Crystal structures of Y41H and Y41F mutants of gene V protein from Ff phage suggest possible protein-protein interactions in the GVP-ssDNA complex.
Gene V protein (GVP) encoded by the filamentous phage Ff (M13, fl, fd) is a homodimeric protein of 87 amino acids that binds to single-stranded DNA (ssDNA) nonspecifically and cooperatively. The structure (monoclinic C2 form) of the wild-type protein has been determined and refined at 1.8-A resolution [Skinner et al. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 2071-2075]. The monomer structure consists of a somewhat distorted five-stranded beta-barrel core with three prominent loops: a DNA-binding loop, a dyad loop, and a dimer contact loop. The amino acid residue at position 41 plays an important role in the dimer-dimer interactions of the protein-ssDNA complex. Two Y41 mutant structures have been studied by X-ray crystallography. The Y41F GVP structure has been refined to an R-factor of 0.180 at 2.2-A resolution and is very similar to the wild-type (wt) structure (rmsd of all C alpha atoms = 0.30 A). In contrast, Y41H GVP forms a new crystal lattice in the space group P2(1)2(1)2(1) with a = 77.18 A, b = 84.17 A, and c = 28.62 A. Its structure has been solved by the molecular replacement method and refined to an R-factor of 0.170 at 2.5-A resolution. The two monomers of Y41H are crystallographically independent, and their structures remain similar to wt-GVP but with significant differences, particularly in the DNA-binding hairpin region. In both crystals, the loop (residues 36-43) that contains the Y41 residue is involved in the crystal dimer packings but in a different manner. The dimer-dimer contacts found in the wt-GVP crystal may be important for GVP aggregation in the absence of DNA. In the presence of DNA, the dimer-dimer contacts may switch to the type found in the Y41H crystal, allowing the GVP-ssDNA complex to form cooperatively. A model of the complex, consistent with existing biochemical and biophysical data, has been constructed from those crystal packing data.


Evaluation of NASA Deep Blue/SOAR aerosol retrieval algorithms applied to AVHRR measurements.
The Deep Blue (DB) and Satellite Ocean Aerosol Retrieval (SOAR) algorithms have previously been applied to observations from sen-sors like the Moderate Resolution Imaging Spectroradiometers (MODIS) and Sea-viewing Wide Field-of-view Sensor (SeaWiFS) to provide records of mid-visible aerosol optical depth (AOD) and related quantities over land and ocean surfaces respectively. Recently, DB and SOAR have also been applied to Ad-vanced Very High Resolution Radiometer (AVHRR) observations from several platforms (NOAA11, NOAA14, and NOAA18), to demonstrate the potential for extending the DB and SOAR AOD records. This study provides an evaluation of the initial version (V001) of the resulting AVHRR-based AOD data set, including validation against Aerosol Robotic Network (AERONET) and ship-borne observations, and comparison against both other AVHRR AOD Research (GESTAR), Universities Space Research Association. records and MODIS/SeaWiFS products at select long-term AERONET sites. Although it is difficult to distil error characteristics into a simple expression, the results suggest that one standard deviation confidence intervals on retrieved AOD of ±(0.03+15%) over water and ±(0.05+25%) over land represent the typical level of uncertainty, with a tendency towards negative biases in high-AOD conditions, caused by a combination of algorithmic assumptions and sensor calibration issues. Most of the available validation data are for NOAA18 AVHRR, although performance appears to be similar for the NOAA11 and NOAA14 sensors as well.


Geopolitical factors and mental health I.
Natural and man-made disasters carry with them major burden and very often the focus is on immediate survival and management of resulting infectious diseases. The impact of disasters directly and indirectly on the well-being and mental health of those affected often gets ignored. The reasons are often stigma and lack of attention to mental health consequences. In addition, often the focus is on preventing the spread of infectious diseases such as waterborne or airborne diseases. This is further complicated by the fact that often aid agencies in offering aid tend to focus on communicable diseases and not on mental health of populations. This focus may reflect easily to measure outcomes in comparison with mental illnesses as the global burden of disease is likely to increase in the next few decades. There is an urgent need to apply the principles of social justice on social and health care policies, which will lead to elimination of stigma. In this article, we propose that the impact of mental illness as a result of disasters needs to be taken seriously in any planning and delivery of relief. Mental health is likely to be affected both directly and indirectly as a result of disasters and also likely to be influenced by ongoing factors such as poor housing, overcrowding and other social determinants. In addition to deliver equity between physical and mental illnesses, appropriate and adequate resources are needed so that identifiable needs can be met with clear outcomes.


Metalo components exhibiting significant anticancer and antibacterial properties: a novel sandwich-type like polymeric structure.
Four new dicyanoargentate(I)-based complexes 1-4 were synthesized from certain metal ions with a tetradentate ligand [N, N-bis (2-hydroxyethyl) -ethylenediamine; N-bishydeten] and determined by diverse procedures (elemental, thermal, FT-IR, ESI-MS for 1-3 and, magnetic susceptibility and EPR for 1, and 2) including crystal analysis of 4. The crystal method revealed that complex 4 has a sandwich-type like polymeric chemical structure with layers formed by [Cd(N-bishydeten)2]2+ cations and [Ag(CN)2]- anions. The complexes were further characterized by fluorescence and UV spectroscopy to determine their physicochemical features. The complexes displayed a DNA binding activity within the same range as found for cisplatin, in addition to their strong stability in the presence of the physiological buffer system. The complexes were also investigated for pharmacological properties like interaction with DNA/Bovine serum albumin, anticancer and antibacterial activities. Physicochemical studies of DNA with the complexes suggested that the interaction mode between them are possibly both intercalative and groove binding types. These spectroscopic measurements also show that there may be a binding tendency between BSA and the complexes via hydrogen or Van der Waals bonds. The viability tests demonstrated that all the complexes exhibited antibacterial (1-4) and anticancer effects (2-4) toward ten diverse bacterial strains and three tumor cells (HT-29 colon adenocarcinoma, HeLa cervical cancer, and C6 glioma), respectively.


The Ebola-effect in Guinea 2014-15: Tangled trends of malaria care in children under-five.
The 2014-15 Ebola outbreak in West Africa was disruptive for the general health services in the affected countries. This study assessed the impact of the outbreak on the reported number and management of malaria in children under-five in rural Guinea. A retrospective cross-sectional study was conducted in nineteen health centres in two rural, malaria-endemic health districts, one at the epicentre of the outbreak (Guéckédou) and one (Koubia) spared by Ebola. Routine surveillance data at health facility level were compared over similar periods of high malaria transmission in both districts before, during and after the outbreak. There were significant declines in the number of visits during the Ebola outbreak (3,700) in Guéckédou compared to before (4,616) and after it (4,195), while this trend remained more stable within the three periods for Koubia. Differences were nonetheless significant in both districts (p<0.001). In 2014, during the peak of the outbreak, the overall number of malaria cases treated exceeded the number of confirmed malaria cases in Guéckédou. There were decreases in antimalarial treatment provision in August and November 2014. In contrast, during 2015 and 2016, the proportion of malaria positive cases and those treated were closely aligned. During the peak of the Ebola outbreak, there was a significant decrease in oral antimalarial drug administration, which corresponded to an increase in injectable antimalarial treatments. Stock-outs in rapid diagnostic tests were evident and prolonged in Guéckédou during the outbreak, while more limited in Koubia. The Ebola outbreak of 2014-15 in Guinea had a significant impact on the admission and management of malaria in children under-five. This study identifies potential challenges in the delivery of care for those at highest risk for malaria mortality during an Ebola outbreak and the need to improve preparedness strategies pre-Ebola and health systems recovery post-Ebola.


Body image during pregnancy: an evaluation of the suitability of the Body Attitudes Questionnaire.
Available data suggest that body dissatisfaction is common during pregnancy and may even be a precursor to post-natal depression. However, in order to accurately identify at-risk women, it is essential to first establish that body image measures function appropriately in pregnant populations. Our study examines the suitability of the Body Attitudes Questionnaire (BAQ) for measuring body dissatisfaction among pregnant women by comparing the psychometric functioning of the BAQ: (1) across key phases of pregnancy, and (2) between pregnant and non-pregnant women. A total of 176 pregnant women from Melbourne, Victoria filled out a questionnaire battery containing demographic questions and the Body Attitudes Questionnaire at 16, 24, and 32 weeks during pregnancy. A comparison group of 148 non-pregnant women also completed the questionnaire battery at Time 1. Evaluations of the psychometric properties of the BAQ consisted of a series of measurement invariance tests conducted within a structural equation modelling framework. Although the internal consistency and factorial validity of the subscales of the BAQ were established across time and also in comparisons between pregnant and non-pregnant women, measurement invariance tests showed non-invariant item intercepts across pregnancy and also in comparison with the non-pregnant subgroup. Inspection of modification indices revealed a complex, non-uniform pattern of differences in item intercepts across groups. Collectively, our findings suggest that comparisons of body dissatisfaction between pregnant and non-pregnant women (at least based on the BAQ) are likely to be conflated by differential measurement biases that serve to undermine attempts to accurately assess level of body dissatisfaction. Researchers should be cautious in assessments of body dissatisfaction among pregnant women until a suitable measure has been established for use in this population. Given the fact that body dissatisfaction is often associated with maladaptive behaviours, such as unhealthy eating and extreme weight loss behaviours, and with ante-and post-natal depression, that have serious negative implications for women's health and well-being, and potentially also for the unborn foetus during pregnancy, developing a suitable body image screening tool, specific to the perinatal period is clearly warranted.


Conformational requirement for lysine hydroxylation in collagen. Structural studies on synthetic peptide substrates of lysyl hydroxylase.
An attempt has been made to understand the conformational determinants that govern the hydroxylation of selected lysyl residues in the nascent collagen molecule by lysyl hydroxylase (EC 220.127.116.11). A series of peptide substrates of the enzyme, ranging in length from 3 to 12 residues, were synthesized. These included: tert-butyloxylcarbonyl (t-Boc)-Ile-Lys-Gly; Boc-Ala-Lys-Gly; N-acetyl-Ala-Lys-Gly-Ser; Hyp-Gly-Pro-Lys-Gly-Glu; Leu-Hyp-Gly-Ala-Lys-Gly-Glu; Gly-Phe-Hyp-Gly-Leu-Hyp-Gly-Ala-Lys-Gly-Glu; (Hyp-Gly-Pro-Lys-Gly-Glu)2; and Ala-Arg-Gly-Ile-Lys-Gly-Ile-Arg-Gly-Phe-Ser-Gly. The conformational features of these peptides were studied by spectroscopic methods so as to relate this information with the kinetic parameters for the interaction of these peptides with purified lysyl hydroxylase. Spectroscopic data, supported by conformational energy calculations, indicated that the tripeptides t-Boc-Ile-Lys-Gly and t-Boc-Ala-Lys-Gly adopt a gamma-turn structure in water and trifluoroethanol with Lys in the second position of the turn. In the tetra- and larger peptides two structures, the beta-turn and a polyproline-II (PP-II) type extended conformation, were identified. The proportions of these two structures in a given peptide depended on the polarity of the solvent. All of the peptides were hydroxylated by lysyl hydroxylase isolated from chicken embryos. In contrast, a control peptide, t-Boc-Ala-Gly-Lys which adopted a beta-turn with Lys at the end of the turn, was not hydroxylated. Competitive inhibition of the hydroxylation of protocollagen by some of the peptides showed a common binding site for these substrates in the enzyme's active site. Kinetic data on the peptides indicated improved hydroxylation rate (higher Vmax) in peptides having relatively higher beta-turn content and improved binding (lower Km) in peptides with higher content of the PP-II structure. The efficacy of the substrate was also governed by its chain length. These data suggest that the conformational criterion for lysine hydroxylation in collagen-related peptides is the presence of a "bent" structure, such as the gamma- or beta-turn at the catalytic site of lysyl hydroxylase and an "extended" PP-II type structure at the binding site(s) of the enzyme's active site. This suggestion also provides a conformational rationale for earlier observations on the substrate specificity of lysyl hydroxylase.


Survey of the year 2008: applications of isothermal titration calorimetry.
Isothermal titration calorimetry (ITC) is a fast, accurate and label-free method for measuring the thermodynamics and binding affinities of molecular associations in solution. Because the method will measure any reaction that results in a heat change, it is applicable to many different fields of research from biomolecular science, to drug design and materials engineering, and can be used to measure binding events between essentially any type of biological or chemical ligand. ITC is the only method that can directly measure binding energetics including Gibbs free energy, enthalpy, entropy and heat capacity changes. Not only binding thermodynamics but also catalytic reactions, conformational rearrangements, changes in protonation and molecular dissociations can be readily quantified by performing only a small number of ITC experiments. In this review, we highlight some of the particularly interesting reports from 2008 employing ITC, with a particular focus on protein interactions with other proteins, nucleic acids, lipids and drugs. As is tradition in these reviews we have not attempted a comprehensive analysis of all 500 papers using ITC, but emphasize those reports that particularly captured our interest and that included more thorough discussions we consider exemplify the power of the technique and might serve to inspire other users.


Robotic lobectomy: flattening the learning curve.
Early experience with robotic technology in pulmonary resection has emphasized a steep learning curve. We initiated a robotic thoracic surgical program with the goal of minimizing complications, operative times, and hospital stays. We implemented robotic lobe resections at our institution with the intent of performing an operationally analogous procedure to that of the open technique. Specifically, we used single docking of the robotic cart, innovative retraction, single interspace port placement, and dockings specific to the resected lobe. We reviewed outcomes for patients undergoing robotic lobectomy at our institution. Data is presented as mean ± standard deviation. 20 patients (69 ± 12 years) underwent robotic lobe resections. American Joint Committee on Cancer staging for 14 patients undergoing resections for non-small cell lung cancers were Stage I (10), Stage II (2), and Stage III (2). Operative times for 20 patients undergoing robotic lobectomies were 203 ± 53 min. Median postoperative hospital stay was 3 days. Conversions to open procedures were required in two patients secondary to failure to progress (1) and bleeding (1). Complications occurred in four (20%) patients and included atelectasis (2), myocardial infarction (1), and atrial fibrillation (1). No fatalities occurred. The perception that robotic pulmonary resection involves a steep learning curve may not be universally accurate; our operative times and hospital stays are consistent with those reported by established programs. For surgeons experienced in open and thoracoscopic lobectomy, appropriate patient selection coupled with the specific robotic techniques described may flatten the learning curve.


The dynamics of population models with distributed maturation periods.
An integro-differential equation for the dynamics of a subpopulation of adults in a closed system where only the adults compete and where there is a distribution of maturation periods is described. We show how the careful choice of a general weighting function based on the gamma distribution with a shift in origin enables us to characterize adequately some observed maturation-period distributions, and also makes local stability and numerical analyses straightforward. Using these results we examine the progression in the behavior of the distributed-delay model as the distribution is narrowed toward the limit of a discrete delay. We conclude that while local stability properties approach those of the limiting equation very rapidly, the persistent fluctuation behavior converges more slowly, with the dominant period and maximum amplitude being least affected by the details of the distribution, and the fine structure of solutions being most sensitive. Finally, we examine the consequences for population modeling, and using several examples of insect populations, conclude that although quite often a full maturation-period distribution should be incorporated in a given model, in many cases a discrete-delay approximation will suffice.


Signals for stress erythropoiesis are integrated via an erythropoietin receptor-phosphotyrosine-343-Stat5 axis.
Anemia due to chronic disease or chemotherapy often is ameliorated by erythropoietin (Epo). Present studies reveal that, unlike steady-state erythropoiesis, erythropoiesis during anemia depends sharply on an Epo receptor-phosphotyrosine-343-Stat5 signaling axis. In mice expressing a phosphotyrosine-null (PY-null) Epo receptor allele (EpoR-HM), severe and persistent anemia was induced by hemolysis or 5-fluorouracil. In short-term transplantation experiments, donor EpoR-HM bone marrow cells also failed to efficiently repopulate the erythroid compartment. In each context, stress erythropoiesis was rescued to WT levels upon the selective restoration of an EpoR PY343 Stat5-binding site (EpoR-H allele). As studied using a unique primary culture system, EpoR-HM erythroblasts exhibited marked stage-specific losses in Epo-dependent growth and survival. EpoR-H PY343 signals restored efficient erythroblast expansion, and the selective Epo induction of the Stat5 target genes proviral integration site-1 (Pim-1) and oncostatin-M. Bcl2-like 1 (Bcl-x), in contrast, was not significantly induced via WT-EpoR, EpoR-HM, or EpoR-H alleles. In Kit+ CD71+ erythroblasts, EpoR-PY343 signals furthermore enhanced SCF growth effects, and SCF modulation of Pim-1 kinase and oncostatin-M expression. In maturing Kit- CD71+ erythroblasts, oncostatin-M exerted antiapoptotic effects that likewise depended on EpoR PY343-mediated events. Stress erythropoiesis, therefore, requires stage-specific EpoR-PY343-Stat5 signals, some of which selectively bolster SCF and oncostatin-M action.


Calculating expected DNA remnants from ancient founding events in human population genetics.
Recent advancements in sequencing and computational technologies have led to rapid generation and analysis of high quality genetic data. Such genetic data have achieved wide acceptance in studies of historic human population origins and admixture. However, in studies relating to small, recent admixture events, genetic factors such as historic population sizes, genetic drift, and mutation can have pronounced effects on data reliability and utility. To address these issues we conducted genetic simulations targeting influential genetic parameters in admixed populations. We performed a series of simulations, adjusting variable values to assess the affect of these genetic parameters on current human population studies and what these studies infer about past population structure. Final mean allele frequencies varied from 0.0005 to over 0.50, depending on the parameters. The results of the simulations illustrate that, while genetic data may be sensitive and powerful in large genetic studies, caution must be used when applying genetic information to small, recent admixture events. For some parameter sets, genetic data will not be adequate to detect historic admixture. In such cases, studies should consider anthropologic, archeological, and linguistic data where possible.


The Microbiota Promotes Arterial Thrombosis in Low-Density Lipoprotein Receptor-Deficient Mice.
Atherosclerotic plaque development depends on chronic inflammation of the arterial wall. A dysbiotic gut microbiota can cause low-grade inflammation, and microbiota composition was linked to cardiovascular disease risk. However, the role of this environmental factor in atherothrombosis remains undefined. To analyze the impact of gut microbiota on atherothrombosis, we rederived low-density lipoprotein receptor-deficient (Ldlr-/- ) mice as germfree (GF) and kept these mice for 16 weeks on an atherogenic high-fat Western diet (HFD) under GF isolator conditions and under conventionally raised specific-pathogen-free conditions (CONV-R). In spite of reduced diversity of the cecal gut microbiome, caused by atherogenic HFD, GF Ldlr-/- mice and CONV-R Ldlr-/- mice exhibited atherosclerotic lesions of comparable sizes in the common carotid artery. In contrast to HFD-fed mice, showing no difference in total cholesterol levels, CONV-R Ldlr-/- mice fed control diet (CD) had significantly reduced total plasma cholesterol, very-low-density lipoprotein (VLDL), and LDL levels compared with GF Ldlr-/- mice. Myeloid cell counts in blood as well as leukocyte adhesion to the vessel wall at the common carotid artery of GF Ldlr-/- mice on HFD were diminished compared to CONV-R Ldlr-/- controls. Plasma cytokine profiling revealed reduced levels of the proinflammatory chemokines CCL7 and CXCL1 in GF Ldlr-/- mice, whereas the T-cell-related interleukin 9 (IL-9) and IL-27 were elevated. In the atherothrombosis model of ultrasound-induced rupture of the common carotid artery plaque, thrombus area was significantly reduced in GF Ldlr-/- mice relative to CONV-R Ldlr-/- mice. Ex vivo, this atherothrombotic phenotype was explained by decreased adhesion-dependent platelet activation and thrombus growth of HFD-fed GF Ldlr-/- mice on type III collagen.IMPORTANCE Our results demonstrate a functional role for the commensal microbiota in atherothrombosis. In a ferric chloride injury model of the carotid artery, GF C57BL/6J mice had increased occlusion times compared to colonized controls. Interestingly, in late atherosclerosis, HFD-fed GF Ldlr-/- mice had reduced plaque rupture-induced thrombus growth in the carotid artery and diminished ex vivo thrombus formation under arterial flow conditions.


Nutrimetabonomics:applications for nutritional sciences, with specific reference to gut microbial interactions.
Understanding the role of the diet in determining human health and disease is one major objective of modern nutrition. Mammalian biocomplexity necessitates the incorporation of systems biology technologies into contemporary nutritional research. Metabonomics is a powerful approach that simultaneously measures the low-molecular-weight compounds in a biological sample, enabling the metabolic status of a biological system to be characterized. Such biochemical profiles contain latent information relating to inherent parameters, such as the genotype, and environmental factors, including the diet and gut microbiota. Nutritional metabonomics, or nutrimetabonomics, is being increasingly applied to study molecular interactions between the diet and the global metabolic system. This review discusses three primary areas in which nutrimetabonomics has enjoyed successful application in nutritional research: the illumination of molecular relationships between nutrition and biochemical processes; elucidation of biomarker signatures of food components for use in dietary surveillance; and the study of complex trans-genomic interactions between the mammalian host and its resident gut microbiome. Finally, this review illustrates the potential for nutrimetabonomics in nutritional science as an indispensable tool to achieve personalized nutrition.


Acute liver failure due to iron overdose in an adult.
The vast majority of acute iron toxicity cases occur in children less than 5 years of age. Moreover, clinical hepatic injury is uncommon with most symptoms stemming from the intestinal tract (eg, nausea, vomiting, diarrhea). Therefore, physicians, particularly those who do not routinely treat pediatric patients, are often unfamiliar with hepatotoxicity related to iron overdose. Nevertheless, hepatotoxicity caused by acute iron poisoning is associated with a high mortality rate. We report a case of severe hepatic injury in an adult who overdosed on iron tablets with suicidal intent. Tests for other hepatotoxins (eg, acetaminophen), hepatatrophic viruses, and other causes of acute liver injury were negative. Although peak serum iron level (340 microg/dL) was significantly lower than that reported to cause hepatotoxicity (>1,700 microg/dL), rapid and significant elevations in aminotransferases (>4,000 U/L), total bilirubin (5 mg/dL), and prothrombin time (50 seconds) occurred within 48 hours. Treatment with deferoxamine was prompt and followed by empiric N-acetylcysteine once liver injury was apparent. The patient was minimally symptomatic and she eventually had a full recovery.


Role of family physicians in hospitals. Did it change between 1977 and 1997?
To investigate whether hospital activities and attitudes toward hospitals of members of an urban family medicine department changed between 1977 and 1997. To explore whether these activities and attitudes are different among fee-for-service (FFS) and non-FFS physicians in 1997. Cross-sectional surveys by interview (1977) and self-administered questionnaire (1997). Community-based family practices in Hamilton, Ont. In 1977, 88 of 89 (98.9%) and, in 1997, 66 of 88 (75.0%) members of the Department of Family Medicine at St Joseph's Hospital in Hamilton. Perceived reasons for involvement in hospital work; time spent and main activities in hospital; use of hospital privileges; attitudes toward family physicians' role in hospital, hospital work, and the Department of Family Medicine; perceptions of patients', consultants', and hospital administrators' attitudes toward family physicians' role in hospitals. In 1977 and 1997, patient care and continuing education remained key reasons for doing hospital work. In 1997, however, respondents spent a mean of 3 hours less per week in hospital; used the hospital less often for procedures, meetings, and teaching; and assumed less responsibility for their patients' in-hospital care. While perceptions of hospital work changed over the years, most physicians continued to see a need and have a desire to remain involved in hospitals. Fee-for-service and non-FFS physicians held different opinions on the needs of both hospitalized patients and family physicians. Although physicians' hospital activities and attitudes changed between 1997 and 1997, most continued to see a need and have a desire to remain involved in hospitals.


Dynamic aspects of musical imagery.
Auditory imagery can represent many aspects of music, such as the starting pitches of a tune or the instrument that typically plays it. In this paper, I concentrate on more dynamic, or time-sensitive aspects of musical imagery, as demonstrated in two recently published studies. The first was a behavioral study that examined the ability to make emotional judgments about both heard and imagined music in real time. The second was a neuroimaging study on the neural correlates of anticipating an upcoming tune, after hearing a cue tune. That study found activation of several sequence-learning brain areas, some of which varied with the vividness of the anticipated musical memory. Both studies speak to the ways in which musical imagery allows us to judge temporally changing aspects of the represented musical experience. These judgments can be quite precise, despite the complexity of generating the rich internal representations of imagery.


Bidirectional associations between COVID-19 and psychiatric disorder: retrospective cohort studies of 62 354 COVID-19 cases in the USA.
Adverse mental health consequences of COVID-19, including anxiety and depression, have been widely predicted but not yet accurately measured. There are a range of physical health risk factors for COVID-19, but it is not known if there are also psychiatric risk factors. In this electronic health record network cohort study using data from 69 million individuals, 62 354 of whom had a diagnosis of COVID-19, we assessed whether a diagnosis of COVID-19 (compared with other health events) was associated with increased rates of subsequent psychiatric diagnoses, and whether patients with a history of psychiatric illness are at a higher risk of being diagnosed with COVID-19. We used the TriNetX Analytics Network, a global federated network that captures anonymised data from electronic health records in 54 health-care organisations in the USA, totalling 69·8 million patients. TriNetX included 62 354 patients diagnosed with COVID-19 between Jan 20, and Aug 1, 2020. We created cohorts of patients who had been diagnosed with COVID-19 or a range of other health events. We used propensity score matching to control for confounding by risk factors for COVID-19 and for severity of illness. We measured the incidence of and hazard ratios (HRs) for psychiatric disorders, dementia, and insomnia, during the first 14 to 90 days after a diagnosis of COVID-19. In patients with no previous psychiatric history, a diagnosis of COVID-19 was associated with increased incidence of a first psychiatric diagnosis in the following 14 to 90 days compared with six other health events (HR 2·1, 95% CI 1·8-2·5 vs influenza; 1·7, 1·5-1·9 vs other respiratory tract infections; 1·6, 1·4-1·9 vs skin infection; 1·6, 1·3-1·9 vs cholelithiasis; 2·2, 1·9-2·6 vs urolithiasis, and 2·1, 1·9-2·5 vs fracture of a large bone; all p<0·0001). The HR was greatest for anxiety disorders, insomnia, and dementia. We observed similar findings, although with smaller HRs, when relapses and new diagnoses were measured. The incidence of any psychiatric diagnosis in the 14 to 90 days after COVID-19 diagnosis was 18·1% (95% CI 17·6-18·6), including 5·8% (5·2-6·4) that were a first diagnosis. The incidence of a first diagnosis of dementia in the 14 to 90 days after COVID-19 diagnosis was 1·6% (95% CI 1·2-2·1) in people older than 65 years. A psychiatric diagnosis in the previous year was associated with a higher incidence of COVID-19 diagnosis (relative risk 1·65, 95% CI 1·59-1·71; p<0·0001). This risk was independent of known physical health risk factors for COVID-19, but we cannot exclude possible residual confounding by socioeconomic factors. Survivors of COVID-19 appear to be at increased risk of psychiatric sequelae, and a psychiatric diagnosis might be an independent risk factor for COVID-19. Although preliminary, our findings have implications for clinical services, and prospective cohort studies are warranted. National Institute for Health Research.


Oxaliplatin, irinotecan and cetuximab in advanced gastric cancer. A multicenter phase II trial (Gastric-2) of the Arbeitsgemeinschaft Medikamentose Tumortherapie (AGMT).
Patients suffering from advanced gastric cancer still have a poor prognosis and treatment options are limited. In our previous phase II trial (AGMT-Gastric-1), we showed that the combination of oxaliplatin and irinotecan was well tolerated and effective. The same chemotherapy regimen was now tested in combination with cetuximab in a multicenter phase II trial. Oxaliplatin at 85 mg/m(2) biweekly and irinotecan at 125 mg/m(2) biweekly were combined with cetuximab at 400 mg/m(2) loading dose and subsequent weekly infusions of 250 mg/m(2). Fifty-one patients with histologically proven unresectable and/or metastatic gastric adenocarcinoma were treated in the first line setting. The median age was 62 years. A single metastatic site was found in 24 patients, 27 patients had multiple metastatic sites. Frequently reported adverse events (in more than 20% of patients) were predominantly grade 1 or 2 and included neutropenia (35%), thrombocytopenia (33%), anemia (73%), nausea (45%), diarrhea (57%), alopecia (22%), and fatigue (37%). Grade 3/4 toxicities included neutropenia in 9/1 patients., thrombocytopenia in 1/0 patients, anemia in 3/1 patients, nausea in 2/0 patients, and diarrhea in 7/2 patients. Sensory neuropathy occurred mostly as grade 1 and 2 in 37% of patients, grade 3 neurotoxicity was observed in 7 patients. Acne-like rash grades 1/2/3/4 were reported in 31%/20%/6%/2% of patients respectively. Thirteen patients discontinued the study due to neutropenia (n=5), nausea/vomiting (n=1), diarrhea (n=1), toxic colon (n=2), and allergic reaction to cetuximab at first (n=2), second (n=1) or third infusion (n=1). Thirty-five patients were assessable for response, with 1 patient (3%) showing a complete response, 21 patients (60%) a partial response, 7 patients (20%) a stable disease, and 6 patients (17%) a progressive disease respectively. The median time to progression was 24.8 weeks, median overall survival was 38.1 weeks. All patients tested had a wild type KRAS status. The combination of oxaliplatin and irinotecan with cetuximab is safe and its action established in advanced gastric cancer.


[Systematic review and meta-analysis of antibiotic prophylaxis in abdominal hysterectomy].
The objective of the present study was to assess the available evidence regarding antibiotic prophylaxis taking the case of abdominal hysterectomy, as an aid to decisions related to coverage and to the development of evidence-based clinical guidelines. Using a previously elaborated protocol, the pertinence and quality of double-blind, randomized, placebo-controlled trials were examined. Clinical heterogeneity among studies was also analyzed. The studies were found to be homogeneous, considering p > 0.10 as the significance level for rejecting heterogeneity. Combination of the 16 selected studies resulted in a summary rate ratio of 0.49 (95%CI: 0.41-0.59), i.e., efficacy of 51%, and in a summary rate difference of 11% (95%CI: 8-14), utilizing the fixed effects model. Results according to the random effects model were very similar. Exploratory analysis of subgroups of single versus multiple doses did not indicate more benefit for multiple dose interventions. Evidence from comparative trials is insufficient to consider third-generation cephalosporins more efficacious than first-generation ones.


Is duodenojejunal anastomosis to the left of the superior mesenteric vessels a feasible option for tumors of the angle of Treitz?
Tumors of the angle of Treitz are a rare entity. Only 3%-5% of gastrointestinal stromal tumors (GISTs) occur at the level of the duodenum, and their location at the duodenojejunal junction is very uncommon. Surgery is the treatment of choice, while adjuvant medical therapy is used on the basis of the degree of radicality of the excision and the tumor's proliferative profile. These factors primarily influence the prognosis. Due to the frailty of the vascular viability of the left duodenum, which can be injured during surgery, it is generally recommended to perform digestive reconstruction at the level of the right portion of the duodenum. We here report the case of a patient with a large GIST located at the duodenojejunal junction behind the ligament of Treitz. We found reconstructive digestive anastomosis at the level of the third part of the duodenum to be a safe procedure.


New tyrosine kinase inhibitors in chronic myeloid leukemia.
The deregulated activity of BCR-ABL tyrosine kinase originating from the t(9;22) chromosomal translocation has been shown to be necessary and sufficient for the transformed phenotype of chronic myeloid leukemia (CML) cells. This peculiarity has paved the way for the development of novel therapies specifically targeting the BCR-ABL gene product. The first BCR-ABL inhibitor to come into use in clinical practice, imatinib mesylate, is now the first-choice treatment for all newly diagnosed CML patients, but the initial striking efficacy of this drug has been overshadowed by the development of clinical resistance. The most common mechanisms of resistance include (i) BCR-ABL overexpression, and (ii) BCR-ABL kinase domain mutations disrupting critical contact points between imatinib and BCR-ABL or inducing a transition to a conformation to which imatinib is unable to bind. Several approaches to overcoming resistance have been studied both in vitro and in vivo. They include dose escalation of imatinib, the combination of imatinib with chemotherapeutic drugs, alternative BCR-ABL inhibitors, and inhibitors of kinases acting downstream of BCR-ABL such as Src kinases. Various novel tyrosine kinase inhibitors (TKI) have been synthesized and have now reached the pre-clinical or clinical phase. This review highlights the development of new TKI as specific molecularly targeted therapy and as the principal mechanisms for overcoming imatinib resistance.


Recreational Cannabis Legalization in the US and Maternal Use during the Preconception, Prenatal, and Postpartum Periods.
In the United States (US), recreational cannabis use is on the rise. Since 2011, 11 states and the District of Columbia have legalized cannabis for adult recreational use. As additional states consider legalizing, there is an urgent need to assess associations between recreational cannabis legalization and maternal use in the preconception, prenatal, and postpartum periods-all critical windows for maternal and child health. Using cross-sectional data from the 2016 Pregnancy Risk Assessment Monitoring System, we assessed associations between state cannabis legalization and self-reported maternal cannabis use. Using logistic regression, we estimated the adjusted prevalence ratio (PR) of cannabis use during the preconception, prenatal, and postpartum period for women delivering a live-born infant in three states that had legalized recreational cannabis (Alaska, Colorado, and Washington) and three states that had not legalized (Maine, Michigan, and New Hampshire) by 2016. Our final sample size was 7258 women. We utilized 95% confidence intervals (CI) and a significance level of alpha = 0.05. After adjustment for potential confounders, women who resided in states with legalized recreational cannabis were significantly more likely to use cannabis during the preconception (PR 1.52; 95%CI ranging from 1.28-1.80; p < 0.001), prenatal (PR 2.21; 95% CI ranging from 1.67-2.94; p < 0.001), and postpartum (PR 1.73; 95%CI ranging from 1.30-2.30; p < 0.001) periods, compared to women who resided in states without legalized recreational cannabis. Although evidence about the effect of marijuana use during these periods is nascent, these findings show potential for increased incidence of child exposure to cannabis. Longitudinal research is needed to assess immediate and sustained impacts of maternal use before and after state legalization of recreational cannabis.


Essential newborn care practice at four primary health facilities in conflict affected areas of Bossaso, Somalia: a cross-sectional study.
Newborn mortality is increasingly concentrated in contexts of conflict and political instability. However, there are limited guidelines and data on the availability and quality of newborn care in conflict settings. In 2016, an interagency collaboration developed the Newborn Health in Humanitarian Settings Field Guide- Interim version (Field Guide). In this study, we sought to understand the baseline availability and quality of essential newborn care in Bossaso, Somalia as part of an investigation to determine the feasibility and effectiveness of the Field Guide in improving newborn care in humanitarian settings. A cross-sectional study was conducted at four purposely selected health facilities serving internally displaced persons affected by conflict in Bossaso. Essential newborn care practice and patient experience with childbirth care received at the facilities were assessed via observation of clinical practice during childbirth and the immediate postnatal period, and through postnatal interviews of mothers. Descriptive statistics and logistic regression were employed to summarize and examine variation by health facility. Of the 332 pregnant women approached, 253 (76.2%) consented and were enrolled. 97.2% (95% CI: 94.4, 98.9) had livebirths and 2.8% (95% CI: 1.1, 5.6) had stillbirths. The early newborn mortality was 1.7% (95% CI: 0.3, 4.8). Nearly all [95.7%, (95% CI: 92.4, 97.8)] births were attended by skilled health worker. Similarly, 98.0% (95% CI: 95.3, 99.3) of newborns received immediate drying, and 99.2% (95% CI: 97.1, 99.9) had delayed bathing. Few [8.6%, (95% CI: 5.4, 12.9)] received immediate skin-to-skin contact and the practice varied significantly by facility (p < 0.001). One-third of newborns [30.1%, (95% CI: 24.4, 36.2)] received early initiation of breastfeeding and there was significant variation by facility (p < 0.001). While almost all [99.2%, (95% CI: 97.2, 100)] service providers wore gloves while attending births, handwashing was not as common [20.2%, (95% CI: 15.4, 25.6)] and varied by facility (p < 0.001). Nearly all [92%, (95% CI: 86.9, 95.5)] mothers were either very happy or happy with the childbirth care received at the facility. Essential newborn care interventions were not universally available. Quality of care varied by health facility and type of intervention. Training and supervision using the Field Guide could improve newborn outcomes.


Effects of vpu start-codon mutations on human immunodeficiency virus type 1 replication in macrophages.
The human immunodeficiency virus type 1 (HIV-1) vpu protein increases the release of virus particles from infected cells. Mutations that abrogate vpu function have a profound effect on HIV-1 replication in primary macrophage cultures. About 1.24 % of primary isolates in the HIV databases have vpu start-codon mutations. In addition, the envelope of the AD8 isolate was reported to compensate for the lack of vpu, whilst the YU-2 virus (cloned directly from the brain tissue of an infected individual) is macrophage-tropic, despite having a vpu start-codon mutation. These observations raise the possibility that envelopes evolve to compensate for the loss of vpu function in vivo. Chimeric vpu+ and vpu- replication-competent clones were constructed that contained the envelopes of SF162, AD8 or YU-2. Macrophages were infected with these chimeras and virus release was measured over time by a reverse transcriptase ELISA. It was found that vpu-deficient chimeras carrying AD8 and YU-2 envelopes were consistently released at lower levels than their wild-type (wt) vpu counterparts, indicating that these envelopes did not compensate for the lack of vpu. Non-chimeric vpu+ and vpu- AD8 and YU-2 followed similar patterns, although replication by vpu-deficient AD8 was variable, with virion release reaching 60 % of that recorded for AD8 with a wt vpu. In summary, no evidence was found that the AD8 or YU-2 envelopes can compensate for the lack of vpu for replication in macrophages.


Water transport confined in graphene oxide channels through the rarefied effect.
Understanding the mechanism of water transport inside an interlayer between graphene-based plates has tremendous value for theoretical studies and industrial applications. The fluid flow confined in nano-scaled spaces experiences a slip velocity near the wall, which is significantly different to that of bulk water. Here we propose a model combining classic hydrodynamics with kinetic theory to depict the dependency of the slip effect on the oxide concentration of valley plates. The influence of oxidized graphene on water flow is a comprehensive result of a slipped boundary, and depends on both the diffuse reflection coefficient of the wall, and the shrunken effective passageway caused by the electrostatic interactions between the oxidized surface and the water molecules. The former effect enhances the water flow, which reduces with increasing oxide concentration, while the latter effect inhibits water flow. We examine the diffuse reflection coefficient and the shrunken effective passageway at different oxide concentrations of the GO sheets by molecular dynamics simulations, and we quantitively predict the flux relationship at various concentrations. This work provides a molecular insight into transport processes of confined water and a useful guideline for the design of perfect graphene-derived membranes for desalination.


Thoracoscopy: present diagnostic and therapeutic indications.
Thoracoscopy is increasingly being used for diagnosis and treatment of pleuropulmonary disease. The recent revival was made possible by the tremendous advances in endoscopic technology. The main requirements for diagnostic purposes are rigid telescopes and forceps, and for interventional thoracoscopy scissors, staplers and a video recorder. The procedure can be performed either under local or general anaesthesia, with or without double lumen intubation, after inducing an artificial pneumothorax. At the end of the procedure, a chest tube should always be inserted, even if only for a few minutes until the lung re-expands. Main diagnostic indications are pleural effusions, pneumothorax and diffuse lung disease. Main therapeutic indications are pleurodesis by talcage in effusion and pneumothorax and a variety of diseases of the lung, the pleura and the mediastinum, where thoracotomy may be replaced by video-assisted thoracoscopy. The well-known indications of the past remain a domain of pneumologists, whereas minimal invasive thoracotomy is the task of thoracic surgeons. For some indications no sharp line has to be drawn, provided the facilities and skills are present, including those for the management of complications.


Stem rust resistance in wheat is suppressed by a subunit of the mediator complex.
Stem rust is an important disease of wheat that can be controlled using resistance genes. The gene SuSr-D1 identified in cultivar 'Canthatch' suppresses stem rust resistance. SuSr-D1 mutants are resistant to several races of stem rust that are virulent on wild-type plants. Here we identify SuSr-D1 by sequencing flow-sorted chromosomes, mutagenesis, and map-based cloning. The gene encodes Med15, a subunit of the Mediator Complex, a conserved protein complex in eukaryotes that regulates expression of protein-coding genes. Nonsense mutations in Med15b.D result in expression of stem rust resistance. Time-course RNAseq analysis show a significant reduction or complete loss of differential gene expression at 24 h post inoculation in med15b.D mutants, suggesting that transcriptional reprogramming at this time point is not required for immunity to stem rust. Suppression is a common phenomenon and this study provides novel insight into suppression of rust resistance in wheat.


Current perspectives in the management of hyponatremia: prevention of CPM.
Central pontine myelinolysis represents a relatively contemporary neurologic entity in which an imbalance of water relative to alterations in the body's electrolyte levels induces characteristic demyelination in the central part of the basis pontis as well as extrapontine sites. The clinical scenario is typically one of chronic hyponatremia followed by a rapid correction to normonatremic or hypernatremic levels. This seemingly innocuous series of events can result in fatal consequences. Better understanding of the pathophysiology of this disorder provides avenues for clinical management, including use of steroids and organic osmoles, such as myoinositol. More recently, the introduction of a newer class of pharmacologic agents - the vasopressin receptor antagonists, known as vaptans, which induce an excretion of increased amounts of water without altered sodium or potassium excretion - is of particular interest. This review addresses classic approaches to the management of this disorder along with a discussion of newer pharmacologic agents that may become important therapeutic interventions in the clinical management of the osmotic myelinolysis syndrome.


Telephone advice nursing services in a US health maintenance organization.
We studied telephone advice nursing (TAN) in the US. We recorded 4269 TAN calls in four regions served by Kaiser Permanente, a health maintenance organization. A call description form was used to record information regarding both calls and callers. The mean call length was 5.9 min (SD 3.6, range 0.3-35.8); 300 calls lasted less than 2 min. The mean call length differed significantly across regions, from 4.4 min in Hawaii to 8.7 min in Southern California. Calls to call centres lasted an average of 6.5 min (SD 3.8), compared with 4.2 min (SD 2.9) for those to medical offices. These differences were significant. Although 42% of calls required some further medical management, only 18% (n = 754) resulted in an urgent disposition, and only 16% (n = 121; 3% of all calls) of the urgent dispositions involved referral to emergency services. The likelihood of urgent disposition varied significantly by region. Callers generally used the TAN services for the right reasons, that is, with questions or concerns that could be reasonably handled by telephone advice nurses.


A novel link between Campylobacter jejuni bacteriophage defence, virulence and Guillain-Barré syndrome.
Guillain-Barré syndrome (GBS) is a post-infectious disease in which the human peripheral nervous system is affected after infection by specific pathogenic bacteria, including Campylobacter jejuni. GBS is suggested to be provoked by molecular mimicry between sialylated lipooligosaccharide (LOS) structures on the cell envelope of these bacteria and ganglioside epitopes on the human peripheral nerves, resulting in autoimmune-driven nerve destruction. Earlier, the C. jejuni sialyltransferase (Cst-II) was found to be linked to GBS and demonstrated to be involved in the biosynthesis of the ganglioside-like LOS structures. Apart from a role in pathogenicity, we report here that Cst-II-generated ganglioside-like LOS structures confer efficient bacteriophage resistance in C. jejuni. By bioinformatic analysis, it is revealed that the presence of sialyltransferases in C. jejuni and other potential GBS-related pathogens correlated significantly with the apparent degeneration of an alternative anti-virus system: type II Clusters of Regularly Interspaced Short Palindromic Repeat and associated genes (CRISPR-Cas). Molecular analysis of the C. jejuni CRISPR-Cas system confirmed the bioinformatic investigation. CRISPR degeneration and mutations in the cas genes cas2, cas1 and csn1 were found to correlate with Cst-II sialyltransferase presence (p < 0.0001). Remarkably, type II CRISPR-Cas systems are mainly found in mammalian pathogens. To study the potential involvement of this system in pathogenicity, we inactivated the type II CRISPR-Cas marker gene csn1, which effectively reduced virulence in primarily cst-II-positive C. jejuni isolates. Our findings indicate a novel link between viral defence, virulence and GBS in a pathogenic bacterium.


Developing a model for embedded palliative care in a cancer clinic.
Describe the development and key features of a model for embedded palliative care (PC) for patients with advanced kidney cancer or melanoma seen in a cancer clinic. Retrospective chart review of patients following an initial phase and then a prospective review following the implementation of a model for embedded PC. In the initial phase, 18 patients were seen for a total of 53 visits; 78% were seen more than once, with a mean of three visits per patient. In the model phase, 46 patients were seen for a total of 163 visits; 74% were seen more than once, with a mean of 3.5 visits. Demographics were similar between the two groups. Content of the first PC visit in the initial and model phases was symptom management (61% and 57%), psychosocial support/relationship building (28% and 35%) and advance care planning/decision-making support (11% and 8%), respectively. The initial phase demonstrated acceptability and feasibility of a model for embedded PC for patients and the oncology team. Establishment of specific eligibility criteria and screening to identify eligible patients in the model phase led to an increased uptake of PC for patients with advanced kidney cancer and melanoma in a cancer clinic.


Angiographic outcomes in the PLATO Trial (Platelet Inhibition and Patient Outcomes).
The PLATO (Platelet Inhibition and Patient Outcomes) angiographic substudy sought to compare the efficacy of ticagrelor versus clopidogrel with respect to angiographic outcomes before and after PCI in the setting of acute coronary syndrome. Greater platelet inhibition has been associated with improved angiographic outcomes before and after percutaneous coronary intervention (PCI). Therefore, it was hypothesized that treatment with ticagrelor, which achieves more rapid, higher, and more consistent platelet inhibition, would be associated with improved angiographic outcomes when compared with those of clopidogrel treatment. The angiographic cohort consists of 2,616 patients drawn from the 18,624-patient PLATO trial. Clopidogrel naïve or pre-treated patients were randomized to 180 mg of ticagrelor or 300 mg of clopidogrel (75 mg for clopidogrel pre-treated patients). PCI patients were administered, as per treatment group: 1) an additional 90 mg of ticagrelor if >24 h following the initial loading dose; or 2) an optional further 300 mg of clopidogrel or placebo (total 600 mg) prior to PCI. The substudy primary endpoint was the incidence of post-PCI TIMI (Thrombolysis In Myocardial Infarction) myocardial perfusion grade 3 (TMPG 3) among patients who received a study drug prior to PCI. In total, 21.3% of patients were pretreated with clopidogrel prior to randomization. There was a short time interval between randomization and PCI (median: 0.68 [interquartile range (IQR): 0.30 to 2.21] h) among all patients. Post-PCI TMPG 3 was similar between the ticagrelor and clopidogrel groups (47.1% vs. 46.9%; p = 0.96). Likewise, the following pre-PCI outcomes were similar in the ticagrelor and clopidogrel groups, respectively: TMPG 3 (30.5% vs. 31.2%), TIMI flow grade 3 (37.1% vs. 39.3%), corrected TIMI frame count (median: 100 vs. 71 frames), TIMI thrombus grade 0 (24.1% vs. 27.6%), minimum lumen diameter (median: 0.3 [IQR: 0.0 to 0.6] vs. 0.3 [IQR: 0.0 to 0.6] mm) and percentage of diameter stenosis (median: 89 [IQR: 78 to 100] vs. 89 [IQR: 77 to 100]). Neither coronary flow nor myocardial perfusion, evaluated on coronary angiograms performed before or following PCI procedures within a few hours after the start of oral antiplatelet treatment in the setting of acute coronary syndromes, demonstrated a difference with ticagrelor versus clopidogrel. (A Comparison of Ticagrelor [AZD6140] and Clopidogrel in Patients With Acute Coronary Syndrome [PLATO]; NCT00391872).


Minocycline prevents osmotic demyelination syndrome by inhibiting the activation of microglia.
Rapid correction of chronic hyponatremia can lead to osmotic demyelination syndrome (ODS), a severe demyelination disease. The microglia that accumulate in the demyelinative lesions may play a detrimental role in the pathogenesis of ODS by producing proinflammatory cytokines, suggesting that they may be a target for therapeutic intervention. Here, we investigated whether minocycline, a selective and potent inhibitor of microglial activation, could protect against ODS in rats. We induced hyponatremia by liquid diet feeding and dDAVP infusion. Rapid correction of the hyponatremia 7 days later resulted in neurologic impairment with severe demyelinative lesions. Activated microglia accumulated at the site of demyelination. Treatment with minocycline within 24 hours of rapid correction, however, was protective: rats exhibited minimal neurologic impairment, and survival improved. Histologic analysis showed that minocycline inhibited demyelination and suppressed the accumulation of microglia at the site of demyelination. Real-time RT-PCR and immunohistochemical analyses showed that minocycline inhibited the activity of microglia and the expression of inflammatory cytokines (e.g. IL-1β, inducible nitric-oxide synthase, and TNF-α), monocyte chemoattractant protein-1, and matrix metalloproteinase-12 in microglia. These results demonstrate that minocycline can protect against ODS by inhibiting the activation and accumulation of microglia at the site of demyelinative lesions, suggesting its possible use in clinical practice.


Effect of phenylisothiocyanate on organic cation transport in opossum kidney cells.
In this study tetraethylammonium (TEA) was used as a model compound to determine the effect of the lysine selective modifying reagent, phenylisothiocyanate (PITC), on organic cation transport across the brush-border membrane of opossum kidney (OK) cells. TEA uptake in OK cell monolayers treated with PITC was reduced in a time- and concentration- dependent manner (IC50, 200 microM). Two lines of evidence suggested that amine rather than sulfhydryl residues were being modified by PITC. First, treatment of the cells with PITC at pH 8.5 was significantly more effective in reducing TEA uptake than PITC treatment at pH 7.4. Secondly, dithiothreitol (10 mM) could not reverse the effect of PITC on TEA uptake. Preincubation of OK cell monolayers with PITC (50 microM) resulted in a significant increase in the Km of TEA (control cells: 81 +/- 10 microM; treated cells: 165 +/- 54 microM), whereas the Vmax was unaffected. Incubation of the cells with quinine or unlabeled TEA significantly altered the inhibitory effects of PITC on TEA uptake. At low concentrations, quinine protected the transporter from the inhibitory effects of PITC, whereas at higher concentrations (> 2.5 microM) quinine enhanced PITC inhibition of TEA uptake. Other organic cations that are substrates for the transporter including cimetidine and N1-methylnicotinamide did not alter the effects of PITC on TEA transport. Collectively, these data suggest that covalent modification of amine groups reduces the transport of TEA in OK cells by decreasing the affinity of TEA to the transporter.(ABSTRACT TRUNCATED AT 250 WORDS)


A highly compliant protein native state with a spontaneous-like mechanical unfolding pathway.
The mechanical properties of proteins and their force-induced structural changes play key roles in many biological processes. Previous studies have shown that natively folded proteins are brittle under tension, unfolding after small mechanical deformations, while partially folded intermediate states, such as molten globules, are compliant and can deform elastically a great amount before crossing the transition state barrier. Moreover, under tension proteins appear to unfold through a different sequence of events than during spontaneous unfolding. Here, we describe the response to force of the four-α-helix acyl-CoA binding protein (ACBP) in the low-force regime using optical tweezers and ratcheted molecular dynamics simulations. The results of our studies reveal an unprecedented mechanical behavior of a natively folded protein. ACBP displays an atypical compliance along two nearly orthogonal pulling axes, with transition states located almost halfway between the unfolded and folded states. Surprisingly, the deformability of ACBP is greater than that observed for the highly pliant molten globule intermediate states. Furthermore, when manipulated from the N- and C-termini, ACBP unfolds by populating a transition state that resembles that observed during chemical denaturation, both for structure and position along the reaction coordinate. Our data provide the first experimental evidence of a spontaneous-like mechanical unfolding pathway of a protein. The mechanical behavior of ACBP is discussed in terms of topology and helix propensity.


Effects on human peripheral lymphocytes of in vivo administration of 9-beta-D-arabinofuranosyl-2-fluoroadenine-5'-monophosphate (NSC 312887), a new purine antimetabolite.
9-beta-D-Arabinofuranosyl-2-fluoroadenine-5'-monophosphate (NSC 312887) (2-F-ara-AMP) is a new purine antimetabolite with documented preclinical activity against a number of animal tumors. Data from in vitro studies and preclinical animal toxicology trials indicated that 2-F-ara-AMP might be lymphocytotoxic. We studied effects of 2-F-ara-AMP on peripheral lymphocytes of patients receiving the agent in a Phase I clinical trial. Eleven patients received 13 courses given by i.v. bolus daily for 5 days. Mononuclear cells were isolated, and lymphocyte subsets were quantitated by immunofluorescence and flow cytometry 1 day before treatment and 4 hr after the final infusion. Lymphocytopenia developed rapidly (median time to nadir, 6 days) and was reversible. Standard leukocyte counts, differential counts, and percentages of isolated mononuclear cells reactive with monoclonal antibodies were used to calculate numbers of peripheral cells in each major lymphocyte subpopulation. Total T-lymphocyte counts fell during all treatment courses, with calculated mean absolute T-cell counts decreasing by 90%. Decreases were observed in all major T-lymphocyte subsets. By contrast, calculated B-lymphocyte counts decreased an average of 50% and were noted to increase during two treatment cycles. We also compared in vitro recoveries of cells from each major lymphocyte subpopulation before and after administration of 2-F-ara-AMP. Recoveries of total mononuclear cells, total T-cells, and non-T-, non-B-cells all were reduced substantially by 2-F-ara-AMP, but B-cell recovery was not reduced. These in vivo data, the first in human subjects, are in agreement with in vitro studies of halopurine nucleotide analogues which have demonstrated that T-cells are more sensitive than are B-cells to the cytotoxic effects of these compounds.


Determining the pattern and prevalence of alcohol consumption in pregnancy by measuring biomarkers in meconium.
To investigate the feasibility of determining the pattern and prevalence of alcohol consumption in pregnancy by measuring ethanol biomarkers in meconium. Population-based observational study. Inner-city maternity unit in Scotland, UK. Random sample of singleton infants delivered after 36 completed weeks' gestation. Fatty acid ethyl esters (FAEEs) and ethyl glucuronide (EtG) in meconium were measured by liquid chromatography-mass spectroscopy. Samples were frozen at -20°C before analysis. Results were compared anonymously with demographic data including maternal age, parity, smoking, ethnicity and postcode and with infant gestation, birth weight and head circumference. Written informed consent was obtained from all subjects. 235 samples of meconium were analysed (70% of eligible babies). Only four (1%) of mothers declined to participate. FAAEs were detected in all, including four samples below the limit of quantification (10 ng/g). 98 (42%) samples had FAEE concentrations >600 ng/g. EtG was detectable in 93 (40%) samples; in 35 (15%) EtG concentration was >30 ng/g. No mother reported heavy alcohol consumption in pregnancy. FAAE concentration correlated with EtG (Pearson's coefficient; p<0.001). There was no association between either biomarker and maternal age, parity, smoking, ethnicity or postcode, or infant gestation, birth weight or head circumference. Measurement of ethanol biomarkers in meconium is a feasible tool for determining the pattern and prevalence of alcohol consumption in pregnancy. Data suggest that at least 15% of pregnant women in the west of Scotland are consuming significant quantities of alcohol during latter pregnancy.


Ecological generalism drives hyperdiversity of secondary metabolite gene clusters in xylarialean endophytes.
Although secondary metabolites are typically associated with competitive or pathogenic interactions, the high bioactivity of endophytic fungi in the Xylariales, coupled with their abundance and broad host ranges spanning all lineages of land plants and lichens, suggests that enhanced secondary metabolism might facilitate symbioses with phylogenetically diverse hosts. Here, we examined secondary metabolite gene clusters (SMGCs) across 96 Xylariales genomes in two clades (Xylariaceae s.l. and Hypoxylaceae), including 88 newly sequenced genomes of endophytes and closely related saprotrophs and pathogens. We paired genomic data with extensive metadata on endophyte hosts and substrates, enabling us to examine genomic factors related to the breadth of symbiotic interactions and ecological roles. All genomes contain hyperabundant SMGCs; however, Xylariaceae have increased numbers of gene duplications, horizontal gene transfers (HGTs) and SMGCs. Enhanced metabolic diversity of endophytes is associated with a greater diversity of hosts and increased capacity for lignocellulose decomposition. Our results suggest that, as host and substrate generalists, Xylariaceae endophytes experience greater selection to diversify SMGCs compared with more ecologically specialised Hypoxylaceae species. Overall, our results provide new evidence that SMGCs may facilitate symbiosis with phylogenetically diverse hosts, highlighting the importance of microbial symbioses to drive fungal metabolic diversity.


Adsorption of Cd, Cu and Zn from aqueous solutions onto ferronickel slag under different potentially toxic metal combination.
Adsorption characteristics of potentially toxic metals in single- and multi-metal forms onto ferronickel slag were evaluated. Competitive sorption of metals by ferronickel slag has never been reported previously. The maximum adsorption capacities of toxic metals on ferronickel were in the order of Cd (10.2 mg g(-1)) > Cu (8.4 mg g(-1)) > Zn (4.4 mg g(-1)) in the single-metal adsorption isotherm and Cu (6.1 mg g(-1)) >> Cd (2.3 mg g(-1)) > Zn (0.3 mg g(-1)) in the multi-metal adsorption isotherm. In comparison with single-metal adsorption isotherm, the reduction rates of maximum toxic metal adsorption capacity in the multi-metal adsorption isotherm were in the following order of Zn (93%) > Cd (78%) >> Cu (27%). The Freundlich isotherm provides a slightly better fit than the Langmuir isotherm equation using ferronickel slag for potentially toxic metal adsorption. Multi-metal adsorption behaviors differed from single-metal adsorption due to competition, based on data obtained from Freundlich and Langmuir adsorption models and three-dimensional simulation. Especially, Cd and Zn were easily exchanged and substituted by Cu during multi-metal adsorption. Further competitive adsorption studies are necessary in order to accurately estimate adsorption capacity of ferronickel slag for potentially toxic metals in natural environments.


Macrodontia, shovel-shaped incisors, and multituberculism: probable Ekman-Westborg-Julin trait.
Multiple macrodontia is a rare finding and is defined as a condition in which a tooth is significantly larger than normal. Macrodontia may occur as an isolated finding, part of a group of dental anomalies, or as a component of a syndrome with multiple oral and systemic manifestations. The purpose of this paper was to report a case of macrodontia affecting all permanent teeth and exhibiting shovel-shaped maxillary and mandibular incisors and multituberculate molars and premolars. Some or all of this patient's characteristics have been reported in both males and females, with a ratio of 5:2. No inheritance pattern has been established, as these traits have generally occurred spontaneously. As more individuals are identified and as molecular techniques continue to advance, it is probable that a gene or genes responsible for macrodontia and the associated traits will be identified.


Zinc Acetate Potentiates the Action of Tosufloxacin against Escherichia coli Biofilm Persisters.
Formation of bacterial biofilms is a major health threat due to their high levels of tolerance to multiple antibiotics and the presence of persisters responsible for infection relapses. We previously showed that a combination of starvation and induction of SOS response in biofilm led to increased levels of persisters and biofilm tolerance to fluoroquinolones. In this study, we hypothesized that inhibition of the SOS response may be an effective strategy to target biofilms and fluoroquinolone persister cells. We tested the survival of Escherichia coli biofilms to different classes of antibiotics in starved and nonstarved conditions and in the presence of zinc acetate, a SOS response inhibitor. We showed that zinc acetate potentiates, albeit moderately, the activity of fluoroquinolones against E. coli persisters in starved biofilms. The efficacy of zinc acetate to increase fluoroquinolone activity, particularly that of tosufloxacin, suggests that such a combination may be a potential strategy for treating biofilm-related bacterial infections.


Taxonomy and conservation of grassland earless dragons: new species and an assessment of the first possible extinction of a reptile on mainland Australia.
Taxonomic research is of fundamental importance in conservation management of threatened species, providing an understanding of species diversity on which management plans are based. The grassland earless dragon lizards (Agamidae: Tympanocryptis) of southeastern Australia have long been of conservation concern but there have been ongoing taxonomic uncertainties. We provide a comprehensive taxonomic review of this group, integrating multiple lines of evidence, including phylogeography (mtDNA), phylogenomics (SNPs), external morphology and micro X-ray CT scans. Based on these data we assign the lectotype of T. lineata to the Canberra region, restrict the distribution of T. pinguicolla to Victoria and name two new species: T. osbornei sp. nov. (Cooma) and T. mccartneyi sp. nov. (Bathurst). Our results have significant conservation implications. Of particular concern is T. pinguicolla, with the last confident sighting in 1969, raising the possibility of the first extinction of a reptile on mainland Australia. However, our results are equivocal as to whether T. pinguicolla is extant or extinct, emphasizing the immediate imperative for continued surveys to locate any remaining populations of T. pinguicolla. We also highlight the need for a full revision of conservation management plans for all the grassland earless dragons.


Proposal for electro-optic multiplier based on dual transverse electro-optic Kerr effect.
A novel electro-optic multiplier is proposed, which can perform voltage multiplication operation by use of the Kerr medium exhibiting dual transverse electro-optic Kerr effect. In this kind of Kerr medium, electro-optic phase retardation is proportional to the square of its applied electric field, and orientations of the field-induced birefringent axes are only related to the direction of the field. Based on this effect, we can design an electro-optic multiplier by selecting the crystals of 6/mmm, 432, and m3m classes and isotropic Kerr media such as glass. Simple calculation demonstrates that a kind of glass-ceramic material with a large Kerr constant can be used for the design of the proposed electro-optic multiplier.


Study Comparing Vein Integrity and Clinical Outcomes in Open Vein Harvesting and 2 Types of Endoscopic Vein Harvesting for Coronary Artery Bypass Grafting: The VICO Randomized Clinical Trial (Vein Integrity and Clinical Outcomes).
Current consensus statements maintain that endoscopic vein harvesting (EVH) should be standard care in coronary artery bypass graft surgery, but vein quality and clinical outcomes have been questioned. The VICO trial (Vein Integrity and Clinical Outcomes) was designed to assess the impact of different vein harvesting methods on vessel damage and whether this contributes to clinical outcomes after coronary artery bypass grafting. In this single-center, randomized clinical trial, patients undergoing coronary artery bypass grafting with an internal mammary artery and with 1 to 4 vein grafts were recruited. All veins were harvested by a single experienced practitioner. We randomly allocated 300 patients into closed tunnel CO2 EVH (n=100), open tunnel CO2 EVH (n=100), and traditional open vein harvesting (n=100) groups. The primary end point was endothelial integrity and muscular damage of the harvested vein. Secondary end points included clinical outcomes (major adverse cardiac events), use of healthcare resources, and impact on health status (quality-adjusted life-years). The open vein harvesting group demonstrated marginally better endothelial integrity in random samples (85% versus 88% versus 93% for closed tunnel EVH, open tunnel EVH, and open vein harvesting; P<0.001). Closed tunnel EVH displayed the lowest longitudinal hypertrophy (1% versus 13.5% versus 3%; P=0.001). However, no differences in endothelial stretching were observed between groups (37% versus 37% versus 31%; P=0.62). Secondary clinical outcomes demonstrated no significant differences in composite major adverse cardiac event scores at each time point up to 48 months. The quality-adjusted life-year gain per patient was 0.11 (P<0.001) for closed tunnel EVH and 0.07 (P=0.003) for open tunnel EVH compared with open vein harvesting. The likelihood of being cost-effective, at a predefined threshold of £20 000 per quality-adjusted life-year gained, was 75% for closed tunnel EVH, 19% for open tunnel EVH, and 6% for open vein harvesting. Our study demonstrates that harvesting techniques affect the integrity of different vein layers, albeit only slightly. Secondary outcomes suggest that histological findings do not directly contribute to major adverse cardiac event outcomes. Gains in health status were observed, and cost-effectiveness was better with closed tunnel EVH. High-level experience with endoscopic harvesting performed by a dedicated specialist practitioner gives optimal results comparable to those of open vein harvesting. URL: https://www.isrctn.com. International Standard Randomised Controlled Trial Registry Number: 91485426.


Single Amino Acid Substitutions at Specific Positions of the Heptad Repeat Sequence of Piscidin-1 Yielded Novel Analogs That Show Low Cytotoxicity and In Vitro and In Vivo Antiendotoxin Activity.
Piscidin-1 possesses significant antimicrobial and cytotoxic activities. To recognize the primary amino acid sequence(s) in piscidin-1 that could be important for its biological activity, a long heptad repeat sequence located in the region from amino acids 2 to 19 was identified. To comprehend the possible role of this motif, six analogs of piscidin-1 were designed by selectively replacing a single isoleucine residue at a d (5th) position or at an a (9th or 16th) position with either an alanine or a valine residue. Two more analogs, namely, I5F,F6A-piscidin-1 and V12I-piscidin-1, were designed for investigating the effect of interchanging an alanine residue at a d position with an adjacent phenylalanine residue and replacing a valine residue with an isoleucine residue at another d position of the heptad repeat of piscidin-1, respectively. Single alanine-substituted analogs exhibited significantly reduced cytotoxicity against mammalian cells compared with that of piscidin-1 but appreciably retained the antibacterial and antiendotoxin activities of piscidin-1. All the single valine-substituted piscidin-1 analogs and I5F,F6A-piscidin-1 showed cytotoxicity greater than that of the corresponding alanine-substituted analogs, antibacterial activity marginally greater than or similar to that of the corresponding alanine-substituted analogs, and also antiendotoxin activity superior to that of the corresponding alanine-substituted analogs. Interestingly, among these peptides, V12I-piscidin-1 showed the highest cytotoxicity and antibacterial and antiendotoxin activities. Lipopolysaccharide (12 mg/kg of body weight)-treated mice, further treated with I16A-piscidin-1, the piscidin-1 analog with the highest therapeutic index, at a single dose of 1 or 2 mg/kg of body weight, showed 80 and 100% survival, respectively. Structural and functional characterization of these peptides revealed the basis of their biological activity and demonstrated that nontoxic piscidin-1 analogs with significant antimicrobial and antiendotoxin activities can be designed by incorporating single alanine substitutions in the piscidin-1 heptad repeat.


A genome-wide association study confirms PNPLA3 and identifies TM6SF2 and MBOAT7 as risk loci for alcohol-related cirrhosis.
Alcohol misuse is the leading cause of cirrhosis and the second most common indication for liver transplantation in the Western world. We performed a genome-wide association study for alcohol-related cirrhosis in individuals of European descent (712 cases and 1,426 controls) with subsequent validation in two independent European cohorts (1,148 cases and 922 controls). We identified variants in the MBOAT7 (P = 1.03 × 10(-9)) and TM6SF2 (P = 7.89 × 10(-10)) genes as new risk loci and confirmed rs738409 in PNPLA3 as an important risk locus for alcohol-related cirrhosis (P = 1.54 × 10(-48)) at a genome-wide level of significance. These three loci have a role in lipid processing, suggesting that lipid turnover is important in the pathogenesis of alcohol-related cirrhosis.


Construction and manipulation of a new Kaposi's sarcoma-associated herpesvirus bacterial artificial chromosome clone.
Efficient genetic modification of herpesviruses such as Kaposi's sarcoma-associated herpesvirus (KSHV) has come to rely on bacterial artificial chromosome (BAC) technology. In order to facilitate this approach, we generated a new KSHV BAC clone, called BAC16, derived from the rKSHV.219 virus, which stems from KSHV and Epstein-Barr virus-coinfected JSC1 primary effusion lymphoma (PEL) cells. Restriction enzyme and complete sequencing data demonstrate that the KSHV of JSC1 PEL cells showed a minimal level of sequence variation across the entire viral genome compared to the complete genomic sequence of other KSHV strains. BAC16 not only stably propagated in both Escherichia coli and mammalian cells without apparent genetic rearrangements, but also was capable of robustly producing infectious virions (∼5 × 10(7)/ml). We also demonstrated the utility of BAC16 by generating deletion mutants of either the K3 or K5 genes, whose products are E3 ligases of the membrane-associated RING-CH (MARCH) family. While previous studies have shown that individual expression of either K3 or K5 results in efficient downregulation of the surface expression of major histocompatibility complex class I (MHC-I) molecules, we found that K5, but not K3, was the primary factor critical for the downregulation of MHC-I surface expression during KSHV lytic reactivation or following de novo infection. The data presented here demonstrate the utility of BAC16 for the generation and characterization of KSHV knockout and mutant recombinants and further emphasize the importance of functional analysis of viral genes in the context of the KSHV genome besides the study of individual gene expression.


Ageism and equality.
This paper rebuts suggestions made by Littlejohns et al that NICE is not ageist by analysing the concept of ageism. It recognises the constraints that finite resources impose on decision making bodies such as NICE and then makes a number of positive suggestions as to how NICE might more effectively and more justly intervene in the allocation of scarce resources for health.


Identification of conserved residues contributing to the activities of adenovirus DNA polymerase.
Adenovirus codes for a DNA polymerase that is a member of the DNA polymerase alpha family and uses a protein primer for initiation of DNA synthesis. It contains motifs characteristic of a proofreading 3'-5'-exonuclease domain located in the N-terminal region and several polymerase motifs located in the C-terminal region. To determine the role of adenovirus DNA polymerase in DNA replication, 22 site-directed mutations were introduced into the conserved DNA polymerase motifs in the C-terminal region of adenovirus DNA polymerase and the mutant forms were expressed in insect cells using a baculovirus expression system. Each mutant enzyme was tested for DNA binding activity, the ability to interact with pTP, DNA polymerase catalytic activity, and the ability to participate in the initiation of adenovirus DNA replication. The mutant phenotypes identify functional domains within the adenovirus DNA polymerase and allow discrimination between the roles of conserved residues in the various activities carried out by the protein. Using the functional data in this study and the previously published structure of the bacteriophage RB69 DNA polymerase (J. Wang et al., Cell 89:1087-1099, 1997), it is possible to envisage how the conserved domains in the adenovirus DNA polymerase function.


Prevention of exercise-related injuries and adverse events in patients with type 2 diabetes.
Physical activity is widely recommended as an essential non-pharmacological therapeutic strategy to the prevention and control of type 2 diabetes and cardiovascular risk. Microvascular and macrovascular complications associated with the natural progression of the disease and typical age and anthropometric profile of individuals with type 2 diabetes may expose these patients to an increased risk of injury and acute adverse events during exercise. These injuries and adverse events can lead to fear of new injury and consequent physical inactivity. Preventative measures are essential to reduce risk, increase safety and avoid the occurrence of exercise-related injuries in people with type 2 diabetes. This population can exercise safely if certain precautions are taken and if exercise is adapted to complications and contraindications of each individual. Conditions such as diabetic foot, diabetic retinopathy, diabetic nephropathy, diabetic autonomic neuropathy, cardiovascular risk factors, musculoskeletal disorders, hypoglycaemia, hyperglycaemia, dehydration and interactions between medication and exercise should be taken into consideration when prescribing exercise.


The immunological synapse and the actin cytoskeleton: molecular hardware for T cell signaling.
The actin cytoskeleton seems to play two critical roles in the activation of T cells. One of these roles is T cell shape development and movement, including formation of the immunological synapse. The other is the formation of a scaffold for signaling components. This review focuses on the recent convergence of cell biology and immunology studies to explain the role of the actin cytoskeleton in creating the molecular basis for immunological synapse formation and T cell signaling.


Use of Intravenous Gadolinium-based Contrast Media in Patients with Kidney Disease: Consensus Statements from the American College of Radiology and the National Kidney Foundation.
Inaugural consensus statements were developed and endorsed by the American College of Radiology (ACR) and the National Kidney Foundation to improve and standardize the care of patients with kidney disease who have indication(s) to receive ACR-designated group II or group III intravenous gadolinium-based contrast media (GBCM). The risk of nephrogenic systemic fibrosis (NSF) from group II GBCM in patients with advanced kidney disease is thought to be very low (zero events following 4931 administrations to patients with estimated glomerular filtration rate [eGFR] <30 mL/min per 1.73 m2; upper bounds of the 95% confidence intervals: 0.07% overall, 0.2% for stage 5D chronic kidney disease [CKD], 0.5% for stage 5 CKD and no dialysis). No unconfounded cases of NSF have been reported for the only available group III GBCM (gadoxetate disodium). Depending on the clinical indication, the potential harms of delaying or withholding group II or group III GBCM for an MRI in a patient with acute kidney injury or eGFR less than 30 mL/min per 1.73 m2 should be balanced against and may outweigh the risk of NSF. Dialysis initiation or alteration is likely unnecessary based on group II or group III GBCM administration. This article is a simultaneous joint publication in Radiology and Kidney Medicine. The articles are identical except for stylistic changes in keeping with each journal's style. Either version may be used in citing this article.


Impact of impaired fractional flow reserve after coronary interventions on outcomes: a systematic review and meta-analysis.
FFR is routinely used to guide percutaneous coronary interventions (PCI). Visual assessment of the angiographic result after PCI has limited efficacy. Even when the angiographic result seems satisfactory FFR after a PCI might be useful for identifying patients with a suboptimal interventional result and higher risk for poor clinical outcome who might benefit from additional procedures. The aim of this meta-analysis was to investigate available data of studies that examined clinical outcomes of patients with impaired vs. satisfactory fractional flow reserve (FFR) after percutaneous coronary interventions (PCI). This meta-analysis was carried out according to the Cochrane Handbook for Systematic Reviews. The Mantel-Haenszel method using the fixed-effect meta-analysis model was used for combining the results. Studies were identified by searching the literature through mid-January, 2016, using the following search terms: fractional flow reserve, coronary circulation, after, percutaneous coronary intervention, balloon angioplasty, stent implantation, and stenting. Primary endpoint was the rate of major adverse cardiac events (MACE). Secondary endpoints included rates of death, myocardial infarction (MI), repeated revascularisation. Eight relevant studies were found including a total of 1337 patients. Of those, 492 (36.8 %) had an impaired FFR after PCI, and 853 (63.2 %) had a satisfactory FFR after PCI. Odds ratios indicated that a low FFR following PCI was associated with an impaired outcome: major adverse cardiac events (MACE, OR: 4.95, 95 % confidence interval [CI]: 3.39-7.22, p <0.001); death (OR: 3.23, 95 % CI: 1.19-8.76, p = 0.022); myocardial infarction (OR: 13.83, 95 % CI: 4.75-40.24, p <0.0001) and repeated revascularisation (OR: 4.42, 95 % CI: 2.73-7.15, p <0.0001). Compared to a satisfactory FFR, a persistently low FFR following PCI is associated with a worse clinical outcome. Prospective studies are needed to identify underlying causes, determine an optimal threshold for post-PCI FFR, and clarify whether simple additional procedures can influence the post-PCI FFR and clinical outcome.


Comprehensive functional characterization of SGCB coding variants predicts pathogenicity in limb-girdle muscular dystrophy type R4/2E.
Genetic testing is essential for patients with a suspected hereditary myopathy. More than 50% of patients clinically diagnosed with a myopathy carry a variant of unknown significance in a myopathy gene, often leaving them without a genetic diagnosis. Limb-girdle muscular dystrophy (LGMD) type R4/2E is caused by mutations in β-sarcoglycan (SGCB). Together, β-, α-, γ-, and δ-sarcoglycan form a 4-protein transmembrane complex (SGC) that localizes to the sarcolemma. Biallelic loss-of-function mutations in any subunit can lead to LGMD. To provide functional evidence for the pathogenicity of missense variants, we performed deep mutational scanning of SGCB and assessed SGC cell surface localization for all 6,340 possible amino acid changes. Variant functional scores were bimodally distributed and perfectly predicted pathogenicity of known variants. Variants with less severe functional scores more often appeared in patients with slower disease progression, implying a relationship between variant function and disease severity. Amino acid positions intolerant to variation mapped to points of predicted SGC interactions, validated in silico structural models, and enabled accurate prediction of pathogenic variants in other SGC genes. These results will be useful for clinical interpretation of SGCB variants and improving diagnosis of LGMD; we hope they enable wider use of potentially life-saving gene therapy.


Feasibility and Efficacy of Presurgical Exercise in Survivors of Rectal Cancer Scheduled to Receive Curative Resection.
Localized rectal carcinoma is invasive, with surgical resection the standard treatment. The aim of this study was to determine the feasibility of a supervised presurgical exercise intervention in patients with rectal cancer prior to rectal resection. Twelve patients volunteered to undertake twice-weekly aerobic and resistance exercise for ∼16 weeks prior to surgery. At baseline, presurgery, and ∼8 weeks postsurgery, muscle strength and physical performance, body composition, quality of life, and fatigue were assessed. Ten patients completed training, with 80% completing more than one-half of the exercise sessions. Muscle strength improved 9% to 29% at presurgery, although this was not statistically significant, and declined postsurgery (P < .05). Importantly, postsurgery strength levels were comparable with pretraining levels. Lean mass was preserved at presurgery despite neoadjuvant chemoradiation treatment, whereas postsurgery lean mass decreased (P < .05) compared with baseline (-3.2 ± 5.4 kg) and presurgery (-3.7 ± 5.4 kg). There were no substantial changes in quality of life or fatigue. Presurgical exercise is feasible, leading to modest improvements in some outcomes despite chemoradiation treatment. The detrimental effects of surgery were evident, especially in relation to lean mass. As such, exercise may facilitate recovery by enhancing presurgery physical reserve capacity, thereby providing a buffer to declines following surgery.


Randomized pilot trial of a synbiotic dietary supplement in chronic HIV-1 infection.
Infection with HIV-1 results in marked immunologic insults and structural damage to the intestinal mucosa, including compromised barrier function. While the development of highly active antiretroviral therapy (HAART) has been a major advancement in the treatment of HIV-1 infection, the need for novel complementary interventions to help restore intestinal structural and functional integrity remains unmet. Known properties of pre-, pro-, and synbiotics suggest that they may be useful tools in achieving this goal. This was a 4-week parallel, placebo-controlled, randomized pilot trial in HIV-infected women on antiretroviral therapy. A synbiotic formulation (Synbiotic 2000®) containing 4 strains of probiotic bacteria (10(10) each) plus 4 nondigestible, fermentable dietary fibers (2.5 g each) was provided each day, versus a fiber-only placebo formulation. The primary outcome was bacterial translocation. Secondary outcomes included the levels of supplemented bacteria in stool, the activation phenotype of peripheral T-cells and monocytes, and plasma levels of C-reactive protein and soluble CD14. Microbial translocation, as measured by plasma bacterial 16S ribosomal DNA concentration, was not altered by synbiotic treatment. In contrast, the synbiotic formulation resulted in significantly elevated levels of supplemented probiotic bacterial strains in stool, including L. plantarum and P. pentosaceus, with the colonization of these two species being positively correlated with each other. T-cell activation phenotype of peripheral blood lymphocytes showed modest changes in response to synbiotic exposure, with HLA-DR expression slightly elevated on a minor population of CD4+ T-cells which lack expression of HLA-DR or PD-1. In addition, CD38 expression on CD8+ T-cells was slightly lower in the fiber-only group. Plasma levels of soluble CD14 and C-reactive protein were unaffected by synbiotic treatment in this study. Synbiotic treatment for 4 weeks can successfully augment the levels of probiotic species in the gut during chronic HIV-1 infection. Associated changes in microbial translocation appear to be absent, and markers of systemic immune activation appear largely unchanged. These findings may help inform future studies aimed at testing pre- and probiotic approaches to improve gut function and mucosal immunity in chronic HIV-1 infection. Clinical Trials.gov: NCT00688311.


Considerations and consequences of allowing DNA sequence data as types of fungal taxa.
Nomenclatural type definitions are one of the most important concepts in biological nomenclature. Being physical objects that can be re-studied by other researchers, types permanently link taxonomy (an artificial agreement to classify biological diversity) with nomenclature (an artificial agreement to name biological diversity). Two proposals to amend the International Code of Nomenclature for algae, fungi, and plants (ICN), allowing DNA sequences alone (of any region and extent) to serve as types of taxon names for voucherless fungi (mainly putative taxa from environmental DNA sequences), have been submitted to be voted on at the 11th International Mycological Congress (Puerto Rico, July 2018). We consider various genetic processes affecting the distribution of alleles among taxa and find that alleles may not consistently and uniquely represent the species within which they are contained. Should the proposals be accepted, the meaning of nomenclatural types would change in a fundamental way from physical objects as sources of data to the data themselves. Such changes are conducive to irreproducible science, the potential typification on artefactual data, and massive creation of names with low information content, ultimately causing nomenclatural instability and unnecessary work for future researchers that would stall future explorations of fungal diversity. We conclude that the acceptance of DNA sequences alone as types of names of taxa, under the terms used in the current proposals, is unnecessary and would not solve the problem of naming putative taxa known only from DNA sequences in a scientifically defensible way. As an alternative, we highlight the use of formulas for naming putative taxa (candidate taxa) that do not require any modification of the ICN.


Coordinate expression of the murine Hox-5 complex homoeobox-containing genes during limb pattern formation.
The homoebox-containing genes of the Hox-5 complex are expressed in different but overlapping domains in limbs during murine development. The more 5' the position of these genes in the complex, the later and more distal is their expression. Antero-posterior differences are also observed. A model is proposed that accounts for the establishment of these expression domains in relation to the existence of a morphogen released by the zone of polarizing activity. Comparison of these observations with the expression patterns of the genes of Hox complexes in the early embryo suggests that similar molecular mechanisms are involved in the positional signalling along the axes of both the embryonic trunk and the fetal limbs.


Quantifying past and present connectivity illuminates a rapidly changing landscape for the African elephant.
There is widespread concern about impacts of land-use change on connectivity among animal and plant populations, but those impacts are difficult to quantify. Moreover, lack of knowledge regarding ecosystems before fragmentation may obscure appropriate conservation targets. We use occurrence and population genetic data to contrast connectivity for a long-lived mega-herbivore over historical and contemporary time frames. We test whether (i) historical gene flow is predicted by persistent landscape features rather than human settlement, (ii) contemporary connectivity is most affected by human settlement and (iii) recent gene flow estimates show the effects of both factors. We used 16 microsatellite loci to estimate historical and recent gene flow among African elephant (Loxodonta africana) populations in seven protected areas in Tanzania, East Africa. We used historical gene flow (FST and G'ST ) to test and optimize models of historical landscape resistance to movement. We inferred contemporary landscape resistance from elephant resource selection, assessed via walking surveys across ~15 400 km(2) of protected and unprotected lands. We used assignment-based recent gene flow estimates to optimize and test the contemporary resistance model, and to test a combined historical and contemporary model. We detected striking changes in connectivity. Historical connectivity among elephant populations was strongly influenced by slope but not human settlement, whereas contemporary connectivity was influenced most by human settlement. Recent gene flow was strongly influenced by slope but was also correlated with contemporary resistance. Inferences across multiple timescales can better inform conservation efforts on large and complex landscapes, while mitigating the fundamental problem of shifting baselines in conservation.