lymphoscintigraphy and SPECT/CT were carried out. We evaluated the number of SLNs in lymphoscintigraphy, SPECT/CT, and intraoperative histology of SLN and non-SLN as well as the impact of these results to therapeutic approach. Between January 2009 and December 2012, the SLN technique was used for 7 of 11 patients treated due to vaginal cancer. Detection rate was 100% (7/7). Lymphoscintigraphy and SPECT/CT showed at least one SLN in each patient. Lymphoscintigraphy detected 2.6 SLNs (range, 2-4 SLNs) per patient compared to 4.3 SLNs (range, 2-8 SLNs) in SPECT/CT (P = 0.053). Sentinel lymph nodes were detected in all patients during surgery with a mean number of 4.3 (range, 1-5). Pelvic SLNs were detected in all 6 patients with infiltration of middle or proximal vaginal third (100%). If the distal vaginal third was additional (3/7 patients) or exclusively (1/7 patients) infiltrated, the inguinal SLN detection rate was 33% and 100%, respectively. All patients with nodal metastases had at least one SLN positive for tumor. There were no false negatives. In 2 (29%) of 7 patients, treatment approach was modified owing to affected SLN. The SLN technique was favorably used in vaginal cancer in this series. It assists in identifying an inguinal and/or pelvic lymphatic drainage. When performed accurately (technetium Tc 99m nanocolloid, lymphoscintigraphy and/or SPECT/CT, blue dye), this technique predicts regional nodal status. This allows tumor stage-adjusted therapy. Single photon emission computed tomography/CT improves preoperative planning and facilitates detection, thus enhancing the clinical value of the SLN technique and improving the oncologic safety of surgery.


Spices for Prevention and Treatment of Cancers.
Spices have been widely used as food flavorings and folk medicines for thousands of years. Numerous studies have documented the antioxidant, anti-inflammatory and immunomodulatory effects of spices, which might be related to prevention and treatment of several cancers, including lung, liver, breast, stomach, colorectum, cervix, and prostate cancers. Several spices are potential sources for prevention and treatment of cancers, such as Curcuma longa (tumeric), Nigella sativa (black cumin), Zingiber officinale (ginger), Allium sativum (garlic), Crocus sativus (saffron), Piper nigrum (black pepper) and Capsicum annum (chili pepper), which contained several important bioactive compounds, such as curcumin, thymoquinone, piperine and capsaicin. The main mechanisms of action include inducing apoptosis, inhibiting proliferation, migration and invasion of tumors, and sensitizing tumors to radiotherapy and chemotherapy. This review summarized recent studies on some spices for prevention and treatment of cancers, and special attention was paid to bioactive components and mechanisms of action.


Hydrolysed formula and risk of allergic or autoimmune disease: systematic review and meta-analysis.
To determine whether feeding infants with hydrolysed formula reduces their risk of allergic or autoimmune disease. Systematic review and meta-analysis, as part of a series of systematic reviews commissioned by the UK Food Standards Agency to inform guidelines on infant feeding. Two authors selected studies by consensus, independently extracted data, and assessed the quality of included studies using the Cochrane risk of bias tool. Medline, Embase, Web of Science, CENTRAL, and LILACS searched between January 1946 and April 2015. Prospective intervention trials of hydrolysed cows' milk formula compared with another hydrolysed formula, human breast milk, or a standard cows' milk formula, which reported on allergic or autoimmune disease or allergic sensitisation. 37 eligible intervention trials of hydrolysed formula were identified, including over 19,000 participants. There was evidence of conflict of interest and high or unclear risk of bias in most studies of allergic outcomes and evidence of publication bias for studies of eczema and wheeze. Overall there was no consistent evidence that partially or extensively hydrolysed formulas reduce risk of allergic or autoimmune outcomes in infants at high pre-existing risk of these outcomes. Odds ratios for eczema at age 0-4, compared with standard cows' milk formula, were 0.84 (95% confidence interval 0.67 to 1.07; I(2)=30%) for partially hydrolysed formula; 0.55 (0.28 to 1.09; I(2)=74%) for extensively hydrolysed casein based formula; and 1.12 (0.88 to 1.42; I(2)=0%) for extensively hydrolysed whey based formula. There was no evidence to support the health claim approved by the US Food and Drug Administration that a partially hydrolysed formula could reduce the risk of eczema nor the conclusion of the Cochrane review that hydrolysed formula could allergy to cows' milk. These findings do not support current guidelines that recommend the use of hydrolysed formula to prevent allergic disease in high risk infants. PROSPERO CRD42013004252.


Mutation in human desmoplakin domain binding to plakoglobin causes a dominant form of arrhythmogenic right ventricular cardiomyopathy.
Arrhythmogenic right ventricular cardiomyopathy (ARVD/C) is a genetically heterogeneous disease characterized by progressive degeneration of the right ventricular myocardium and increased risk of sudden death. Here, we report on a genome scan in one Italian family in which the disease appeared unlinked to any of the six different ARVD loci reported so far; we identify a mutation (S299R) in exon 7 of desmoplakin (DSP), which modifies a putative phosphorylation site in the N-terminal domain binding plakoglobin. It is interesting that a nonsense DSP mutation was reported elsewhere in the literature, inherited as a recessive trait and causing a biventricular dilative cardiomyopathy associated with palmoplantar keratoderma and woolly hairs. Therefore, different DSP mutations might produce different clinical phenotypes, with different modes of inheritance.


Early post-treatment risk stratification of differentiated thyroid cancer: comparison of three high-sensitive Tg assays.
To assess the diagnostic performance of three high-sensitive assays in a cohort of TgAb-negative and TgAb-positive differentiated thyroid cancer (DTC) patients. Retrospective study on prospectively selected DTC patients. Serum samples from 154 DTC patients were obtained 6-12 months after radioiodine ablation and tested by Beckman, Roche, BRAHMS Tg and TgAb assays, respectively. Receiver operating characteristics curves for Tg were plotted using outcome over time as benchmark and assay-specific Tg thresholds were obtained for TgAb-negative and TgAb-positive patients. The frequency of positive TgAb was 21, 20 and 20% for Beckman, Roche and BRAHMS, respectively. In TgAb-negative patients, clinical sensitivities and specificities of 100% and 85-95%, respectively, were observed across all assays. In TgAb-positive patients, clinical sensitivities and specificities of 80-100% and 92-96%, respectively, were observed using lower thresholds than in patients without TgAb. Adopting appropriate thresholds, lower than those for TgAb-negative patients, is possible to reliably follow TgAb-positive patients using highly sensitive Tg assays.


Xenopus ADAM 13 is a metalloprotease required for cranial neural crest-cell migration.
Cranial neural-crest (CNC) cells originate from the lateral edge of the anterior neuroepithelium and migrate to form parts of the peripheral nervous system, muscles, cartilage, and bones of the face. Neural crest-cell migration involves the loss of adhesion from the surrounding neuroepithelium and a corresponding increase in cell adhesion to the extracellular matrix (ECM) present in migratory pathways. While proteolytic activity is likely to contribute to the regulation of neural crest-cell adhesion and migration, the role of a neural crest-specific protease in these processes has yet to be demonstrated. We previously showed that CNC cells express ADAM 13, a cell surface metalloprotease/disintegrin. Proteins of this family are known to act in cell-cell adhesion and as sheddases. ADAMs have also been proposed to degrade the ECM, but this has not yet been shown in a physiological context. Using a tissue transplantation technique, we show that Xenopus CNC cells overexpressing wild-type ADAM 13 migrate along the same hyoid, branchial, and mandibular pathways used by normal CNC cells. In contrast, CNC cell grafts that express protease-defective ADAM 13 fail to migrate along the hyoid and branchial pathways. In addition, ectopic expression of wild-type ADAM 13 results in a gain-of-function phenotype in embryos, namely the abnormal positioning of trunk neural-crest cells. We further show that explanted embryonic tissues expressing wild-type, but not protease-defective, ADAM 13 display decreased cell-matrix adhesion. Purified ADAM 13 can cleave fibronectin, and tissue culture cells that express wild-type, but not protease-defective, ADAM 13 can remodel a fibronectin substrate. Our findings support the hypothesis that the protease activity of ADAM 13 plays a critical role in neural crest-cell migration along defined pathways. We propose that the ADAM 13-dependent modification of ECM and/or other guidance molecules is a key step in the directed migration of the CNC.


Ten hub genes associated with progression and prognosis of pancreatic carcinoma identified by co-expression analysis.
Since the five-year survival rate is less than 5%, pancreatic ductal adenocarcinoma (PDAC) remains the 4th cause of cancer-related death. Although PDAC has been repeatedly researched in recent years, it is still predicted to be the second leading cause of cancer death by year 2030. In our study, the differentially expressed genes in dataset GSE62452 were used to construct a co-expression network by WGCNA. The yellow module related to grade of PDAC was screened. Combined with co-expression network and PPI network, 36 candidates were screened. After survival and regression analysis by using GSE62452 and TCGA dataset, we identified 10 real hub genes (CCNA2, CCNB1, CENPF, DLGAP5, KIF14, KIF23, NEK2, RACGAP1, TPX2 and UBE2C) tightly related to progression of PDAC. According to Oncomine database and The Human Protein Atlas (HPA), we found that all real hub genes were overexpressed in pancreatic carcinoma compared with normal tissues on transcriptional and translational level. ROC curve was plotted and AUC was calculated to distinguish recurrent and non-recurrent PDAC and every AUC of the real hub gene was greater than 0.5. Finally, functional enrichment analysis and gene set enrichment (GSEA) was performed and both of them showed the cell cycle played a vital role in PDAC.


Cloning and characterization of a novel Cry1A toxin from Bacillus thuringiensis with high toxicity to the Asian corn borer and other lepidopteran insects.
A novel cry1A was cloned from Bacillus thuringiensis strain BT8 and expressed in the B. thuringiensis acrystalliferous mutant HD73(-). The gene, designated cry1Ah1, encoded a protein with a molecular weight of 134 kDa. Reverse transcriptase-PCR and Western blotting showed that Cry1Ah was expressed in the host strain BT8. The toxin expressed in HD73(-) exhibited high toxicity against lepidopteran larvae of Ostrinia furnacalis, Helicoverpa armigera, Chilo suppressalis, and Plutella xylostella. The 50% lethal concentrations (LC(50)s) were 0.05, 1.48, 0.98 microg g(-1) and 1.52 microg mL(-1), respectively. The LC(50)s of Cry1Ah were significantly lower than that of Cry1Ac for H. armigera, C. suppressalis, and O. furnacalis, and lower than that of Cry1Ab and Cry1Ie for Ostrinia furnacalis. The high toxicity against a range of pest species makes this novel toxin a potential candidate for insect biocontrol.


Generic Meal Patterns Identified by Latent Class Analysis: Insights from NANS (National Adult Nutrition Survey).
Nutritional data reduction methods are widely applied in nutrition epidemiology in order to classify individuals into meaningful groups with similar dietary patterns. To date, none of the existing studies have applied latent class analysis to examine dietary patterns which include meal types consumed throughout a day. We investigated main meal patterns followed on weekend and weekdays, and evaluated their associations with cardio-metabolic biomarkers. The analyses were performed within the NANS (National Adult Nutrition Survey) a cross-sectional national food consumption survey of 1500 nationally representative Irish adults. A total number of seven dietary patterns were identified using latent class analysis. The typical meal pattern followed by the majority of the population was characterized by consumption of cereal or toast for breakfast, skipping or consuming a sandwich for light meal, and meat or fish with potatoes, pasta or vegetables for the main meal. Eating patterns differed on weekends, and those participants who consumed meat and eggs for breakfast instead of breakfast cereal and skipped light meal were more likely to have an unhealthier dietary pattern, a higher diastolic blood pressure, and increased serum ferritin. The application of data reduction techniques to simplify the multifaceted nature of dietary data is a useful approach to derive patterns, which might shed further light on the typical dietary patterns followed by populations.