Silencing of GSTP1, a prostate cancer prognostic gene, by the estrogen receptor-β and endothelial nitric oxide synthase complex.
We recently identified in prostate tumors (PCa) a transcriptional prognostic signature comprising a significant number of genes differentially regulated in patients with worse clinical outcome. Induction of up-regulated genes was due to chromatin remodeling by a combinatorial complex between estrogen receptor (ER)-β and endothelial nitric oxide synthase (eNOS). Here we show that this complex can also repress transcription of prognostic genes that are down-regulated in PCa, such as the glutathione transferase gene GSTP1. Silencing of GSTP1 is a common early event in prostate carcinogenesis, frequently caused by promoter hypermethylation. We validated loss of glutathione transferase (GST) P1-1 expression in vivo, in tissue microarrays from a retrospective cohort of patients, and correlated it with decreased disease-specific survival. Furthermore, we show that in PCa cultured cells ERβ/eNOS causes GSTP1 repression by being recruited at estrogen responsive elements in the gene promoter with consequential remodeling of local chromatin. Treatment with ERβ antagonist or its natural ligand 5α-androstane-3β,17β-diol, eNOS inhibitors or ERβ small interference RNA abrogated the binding and reversed GSTP1 silencing, demonstrating the direct involvement of the complex. In vitro, GSTP1 silencing by ERβ/eNOS was specific for cells from patients with worse clinical outcome where it appeared the sole mechanism regulating GSTP1 expression because no promoter hypermethylation was present. However, in vivo chromatin immunoprecipitation assays on fresh PCa tissues demonstrated that silencing by ERβ/eNOS can coexist with promoter hypermethylation. Our findings reveal that the ERβ/eNOS complex can exert transcriptional repression and suggest that this may represent an epigenetic event favoring inactivation of the GSTP1 locus by methylation. Moreover, abrogation of ERβ/eNOS function by 3β-adiol emphasizes the significance of circulating or locally produced sex steroid hormones or their metabolites in PCa biology with relevant clinical/therapeutic implications.


Functional liquid structures by emulsification of graphene and other two-dimensional nanomaterials.
Pickering emulsions stabilised with nanomaterials provide routes to a range of functional macroscopic assemblies. We demonstrate the formation and properties of water-in-oil emulsions prepared through liquid-phase exfoliation of graphene. Due to the functional nature of the stabiliser, the emulsions exhibit conductivity due to inter-particle tunnelling. We demonstrate a strain sensing application with a large gauge factor of ∼40; the highest reported in a liquid. Our methodology can be applied to other two-dimensional layered materials opening up applications such as energy storage materials, and flexible and printable electronics.


Prevalence of laparoscopic surgical treatment and its clinical outcomes in patients with familial adenomatous polyposis in Japan.
Laparoscopic surgery is becoming the preferred technique for most colorectal interventions. This study aimed to clarify the time trend of surgical treatment for familial adenomatous polyposis (FAP) and its relevance to clinical outcomes in Japan over a 13-year period. This was a multicenter retrospective cohort study comprising 23 specialist institutions for colorectal disease and a cohort of 282 FAP patients who underwent total colectomy or proctocolectomy during 2000-2012. Patient clinical backgrounds and surgical outcomes were compared between the first and second halves of the study period. The proportion of surgical types adopted over the entire study period was 46, 21, 30, and 3 % for ileoanal anastomosis (IAA), ileoanal canal anastomosis, ileorectal anastomosis, and permanent ileostomy, respectively. FAP patients undergoing laparoscopic surgery have increased since 2008 and reached 74 % in the past 3 years. In particular, the number of patients undergoing laparoscopic proctocolectomy with IAA increased approximately four-fold from the first to the second half of the study period. A laparoscopic approach was increasingly used in patients with coexisting colorectal malignancies. Despite this trend, surgical results of the laparoscopic approach between the two study periods showed similar morbidity, pouch operation and stoma closure completion rates. No postoperative mortality was observed in this series, and laparoscopic surgery was comparable to open surgery in terms of stoma closure rate, incidence of intra-abdominal/abdominal desmoid tumors, and postoperative survival rate in both study periods. Laparoscopic approach is increasingly being adopted for prophylactic FAP surgery in Japan and may provide clinically acceptable practical outcomes.


Dairy consumption and the risk of 15-year cardiovascular disease mortality in a cohort of older Australians.
The effects of habitual dairy consumption and the risk of 15-year cardiovascular disease (CVD) mortality in a cohort of older Australians were investigated. Participants (n = 2900) completed a validated 145-item semi-quantitative food frequency questionnaire. Cox proportional hazards regression models were used to investigate associations between tertiles of the dairy consumption, including low/reduced fat dairy, whole fat dairy and their ratio (ratio(LF/WF)), and risk of mortality from coronary heart disease (CHD), stroke or combined CVD. There were 548 recorded cases of CVD mortality in this cohort. For total dairy intake, a reduction in risk of CVD was only seen in tertile 2 (adjusted hazard ratio, AHR: 0.71; 95% CI: 0.55-0.93), and for CHD both tertile 2 and tertile 3 were associated with a reduced risk (both with AHR: 0.71). However there were no linear trends between total dairy consumption and any of the three outcomes. There were no associations or trends between low/reduced fat dairy or whole fat dairy consumption, or ratio(LF/WF) and any of the three outcomes in the fully adjusted model (all p > 0.05). This study found no consistent association between baseline consumption of dairy foods and the risk of CHD, stroke and combined CVD mortality.


Do Direct Survivors of Terrorism Remaining in the Disaster Community Show Better Long-Term Outcome than Survivors Who Relocate?
Little is known about whether, compared to terrorism survivors who relocated to another area, better long-term outcome occurs in terrorism survivors who remain in the community, which may offer social support and formal services as well as ongoing trauma reminders and adversities. A cross-sectional telephone survey of OKC bombing survivors 19 years later assessed current symptoms of PTSD, anxiety and depression; posttraumatic growth; life satisfaction; medical conditions; alcohol use and smoking. We interviewed 138 survivors-114 (82.6%) remaining in OKC area and 24 (17.4%) relocated. Remaining survivors had higher PTS, anxiety and depression and lower posttraumatic growth scores than relocated survivors, and more remaining survivors disagreed with being satisfied with life, with differences not statistically significant. Groups did not differ in major medical problems except heart disease, not significantly different after adjusting for gender. Groups did not differ significantly in smoking or alcohol use. Contrary to expectations, remaining within the community after terrorism was not associated with better long-term psychological or medical outcome. Possible factors relevant to the literature are discussed.


Regenerative PbS and CdS quantum dot sensitized solar cells with a cobalt complex as hole mediator.
Metal sulfide (PbS and CdS) quantum dots (QDs) were prepared over mesoporous TiO2 films by improved successive ionic layer adsorption and reaction (SILAR) processes. The as-prepared QD-sensitized electrodes were combined with a cobalt complex redox couple [Co(o-phen)3]2+/3+ to make a regenerative liquid-type photovoltaic cell. The optimized PbS QD-sensitized solar cells exhibited promising incident photon-to-current conversion efficiency (IPCE) of over 50% and an overall conversion efficiency of 2% at 0.1 sun in a regenerative mode. The overall photovoltaic performance of the PbS QD-sensitized cells was observed to be dependent on the final turn of the SILAR process, giving a better result when the final deposition was Pb2+, not S2-. However, in the case of CdS QD-sensitized cells, S2- termination was better than that of Cd2+. The cobalt complex herein used as a regenerative redox couple was found to be more efficient in generating photocurrents from PbS QD cells than the typical hole scavenger Na2S in a three-electrode configuration. The CdS-sensitized cell with this redox mediator also showed better defined current-voltage curves and an IPCE reaching 40%.


A physical action potential generator: design, implementation and evaluation.
The objective was to develop a physical action potential generator (Paxon) with the ability to generate a stable, repeatable, programmable, and physiological-like action potential. The Paxon has an equivalent of 40 nodes of Ranvier that were mimicked using resin embedded gold wires (Ø = 20 μm). These nodes were software controlled and the action potentials were initiated by a start trigger. Clinically used Ag-AgCl electrodes were coupled to the Paxon for functional testing. The Paxon's action potential parameters were tunable using a second order mathematical equation to generate physiologically relevant output, which was accomplished by varying the number of nodes involved (1-40 in incremental steps of 1) and the node drive potential (0-2.8 V in 0.7 mV steps), while keeping a fixed inter-nodal timing and test electrode configuration. A system noise floor of 0.07 ± 0.01 μV was calculated over 50 runs. A differential test electrode recorded a peak positive amplitude of 1.5 ± 0.05 mV (gain of 40x) at time 196.4 ± 0.06 ms, including a post trigger delay. The Paxon's programmable action potential like signal has the possibility to be used as a validation test platform for medical surface electrodes and their attached systems.


Antisense suppression of 2-cysteine peroxiredoxin in Arabidopsis specifically enhances the activities and expression of enzymes associated with ascorbate metabolism but not glutathione metabolism.
The aim of this study was to characterize the effect of decreased 2-cysteine peroxiredoxin (2-CP) on the leaf anti-oxidative system in Arabidopsis. At three stages of leaf development, two lines of transgenic Arabidopsis mutants with decreased contents of chloroplast 2-CP were compared with wild type and a control line transformed with an empty vector. Glutathione contents and redox state were similar in all plants, and no changes in transcript levels for enzymes involved in glutathione metabolism were observed. Transcript levels for chloroplastic glutathione peroxidase were much lower than those for 2-CP, and both cytosolic and chloroplastic glutathione peroxidase were not increased in the mutants. In contrast, the foliar ascorbate pool was more oxidized in the mutants, although the difference decreased with plant age. The activities of thylakoid and stromal ascorbate peroxidase and particularly monodehydroascorbate reductase were increased as were transcripts for these enzymes. No change in dehydroascorbate reductase activity was observed, and effects on transcript abundance for glutathione reductase, catalase, and superoxide dismutase were slight or absent. The results demonstrate that 2-CP forms an integral part of the anti-oxidant network of chloroplasts and is functionally interconnected with other defense systems. Suppression of 2-CP leads to increased expression of other anti-oxidative genes possibly mediated by increased oxidation state of the leaf ascorbate pool.


Waldenström macroglobulinemia caused by extranodal marginal zone B-cell lymphoma: a report of six cases.
Waldenström macroglobulinemia (WM) and its associated hyperviscosity syndrome (HVS) are generally caused by lymphoplasmacytoid lymphoma or other small B-cell lymphoproliferative disorders. WM associated with extranodal marginal zone B-cell-mucosa-associated lymphoid tissue lymphoma (EMZL/MALT-type) has not been emphasized. We describe 4 men and 2 women (age, 40-79 years) with clinical and laboratory manifestations of WM and EMZL/MALT-type involving one or more sites: lung, pericardium/pleura, ocular adnexa, nasopharynx, minor salivary gland, glossopharyngeal fold, skin, and stomach. The following immunophenotypic patterns were observed: CD20+, 6; CD43+, 3; kappa light chain restriction, 5; and lambda light chain restriction, 1. All were negative for CD5, CD10, and cyclin D1 expression. A clonal paraproteinemia was present in each (IgM kappa, 4; IgM lambda, 1; biclonal IgM kappa/IgA kappa, 1). All 4 patients tested had elevated plasma viscosity; clinical HVS occurred in 3, and 2 required emergency plasmapheresis. These findings suggest that EMZL/MALT-type can cause WM and that the laboratory evaluation of EMZL/MALT-type should include serum protein electrophoresis/immunofixation, and plasma viscosity measurements and urine immunofixation in select cases. EMZL/MALT-type should be considered in the differential diagnosis in patients with clinicopathologic features of WM.


Diagnostic accuracy of the appearance of Nigrosome-1 on magnetic resonance imaging in Parkinson's disease: A systematic review and meta-analysis.
There is currently no definitive diagnostic test for Parkinson's disease (PD) and the current diagnostic procedure primarily relies on clinical manifestations. A hypointense appearance of nigrosome-1 (or absence of the "swallow tail" sign) on magnetic resonance imaging (MRI) has been proposed as a biomarker for PD. This meta-analysis examined the diagnostic accuracy of the appearance of nigrosome-1 on Magnetic Resonance Imaging (MRI) in differentiating idiopathic PD patients from healthy adults. Databases (MEDLINE, Embase, Scopus) were searched from 2012 (first publication of nigrosome-1 MRI scans) up until September 2019. Two researchers independently screened all titles and abstracts to identify studies that met the inclusion criteria and extracted relevant articles in a uniform manner. Two authors independently extracted data and assessed the risk of bias using a customized QUADAS-2 tool. Pooled sensitivity and specificity were calculated using a hierarchical summary receiver operating characteristic approach, as were positive and negative likelihood ratios. Nineteen studies containing a total of 1508 participants (903 idiopathic PD patients and 605 healthy controls) were included. The overall sensitivity and specificity were 0.94 (95%CI, 0.93-0.96) and 0.90 (95%CI, 0.88-0.92), respectively. The likelihood ratios for positive and negative test results were 9.72 (95%CI, 5.58-16.04) and 0.08 (95%CI, 0.05-0.12). The pooled area under the receiver operating characteristics curve (AUC) in the diagnosis of idiopathic PD was 0.98. Visual assessment of the nigrosome-1 appearance, at 3 or 7T, yields excellent diagnostic accuracy for differentiating idiopathic PD from healthy adults.


Perioperative management of ventriculoperitoneal shunts during abdominal surgery.
Patients with ventriculoperitoneal shunts (VPSs) inserted for a variety of disorders may subsequently undergo gastrointestinal or urologic operations, and surgeons must determine the appropriate perioperative management to minimize the risk for shunt malfunction or infection. There is currently no established set of guidelines for this scenario. The objective of this study was to determine the risks and standard of practice for patients with VPSs undergoing abdominal surgery. A retrospective review of the charts of patients with VPSs who underwent abdominal or urologic surgery at the Stanford University Medical Center between 1995 and 2003 was performed. Data regarding type of abdominal surgery, level of contamination, choice of antibiotic therapy, perioperative management of the VPS, and outcomes were obtained. Twenty-six patient charts were reviewed, for a total of 39 operations (5 urologic, 23 upper gastrointestinal, and 11 lower gastrointestinal). Of these, 3 were clean, 34 were clean-contaminated, and 2 were dirty operations. Seven cases were laparoscopic, whereas 32 were open. Thirty-four cases required opening the bowel or urologic system. No patient had preoperative shunt externalization. All except one patient received pre- and postoperative antibiotics, but the duration and type of antibiotics were widely variable. The remaining patient had an inguinal hernia repair and received only one preoperative dose of cephalexin. Purulent fluid was found in 2 cases. One VPS found lying in purulent material next to an anastomotic leak was externalized and subsequently revised. However, in another patient, a VPS found lying next to a purulent jejunal tear was not externalized. This patient returned 2 months later with a VPS malfunction. In the remaining 35 cases, no VPS infection or malfunction was noted over 2 to 10 years of follow-up. The data suggest that there is minimal risk for VPS malfunction or infection among patients undergoing routine clean and clean-contaminated abdominal and urologic surgeries. Patients with VPSs undergoing these operations do not need externalization of their shunt. None of the patients in this study had a contaminated procedure. For dirty procedures, surgeons should opt to externalize the shunt. Future studies will aim to better standardize the perioperative management of VPSs during abdominal surgery.


Axonal signals regulate the differentiation of non-myelin-forming Schwann cells: an immunohistochemical study of galactocerebroside in transected and regenerating nerves.
Little is known about the factors involved in directing and maintaining the divergent differentiation of the 2 major Schwann cell variants, myelin and non-myelin-forming cells, in peripheral nerves. There is strong evidence that the differentiation of myelin-forming cells depends critically on cell-cell signaling through contact with appropriate axons. In this paper we ask whether this remarkable dependence of the Schwann cell on axonal contact for full differentiation is unique to those cells that form myelin or whether axonal signaling is also an important factor in the differentiation of non-myelin-forming Schwann cells. Sciatic nerves or cervical sympathetic trunks of adult rats were either transected or crushed and the axons allowed to degenerate and, in the case of crushed nerves, to regenerate into the distal stump for periods of time varying from 2 d to 9 weeks. The distal stump of the nerve was excised at specific times, the Schwann cells dissociated and immunolabeled with antibodies to galactocerebroside. In the sciatic nerve, which contains a mixture of non-myelin-forming and myelin-forming Schwann cells, transection resulted in a loss of galactocerebroside expression from the surface of all the Schwann cells in the distal stump over a 9 week period, irrespective of their original phenotype. In crushed sciatic nerves, where axons were allowed to regrow into the distal stumps, the number of Schwann cells expressing immunohistochemically detectable quantities of galactocerebroside in the stump declined over the first 3 weeks, but by 9 weeks after crush the total percentage of galactocerebroside-positive cells in the nerve had risen to control levels.(ABSTRACT TRUNCATED AT 250 WORDS)


Spectral beam combining of a 980 nm tapered diode laser bar.
We demonstrate spectral beam combining of a 980 nm tapered diode laser bar. The combined beam from 12 tapered emitters on the bar yielded an output power of 9.3 W at 30 A of operating current. An M2 value of 5.3 has been achieved along the slow axis. This value is close to that of a free running single tapered emitter on the bar at the same current level. The overall spectral beam combining efficiency was measured to be 63%.


Cloning and functional characteristics of murine large granular lymphocyte-1: a member of the Ly-49 gene family (Ly-49G2)
Large granular lymphocyte (LGL) 1 is a cell surface glycoprotein expressed on a subset (50%) of C57BL/6 natural killer (NK) cells. Immunoprecipitation experiments reveal that the LGL-1 protein exists as a disulfide-linked 40-kD homodimer. Functional studies of LGL-1+ cells indicate that selected H-2d target cells are not lysed efficiently by these interleukin (IL)-2-cultured NK cells. These findings suggested that LGL-1 may be a member of the Ly-49 gene family. Here we report the molecular cloning of the LGL-1 cDNA from a severe combined immunodeficient-adherent lymphokine-activated killer cell library transfected into Cos-7 cells and find LGL-1 to be homologous to the Ly-49 gene at both the nucleotide (85%) and amino acid levels (73%). Sequencing of our LGL-1 cDNA has revealed it to be nearly identical to the Ly-49G2 cDNA recently isolated by cross-hybridization with an Ly-49 probe. LGL-1 represents a type II transmembrane protein of 267 amino acids with its carboxyl end exposed extracellularly. The LGL-1 protein contains 11 highly conserved cysteine residues and a 25-amino acid transmembrane region. Southern blot analysis demonstrates that there are a number of homologous genes in mouse DNA that hybridize strongly to LGL-1. Northern analyses using poly A+ RNA from LGL-1+ NK cells indicate that LGL-1 is expressed as a 1.4 kb mRNA. Two-color flow cytometry analysis (FCA) of C57BL/6 splenic NK cells demonstrates that LGL-1 and Ly-49 label overlapping subsets of cells. FCA identifies four subsets of NK cells as defined by LGL-1 versus Ly-49 staining. We have sorted these individual subsets, expanded them in IL-2, and performed cytotoxicity experiments to determine their target cell profiles in relation to class I expression. Results of these studies are complex, but indicate that Ly-49 may not be the only molecule that recognizes class I as an inhibitory signal for cytotoxicity. LGL-1+ cells also fail to lyse several H-2d-expressing tumor targets and concanavalin A lymphoblasts from BALB/c but not C57BL/6 mice. This inhibition of lysis by LGL-1+ NK cells is negated by addition of monoclonal antibody (mAb) 4D11 that recognizes the LGL-1 protein. When mAbs to the class I molecules H-2Dd and H-2Ld (alpha 1 alpha 2 domains only) are added to cytotoxicity assays, LGL-1+ cells lyse H-2d targets very effectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Evidence-based medicine: quality and comparability of clinical trials investigating the efficacy of prostaglandin F(2α) for the treatment of bovine endometritis.
The objective of this study was to evaluate the quality and comparability of published literature, and to summarize the effect of prostaglandin F(2α) (PGF(2α)) for the treatment of endometritis. It has been postulated that there is a dearth of high-level evidence-based research results in veterinary medicine. Also, there is a marked variation in the quality of studies in veterinary and animal science. Post-partum uterine infections occur commonly in dairy cattle and are reported to have a negative impact on reproductive performance. A comprehensive literature search was conducted utilizing online databases revealing a total of 2723 references. After applying specific exclusion criteria, a total of 68 trials were eligible for further analysis. These articles were evaluated utilizing specific parameters listed in an evaluation form such as randomization and the involvement of control groups. The analysis revealed that more than half of the trials (51·5%) were at least 20 years old. Furthermore, we found that about one third (36·8%) of all trials were controlled and randomized, while 3 of those (4·4%) were also blinded. Of those trials which calculated a calving-to-conception interval (n=30), 50% of the authors claimed an improvement, which was statistically significant in 23·3% of the cases. We conclude that there is a wide discrepancy between research results investigating the efficacy of PGF(2α).


Structure-function relationships in helix-bundle channels probed via total chemical synthesis of alamethicin dimers: effects of a Gln7 to Asn7 mutation.
Alamethicin channels are prototypical helix bundles that may serve as tractable models for more complex protein ion channels. Solid-phase peptide synthesis of alamethicin analogues using FMOC-amino acid fluorides followed by chemical dimerization of these peptides facilitates structure-function studies of particular channel states in bilayer membranes. State 3 in particular, tentatively assigned to a hexameric helix bundle, is sufficiently long-lived that current-voltage measurements can be made during the lifetime of an individual channel opening. Molecular models of hexameric helix bundles, generated using restrained molecular dynamics with simulated annealing, indicate that a Gln7-->Asn7 (Q7-->N7) mutation will increase channel diameter locally. Experimentally, the conductance of state 3 of the N7-alm channel is found to be larger than that of the Q7-alm channel when ion flow is in the usual direction (cations entering the C-terminal end of the channel). When ion flow is in the opposite direction, no difference in the conductances of state 3 of Q7 and state 3 of N7 channels is observed. These results indicate that the effect of a change in pore diameter at position 7 is dependent on the magnitude of other barriers to permeation and that these barriers are voltage-dependent.


α3β4 nicotinic acetylcholine receptors in the medial habenula modulate the mesolimbic dopaminergic response to acute nicotine in vivo.
Habenulo-interpeduncular nicotinic receptors, particularly those containing α3, β4 and α5 subunits, have recently been implicated in the reinforcing effects of nicotine. Our laboratory has shown that injection of α3β4 nicotinic receptor antagonists into the medial habenula (MHb) decreases self-administration of multiple abused drugs, including nicotine (Glick et al., 2006, 2008; 2011). However, it is unclear whether blockade of MHb nicotinic receptors has a direct effect on mesolimbic dopamine. Here, we performed in vivo microdialysis in female rats. Microdialysis probes were implanted into the nucleus accumbens (NAcc) and α3β4 nicotinic receptor antagonists (18-methoxycoronaridine; 18-MC or α-conotoxin AuIB; AuIB), were injected into the ipsilateral MHb, just prior to systemic nicotine (0.4 mg/kg, s.c.). Dialysate samples were collected before and after drug administration and levels of extracellular dopamine and its metabolites were measured using HPLC. Acute nicotine administration increased levels of extracellular dopamine and its metabolites in the NAcc. Pre-treatment with intra-habenular AuIB or 18-MC prevented nicotine-induced increases in accumbal dopamine. Neither drug had an effect on nicotine-induced increases in dopamine metabolites, suggesting that α3β4 receptors do not play a role in dopamine metabolism. The effect of intra-habenular blockade of α3β4 receptors on NAcc dopamine was selective for acute nicotine: neither AuIB nor 18-MC prevented increases in NAcc dopamine stimulated by acute d-amphetamine or morphine. These results suggest the mesolimbic response to acute nicotine, but not to acute administration of other drugs of abuse, is directly modulated by α3β4 nicotinic receptors in the MHb, and emphasize a critical role for habenular nicotinic receptors in nicotine's reinforcing effects.


Homologous monoclonal antibodies specific for alpha 1-3 dextran stimulate polyclonal proliferation of Lyb-5+ B cells.
In this paper we describe the polyclonal blast response induced in nonimmune murine splenocytes by two homologous alpha 1-3 dextran-binding myeloma protein, J558 and MOPC104E. This stimulation appears to be independent of MHC or IgCh gene complex control, and proceeds entirely in a T-dependent fashion. The responding B cell population appears to belong to the more mature Lyb-5+ subset. This response was elicited by several independently prepared batches of J558 and MOPC104E, each of which was conclusively shown to be free of endotoxin contamination. Experiments are presented that suggest that this stimulation is being mediated via the Fab portion of these two myeloma proteins, in particular, a shared IdX, rather than through their Fc.


Universal scaling rules predict evolutionary patterns of myogenesis in species with indeterminate growth.
Intraspecific phenotypic variation is ubiquitous and often associated with resource exploitation in emerging habitats. For example, reduced body size has evolved repeatedly in Arctic charr (Salvelinus alpinus L.) and threespine stickleback (Gasterosteus aculeatus L.) across post-glacial habitats of the Northern Hemisphere. Exploiting these models, we examined how body size and myogenesis evolve with respect to the 'optimum fibre size hypothesis', which predicts that selection acts to minimize energetic costs associated with ionic homeostasis by optimizing muscle fibre production during development. In eight dwarf Icelandic Arctic charr populations, the ultimate production of fast-twitch muscle fibres (FN(max)) was only 39.5 and 15.5 per cent of that in large-bodied natural and aquaculture populations, respectively. Consequently, average fibre diameter (FD) scaled with a mass exponent of 0.19, paralleling the relaxation of diffusional constraints associated with mass-specific metabolic rate scaling. Similar reductions in FN(max) were observed for stickleback, including a small-bodied Alaskan population derived from a larger-bodied oceanic stock over a decadal timescale. The results suggest that in species showing indeterminate growth, body size evolution is accompanied by strong selection for fibre size optimization, theoretically allowing resources saved from ionic homeostasis to be allocated to other traits affecting fitness, including reproduction. Gene flow between small- and large-bodied populations residing in sympatry may counteract the evolution of this trait.


Herpetic esophagitis: a case report on an immunocompetent adolescent.
Herpetic esophagitis in immunocompetent individuals is a rare entity that should be suspected clinically by an acute onset of symptoms, and without apparent cause of a symptomatic triad consisting on odynophagia, heartburn and fever. Its occurrence may be due to reactivation of a previous infection or less often a primary infection. Herpes simplex type 1 is the most common cause. Upper endoscopy establishes the diagnosis of suspicion of herpetic esophagitis. It also allows to take multiple biopsy samples and viral culture, leading to a definitive diagnosis. The severity of symptoms is related to the degree of oesophageal involvement. In immunocompromised patients treatment is indicated with acyclovir, but the indication in immunocompetent patients is controversial because the process is time, limited with a low probability of complications. We present a case of acute herpetic esophagitis in an immunocompetent host that debuted acutely with severe upper gastrointestinal tract symptoms, associated with an insidious and nonspecific onset of flu-like symptoms. Endoscopic findings showed a severe involvement in the lower third of the oesophageal mucosa.


Utility of the antigen-specific interferon-gamma assay for the management of tuberculosis.
The tuberculin skin test, now over a century old, is prone to reader variability, and outcomes are influenced by cross-reactivity with environmental mycobacteria, previous bacillus Calmette-Guerin (BCG) vaccination, and anergy in immunosuppressed individuals. More recently, T-cell-based interferon-gamma responses to Mycobacterium tuberculosis - specific antigens have been investigated for their role in diagnosing latent tuberculosis infection. We review the evidence supporting the utility of the interferon-gamma assay for the diagnosis of latent tuberculosis infection (LTBI) in low-prevalence countries. We discuss the principle of the test, technical factors related to performance, and its utility in active tuberculosis, in specialised subgroups such as immunocompromised patients, and its applicability in developing countries. Compared with the tuberculin skin test, the antigen-specific interferon-gamma assay, when used in a standardised protocol (overnight incubation assay using a combination of two antigens) for the diagnosis of LTBI, has greater specificity in BCG-vaccinated individuals, displays a stronger association with exposure, and is less biased by environmental mycobacteria such as Mycobacterium avium. Prospective studies are required, however, to confirm that treating LTBI, as defined by the interferon-gamma assay, will reduce the tuberculosis burden in low-prevalence countries and whether interferon-gamma responses are predictive of those who have a high risk of progression to active tuberculosis. Further studies are required to address the utility of the interferon-gamma assay in specialised subgroups of patients, in developing countries, and as a marker of disease activity.


Molecular mechanisms of homologous desensitization and internalization of muscarinic receptors in primary cultures of neonatal corticostriatal neurons.
Homologous desensitization of muscarinic acetylcholine receptors (mAChR) was studied using primary cultures of corticostriatal neurons from neonatal rats. Prolonged incubation with carbachol attenuated phospholipase C responsiveness to muscarinic agonists and decreased the number of cell surface mAChR, as measured by binding of N-[3H] methylscopolamine to neuronal monolayers. When neurons were exposed to carbachol for 15 min, 40% of the mAChR lost from the membrane domain was recovered in the cytosol; a decrease of the total neuronal receptors was detected following an incubation with the agonist lasting longer than 15 min. Both 8-Br-cyclic AMP and forskolin neither affected N-[3H]methylscopolamine binding to cell monolayers or did they prevent the agonist-mediated mAChR desensitization. 8-Br-cyclic GMP also failed to decrease mAChR number. Pertussis toxin failed to prevent the homologous desensitization of mAChR under conditions that blocked the agonist-mediated inhibition of forskolin-stimulated cyclic AMP formation. The phorbol ester 12-O-tetradecanoyl-phorbol-12, 13-acetate induced a concentration-dependent decrease of N-[3H]methylscopolamine binding to neuronal monolayers. However, the protein kinase C inhibitors sphingosine and the ganglioside monosialosyl-gangliotetraglicosylceramide inhibited the 12-O-tetradecanoyl-phorbol-12,13-acetate-induced but not the agonist-induced desensitization of mAChRs. Furthermore, incubation with muscarinic agonists failed to translocate protein kinase C from cytosol to plasma membranes, as measured by binding of the phorbol ester [3H]-4-beta-phorbol-12,13-dibutyrate to neuronal monolayers. In corticostriatal neurons the agonist-induced desensitization and internalization of mAChR involves neither protein kinase C and protein kinase A activation nor changes in cyclic GMP and cyclic AMP content.


Enhancing the detection of injuries and near-misses among patient care staff in a large pediatric hospital.
Objectives Compared to other industries, healthcare has one of the highest rates of non-fatal occupational injury/illness. Evidence indicates these rates are underestimated, highlighting the need for improved injury surveillance. This study aims to demonstrate the feasibility of integrating active data collection in a passive injury surveillance system to improve detection of injuries in a healthcare establishment. Methods Using digital voice recorders (DVR), pediatric healthcare providers prospectively recorded events throughout their shift for two weeks. This sample-based active injury surveillance was then integrated into an institutional surveillance system (ISS) centered on passive data collection initiated by employee reports. Results Injuries reported using DVR during two-week intervals from February 2014 to July 2015 were 40.7 times more frequent than what would be expected on the basis of the usual ISS reports. Psychological injuries (eg, stress, conflict) and near-misses were captured at a rate of 16.1 per 1000 days [95% confidence interval (CI) 14.1-18.3] and 35.6 per 1000 days (95% CI 32.7-38.8), respectively. Finally, 68% (95% CI 65-72%) of participants preferred using DVR either as an alternative or complement to the existing ISS. Conclusions This study showed that it is feasible to improve injury surveillance in a healthcare establishment by integrating active data collection based on voice recording within a passive injury surveillance system. Enhanced surveillance provides richer information that can guide the development of effective injury prevention strategies.


Self-reported dry eye disease across refractive modalities.
To compare self-reported dry eye disease across contact lens wearers, spectacles wearers, and clinical emmetropes (i.e., those not requiring refractive correction). The survey included two symptom questions (dryness and light sensitivity) that inquired about frequency and intensity at three times of day (morning, afternoon, and evening) and a self-perception question (i.e., Do you think you have dry eyes?). Dryness and light sensitivity scales were then calculated, summed, and scored, providing a dry eye classification. Logistic regression (binary and multinomial) and analysis of covariance were used to examine the relation between mode of refractive correction and dry eye status, frequency of symptoms, and diurnal change in symptom intensity. Overall, 893 surveys were completed, and the age-adjusted frequency of dry eye in the sample was 28.7%, with 3.5% of the sample reporting severe symptoms (at least grade 4 of a possible 5 for both symptoms). Contact lens wearers were most likely to report dry eye disease (52.3%), followed by spectacle wearers (23.9%) and clinical emmetropes (7.1%). Adjustment for age and gender showed that contact lens (adjusted odds ratio = 12.37, 95% confidence interval = 7.55-20.26) and spectacle wearers (adjusted odds ratio = 2.06, 95% confidence interval = 1.12-3.80) were more likely than emmetropes to report dry eye problems. After adjustment for age and gender, contact lens wearers were shown to be more likely to experience frequent symptoms and an increase in symptoms throughout the day (F = 51.4, P < 0.0001). The frequency of self-reported dry eye is high, especially in contact lens wearers.


Salmonella typhimurium phage types from human salmonellosis in Denmark 1988-1993.
A total of 989 isolates of Salmonella enterica ssp. enterica serovar Typhimurium from cases of human salmonellosis were investigated by phage typing. The isolates comprised all isolates recovered during the month of August in each of the years from 1988 to 1993. Phage typing assigned 82.6% of the strains to 36 different definitive types, 11.9% of the strains belonged to types of unknown lysis pattern (RDNC), and 5.5% could not be typed by the phages used (NT). Three phage types (12, 66 and 110) made up approximately 50% of the isolates in each of the years investigated. During the period in question these types showed major changes in prevalence: phage type 12 increased from 4.0% in 1988 to a maximum of 55.2% in 1992, and phage type 66 and phage type 110 were reduced from 40.1% and 27.8% to a minimum of 3.9% and 4.8% in 1993 and 1992, respectively. The increasing prevalence of phage type 12 among isolates from human salmonellosis most likely reflects the increasing significance of pork as a source of human salmonellosis. The reduction in phage types 66 and 110 is paralleled by a reduction in S. Typhimurium in Danish poultry. Some phage types were associated with travel, namely 17, 193 and 194. It is concluded that phage typing, although here performed retrospectively, produces valuable epidemiological information regarding changes in the relative importance of different sources of infection in humans. It is suggested that phage typing be performed prospectively on both human and animal S. Typhimurium isolates in Denmark.


Using the Natural Experiment Study Design to Evaluate the Effect of a Change in Doctor's Roster on Patient Flow in an Emergency Department.
The effect of changes in doctors' rosters is rarely subjected to scientific evaluation. We describe how a natural experiment (NE) study design can be used to evaluate if a managerial decision about doctors' rosters has an effect on patient flow in an emergency department (ED). We hypothesized that an extra doctor each morning from 6 a.m. (i.e., a modified "casino shift") might improve the productivity of a hospital's ED. This was an NE observational study using data on patient flow in the ED of Zealand University Hospital, Denmark, between April 1, 2016, and April 1, 2017. We compared days on which the 6 a.m. emergency physician called in sick (case days) with data from the same weekday a week later where staffing was as scheduled (control days). Patient caseload did not did differ significantly on days with and without the extra doctor from 6 a.m. (measured by number of admissions, triage scores and mean patient age). Door-to-doctor time was 70 minutes (mean, standard deviation [SD], 49) on days without the extra doctor and 56 minutes (mean, SD 41) on days with the early-morning doctor present (p > 0.05). ED length of stay was 250 minutes (mean, SD 119) on days without the extra doctor and 209 minutes (mean, SD 109) on days with the early-morning doctor present (p > 0.05). In our setting, an extra doctor in the ED from 6 a.m. did not change patient flow. These results suggest that the workflow in the ED should be viewed as a connected supply chain. The study also demonstrates that a natural experiment study design can be used to evaluate ED managerial decisions.


Documentation of neurovascular status in supracondylar fractures and the development of an assessment proforma.
Supracondylar fractures are associated with a high incidence of neurovascular complications. Comprehensive clinical evaluation is a necessity when children with these injuries present to the emergency department. Neurovascular assessment can be difficult due to pain, anxiety and the young age of these patients; however, it is crucial findings are well documented to identify patients requiring urgent surgical intervention, in addition to allowing the neurovascular status to be monitored over time. The aim of this study was to evaluate the preoperative neurovascular documentation in children presenting with displaced supracondylar fractures and devise an emergency department assessment proforma to facilitate comprehensive evaluation. A retrospective case-note review was performed on patients with Gartland grades 2 and 3 supracondylar fractures observed in a 2-year period from July 2008 to July 2010. 137 patients were included; only 12 patients (8.8%) and 19 patients (13.9%), respectively, had a complete preoperative neurological or vascular assessment documented. Regarding the individual nerves, 59 (43.1%) patients had median nerve integrity documented, 55 (40.1%) ulnar nerve and 49 (35.8%) radial nerve integrity documented. Only 18 patients (13.1%) had their anterior interosseous nerve (AIN) function documented. Preoperative documentation of neurovascular status in children with displaced supracondylar fractures was poor. Documentation of AIN examination was particularly poor. The introduction of a proforma (Liverpool upper limb fracture assessment) is proposed to increase documentation of neurovascular assessment and optimise emergency department evaluation of children presenting with upper limb injuries.


Differential effects of GM-CSF and G-CSF on infiltration of dendritic cells during early left ventricular remodeling after myocardial infarction.
Several lines of evidence suggest that the immune activation after myocardial infarction (MI) induces secondary myocardial injury. Although dendritic cells (DC) are potent regulators of immunity, their role in MI is still undetermined. We investigated the effect of DC modulation by CSF on left ventricular (LV) remodeling after MI. MI was induced by ligation of the left coronary artery in male Wistar rats. G-CSF (20 microg/kg/day, MI-G, n = 33), a GM-CSF inducer (romurtide, 200 microg/kg/day, MI-GM, n = 28), or saline (MI-C, n = 55) was administered for 7 days. On day 14, MI-G animals had higher LV max dP/dt and smaller LV dimensions, whereas MI-GM animals had lower LV max dP/dt and larger LV dimensions than did MI-C animals, despite similar infarct size. In MI-C, OX62(+) DC infiltrated the infarcted and border areas, peaking on day 7. Bromodeoxyuridine-positive DC were observed in the border area during convalescence. Infiltration by DC was decreased in MI-G animals and increased in MI-GM animals compared with MI-C (p < 0.05). In the infarcted area, the heat shock protein 70, TLR2 and TLR4, and IFN-gamma expression were reduced in MI-G, but increased in MI-GM in comparison with those in MI-C animals. IL-10 expression was higher in MI-G and lower in MI-GM than in MI-C animals. In conclusion, G-CSF improves and GM-CSF exacerbates early postinfarction LV remodeling in association with modulation of DC infiltration. Suppression of DC-mediated immunity could be a new strategy for the treatment of LV remodeling after MI.


Performance-based incentives to improve health status of mothers and newborns: what does the evidence show?
Performance-based incentives (PBIs) aim to counteract weak providers' performance in health systems of many developing countries by providing rewards that are directly linked to better health outcomes for mothers and their newborns. Translating funding into better health requires many actions by a large number of people. The actions span from community to the national level. While different forms of PBIs are being implemented in a number of countries to improve health outcomes, there has not been a systematic review of the evidence of their impact on the health of mothers and newborns. This paper analyzes and synthesizes the available evidence from published studies on the impact of supply-side PBIs on the quantity and quality of health services for mothers and newborns. This paper reviews evidence from published and grey literature that spans PBI for public-sector facilities, PBI in social insurance reforms, and PBI in NGO contracting. Some initiatives focus on safe deliveries, and others reward a broader package of results that include deliveries. The Evidence Review Team that focused on supply-side incentives for the US Government Evidence Summit on Enhancing Provision and Use of Maternal Health Services through Financial Incentives, reviewed published research reports and papers and added studies from additional grey literature that were deemed relevant. After collecting and reviewing 17 documents, nine studies were included in this review, three of which used before-after designs; four included comparison or control groups; one applied econometric methods to a five-year time series; and one reported results from a large-scale impact evaluation with randomly-assigned intervention and control facilities. The available evidence suggests that incentives that reward providers for institutional deliveries result in an increase in the number of institutional deliveries. There is some evidence that the content of antenatal care can improve with PBI. We found no direct evidence on the impact of PBI on neonatal health services or on mortality of mothers and newborns, although intention of the study was not to document impact on mortality. A number of studies describe approaches to rewarding quality as well as increases in the quantities of services provided, although how quality is defined and monitored is not always clear. Because incentives exist in all health systems, considering how to align the incentives of the many health workers and their supervisors so that they focus efforts on achieving health goals for mothers and newborns is critical if the health system is to perform more effectively and efficiently. A wide range of PBI models is being developed and tested, and there is still much to learn about what works best. Future studies should include a larger focus on rewarding quality and measuring its impact. Finally, more qualitative research to better understand PBI implementation and how various incentive models function in different settings is needed to help practitioners refine and improve their programmes.


Spontaneous Up states in vitro: a single-metric index of the functional maturation and regional differentiation of the cerebral cortex.
Understanding the development and differentiation of the neocortex remains a central focus of neuroscience. While previous studies have examined isolated aspects of cellular and synaptic organization, an integrated functional index of the cortical microcircuit is still lacking. Here we aimed to provide such an index, in the form of spontaneously recurring periods of persistent network activity -or Up states- recorded in mouse cortical slices. These coordinated network dynamics emerge through the orchestrated regulation of multiple cellular and synaptic elements and represent the default activity of the cortical microcircuit. To explore whether spontaneous Up states can capture developmental changes in intracortical networks we obtained local field potential recordings throughout the mouse lifespan. Two independent and complementary methodologies revealed that Up state activity is systematically modified by age, with the largest changes occurring during early development and adolescence. To explore possible regional heterogeneities we also compared the development of Up states in two distinct cortical areas and show that primary somatosensory cortex develops at a faster pace than primary motor cortex. Our findings suggest that in vitro Up states can serve as a functional index of cortical development and differentiation and can provide a baseline for comparing experimental and/or genetic mouse models.


Key considerations for the experimental training and evaluation of cancer odour detection dogs: lessons learnt from a double-blind, controlled trial of prostate cancer detection.
Cancer detection using sniffer dogs is a potential technology for clinical use and research. Our study sought to determine whether dogs could be trained to discriminate the odour of urine from men with prostate cancer from controls, using rigorous testing procedures and well-defined samples from a major research hospital. We attempted to train ten dogs by initially rewarding them for finding and indicating individual prostate cancer urine samples (Stage 1). If dogs were successful in Stage 1, we then attempted to train them to discriminate prostate cancer samples from controls (Stage 2). The number of samples used to train each dog varied depending on their individual progress. Overall, 50 unique prostate cancer and 67 controls were collected and used during training. Dogs that passed Stage 2 were tested for their ability to discriminate 15 (Test 1) or 16 (Tests 2 and 3) unfamiliar prostate cancer samples from 45 (Test 1) or 48 (Tests 2 and 3) unfamiliar controls under double-blind conditions. Three dogs reached training Stage 2 and two of these learnt to discriminate potentially familiar prostate cancer samples from controls. However, during double-blind tests using new samples the two dogs did not indicate prostate cancer samples more frequently than expected by chance (Dog A sensitivity 0.13, specificity 0.71, Dog B sensitivity 0.25, specificity 0.75). The other dogs did not progress past Stage 1 as they did not have optimal temperaments for the sensitive odour discrimination training. Although two dogs appeared to have learnt to select prostate cancer samples during training, they did not generalise on a prostate cancer odour during robust double-blind tests involving new samples. Our study illustrates that these rigorous tests are vital to avoid drawing misleading conclusions about the abilities of dogs to indicate certain odours. Dogs may memorise the individual odours of large numbers of training samples rather than generalise on a common odour. The results do not exclude the possibility that dogs could be trained to detect prostate cancer. We recommend that canine olfactory memory is carefully considered in all future studies and rigorous double-blind methods used to avoid confounding effects.


High rates of diabetes reversal in newly diagnosed Asian Indian young adults with type 2 diabetes mellitus with intensive lifestyle therapy.
There are variable reports on the reversibility of type 2 diabetes mellitus (type 2 DM) with higher rates among younger patients with short duration of diabetes. Hence, we studied the reversibility of diabetes among young adults with newly diagnosed type 2 DM. This prospective study included 32 patients with newly diagnosed type 2 DM. All type 2 DM patients were initially treated with intensive lifestyle therapy (ILT) (low-calorie diet [1500 kcal/day] and brisk walking for 1 h/day]). Four patients who with HbA1C <9.0% were treated with ILT alone. Except for three patients with concomitant infections who were treated with insulin, remaining 25 patients with HbA1C ≥9.0% were treated with metformin (1000-2000 g) in addition to ILT. When fasting plasma glucose was <126 mg/dl or HbA1C was <6.5% antidiabetic drug dose was reduced or stopped. The patients were followed for a minimum period of 2 years. Reversal/remission rates at 3 months, 1 year, and 2 years were 24 (75%), 24 (75%), and 22 (68.75%), respectively. Seventeen (53.1%) patients achieved complete reversal and seven (21.9%) patients achieved partial reversal at 3 months. Rates of complete and partial remission at 1 year were 50% and 25% and at 2 years were 46.9% and 21.9%, respectively. Young adults with newly diagnosed type 2 DM have high rates of diabetes reversal and should receive ILT to achieve reversal of diabetes.


Maitotoxin stimulates steroid and peptide hormone secretion by swine luteal tissue.
The impact of calcium influx on the secretion of steroidal and nonsteroidal ovarian products was tested using a novel marine toxin, maitotoxin, a putative activator of endogenous calcium channels. In incubations of swine luteal minces in balanced salt solution, maitotoxin induced dose-dependent (3-12.5 ng/ml) and time-dependent (15-60 min) secretion of progesterone and relaxin. The magnitude of these stimulatory effects approached that observed for luteinizing hormone (LH) and was explicitly dependent on the presence of extracellular calcium ions. Maitotoxin's actions were not significantly impeded by substitution of choline for sodium ions in the incubation medium. Moreover, measurements of tissue calcium content by atomic absorption spectrometry indicated that maitotoxin augmented the total cellular content of calcium by 44 +/- 3.1% within 1 min. This time course was distinct from that observed for LH. The specificity of these actions on the calcium pathway was further suggested by the absence of any measurable effects of maitotoxin on luteal cAMP generation. We conclude that maitotoxin transiently increases luteal calcium content without altering cAMP generation and elicits concomitant release of both a steroidal (progesterone) and nonsteroidal (relaxin) product of the corpus luteum. Such actions are dependent on extracellular calcium but not sodium ions. These observations provide evidence for a calcium-dependent stimulatory pathway that activates both steroidal and nonsteroidal mechanisms of secretion in the swine corpus luteum.


Baculovirus-Free SARS-CoV-2 Virus-like Particle Production in Insect Cells for Rapid Neutralization Assessment.
Virus-like particles (VLPs) resemble authentic virus while not containing any genomic information. Here, we present a fast and powerful method for the production of SARS-CoV-2 VLP in insect cells and the application of these VLPs to evaluate the inhibition capacity of monoclonal antibodies and sera of vaccinated donors. Our method avoids the baculovirus-based approaches commonly used in insect cells by employing direct plasmid transfection to co-express SARS-CoV-2 envelope, membrane, and spike protein that self-assemble into VLPs. After optimization of the expression plasmids and vector ratios, VLPs with an ~145 nm diameter and the typical "Corona" aura were obtained, as confirmed by nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Fusion of the membrane protein to GFP allowed direct quantification of binding inhibition to angiotensin II-converting enzyme 2 (ACE2) on cells by therapeutic antibody candidates or sera from vaccinated individuals. Neither VLP purification nor fluorescent labeling by secondary antibodies are required to perform these flow cytometric assays.


Modeling the effect of chronic schistosomiasis on childhood development and the potential for catch-up growth with different drug treatment strategies promoted for control of endemic schistosomiasis.
In areas endemic for schistosomiasis having limited healthcare, targeted drug treatment of school-age children is recommended for control of Schistosoma-associated morbidity. However, optimal timing, number, and frequency of treatments are not established. Because longitudinal studies of long-term impact of treatment are few, for current policy considerations we performed quantitative simulation (based on calibrated modeling of Schistosoma-associated disease formation) to project the impact of different school-age treatment regimens. Using published efficacy data from targeted programs, combined with age-specific risk for growth retardation and reinfection, we examined the likely impact of different strategies for morbidity prevention. Results suggest the need for early, repeated treatment through primary school years to optimally prevent the disabling sequelae of stunting and undernutrition. Dynamics of infection/reinfection during childhood and adolescence, combined with early treatment effects against reversible infection-associated morbidities, create a need for aggressive retreatment of preadolescents to achieve optimal suppression of morbidity where drug-based control is used.


The impacts of dwelling conditions on older persons' psychological well-being in Hong Kong: the mediating role of residential satisfaction.
About 11% of Hong Kong's population of 7 million people are aged 65 and over and many of them live in old urban areas. Many of these areas have been subjected to urban redevelopment and some of the residents have been relocated to newer estates in peripheral new towns. Previous studies have focused on the challenges the urban environment has placed on older persons in terms of capability to cope with the demands that the environment places upon them. This paper suggests that dwelling conditions can act as stressors and become contributing factors that impact on older persons' residential satisfaction and psychological well-being (subjective well-being). This study examines the role of residential satisfaction (satisfaction with dwelling unit, estate and district) in mediating the effects of dwelling conditions (interior environment and exterior environment) on psychological well-being. A sample of older persons was recruited from a sampling frame of 16 urban sub-areas located in old urban areas and new towns. 518 older persons (224 males, 294 females) aged 60 and over were interviewed and the findings indicated that residential satisfaction was determined by assessment of both the interior environment and the exterior environment, although these were appraised differently. The interior environment had a greater impact on residential satisfaction than the exterior environment. It appeared that environmental dwelling conditions mainly affected older persons' psychological well-being indirectly and, hence, probably influenced their opportunities for successful ageing. However, subsequent tests revealed that dwelling conditions had no direct impact on psychological well-being. In light of these findings, it is proposed that the role of environmental factors and their relation to older persons' psychological well-being depends on the extent to which a person's expectations of residential satisfaction are met. Some implications of these findings for local housing and social care policy are discussed.


Sporadic and syndromic hyperplastic polyps and serrated adenomas of the colon: classification, molecular genetics, natural history, and clinical management.
There is now strong evidence for an alternative pathway of colorectal carcinogenesis implicating hyperplastic polyps and serrated adenomas. This article briefly reviews the evidence for this serrated pathway, provides diagnostic criteria for clinically significant hyperplastic polyps and allied serrated polyps, and suggests how this information may be translated into safe, effective guidelines for colonoscopy-based colon cancer prevention. Consideration also is given to the definition and management of hyperplastic polyposis syndrome. The currently proposed management plan for serrated polyps is tentative because of incomplete knowledge of the nature and behavior of these polyps. This article highlights key areas warranting further research.


Basic FGF and TGF-beta 1 influence commitment to melanogenesis in neural crest-derived cells of avian embryos.
In previous studies, we showed that neural crest (NC)-derived cells from embryonic quail dorsal root ganglia (DRG) and peripheral nerve (PN), which do not normally give rise to melanocytes, become committed to melanogenesis following treatment in culture with the phorbol ester drug 12-O-tetradecanoyl phorbol-13-acetate (TPA). These and other observations support the notion that melanocytes and Schwann cells are derived from a common bipotent intermediate in the neural crest lineage--the melanocyte/Schwann cell progenitor. In this study, we test the possibility that peptide growth factors found in the embryonic environment might act similarly to TPA to influence the fates of these cells. DRG and PN explants were cultured in medium supplemented with a variety of growth factors, and then the cultures were examined for the presence of pigment cells. We found that basic fibroblast growth factor (bFGF), but not various other growth factors, induced pigmentation in about 20% of these cultures. When low concentrations of TPA were included in the culture medium, bFGF augmented the TPA-induced pigmentation, significantly increasing the proportion of pigmented cultures. These effects of bFGF were age-dependent, and could be blocked by addition of a bFGF-neutralizing antibody to the culture medium. In contrast to these stimulatory effects of bFGF, transforming growth factor-beta 1 (TGF-beta 1) was found to inhibit the TPA- or bFGF-induced pigmentation of DRG cultures. These data suggest, therefore, that at least some NC-derived cells are responsive to bFGF and TGF-beta 1, and that these growth factors may play an important role in the control of NC cell fate.


Unusual temperature dependence of photosynthetic electron transfer due to protein dynamics.
The initial electron transfer rate and protein dynamics in wild type and five mutant reaction centers from Rhodobacter sphaeroides have been studied as a function of temperature (10-295 K). Detailed kinetic measurements of initial electron transfer in Rhodobacter sphaeroides reaction centers can be quantitatively described by a reaction diffusion formalism at all temperatures from 10 to 295 K. In this model, the time course of electron transfer is determined by the ability of the protein to interconvert between conformations until one is found where the activation energy for electron transfer is near zero. In reaction centers with a free energy for electron transfer similar to wild type, the reaction proceeds at least as fast at cryogenic temperatures as at room temperature. This may be because interconversion between conformations at low temperature is restricted to conformations with near zero activation energy (it is not possible to diffuse away from this region of conformational space). In contrast, mutants with a decreased free energy initially find themselves in conformations unfavorable for electron transfer and require more extensive conformational diffusion to achieve a low activation energy conformation. They therefore undergo electron transfer more slowly at 10 K vs 295 K.


Impact of hospital and surgeon volume on in-hospital mortality from radical cystectomy: data from the health care utilization project.
We determined the influence of hospital and surgeon volume on various outcome parameters after radical cystectomy for bladder cancer. All inpatient discharges after radical cystectomy for bladder cancer (1988 to 1999) from the Health Care Utilization Project-Nationwide Inpatient Sample were included in the analysis. Hospital and individual surgeon volume of discharges per year were separated into terciles. Outcome measures were in-hospital mortality, length of stay (LOS), and inflation adjusted charge per admission. Mortality was compared among hospital volume levels using the Mantel-Haenszel chi-square test while the LOS and charges were compared using ANOVA. Multivariate linear and logistic regression analyses were used to adjust for confounding factors. All the analyses were also performed in 3 different age strata (younger than 50 years, 50 to 69 years and 70 years or more). There were 13,964 patients who underwent radical cystectomy. Overall in-hospital mortality was 408 of 13,964 (2.9%), average LOS was 14 days (+/- SD 10.28) and average charges were 47,146 dollars (+/- SD 45,263 dollars). In-hospital mortality was significantly associated with higher volume particularly for patients older than 50 years. Surgeon volume did not influence in-hospital mortality except for patients in the 50 to 69-year-old age group. Results of multivariate regression analysis demonstrated hospital volume was a significant predictor of in-hospital mortality but this effect was lost when controlling for surgeon volume. LOS was significantly higher for low volume surgeons. High volume hospitals had lower average total charges compared with the low and moderate volume hospitals. Hospital and surgeon volume have a significant impact on in-hospital mortality and LOS after radical cystectomy. Radical cystectomy performed at a higher volume center may result in lower charges and shorter hospital stay while decreasing the likelihood of in-hospital mortality.


Circuitry and plasticity of the dorsal horn--toward a better understanding of neuropathic pain.
Maladaptive plasticity within the dorsal horn (DH) of the spinal cord is a key substrate for development of neuropathic pain following peripheral nerve injury. Advances in genetic engineering, tracing techniques and opto-genetics are leading to a much better understanding of the complex circuitry of the spinal DH and the radical changes evoked in such circuitry by nerve injury. These changes can be viewed at multiple levels including: synaptic remodeling including enhanced excitatory and reduced inhibitory drive, morphological and electrophysiological changes which are observed both to primary afferent inputs as well as DH neurons, and ultimately circuit-level rewiring which leads to altered connectivity and aberrant processing of sensory inputs in the DH. The DH should not be seen in isolation but is subject to important descending modulation from the brainstem, which is further dysregulated by nerve injury. Understanding which changes relate to specific disease-states is essential, and recent work has aimed to stratify patient populations in a mechanistic fashion. In this review we will discuss how such pathophysiological mechanisms may lead to the distressing sensory phenomena experienced by patients suffering neuropathic pain, and the relationship of such mechanisms to current and potential future treatment modalities.


Introducing GUt low-density array (GULDA): a validated approach for qPCR-based intestinal microbial community analysis.
Alterations in the human gut microbiota caused, for example, by diet, functional foods, antibiotics, or occurring as a function of age are now known to be of relevance for host health. Therefore, there is a strong need for methods to detect such alterations in a rapid and comprehensive manner. In the present study, we developed and validated a high-throughput real-time quantitative PCR-based analysis platform, termed 'GUt Low-Density Array' (GULDA). The platform was designed for simultaneous analysis of the change in the abundance of 31 different microbial 16S rRNA gene targets in fecal samples obtained from individuals at various points in time. The target genes represent important phyla, genera, species, or other taxonomic groups within the five predominant bacterial phyla of the gut, Firmicutes, Bacteroidetes, Actinobacteria, Proteobacteria, and Verrucomicrobia and also Euryarchaeota. To demonstrate the applicability of GULDA, analysis of fecal samples obtained from six healthy infants at both 9 and 18 months of age was performed and showed a significant increase over time of the relative abundance of bacteria belonging to Clostridial cluster IV (Clostridia leptum group) and Bifidobacterium bifidum and concurrent decrease in the abundance of Clostridium butyricum and a tendency for decrease in Enterobacteriaceae over the 9-month period.


Risk Factors for Hip Fracture in Older Men: The Osteoporotic Fractures in Men Study (MrOS).
Almost 30% of hip fractures occur in men; the mortality, morbidity, and loss of independence after hip fractures are greater in men than in women. To comprehensively evaluate risk factors for hip fracture in older men, we performed a prospective study of 5994 men, primarily white, age 65+ years recruited at six US clinical centers. During a mean of 8.6 years of 97% complete follow-up, 178 men experienced incident hip fractures. Information on risk factors including femoral neck bone mineral density (FNBMD) was obtained at the baseline visit. Cox proportional hazards models were used to calculate the hazard ratio (HR) with 95% confidence intervals; Fine and Gray models adjusted for competing mortality risk. Older age (≥75 years), low FNBMD, currently smoking, greater height and height loss since age 25 years, history of fracture, use of tricyclic antidepressants, history of myocardial infarction or angina, hyperthyroidism or Parkinson's disease, lower protein intake, and lower executive function were all associated with an increased hip fracture risk. Further adjustment for competing mortality attenuated HR for smoking, hyperthyroidism, and Parkinson's disease. The incidence rate of hip fracture per 1000 person-years (PY) was greatest in men with FNBMD T-scores <-2.5 (white women reference database) who also had 4+ risk factors, 33.4. Men age ≥80 years with 3+ major comorbidities experienced hip fracture at rates of 14.52 versus 0.88 per 1000 PY in men age <70 years with zero comorbidities. Older men with low FNBMD, multiple risk factors, and multimorbidity have a high risk of hip fracture. Many of these assessments can easily be incorporated into routine clinical practice and may lead to improved risk stratification. © 2016 American Society for Bone and Mineral Research.


Ebselen and analogs as inhibitors of Bacillus anthracis thioredoxin reductase and bactericidal antibacterials targeting Bacillus species, Staphylococcus aureus and Mycobacterium tuberculosis.
Bacillus anthracis is the causative agent of anthrax, a disease associated with a very high mortality rate in its invasive forms. We studied a number of ebselen analogs as inhibitors of B. anthracis thioredoxin reductase and their antibacterial activity on Bacillus subtilis, Staphylococcus aureus, Bacillus cereus and Mycobacterium tuberculosis. The most potent compounds in the series gave IC(50) values down to 70 nM for the pure enzyme and minimal inhibitory concentrations (MICs) down to 0.4 μM (0.12 μg/ml) for B. subtilis, 1.5 μM (0.64 μg/ml) for S. aureus, 2 μM (0.86 μg/ml) for B. cereus and 10 μg/ml for M. tuberculosis. Minimal bactericidal concentrations (MBCs) were found at 1-1.5 times the MIC, indicating a general, class-dependent, bactericidal mode of action. The combined bacteriological and enzymological data were used to construct a preliminary structure-activity-relationship for the benzoisoselenazol class of compounds. When S. aureus and B. subtilis were exposed to ebselen, we were unable to isolate resistant mutants on both solid and in liquid medium suggesting a high resistance barrier. These results suggest that ebselen and analogs thereof could be developed into a novel antibiotic class, useful for the treatment of infections caused by B. anthracis, S. aureus, M. tuberculosis and other clinically important bacteria. Furthermore, the high barrier against resistance development is encouraging for further drug development. We have characterized the thioredoxin system from B. anthracis as a novel drug target and ebselen and analogs thereof as a potential new class of antibiotics targeting several important human pathogens.


A SNARE-protein has opposing functions in penetration resistance and defence signalling pathways.
Penetration resistance is often the first line of defence against fungal pathogens. Subsequently induced defences are mediated by the programmed cell death (PCD) reaction pathway and the salicylic acid (SA), jasmonic acid (JA) and ethylene (ET) signalling pathways. We previously demonstrated that full penetration resistance in Arabidopsis against the non-host barley powdery mildew fungus (Blumeria graminis f.sp. hordei) requires the syntaxin SYP121 (PEN1). Here we report that SYP121, together with SYP122, functions as a negative regulator of subsequently induced defence pathways. The SA level in the syntaxin double mutant syp121-1 syp122-1 is dramatically elevated, resulting in necrosis and dwarfism. This phenotype is partially rescued by introducing the SA-signalling mutations eds1-2, eds5-3, sid2-1 and npr1-1 as well as the NahG transgene. These partially rescued triple mutants have an unknown defence to Pseudomonas syringae pv. tomato, and have increased HR-like responses to non-host and host powdery mildew fungi. The HR-like responses cause efficient resistance to the latter. These defence pathways are SA-independent. Furthermore, the JA/ET signalling marker, PDF1.2, is highly upregulated in the triple mutants. Thus SYP121 and SYP122 are negative regulators of PCD, SA, JA and ET pathways through a molecular function distinct from that of SYP121 in penetration resistance. Our data suggest that individual cells preferentially express either penetration resistance or the subsequently induced defences.


Localization of the sinoatrial and atrioventricular nodal region in neonatal and juvenile ovine hearts.
Localization of the components of the cardiac conduction system (CCS) is essential for many therapeutic procedures in cardiac surgery and interventional cardiology. While histological studies provided fundamental insights into CCS localization, this information is incomplete and difficult to translate to aid in intraprocedural localization. To advance our understanding of CCS localization, we set out to establish a framework for quantifying nodal region morphology. Using this framework, we quantitatively analyzed the sinoatrial node (SAN) and atrioventricular node (AVN) in ovine with postmenstrual age ranging from 4.4 to 58.3 months. In particular, we studied the SAN and AVN in relation to the epicardial and endocardial surfaces, respectively. Using anatomical landmarks, we excised the nodes and adjacent tissues, sectioned those at a thickness of 4 μm at 100 μm intervals, and applied Masson's trichrome stain to the sections. These sections were then imaged, segmented to identify nodal tissue, and analyzed to quantify nodal depth and superficial tissue composition. The minimal SAN depth ranged between 20 and 926 μm. AVN minimal depth ranged between 59 and 1192 μm in the AVN extension region, 49 and 980 μm for the compact node, and 148 and 888 μm for the transition to His Bundle region. Using a logarithmic regression model, we found that minimal depth increased logarithmically with age for the AVN (R2 = 0.818, P = 0.002). Also, the myocardial overlay of the AVN was heterogeneous within different regions and decreased with increasing age. Age associated alterations of SAN minimal depth were insignificant. Our study presents examples of characteristic tissue patterns superficial to the AVN and within the SAN. We suggest that the presented framework provides quantitative information for CCS localization. Our studies indicate that procedural methods and localization approaches in regions near the AVN should account for the age of patients in cardiac surgery and interventional cardiology.


Prepubertal unilateral gynecomastia: report of 2 cases.
Prepubertal unilateral gynecomastia is an extremely rare condition. At present, its etiology and management strategy are not well known. Two unrelated prepubertal boys of ages 8 and 9 who presented with complaints of unilateral enlargement of breast tissue are reported. Physical examination, biochemical, hormonal and oncologic work-up findings were normal. Both patients were treated with peripheral liposuction successfully. Histopathological and immunohistochemical examinations showed benign fibroglandular gynecomastia and intensive (3+) estrogen receptor expression in 100% of periductal epithelial cells. Although an extremely rare and generally benign condition, patients with prepubertal unilateral gynecomastia should have a full endocrine and oncologic work-up.


Online identification of viruses.
A computerized animal virus information system is developed in the Sequence Retrieval System (SRS) format. This database is available on the Word Wide Web (WWW) at the site http://bioinfo.ernet.in/www/avis/avis++ +.html. The database has been used to generate large number of identification matrices for each family. The software is developed in C. Unix shell scripts and Hypertext Marked-up Language (HTML) to assign the family to an unknown virus deterministically and to identify the virus probabilistically. It has been shown that such web based virus identification approach provides results with high confidence in those cases where identification matrix uses large number of independent characters. Protein sequence data for animal viruses have been analyzed and oligopeptides specific to each virus family and also specific to each virus species are identified for several viruses. These peptides thus could be used to identify the virus and to assign the virus family with high confidence showing the usefulness of sequence data in virus identification.


Prevalence of human papillomavirus among STD clinic attenders in Jamaica: association of younger age and increased sexual activity.
Human papillomavirus is the major etiologic agent of cervical cancer. Although the incidence of cancer of the cervix is high in Jamaica, the prevalence of human papillomavirus among Jamaican women has not been defined. To estimate the prevalence of human papillomavirus infection and associated risk factors in women attending an STD clinic in Kingston, Jamaica. A cross-sectional survey was done of 202 women attending an STD clinic in Kingston in 1990. Cervical and vaginal cells were collected by lavage, and human papillomavirus genomes were detected in extracted DNA using low-stringency Southern blot hybridization. Fifty-eight (28.7%) women were identified as HPV positive. Prevalence of HPV by age group was 39% in women 15-19 years old, 33% of women 20-24 years old, 31% in women 25-29 years old, and 17% in those 30 years or older. Increasing age was significantly associated with a lower prevalence of human papillomavirus infection (test for trend, P = 0.025). The effect of age was independent of years of sexual activity. Women reporting more than one sexual partner per month on average were found to have a significantly higher HPV prevalence (odds ratio 2.87, 95% confidence Interval 1.29-6.38), as were women who reported more frequent sex (test for trend, P = 0.006). Sexual behavior is associated with risk of human papillomavirus infection. The decrease of human papillomavirus prevalence in older women agrees with other studies that argue in favor of a biological effect, such as increased immunity to human papillomavirus with age. A better understanding of why immunity to human papillomavirus may develop in older women may provide the basis for developing an effective vaccine to prevent cancer of the cervix.


A new approach to untargeted integration of high resolution liquid chromatography-mass spectrometry data.
Because of its high sensitivity and specificity, hyphenated mass spectrometry has become the predominant method to detect and quantify metabolites present in bio-samples relevant for all sorts of life science studies being executed. In contrast to targeted methods that are dedicated to specific features, global profiling acquisition methods allow new unspecific metabolites to be analyzed. The challenge with these so-called untargeted methods is the proper and automated extraction and integration of features that could be of relevance. We propose a new algorithm that enables untargeted integration of samples that are measured with high resolution liquid chromatography-mass spectrometry (LC-MS). In contrast to other approaches limited user interaction is needed allowing also less experienced users to integrate their data. The large amount of single features that are found within a sample is combined to a smaller list of, compound-related, grouped feature-sets representative for that sample. These feature-sets allow for easier interpretation and identification and as important, easier matching over samples. We show that the automatic obtained integration results for a set of known target metabolites match those generated with vendor software but that at least 10 times more feature-sets are extracted as well. We demonstrate our approach using high resolution LC-MS data acquired for 128 samples on a lipidomics platform. The data was also processed in a targeted manner (with a combination of automatic and manual integration) using vendor software for a set of 174 targets. As our untargeted extraction procedure is run per sample and per mass trace the implementation of it is scalable. Because of the generic approach, we envision that this data extraction lipids method will be used in a targeted as well as untargeted analysis of many different kinds of TOF-MS data, even CE- and GC-MS data or MRM. The Matlab package is available for download on request and efforts are directed toward a user-friendly Windows executable.


In vivo study of bacterial adhesion to five types of intraocular lenses.
To determine in vivo behavior of the ability of the Staphylococcus epidermidis strain (American Type Culture Collection [ATCC] 14990) to attach to 120 intraocular lenses (IOLs) made of five different biomaterials: fluorine polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophobic acrylic, and hydrogel. The pig was chosen as an animal model of endophthalmitis, after a bibliographical analysis and a personal study of its aqueous humor composition. Crystalline lenses from 90 domestic pigs were removed aseptically and replaced with previously infected IOLs. The animals were killed 24 hours, 72 hours, and 1 week after implantation of the IOLs. The extent of bacterial binding was then measured by counting. Results were compared with a two-factor analysis of variance (ANOVA 2), confirmed by the Kruskal-Wallis nonparametric test. The extent of bacterial binding (expressed as bound bacteria per area unit) was found to range in increasing order from hydrogel, to fluorine PMMA, to hydrophobic acrylic, to heparinized PMMA, to silicone polymer. Comparison of pairs of materials showed statistically significant differences, except between hydrogel and fluorine PMMA. To the authors' knowledge, no study has been published so far concerning the in vivo evolution of populations of bacteria adhering to different intraocular materials. Bacterial adhesion to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial. Adhesion is also affected by the nature of the surrounding medium. Because of its complexity, the latter appears to be very difficult to model, thus making in vivo studies essential.


Glycol methacrylate (GMA) embedding for light microscopy. II. Immunohistochemical analysis of semithin sections of undecalcified marrow cores.
A routine method allows bone marrow biopsy specimens to be embedded in glycol methacrylate (GMA), a water miscible plastic, and to benefit from the advantages of good morphology with immunoperoxidase detection of a wide range of cellular antigens useful in diagnosing and classifying various haematopoietic disorders. Marrow cores were fixed in cold Bouin's solution, rinsed in cold phosphate buffer, dehydrated in cold methanol, infiltrated and embedded in cold GMA, then polymerised at 4 degrees C. Sections were cut at 2 micron thickness with a Tungsten carbide knife in a Jung's high performance microtome (Autocut). Antigenecity was preserved when drying slides at room temperature but pronase digestion was necessary to re-expose the antigens in bone marrow biopsy sections embedded in GMA. Histostik, a new adhesive, was used to coat the glass slides to prevent section loss during enzyme digestion and immunostaining procedures. This method of adapting plastic embedding to undecalcified marrow cores preserves marrow architecture and cellular details and it can serve as a useful adjunct to analyse the bone marrow from patients with myeloproliferative and lymphoproliferative disorders. This technique may also be applicable in non-haematological malignant conditions which affect the marrow.


Subtracting the sequence bias from partially digested MNase-seq data reveals a general contribution of TFIIS to nucleosome positioning.
TFIIS stimulates RNA cleavage by RNA polymerase II and promotes the resolution of backtracking events. TFIIS acts in the chromatin context, but its contribution to the chromatin landscape has not yet been investigated. Co-transcriptional chromatin alterations include subtle changes in nucleosome positioning, like those expected to be elicited by TFIIS, which are elusive to detect. The most popular method to map nucleosomes involves intensive chromatin digestion by micrococcal nuclease (MNase). Maps based on these exhaustively digested samples miss any MNase-sensitive nucleosomes caused by transcription. In contrast, partial digestion approaches preserve such nucleosomes, but introduce noise due to MNase sequence preferences. A systematic way of correcting this bias for massively parallel sequencing experiments is still missing. To investigate the contribution of TFIIS to the chromatin landscape, we developed a refined nucleosome-mapping method in Saccharomyces cerevisiae. Based on partial MNase digestion and a sequence-bias correction derived from naked DNA cleavage, the refined method efficiently mapped nucleosomes in promoter regions rich in MNase-sensitive structures. The naked DNA correction was also important for mapping gene body nucleosomes, particularly in those genes whose core promoters contain a canonical TATA element. With this improved method, we analyzed the global nucleosomal changes caused by lack of TFIIS. We detected a general increase in nucleosomal fuzziness and more restricted changes in nucleosome occupancy, which concentrated in some gene categories. The TATA-containing genes were preferentially associated with decreased occupancy in gene bodies, whereas the TATA-like genes did so with increased fuzziness. The detected chromatin alterations correlated with functional defects in nascent transcription, as revealed by genomic run-on experiments. The combination of partial MNase digestion and naked DNA correction of the sequence bias is a precise nucleosomal mapping method that does not exclude MNase-sensitive nucleosomes. This method is useful for detecting subtle alterations in nucleosome positioning produced by lack of TFIIS. Their analysis revealed that TFIIS generally contributed to nucleosome positioning in both gene promoters and bodies. The independent effect of lack of TFIIS on nucleosome occupancy and fuzziness supports the existence of alternative chromatin dynamics during transcription elongation.


Molybdenum availability, nitrogen limitation, and phytoplankton growth in natural waters.
Sulfate inhibits molybdate assimilation by phytoplankton, making molybdate less available in seawater than it is in freshwater. As a result, nitrogen fixation and nitrate assimilation, both processes that require molybdenum, may require a greater expenditure of energy in seawater than in freshwater. This may explain in part why coastal marine ecosystems are usually nitrogen limited whereas lakes usually are not. Experimentally increasing the ratio of sulfate to molybdate (i) inhibits molybdate uptake, (ii) slows nitrogen fixation rates, and (iii) slows the growth of organisms that use nitrate as their nitrogen source.


A synthetic biology approach to understanding cellular information processing.
The survival of cells and organisms requires proper responses to environmental signals. These responses are governed by cellular networks, which serve to process diverse environmental cues. Biological networks often contain recurring network topologies called "motifs". It has been recognized that the study of such motifs allows one to predict the response of a biological network and thus cellular behavior. However, studying a single motif in complete isolation of all other network motifs in a natural setting is difficult. Synthetic biology has emerged as a powerful approach to understanding the dynamic properties of network motifs. In addition to testing existing theoretical predictions, construction and analysis of synthetic gene circuits has led to the discovery of novel motif dynamics, such as how the combination of simple motifs can lead to autonomous dynamics or how noise in transcription and translation can affect the dynamics of a motif. Here, we review developments in synthetic biology as they pertain to increasing our understanding of cellular information processing. We highlight several types of dynamic behaviors that diverse motifs can generate, including the control of input/output responses, the generation of autonomous spatial and temporal dynamics, as well as the influence of noise in motif dynamics and cellular behavior.


Systemic and neurologic autoimmune disorders associated with seizures or epilepsy.
In this article, we review the incidence and significance of seizures in well-established autoimmune disorders, including multiple sclerosis (MS), diabetes mellitus, celiac disease, thyroid disease, and systemic lupus erythematosus (SLE). The five following presentations discuss the incidence and possible pathogenesis of epilepsies that are found in these well-known autoimmune conditions. There is a large body of evidence describing the clinical presentation of seizures with MS and SLE, and showing that refractory epilepsy can complicate these already challenging disorders. However, the mechanisms involved are complex and generally not well understood. Neurologic syndromes, including seizure disorders, can also be a feature of celiac disease (CD) or subclinical CD, sometimes associated with cerebral calcification. The association between type-1 diabetes mellitus (T1DM) and epilepsy is unclear and requires more definitive epidemiologic analysis, despite the fact that antibodies to glutamic acid decarboxylase may provide a link between the two conditions. The association between thyroid disorders and encephalopathies, often termed Hashimoto's encephalopathy, is well known but the pathogenic significance of antithyroid antibodies in this condition is still debated. In general, the relationships between autoimmune mechanisms and seizures in these conditions are unclear; the seizures are likely to be caused by a variety of mechanisms, including ischemia, neuronal damage, and specific and nonspecific immunity.


Prevalence of Hearing Impairment in Mahabubnagar District, Telangana State, India.
To estimate the prevalence of hearing impairment in Mahabubnagar district, Telangana state, India. A population-based prevalence survey of hearing impairment was undertaken in 2014. Fifty-one clusters of 80 people aged 6 months and older were selected using probability-proportionate-to-size sampling. A two-stage hearing screening was conducted using otoacoustic emissions on all participants followed by pure-tone audiometry on those aged 4 years and older who failed otoacoustic emissions. Cases of hearing impairment were defined using the World Health Organization definition of disabling hearing impairment: a pure-tone average of thresholds at 500, 1000, 2000, and 4000 Hz of ≥41 dB HL for adults and ≥31 dB HL for children based on the better ear. Possible causes of hearing impairment were ascertained by a certified audiologist. Reported hearing difficulties were also measured in this survey and compared with audiometry results. Three thousand five hundred seventy-three people were examined (response rate 87%), of whom 52% were female. The prevalence of disabling hearing impairment was 4.5% [95% confidence interval (CI) = 3.8 to 5.3). Disabling hearing impairment prevalence increased with age from 0.4% in those aged 4 to 17 years (95% CI = 0.2 to 1.1) to 34.7% (95% CI = 28.7 to 41.1) in those aged older than 65 years. No difference in prevalence was seen by sex. Ear examination suggested that the possible cause of disabling hearing impairment was chronic suppurative otitis media for 6.9% of cases and dry perforation for 5.6% cases. For the vast majority of people with disabling hearing impairment, a possible cause could not be established. The overall prevalence of reported or proxy reported hearing impairment was 2.6% (95% CI = 2.0 to 3.4), and this ranged from 0.6% (95% CI = 0.08 to 4.4) in those aged 0 to 3 years to 14.4% (95% CI = 9.8 to 20.7) in those aged older than 65 years. Disabling hearing impairment in Telangana State is common, affecting approximately 1 in 23 people overall and a third of people aged older than 65 years. These findings suggest that there are a substantial number of individuals with hearing impairment who could potentially benefit from improved access to low-cost interventions.


The G20 and development assistance for health: historical trends and crucial questions to inform a new era.
One of the most important gatherings of the world's economic leaders, the G20 Summit and ministerial meetings, takes place in June, 2019. The Summit presents a valuable opportunity to reflect on the provision and receipt of development assistance for health (DAH) and the role the G20 can have in shaping the future of health financing. The participants at the G20 Summit (ie, the world's largest providers of DAH, emerging donors, and DAH recipients) and this Summit's particular focus on global health and the Sustainable Development Goals offers a unique forum to consider the changing DAH context and its pressing questions. In this Health Policy perspective, we examined trends in DAH and its evolution over time, with a particular focus on G20 countries; pointed to persistent and emerging challenges for discussion at the G20 Summit; and highlighted key questions for G20 leaders to address to put the future of DAH on course to meet the expansive Sustainable Development Goals. Key questions include how to best focus DAH for equitable health gains, how to deliver DAH to strengthen health systems, and how to support domestic resource mobilisation and transformative partnerships for sustainable impact. These issues are discussed in the context of the growing effects of climate change, demographic and epidemiological transitions, and a global political shift towards increasing prioritisation of national interests. Although not all these questions are new, novel approaches to allocating DAH that prioritise equity, efficiency, and sustainability, particularly through domestic resource use and mobilisation are needed. Wrestling with difficult questions in a changing landscape is essential to develop a DAH financing system capable of supporting and sustaining crucial global health goals.


Necrotizing fasciitis.
The author's experience of fourteen patients with necrotizing fasciitis is reviewed. The pathognomonic feature of this condition is an extensive necrosis of subcutaneous tissue caused by a vicious cycle of infection, local ischaemia and reduced host defence mechanisms. The diagnosis can only be confirmed by immediate exploratory incision. The reported mortality of 30-40% reflects the inadaquacy of conservative surgery in the treatment of this serious condition. Mortality can be reduced by early recognition followed by radical excision of the necrotic fascia and overlying skin. The preservation and subsequent use of the excised skin has the advantage of economy in the use of donor areas and reduction in morbidity. Hyperbaric oxygen therapy does not halt the spread of the necrotizing process and is not a substitute for radical surgery.