Structure elucidation and quantification of the reduction products of anticancer Pt(iv) prodrugs by electrochemistry/mass spectrometry (EC-MS).
Pt(iv) prodrugs are a class of promising anticancer agents, which are activated by reduction to the active Pt(ii) species. Consequently, the reduction process is a crucial parameter. Herein, a new approach using electrochemistry (EC) coupled to liquid chromatography (LC) and electrospray ionization-mass spectrometry (ESI-MS) or inductively coupled plasma (ICP)-MS was applied. This enabled getting insights into the differences in the reduction and ligand release of platinum(iv) complexes with varying equatorial core structures.


Concomitant induction of microsomal epoxide hydrolase and UDP-glucuronosyltransferase activities by dipyridine compounds.
The coordinated response of the major rat hepatic phase II xenobiotic-metabolizing enzymes following 3-day exposure to diaryl compounds was investigated. Four diaryl compounds containing heterocyclic nitrogen atoms elevated microsomal epoxide hydrolase activity from 2- to 4-fold. Equivalent compounds lacking the heteroatom, when given in the same dosing regimen (75 mg/kg, ig, daily for 3 days), did not induce this or any other drug-metabolizing enzyme activity. Epoxide hydrolase activity closely paralleled UDP-glucuronosyltransferase activity toward three aglycones: 4-nitrophenol (r = 0.87), morphine (r = 0.84), and 1-naphthol (r = 0.78). There was less correlation (r = 0.60) between epoxide hydrolase activity and both UDP-glucuronosyltransferase activity toward testosterone and cytosolic glutathione S-transferase activity. There was no correlation between microsomal epoxide hydrolase activity and cytochrome P-450 or the monooxygenase reaction (4-nitrophenol hydroxylase) preferentially induced by pyridine-containing compounds. Induction of rat hepatic microsomal epoxide hydrolase activity by some pyridine-containing compounds appears coordinately regulated with glucuronidation rather than oxidation enzymes.


An adolescent with sickle cell anaemia experiencing disease-related complications: priapism and leg ulcer--a management challenge.
Sickle-cell anaemia (SCA) is a multi-system disease, associated with episodes of acute illness and progressive organ damage. Disease severity shows substantial variation and it is often a burden for adolescents. Complications such as leg ulcer and priapism have a significant impact on quality of life. There are still no definitive treatment guidelines available. Considering the embarrassing nature of priapism and the dire consequences for erectile dysfunction, it is important to inform patients, parents and providers about the relationship of SCA to prolonged painful erections. This article will review the pathophysiology and treatment options of SCA focusing the complications of leg ulcers, priapism, cholelithiasis and retinopathy. The case study of a 14-year-old boy is used to present a management challenge of multiple SCA-related complications.


Adult Mental Health Outpatients Who Have Minor Children: Prevalence of Parents, Referrals of Their Children, and Patient Characteristics.
Background: A strong connection exists between parental mental illness and lifetime mental health risk for their children. Thus, it is important to determine, when parents attend for treatment for their illness, the prevalence and characteristics of parents with a mental illness and identify referral actions for their children. Previous studies indicate that 12-45% of adult mental health service patients are parents with minor children. There is a need for studies with larger sample sizes that investigate the prevalence and characteristics of parents, and factors associated with referral actions for their children. Method: Data on 23,167 outpatients was drawn from a national census study across 107 Norwegian adult mental health outpatient clinics during 2 weeks in April 2013. Clinicians identified various socio-demographic characteristics of patients who were parents and referral actions for their children. Results: Eight thousand thirty-five (36%) of outpatients had children under 18 years. Thirty-one percent were provided with referrals for their children and 58% were reported to have children with no need for referral. Three percent were reported to have children with unmet needs who were not referred. There were missing data on children's needs and referral actions for 8% of parents. Patients who care for minor children were more likely to be refugees, and less likely to be single, male, not own a house/apartment, and have a schizophrenia spectrum illness or substance use disorder. Children were more likely to be referred when their parent was single, with no income from paid work, low education, not owning house/apartment, poor family network, long outpatient treatment, and an individual care plan; and less likely for men with a moderate or less severe mental illness. Children were referred to child protection agencies, child and adolescent mental health services and school psychological/pedagogic services. Discussion: The prevalence of outpatients with children is similar to other studies. Referrals were made for children of one third of outpatients with minor children. Needs and referrals of children was unknown for one in ten outpatients. Mental health outpatient clinics must improve procedures to identify parenting status and ascertain and act on children's needs.


Equine fetal adrenal, gonadal and placental steroidogenesis.
Equine fetuses have substantial circulating pregnenolone concentrations and thus have been postulated to provide significant substrate for placental 5α-reduced pregnane production, but the fetal site of pregnenolone synthesis remains unclear. The current studies investigated steroid concentrations in blood, adrenal glands, gonads and placenta from fetuses (4, 6, 9 and 10 months of gestational age (GA)), as well as tissue steroidogenic enzyme transcript levels. Pregnenolone and dehydroepiandrosterone (DHEA) were the most abundant steroids in fetal blood, pregnenolone was consistently higher but decreased progressively with GA. Tissue steroid concentrations generally paralleled those in serum with time. Adrenal and gonadal tissue pregnenolone concentrations were similar and 100-fold higher than those in allantochorion. DHEA was far higher in gonads than adrenals and progesterone was higher in adrenals than gonads. Androstenedione decreased with GA in adrenals but not in gonads. Transcript analysis generally supported these data. CYP17A1 was higher in fetal gonads than adrenals or allantochorion, and HSD3B1 was higher in fetal adrenals and allantochorion than gonads. CYP11A1 transcript was also significantly higher in adrenals and gonads than allantochorion and CYP19 and SRD5A1 transcripts were higher in allantochorion than either fetal adrenals or gonads. Given these data, and their much greater size, the fetal gonads are the source of DHEA and likely contribute more than fetal adrenal glands to circulating fetal pregnenolone concentrations. Low CYP11A1 but high HSD3B1 and SRD5A1 transcript abundance in allantochorion, and low tissue pregnenolone, suggests that endogenous placental pregnenolone synthesis is low and likely contributes little to equine placental 5α-reduced pregnane secretion.


Retrohoming of a Mobile Group II Intron in Human Cells Suggests How Eukaryotes Limit Group II Intron Proliferation.
Mobile bacterial group II introns are evolutionary ancestors of spliceosomal introns and retroelements in eukaryotes. They consist of an autocatalytic intron RNA (a "ribozyme") and an intron-encoded reverse transcriptase, which function together to promote intron integration into new DNA sites by a mechanism termed "retrohoming". Although mobile group II introns splice and retrohome efficiently in bacteria, all examined thus far function inefficiently in eukaryotes, where their ribozyme activity is limited by low Mg2+ concentrations, and intron-containing transcripts are subject to nonsense-mediated decay (NMD) and translational repression. Here, by using RNA polymerase II to express a humanized group II intron reverse transcriptase and T7 RNA polymerase to express intron transcripts resistant to NMD, we find that simply supplementing culture medium with Mg2+ induces the Lactococcus lactis Ll.LtrB intron to retrohome into plasmid and chromosomal sites, the latter at frequencies up to ~0.1%, in viable HEK-293 cells. Surprisingly, under these conditions, the Ll.LtrB intron reverse transcriptase is required for retrohoming but not for RNA splicing as in bacteria. By using a genetic assay for in vivo selections combined with deep sequencing, we identified intron RNA mutations that enhance retrohoming in human cells, but <4-fold and not without added Mg2+. Further, the selected mutations lie outside the ribozyme catalytic core, which appears not readily modified to function efficiently at low Mg2+ concentrations. Our results reveal differences between group II intron retrohoming in human cells and bacteria and suggest constraints on critical nucleotide residues of the ribozyme core that limit how much group II intron retrohoming in eukaryotes can be enhanced. These findings have implications for group II intron use for gene targeting in eukaryotes and suggest how differences in intracellular Mg2+ concentrations between bacteria and eukarya may have impacted the evolution of introns and gene expression mechanisms.


Diphoterine for alkali chemical splashes to the skin at alumina refineries.
Diphoterine is a commercially available amphoteric, hypertonic, chelating solution used to decontaminate and irrigate chemical splashes. The aim of this study was to evaluate the implementation of Diphoterine at three alumina refineries. This is the largest case series reported to date. One hundred eighty cases of alkali splashes to the skin were evaluated clinically. Two groups were compared; those who had applied Diphoterine first and those who had applied water first. There were no signs of chemical burn in 52.9% of the group who applied Diphoterine first compared with 21.4% of the group who applied water first. Only 7.9% of the group who applied Diphoterine first had blisters or more severe signs compared with 23.8% of the group who applied water first. The differences were statistically significant (P < 0.001). After implementation of Diphoterine the "first aid" injury rate for chemical burns fell 24.7% (95% CI 0.5–43.0%). Applying Diphoterine first was associated with significantly better outcomes following alkali skin splashes than applying water first.


Characterization of single-nucleotide variation in Indian-origin rhesus macaques (Macaca mulatta).
Rhesus macaques are the most widely utilized nonhuman primate model in biomedical research. Previous efforts have validated fewer than 900 single nucleotide polymorphisms (SNPs) in this species, which limits opportunities for genetic studies related to health and disease. Extensive information about SNPs and other genetic variation in rhesus macaques would facilitate valuable genetic analyses, as well as provide markers for genome-wide linkage analysis and the genetic management of captive breeding colonies. We used the available rhesus macaque draft genome sequence, new sequence data from unrelated individuals and existing published sequence data to create a genome-wide SNP resource for Indian-origin rhesus monkeys. The original reference animal and two additional Indian-origin individuals were resequenced to low coverage using SOLiD™ sequencing. We then used three strategies to validate SNPs: comparison of potential SNPs found in the same individual using two different sequencing chemistries, and comparison of potential SNPs in different individuals identified with either the same or different sequencing chemistries. Our approach validated approximately 3 million SNPs distributed across the genome. Preliminary analysis of SNP annotations suggests that a substantial number of these macaque SNPs may have functional effects. More than 700 non-synonymous SNPs were scored by Polyphen-2 as either possibly or probably damaging to protein function and these variants now constitute potential models for studying functional genetic variation relevant to human physiology and disease. Resequencing of a small number of animals identified greater than 3 million SNPs. This provides a significant new information resource for rhesus macaques, an important research animal. The data also suggests that overall genetic variation is high in this species. We identified many potentially damaging non-synonymous coding SNPs, providing new opportunities to identify rhesus models for human disease.


Candida glabrata persistence in mice does not depend on host immunosuppression and is unaffected by fungal amino acid auxotrophy.
Candida glabrata has emerged as an important fungal pathogen of humans, causing life-threatening infections in immunocompromised patients. In contrast, mice do not develop disease upon systemic challenge, even with high infection doses. In this study we show that leukopenia, but not treatment with corticosteroids, leads to fungal burdens that are transiently increased over those in immunocompetent mice. However, even immunocompetent mice were not capable of clearing infections within 4 weeks. Tissue damage and immune responses to microabscesses were mild as monitored by clinical parameters, including blood enzyme levels, histology, myeloperoxidase, and cytokine levels. Furthermore, we investigated the suitability of amino acid auxotrophic C. glabrata strains for in vitro and in vivo studies of fitness and/or virulence. Histidine, leucine, or tryptophan auxotrophy, as well as a combination of these auxotrophies, did not influence in vitro growth in rich medium. The survival of all auxotrophic strains in immunocompetent mice was similar to that of the parental wild-type strain during the first week of infection and was only mildly reduced 4 weeks after infection, suggesting that C. glabrata is capable of utilizing a broad range of host-derived nutrients during infection. These data suggest that C. glabrata histidine, leucine, or tryptophan auxotrophic strains are suitable for the generation of knockout mutants for in vivo studies. Notably, our work indicates that C. glabrata has successfully developed immune evasion strategies enabling it to survive, disseminate, and persist within mammalian hosts.


The future of model organisms in human disease research.
Model organisms have played a huge part in the history of studies of human genetic disease, both in identifying disease genes and characterizing their normal and abnormal functions. But is the importance of model organisms diminishing? The direct discovery of disease genes and variants in humans has been revolutionized, first by genome-wide association studies and now by whole-genome sequencing. Not only is it now much easier to directly identify potential disease genes in humans, but the genetic architecture that is being revealed in many cases is hard to replicate in model organisms. Furthermore, disease modelling can be done with increasing effectiveness using human cells. Where does this leave non-human models of disease?


Expressing freedom and taking liberties: the paradoxes of aberrant science.
Complete freedom does not exist, despite people's preparedness to die for it. Scientific freedom is much defended and yet much misunderstood. Scientists have limits imposed on their freedom by the disciplines and discourse communities in which they place themselves. Freedom within these socially constructed constraints needs to be distinguished from taking liberties with the rules and practices that make up these constraints, and validate the activities of special groups within society. Scientists (and the public) perceive taking liberties with science's rules and practices as aberrant science, and they often react punitively. Aberrant science can be broadly examined under four headings: wicked science, naughty science, dysfunctional science, and ideologically unacceptable science. When we examine examples of perceived aberrant science, we find that these categories of "misconduct" are connected and often confused. Scientific freedom needs to be redefined with due regard to current understandings of scientists as human beings facing powerful social pressures to deliver results of a particular kind.


Direct demonstration of insulin receptor internalization. A quantitative electron microscopic study of covalently bound 125I-photoreactive insulin incubated with isolated hepatocytes.
When 125I-insulin is incubated with isolated rodent hepatocytes at 37 degrees C, the ligand initially binds to the plasma membrane of the cell and is subsequently internalized by adsorptive endocytosis. To confirm directly that the insulin receptor is internalized with the ligand, we covalently linked photoreactive 125I-N sigma B29 (azidobenzoyl) insulin to its specific hepatocyte receptor and followed its fate by quantitative electron microscopic autoradiography. We found that the covalently linked photoreactive insulin is internalized by the cell in fashion analogous to the internalization of ordinary 125I-insulin, indicating that, at least under these conditions, the insulin receptor is internalized with the ligand.


Physiological and pathological impact of blood sampling by retro-bulbar sinus puncture and facial vein phlebotomy in laboratory mice.
Retro-bulbar sinus puncture and facial vein phlebotomy are two widely used methods for blood sampling in laboratory mice. However, the animal welfare implications associated with these techniques are currently debated, and the possible physiological and pathological implications of blood sampling using these methods have been sparsely investigated. Therefore, this study was conducted to assess and compare the impacts of blood sampling by retro-bulbar sinus puncture and facial vein phlebotomy. Blood was obtained from either the retro-bulbar sinus or the facial vein from male C57BL/6J mice at two time points, and the samples were analyzed for plasma corticosterone. Body weights were measured at the day of blood sampling and the day after blood sampling, and the food consumption was recorded automatically during the 24 hours post-procedure. At the end of study, cheeks and orbital regions were collected for histopathological analysis to assess the degree of tissue trauma. Mice subjected to facial vein phlebotomy had significantly elevated plasma corticosterone levels at both time points in contrast to mice subjected to retro-bulbar sinus puncture, which did not. Both groups of sampled mice lost weight following blood sampling, but the body weight loss was higher in mice subjected to facial vein phlebotomy. The food consumption was not significantly different between the two groups. At gross necropsy, subcutaneous hematomas were found in both groups and the histopathological analyses revealed extensive tissue trauma after both facial vein phlebotomy and retro-bulbar sinus puncture. This study demonstrates that both blood sampling methods have a considerable impact on the animals' physiological condition, which should be considered whenever blood samples are obtained.


Season- and age-related reproductive changes based on fecal androgen concentrations in male koalas, Phascolarctos cinereus.
The purposes of the present study were to clarify age- and season- related androgen patterns, and to compare the reproductive physiology between Japanese captive koala populations and Australian populations. To measure fecal androgens, feces were collected from male koalas (4.2 to 13.8 years of age) kept in Japanese zoos. Fecal androgens were extracted with methanol from the lyophilized samples and determined by enzyme immunoassay using 4-androstene-3,17-dione antibody. Fecal androgen concentration in male koalas increased after sexual maturation and remained relatively high until old age. In the survey with the Japanese zoo studbook of koalas, copulation (conception) month showed a pyramid shape with a peak in March to June (60.7%) in koalas born and reared in Japanese zoos and from July to April with the highest concentration in September to January (69.7%) in Australian institutes. Japanese zoo koala populations have a characteristic physiological cycle adapted to Japan's seasonal changes. The suitable month of year for copulation or conception in Japan is diametrically opposed to that in Australia. Mean fecal androgen concentrations by month in the males born and reared in Japan indicated annual changes with the highest concentration in May and the lowest value in November. Fecal androgen analysis may be a noninvasive alternative tool to monitor circulating testosterone and may be helpful in understanding reproductive activity and physiology in male koalas.


Estimation of self-absorption effect on aluminum emission in the presence of different noble gases: comparison between thin and thick plasma emission.
Aluminum spectra in the noble gases of helium and argon at initial delay times after plasma formation are numerically calculated. Temporal behavior of plasma emissions up to 200 ns after laser irradiation is investigated. Plasma parameters are computed by coupling the thermal model of laser ablation, hydrodynamic of plasma expansion, and Saha-Eggert equations. A spectrum is constructed from the superposition of 13 strong lines of aluminum and several strong lines of ambient gases. Spectral radiations are superimposed on a continuous emission composed of bremsstrahlung and recombination radiation. The self-absorption effect on plasma radiation at 1 atm gas pressure is studied. In this paper, a comparison between thin and thick aluminum radiation is done. Furthermore, the self-absorption coefficient of each strong line at laser energies of 0.5, 0.7, 0.9, and 1.1 GW/cm(2) is estimated. Results show that at specific laser energy, the self-absorption effect in argon is more significant than in helium. For most of the spectral lines in both noble gases, the self-absorption coefficient will diminish with the delay time. As indicated with passing time, the line widths of the self-absorbed lines will rise. More intense continuous emissions are observed at higher wavelengths, and these radiations will be increased with laser energy.


Enhanced cardiogenesis in embryonic stem cells overexpressing the GATA-4 transcription factor.
GATA-4 is a cardiac-specific member of the GATA family of zinc finger transcription factors. During embryogenesis, GATA-4 expression is detected very early in the cardiogenic area and persists later in the developing heart. Studies have shown that GATA-4 is a potent transcriptional activator of several cardiac muscle-specific genes and a key regulator of the cardiomyocyte gene program. Consistent with a role for GATA-4 in cardiomyocyte formation, inhibition of GATA-4 expression by antisense transcripts interferes with expression of cardiac muscle genes and blocks development of beating cardiomyocytes in P19 embryonic stem cells. In order to better define the function of GATA-4 in cardiogenesis, we have carried out molecular analysis of early stages of cardiomyocyte differentiation in GATA-4-deficient P19 cell lines and in P19 cells stably overexpressing GATA-4. The results indicate that GATA-4 is not required for either endodermal or mesodermal commitment or for initiation of the cardiac pathway. However, in the absence of GATA-4, differentiation is blocked at the precardiac (cardioblasts) stage and cells are lost through extensive apoptosis. In contrast, ectopic expression of GATA-4 in P19 cells accelerates cardiogenesis and markedly increases (over 10-fold) the number of terminally differentiated beating cardiomyocytes following cell aggregation. Together, these findings suggest that, in addition to its role in activation of the cardiac genetic program, GATA-4 may be the nuclear target of inductive and/or survival factors for precardiac cells.


The influence of static magnetic fields on mechanosensitive ion channel activity in artificial liposomes.
The influence of static magnetic fields (SMFs) on the activity of recombinant mechanosensitive ion channels (the bacterial mechanosensitive ion channel of large conductance-MscL) following reconstitution into artificial liposomes has been investigated. Preliminary findings suggest that exposure to 80-mT SMFs does not induce spontaneous MscL activation in the absence of mechanical stimulation. However, SMFs do appear to influence the open probability and single channel kinetics of MscL exposed to negative pipette pressure. Typical responses include an overall reduction in channel activity or an increased likelihood of channels becoming "trapped open" in sub-conducting states following exposure to SMFs. There is a delay in the onset of this effect and it is maintained throughout exposure. Generally, channel activity showed slow or limited recovery following removal of the magnetic field and responses to the magnetic were often reduced or abolished upon subsequent exposures. Pre-exposure of the liposomes to SMFs resulted in reduced sensitivity of MscL to negative pipette pressure, with higher pressures required to activate the channels. Although the mechanisms of this effect are not clear, our initial observations appear to support previous work showing that the effects of SMFs on ion channels may be mediated by changes in membrane properties due to anisotropic diamagnetism of lipid molecules.


Feasibility of a home-based interdisciplinary rehabilitation program for patients with Post-Intensive Care Syndrome: the REACH study.
Survivors of critical illness experience long-term functional challenges, which are complex, heterogeneous, and multifactorial in nature. Although the importance of rehabilitation interventions after intensive care unit (ICU) discharge is universally recognized, evidence on feasibility and effectiveness of home-based rehabilitation programs is scarce and ambiguous. This study investigates the feasibility of an interdisciplinary rehabilitation program designed for patients with Post-Intensive Care Syndrome (PICS) who are discharged home. A mixed method, non-randomized, prospective pilot feasibility study was performed with a 6-month follow-up, comparing the intervention (REACH) with usual care. REACH was provided by trained professionals and included a patient-centered, interdisciplinary approach starting directly after hospital discharge. Primary outcomes were patient safety, satisfaction, adherence, referral need and health care usage. Secondary outcomes, measured at 3 timepoints, were functional exercise capacity, self-perceived health status, health-related quality of life (HRQoL), return to work and psychotrauma. Risk of undernutrition was assessed at baseline. 43 patients with a median mechanical ventilation duration of 8 (IQR:10) days, were included in the study and 79.1% completed 6-month follow-up. 19 patients received the intervention, 23 received usual care. Groups were similar for gender distribution and ICU length of stay. No adverse events occurred. REACH participants showed higher satisfaction with treatment and reported more allied health professional visits, while the usual care group reported more visits to medical specialists. Qualitative analysis identified positive experiences among REACH-professionals related to providing state-of-the-art interventions and sharing knowledge and expertise within an interprofessional network. Similar recovery was seen between groups on all secondary outcomes, but neither group reached reference values for HRQoL at 6 months. Larger return to work rates were seen in the REACH group. Prevalence of undernutrition at hospital discharge was high in both groups (> 80%), warranting the need for careful tuning of physical therapy and nutritional interventions. This study shows that providing early, home-based rehabilitation interventions for patients with PICS-related symptoms is feasible and perceived positively by patients and professionals. When provided in an interdisciplinary collaborative network state of the art, person-centered interventions can be tailored to individual needs potentially increasing patient satisfaction, adherence, and efficacy. Registered in the Dutch Trial register: NL7792: https://www.trialregister.nl/trial/7792 , registered 7-06-2019.


Restricted T cell receptor expression by human T cell clones specific for mycobacterial 65-kDa heat-shock protein: selective in vivo expansion of T cells bearing defined receptors.
We have examined the T cell receptor (TcR) expression of clones specific for epitopes of mycobacterial 65-kDa heat-shock protein (hsp65) in the context of two different HLA molecules, and used this system as a model to assess the selection of T cells responsive to this antigen in vivo. DR3-restricted clones were raised from both the synovial fluid (SF) and peripheral blood (PB) of a patient with reactive arthritis in three separate cloning events. Five of five SF-derived clones tested expressed either V beta 5.2 or a closely related beta chain, V beta 5.6. The alpha chains expressed by V beta 5.2+ and V beta 5.6+ clones were from different families, V alpha 2.4 and V alpha 23.2, respectively. Nine of ten clones derived from two cloning procedures on PB taken 3 years later also expressed either V beta 5.2 or V beta 5.6. This suggests that the TcR repertoire for recognizing this major histocompatibility complex/peptide complex is relatively restricted and favors the use of V beta 5. Conservation of the beta chain third complementarity-determining region (CDR3) sequence was not evident, however. Sequencing alpha and beta chains of representative V beta 5.2+ and V beta 5.6+ PB-derived clones revealed TcR which were identical to those utilized by the SF-derived clones, showing that the repertoire for recognition of this antigen is stable over time. Similar studies of TcR expression were carried out on hsp65-specific, DP4-restricted clones derived from the SF of a patient with rheumatoid arthritis by two independent cloning procedures. There was conservation of alpha chain usage, since all clones expressed a member of the V alpha 1 family, but again CDR3 sequence conservation was not apparent. beta chain usage was not restricted since different clones expressed V beta 6.7, V beta 22.3 and V beta 12. Subtle differences in epitope specificity were detected for two clones with differing TcR. Once more, T cell clones with identical alpha and beta TcR chains were obtained from the separate cloning procedures, suggesting oligoclonalty of T cells with this defined specificity in the patient's SF.


Protocol for the systematic review of the prevention, treatment and public health management of impetigo, scabies and fungal skin infections in resource-limited settings.
Impetigo, scabies, and fungal skin infections disproportionately affect populations in resource-limited settings. Evidence for standard treatment of skin infections predominantly stem from hospital-based studies in high-income countries. The evidence for treatment in resource-limited settings is less clear, as studies in these populations may lack randomisation and control groups for cultural, ethical or economic reasons. Likewise, a synthesis of the evidence for public health control within endemic populations is also lacking. We propose a systematic review of the evidence for the prevention, treatment and public health management of skin infections in resource-limited settings, to inform the development of guidelines for the standardised and streamlined clinical and public health management of skin infections in endemic populations. The protocol has been designed in line with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols statement. All trial designs and analytical observational study designs will be eligible for inclusion. A systematic search of the peer-reviewed literature will include PubMed, Excertpa Medica and Global Health. Grey literature databases will also be systematically searched, and clinical trials registries scanned for future relevant studies. The primary outcome of interest will be the clinical cure or decrease in prevalence of impetigo, scabies, crusted scabies, tinea capitis, tinea corporis or tinea unguium. Two independent reviewers will perform eligibility assessment and data extraction using standardised electronic forms. Risk of bias assessment will be undertaken by two independent reviewers according to the Cochrane Risk of Bias tool. Data will be tabulated and narratively synthesised. We expect there will be insufficient data to conduct meta-analysis. The final body of evidence will be reported against the Grades of Recommendation, Assessment, Development and Evaluation grading system. The evidence derived from the systematic review will be used to inform the development of guidelines for the management of skin infections in resource-limited settings. The evidence derived will be intended for use by clinicians, public health practitioners and policy makers in the treatment of skin infections and the development of skin infection control programmes. The review will identify any gaps in the current evidence to provide direction for future research. PROSPERO CRD42015029453.


Candidacidal effect of fluconazole and chlorhexidine released from acrylic polymer.
To investigate the efficacy and rate of killing of a fluconazole- or chlorhexidine-impregnated polymeric delivery system against fluconazole-susceptible and -resistant Candida albicans and fluconazole-resistant Candida glabrata. Poly(ethyl methacrylate)/tetrahydrofurfuryl methacrylate (PEM/THFM) discs impregnated with chlorhexidine, pure fluconazole (FLCp) or fluconazole from capsules (FLCc) were prepared by substituting a portion of PEM powder with an equivalent amount of each drug. Discs were incubated in sterile water for 1, 3, 7, 14, 21 and 28 days. The amounts of drugs in the leachates were measured spectrophotometrically and their antifungal activity against fluconazole-susceptible (n=1) and fluconazole-resistant (n=2) candidal isolates was determined using a time-kill method and by comparing the released concentrations with the corresponding MICs. Fluconazole and chlorhexidine leached from PEM/THFM polymer for up to 28 days and the released concentrations were fungicidal against all three Candida isolates for at least the first 7 days. Chlorhexidine leachates killed all Candida isolates more rapidly than the two fluconazole formulation leachates throughout the study period. FLCc leachates required longer incubation for 100% killing than FLCp leachates. The proportion of viable C. glabrata dropped more slowly than that of C. albicans with the same MIC. The concentrations of chlorhexidine and fluconazole leached from the PEM/THFM polymer were fungicidal against all Candida isolates, including those resistant to fluconazole, for the first 7 days. Chlorhexidine leachates showed a rapid fungicidal activity for up to 4 weeks, which can be of use in cases with poor response to conventional antifungals.


Diurnal Timing Dependent Alterations in Gut Microbial Composition Are Synchronously Linked to Salt-Sensitive Hypertension and Renal Damage.
Alterations of diurnal rhythms of blood pressure (BP) and reshaping of gut microbiota are both independently associated with hypertension. However, the relationships between biorhythms of BP and gut microbial composition are unknown. We hypothesized that diurnal timing-associated alterations of microbial compositions are synchronous with diurnal rhythmicity, dip in BP, and renal function. To test this hypothesis, Dahl salt-sensitive (S) rats on low- and high-salt diets were examined for time of day effects on gut microbiota, BP, and indicators of renal damage. Major shifts in night and day patterns of specific groups of microbiota were observed between the dark (active) and light (rest) phases, which correlated with diurnal rhythmicity of BP. The diurnal abundance of Firmicutes, Bacteroidetes, and Actinobacteria were independently associated with BP. Discrete bacterial taxa were observed to correlate independently or interactively with one or more of the following 3 factors: (1) BP rhythm, (2) dietary salt, and (3) dip in BP. Phylogenetic Investigation of Communities revealed diurnal timing effects on microbial pathways, characterized by upregulated biosynthetic processes during the active phase of host, and upregulated degradation pathways of metabolites in the resting phase. Additional metagenomics functional pathways with rhythm variations were noted for aromatic amino acid metabolism and taurine metabolism. These diurnal timing dependent changes in microbiota, their functional pathways, and BP dip were associated with concerted effects of the levels of renal lipocalin 2 and kidney injury molecule-1 expression. These data provide evidence for a firm and concerted diurnal timing effects of BP, renal damage, and select microbial communities.


Retinocortical time exhibits spatial selectivity.
We recorded simultaneous electroretinograms and visual evoked potentials to pattern-reversal stimulus in ten normal subjects employing check sizes subtending 7.5, 15, 30, 60, 120, 240 and 480 min of visual arc. The mean peak latency of the pattern electroretinogram (PERG) b wave decreased logarithmically with increasing check size. The mean peak latency of the pattern visual evoked potential (PVEP) P100 component was shortest for 30 min check size and was longer at both smaller and larger check sizes, a phenomenon termed spatial selectivity. Retinocortical time (RCT) was calculated as the latency difference between the PVEP P100 peak and the PERG b wave. The RCT also exhibited spatial selectivity similar to that of the PVEP P100 component; mean RCT was shortest for 30 min check size and was longer at both smaller and larger check sizes. Implications are discussed in light of our current knowledge of the origins of the PERG and PVEP and visual processing of pattern-reversal stimuli. Simultaneous recording of PERGs and PVEPs in patients with disease of the afferent visual pathways employing a large range of check sizes may increase the diagnostic sensitivity of visual electrophysiologic testing.


Biological approaches to global environment change mitigation and remediation.
One of the most pressing and globally recognized challenges is how to mitigate the effects of global environment change brought about by increasing emissions of greenhouse gases, especially CO(2). In this review we evaluate the potential contribution of four biological approaches to mitigating global environment change: reducing atmospheric CO(2) concentrations through soil carbon sequestration and afforestation; reducing predicted increases in global surface temperatures through increasing the albedo of crop plants; and fertilizing the oceans to increase primary productivity and CO(2) drawdown. We conclude that none of these biological approaches are 'magic bullets' capable of reversing environmental changes brought about by increasing emissions of greenhouse gases. However, it is possible that increasing crop albedo and soil carbon sequestration might contribute towards mitigation on a regional scale. In the absence of legally binding international agreements to reduce CO(2) emissions, we propose that: increased efforts are made to identify novel biological mitigatory strategies; further research is conducted to minimise the uncertainties present in all four of the biological approaches described; and pilot-level field work is conducted to examine the feasibility of the most promising strategies. Finally, it is essential to engage with the public concerning strategies for mitigating the effects of climate change because the majority of the biological approaches have effects, quite possibly of a negative nature, on ecosystem services and land usage.


Two-amino acid molecular switch in an epithelial morphogen that regulates binding to two distinct receptors.
Ectodysplasin, a member of the tumor necrosis factor family, is encoded by the anhidrotic ectodermal dysplasia (EDA) gene. Mutations in EDA give rise to a clinical syndrome characterized by loss of hair, sweat glands, and teeth. EDA-A1 and EDA-A2 are two isoforms of ectodysplasin that differ only by an insertion of two amino acids. This insertion functions to determine receptor binding specificity, such that EDA-A1 binds only the receptor EDAR, whereas EDA-A2 binds only the related, but distinct, X-linked ectodysplasin-A2 receptor (XEDAR). In situ binding and organ culture studies indicate that EDA-A1 and EDA-A2 are differentially expressed and play a role in epidermal morphogenesis.


Are commercially-available precontoured anatomical clavicle plating systems offering the purported superior optimum fitting to the clavicle? A cadaveric analysis and review of literature.
The indication for operative treatment of displaced midshaft clavicle fractures remains controversial. However, if plate fixation is considered, implant prominence and skin irritation are the most common causes for re-operation. Low profile implants as well as closely contouring plates to the individual anatomy may reduce these complications. The aim of this study was to compare the fitting accuracy and implant prominence of 3.5mm pelvic reconstruction plates (PRP) with pre-contoured anatomical clavicle plates (PACP) for midshaft clavicle fractures. Three-dimensional data of the largest, median and smallest male and female clavicle of an existing database of 89 cadaveric clavicles were included for analysis. A three-dimensional model of a commercially available PACP was used for digitally positioning of the plate on the segmented clavicles. Three-dimensional printouts of each clavicle were produced and the 3.5mm reconstruction plates were manually bent and positioned by the senior author. Computed tomography scans and three-dimensional reconstructions were then obtained to digitally compare the fitting accuracy and implant prominence. Pelvic reconstruction plates offered superior fitting accuracy and lower implant prominence compared to PACP. The largest difference in implant prominence was observed in large sized female clavicles and measured 3.6mm. Both, the less costly PRP plates and commercially available PACP for midshaft fractures of the clavicle demonstrated a clinically acceptable fitting accuracy. The manually bent pelvic-reconstruction plates demonstrated reduced implant prominence with superior fitting. Hypothetically this might contribute to a reduced rate of reoperation. Level IV cadaveric study.


Meta-analysis shows clinically relevant and long-lasting deterioration in health-related quality of life after esophageal cancer surgery.
The purpose of the study is to (1) estimate the direction, clinical relevance, and duration of health-related quality-of-life (HRQL) change in the first year following esophageal cancer surgery and (2) to assess the robustness of the estimates by subgroup and sensitivity analyses, and an exploration of publication bias. A systematic literature search in MEDLINE, EMBASE, CINAHL, PsychINFO, and CENTRAL to identify randomized and non-randomized studies was performed. We compared the baseline HRQL data with 3-, 6-, 9-, or 12-month follow-ups to estimate the magnitude and duration of HRQL change. These estimates were then classified as trivial, small, medium, or large. Primary outcomes were role functioning, eating, and fatigue. Secondary outcomes were physical and social functioning, dysphagia, pain, and coughing problems. We conducted subgroup analysis for open surgery, open surgery preceded by neoadjuvant therapy, and minimally invasive surgery. Sensitivity analyses assessed the influence of study design, transformation/imputation of the data, and HRQL questionnaire used. We included the data from 15 studies to estimate the change in 28 HRQL outcomes after esophageal cancer surgery. The main analysis showed that patients' social functioning deteriorated. Symptoms of fatigue, pain, and coughing problems increased. These changes lasted for 9-12 months, although some symptoms persisted beyond the first year after surgery. For many other HRQL outcomes, estimates were only robust after subgroup or sensitivity analyses (e.g., role and physical functioning), or remained too heterogeneous to interpret (e.g., eating and dysphagia). Patients will experience a clinically relevant and long-lasting deterioration in HRQL after esophageal cancer surgery. However, for many HRQL outcomes, more and better quality evidence is needed.


CRB1 heterozygotes with regional retinal dysfunction: implications for genetic testing of leber congenital amaurosis.
To test human CRB1 heterozygotes for possible clinical or functional retinal changes and to evaluate whether a patient with Leber congenital amaurosis (LCA) with CRB1 mutations not consistent with previously described CRB1 phenotypes carried a modifier allele in another LCA gene. Seven unrelated heterozygous carriers of CRB1 mutations underwent phenotyping by full eye examinations (indirect ophthalmoscopy and slit lamp biomicroscopy) and functional testing (standard full-field electroretinography [ERG] and multifocal ERG). For genotyping of the LCA patients and their parents, denaturing high-performance liquid chromatography (dHPLC) analyses were performed, followed by sequence analysis of CRB1, followed by sequence analysis of the AIPL1 and CRX genes to identify a putative modifier effect in a patient with an atypical CRB1 phenotype. Reduced full-field ERG b-wave amplitudes were observed with scotopic -2 dB flash (140 microV; P < 0.05), normal full-field cone ERGs, and significant regional retinal dysfunction on mfERG in five of seven carriers of CRB1 mutations. A known AIPL1 mutation (p. R302L) was identified as a potential modifier allele in a patient with LCA carrying two CRB1 mutations and with a prominent maculopathy. In human heterozygotes of CRB1 mutations (parents of offspring with LCA), distinctive regional retinal dysfunctions were found by multifocal ERG measurements that were consistent with the focal histologic abnormalities reported for the two CRB1 knockout mice models. This phenotypic finding may identify CRB1 carriers and point to the causal gene defect in affected LCA offspring, significantly facilitating the molecular diagnostic process. Evidence suggests a modifier allele in AIPL1 in a patient with LCA with prominent atrophic macular lesions and homozygous defects in CRB1.


Fungal-Bacterial Interactions in Health and Disease.
Fungi and bacteria encounter each other in various niches of the human body. There, they interact directly with one another or indirectly via the host response. In both cases, interactions can affect host health and disease. In the present review, we summarized current knowledge on fungal-bacterial interactions during their commensal and pathogenic lifestyle. We focus on distinct mucosal niches: the oral cavity, lung, gut, and vagina. In addition, we describe interactions during bloodstream and wound infections and the possible consequences for the human host.


Global organization of dynamics in oscillatory heterogeneous excitable media.
An oscillatory heterogeneous excitable medium undergoes a transition from periodic target patterns to a bursting rhythm driven by the spontaneous initiation and termination of spiral waves as coupling or density is reduced. We illustrate these phenomena in monolayers of chick embryonic heart cells using calcium-sensitive fluorescent dyes. These results are modeled in a heterogeneous cellular automaton in which the neighborhood of interaction and cell density is modified. Parameters that give rise to bursting rhythms are organized in distinct zones in parameter space, leading to a global organization that should be applicable to the dynamics in a large class of excitable media.


Factors affecting the implementation of evidence-based Progressive Tinnitus Management in Department of Veterans Affairs Medical Centers.
Progressive Tinnitus Management (PTM) is an evidence-based interdisciplinary stepped-care approach to improving quality of life for patients with tinnitus. PTM was endorsed by Department of Veterans Affairs (VA) Audiology leadership in 2009. Factors affecting implementation of PTM are unknown. We conducted a study to: 1) estimate levels of PTM program implementation in VA Audiology and Mental Health clinics across the country; and 2) identify barriers and facilitators to PTM implementation based on the experiences of VA audiologists and mental health providers. We conducted an anonymous, web-based survey targeting Audiology and Mental Health leaders at 144 major VA facilities. Quantitative analyses summarized respondents' facility characteristics and levels of program implementation (full PTM, partial PTM, or no PTM). Qualitative analyses identified themes in factors influencing the implementation of PTM across VA sites. Surveys from 87 audiologists and 66 mental health clinicians revealed that few facilities offered full PTM; the majority offered partial or no PTM. Inductive analysis of the open-ended survey responses identified seven factors influencing implementation of PTM: 1) available resources, 2) service collaboration, 3) prioritization, 4) Veterans' preferences and needs, 5) clinician training, 6) awareness of (evidence-based) options, and 7) perceptions of scope of practice. Results suggest wide variation in services provided, a need for greater engagement of mental health providers in tinnitus care, and an interest among both audiologists and mental health providers in receiving tinnitus-related training. Future research should address barriers to PTM implementation, including methods to: 1) improve understanding among mental health providers of their potential role in tinnitus management; 2) enhance coordination of tinnitus-related care between health care disciplines; and 3) collect empirical data on Veterans' need for and interest in PTM, including delivery by telehealth modalities.


Impact of PDGF-BB on cellular distribution and extracellular matrix in the healing rabbit Achilles tendon three weeks post-operation.
Current methods for tendon rupture repair suffer from two main drawbacks: insufficient strength and adhesion formation, which lead to rerupture and impaired gliding. A novel polymer tube may help to overcome these problems by allowing growth factor delivery to the wound site and adhesion reduction, and by acting as a physical barrier to the surrounding tissue. In this study, we used a bilayered DegraPol® tube to deliver PDGF-BB to the wound site in a full-transection rabbit Achilles tendon model. We then performed histological and immunohistochemical analysis at 3 weeks postoperation. Sustained delivery of PDGF-BB to the healing Achilles tendon led to a significantly more homogenous cell distribution within the healing tissue. Lower cell densities next to the implant material were determined for +PDGF-BB samples compared to -PDGF-BB. PDGF-BB application increased proteoglycan content and reduced alpha-SMA+ areas, clusters of different sizes, mainly vessels. Finally, PDGF-BB reduced collagens I and III in the extracellular matrix. The sustained delivery of PDGF-BB via an electrospun DegraPol® tube accelerated tendon wound healing by causing a more uniform cell distribution with higher proteoglycan content and less fibrotic tissue. Moreover, the application of this growth factor reduced collagen III and alpha-SMA, indicating a faster and less fibrotic tendon healing.


Prices don't drive regional Medicare spending variations.
Per capita Medicare spending is more than twice as high in New York City and Miami than in places like Salem, Oregon. How much of these differences can be explained by Medicare's paying more to compensate for the higher cost of goods and services in such areas? To answer this question, we analyzed Medicare spending after adjusting for local price differences in 306 Hospital Referral Regions. The price-adjustment analysis resulted in less variation in what Medicare pays regionally, but not much. The findings suggest that utilization-not local price differences-drives Medicare regional payment variations, along with special payments for medical education and care for the poor.


In vitro comparison of transfixation and standard full-limb casts for prevention of displacement of a mid-diaphyseal third metacarpal osteotomy site in horses.
To compare transfixation and standard full-limb casts for prevention of in vitro displacement of a mid-diaphyseal third metacarpal osteotomy site in horses. 6 forelimbs from 6 horses euthanatized for reasons not related to the musculoskeletal system. A 30 degrees osteotomy was performed in the mid-diaphysis of the third metacarpal bone. Two 4.5-mm cortical bone screws were placed across the osteotomy site to maintain alignment during casting. Two 6.35-mm Steinmann pins were placed from a lateral-to-medial direction in the distal aspect of the radius. A full-limb cast that incorporated the pins was applied. An extensometer was positioned in the osteotomy site through a window placed in the dorsal aspect of the cast, and after removal of the screws, displacement was recorded while the limb was axially loaded to 5,340 N (1,200 lb). Pins were removed, and the standard full-limb cast was tested in a similar fashion. The transfixation cast significantly reduced displacement across the osteotomy site at 445 N (100 lb), 1,112 N (250 lb), 2,224 N (500 lb), and 4,448 N (1,000 lb), compared with the standard cast. A full-limb transfixation cast provides significantly greater resistance than a standard full-limb cast against axial collapse of a mid-diaphyseal third metacarpal osteotomy site when the bone is placed under axial compression. Placement of full-limb transfixation casts should be considered for the management of unstable fractures of the third metacarpal bone in horses.


First Report of Cherry green ring mottle virus and Cherry necrotic rusty mottle virus in Sweet Cherry (Prunus avium) in Chile and South America.
Cherry green ring mottle virus (CGRMV) infects several Prunus species, while Cherry necrotic rusty mottle virus (CNRMV) has been detected mainly in sweet cherry. In Chile, sweet cherry represents one of the most valuable fruit crops, and the country is the main producer of cherries in the southern hemisphere. In October 2011, leaf samples were collected from 21 trees of cv. Bing in Libertador General Bernardo O'Higgins and Maule regions. Leaves of symptomatic plants showed brown angular necrotic spots, the center of which can drop out giving a shot-hole appearance. Total RNA was extracted by the silica capture method (1). Reverse transcription (RT)-PCR was carried out to test the presence of CGRMV and CNRMV using primer pairs GRM7950/GRM8316 (1), and DetCNR-F (TCCCACCTCAAGTCCTAGCAGAGA) / DetCNR-R (TCATTGCTAATTGCAAAATCCCA). Ten and six samples were tested positive for CGRMV and CNRMV, obtaining 366- and 333-bp fragments, respectively. Mixed infections occurred in five samples. Two sets of primer pairs were designed to amplify a region of the genome which includes the entire coat protein (CP) gene: CGRM-CPF (GGCTGATGAAGAATTTGA-GAAG) and CGRM-CPR (GAGTGGAATTGCAGGGGTTT), and CNRM-CPF (GAGTGTGTGTGAGCTTTCAAGTT) and CNRM-CPR (TTCGCCCCGTGTTGTAAAAC). Amplicons of the expected size of 828 bp (CGRMV) and 892 bp (CNRMV) were obtained from infected samples. Three amplicons for each virus were cloned into pGEM-T and three colonies per cloned fragment were sequenced in both directions. For CNRMV, Chilean isolates CP9754 (GenBank Accession No. KC432619) and CP9956N (KC432621) had 98% for nucleotide identity with isolate JK10 from India (FN546178), while isolate CP9879 (KC432620) had 97% of nucleotide identity. For CGRMV, Chilean isolate CP3359 (KC432616) had 98% identity with isolate HI17 from Poland (JX468873), while isolates CP9731 (KC432617) and CP9956G (KC432618) had 98% and 99% nucleotide identity with isolate ita7 (AF533161) from Italy, respectively. Nucleotide and amino acid sequence identity between Chilean isolates of CGRMV ranged between 94.5% and 99.3%, and from 97.8% to 99.2%, respectively. For Chilean isolates of CNRMV, sequence identity ranged between 98.0% and 98.5% (nucleotide), and from 98.6% to 98.9% (amino acid). Sequence analysis indicated that CGRMV isolates found in Chile belong to group II (3). Detection was confirmed by non-isotopic molecular hybridization. Riboprobes were designed on the basis of a consensus sequence of CP gene and labeled with digoxigenin (2); are complementary to the fragments located from the nucleotide 7415 to 7576 for CGRMV (reference sequence NC 001946.1), and 7475 to 7638 for CNRMV (reference sequence NC 002468.1). The cultivar Bing manifested symptoms only when infected by CNRMV. Results suggest that CNRMV is the cause of symptoms and yield loss observed in Bing, the most important cherry variety cultivated in Chile. To our knowledge, this is the first report of CGRMV and CNRMV infecting sweet cherry in South America. References: (1) M. E. Rott and W. Jelkmann. Eur. J. Plant Pathol. 107:411, 2001. (2) J. A Sánchez-Navarro et al. Plant Pathol. 47:780, 1998. (3) Y. P. Zhang et al. J. Plant. Pathol. 82:49, 2000.


Update on guidelines for management of hypercholesterolemia.
Coronary heart disease (CHD) is a leading cause of morbidity and mortality globally. Hypercholesterolemia is one of the major risk factors for CHD. With the increase in aging populations and progressively sedentary lifestyles, the global burden of CHD is likely to increase in the future despite better preventive strategies. It is vitally important to manage hypercholesterolemia effectively because it is a modifiable risk factor. At present, there are several guidelines with differences in recommendations. In this paper the authors present an update on the guidelines on hypercholesterolemia management. The authors have systematically reviewed guidelines in hypercholesterolemia management and discussed a pragmatic approach to follow the most recent guidelines including guidance from NICE in the primary care setting.


Pharmaceutical pollution of the world's rivers.
Environmental exposure to active pharmaceutical ingredients (APIs) can have negative effects on the health of ecosystems and humans. While numerous studies have monitored APIs in rivers, these employ different analytical methods, measure different APIs, and have ignored many of the countries of the world. This makes it difficult to quantify the scale of the problem from a global perspective. Furthermore, comparison of the existing data, generated for different studies/regions/continents, is challenging due to the vast differences between the analytical methodologies employed. Here, we present a global-scale study of API pollution in 258 of the world's rivers, representing the environmental influence of 471.4 million people across 137 geographic regions. Samples were obtained from 1,052 locations in 104 countries (representing all continents and 36 countries not previously studied for API contamination) and analyzed for 61 APIs. Highest cumulative API concentrations were observed in sub-Saharan Africa, south Asia, and South America. The most contaminated sites were in low- to middle-income countries and were associated with areas with poor wastewater and waste management infrastructure and pharmaceutical manufacturing. The most frequently detected APIs were carbamazepine, metformin, and caffeine (a compound also arising from lifestyle use), which were detected at over half of the sites monitored. Concentrations of at least one API at 25.7% of the sampling sites were greater than concentrations considered safe for aquatic organisms, or which are of concern in terms of selection for antimicrobial resistance. Therefore, pharmaceutical pollution poses a global threat to environmental and human health, as well as to delivery of the United Nations Sustainable Development Goals.


Left mainstem coronary artery ostial stenosis--death after angiography. A report of 5 cases.
Death directly related to selective coronary arteriography in 5 patients with a history of unstable angina pectoris during the period 1975-1985 is reported. Four different cardiologists were involved. A feature common to all the cases was the presence of significant ostial stenosis of the left mainstem coronary artery (LMCA); 2 patients had haemodynamically important obstruction of a dominant right coronary artery (RCA) ostium, while 2 others had total occlusion in the proximal part of a dominant RCA. The RCA in the last case was angiographically normal and non-dominant. Collateral coronary blood flow was fairly sparse in most cases and in 4 left ventricular dysfunction of varying degree was present. All patients developed severe hypotension and electromechanical dissociation after arteriography while still in the cardiac catheterization laboratory. Resuscitation efforts were uniformly unsuccessful. Autopsy on 1 patient demonstrated extensive obstructive coronary atherosclerosis with a massive acute anterior myocardial infarction. Cardiac catheterization poses an extremely high risk for this subgroup of patients with LMCA disease, as does selective coronary arteriography. The possible role of catheter-provoked coronary vasospasm of the LMCA is suggested; a recently introduced soft-tipped cardiovascular catheter may be more appropriate in this setting.


A comparison of the performance on extrinsic and intrinsic cartographic visualizations through correctness, response time and cognitive processing.
The aim of this study was to compare the performance of two bivariate visualizations by measuring response correctness (error rate) and response time, and to identify the differences in cognitive processes involved in map-reading tasks by using eye-tracking methods. The present study is based on our previous research and the hypothesis that the use of different visualization methods may lead to significant cognitive-processing differences. We applied extrinsic and intrinsic visualizations in the study. Participants in the experiment were presented maps which depicted two variables (soil moisture and soil depth) and asked to identify the areas which displayed either a single condition (e.g., "find an area with low soil depth") or both conditions (e.g., "find an area with high soil moisture and low soil depth"). The research sample was composed of 31 social sciences and humanities university students. The experiment was performed under laboratory conditions, and Hypothesis software was used for data collection. Eye-tracking data were collected for 23 of the participants. An SMI RED-m eye-tracker was used to determine whether either of the two visualization methods was more efficient for solving the given map-reading tasks. Our results showed that with the intrinsic visualization method, the participants spent significantly more time with the map legend. This result suggests that extrinsic and intrinsic visualizations induce different cognitive processes. The intrinsic method was observed to generally require more time and led to higher error rates. In summary, the extrinsic method was found to be more efficient than the intrinsic method, although the difference was less pronounced in the tasks which contained two variables, which proved to be better suited to intrinsic visualization.


The need to do better - Are we still letting our patients down and at what cost?
To survey women's views on HRT and alternative therapies and make comparisons with 2007 data. A questionnaire on a UK patient-tailored independent clinician-led website with anonymous responses analysed using descriptive statistics. Answers to survey questions in 2007 and 2014. A total of 1476 responses from 33 countries were obtained. Almost 70% of respondents had used/would consider using HRT. Over the last 5 years, 27.7% felt that their views had changed for the better. Most obtained information from health professionals or the Internet. About 51.1% felt that their family doctor did not recognise the importance of the menopause with one-third feeling resistance to being offered HRT. Compared to 2007, significantly more women were aware of the different risks associated with different types of HRT. More women were able to respond positively to the question asking whether or not they felt able to make an informed choice regarding HRT/alternative therapies. There has been negativity and confusion regarding HRT management since the beginning of the millennium. Our findings suggest that we, as health professionals, continue to let our patients down with poor provision of information, inaccurate or wrong information, or access to the right care. The cost of this is women living with preventable sequelae associated with the menopausal transition with a consequent adverse impact on health and the health economy. The importance of the menopause consultation as part of a life course approach is highlighted as well as the emerging discipline of Health Web Science.


Is there evidence for bacterial transfer via the placenta and any role in the colonization of the infant gut? - a systematic review.
With the important role of the gut microbiome in health and disease, it is crucial to understand key factors that establish the microbial community, including gut colonization during infancy. It has been suggested that the first bacterial exposure is via a placental microbiome. However, despite many publications, the robustness of the evidence for the placental microbiome and transfer of bacteria from the placenta to the infant gut is unclear and hence the concept disputed. Therefore, we conducted a systematic review of the evidence for the role of the placental, amniotic fluid and cord blood microbiome in healthy mothers in the colonization of the infant gut. Most of the papers which were fully assessed considered placental tissue, but some studied amniotic fluid or cord blood. Great variability in methodology was observed especially regarding sample storage conditions, DNA/RNA extraction, and microbiome characterization. No study clearly considered transfer of the normal placental microbiome to the infant gut. Moreover, some studies in the review and others published subsequently reported little evidence for a placental microbiome in comparison to negative controls. In conclusion, current data are limited and provide no conclusive evidence that there is a normal placental microbiome which has any role in colonization of infant gut.


Community health worker interventions for Latinos with type 2 diabetes: a systematic review of randomized controlled trials.
This systematic review aimed to synthesize glucose (HbA1c) outcomes of community health worker (CHW)-delivered interventions for Latinos with type 2 diabetes that were tested in randomized controlled trials and to summarize characteristics of the targeted populations and interventions, including the background, training, and supervision of the CHWs. Searches of PubMed and Google Scholar databases and references from selected articles identified 12 studies that met the inclusion criteria. Of these, seven reported statistically significant improvements in HbA1c. Study participants were largely low-income, female, and Spanish-speaking and had uncontrolled diabetes. The CHWs led the interventions alone, in pairs, or as part of a team. Interventions varied considerably in session time, duration, and number. Most met standards for tailored, high-intensity interventions and half were theory-based. Overall, methodological quality was good but there were inconsistencies in the reporting of key information. Future research should report in greater detail CHW background, training, and supervision; examine factors associated with intervention effectiveness; and provide data on cost and cost-effectiveness.


Surface acoustic waves enhance neutrophil killing of bacteria.
Biofilms are structured communities of bacteria that play a major role in the pathogenicity of bacteria and are the leading cause of antibiotic resistant bacterial infections on indwelling catheters and medical prosthetic devices. Failure to resolve these biofilm infections may necessitate the surgical removal of the prosthetic device which can be debilitating and costly. Recent studies have shown that application of surface acoustic waves to catheter surfaces can reduce the incidence of infections by a mechanism that has not yet been clarified. We report here the effects of surface acoustic waves (SAW) on the capacity of human neutrophils to eradicate S. epidermidis bacteria in a planktonic state and within biofilms. Utilizing a novel fibrin gel system that mimics a tissue-like environment, we show that SAW, at an intensity of 0.3 mW/cm(2), significantly enhances human neutrophil killing of S. epidermidis in a planktonic state and within biofilms by enhancing human neutrophil chemotaxis in response to chemoattractants. In addition, we show that the integrin CD18 plays a significant role in the killing enhancement observed in applying SAW. We propose from out data that this integrin may serve as mechanoreceptor for surface acoustic waves enhancing neutrophil chemotaxis and killing of bacteria.


Reversion of transcriptional repression of Sp1 by 5 aza-2' deoxycytidine restores TGF-beta type II receptor expression in the pancreatic cancer cell line MIA PaCa-2.
The pancreatic cancer cell line, MIA PaCa-2 is not responsive to transforming growth factor beta (TGF-beta) because of a lack of expression of the TGF-beta type II receptor (RII). We show that the lack of RII expression is caused by a deficit of the transcription factor Sp1. Nuclear run-off assays and Western immunoblot showed low levels of transcription and protein levels of Sp1, respectively. Treatment of MIA PaCa-2 cells with the DNA methyl transferase inhibitor, 5-aza-2'-deoxycytidine, resulted in an increase in the rate of Sp1 transcription, in Sp1 protein expression, and in the binding of Sp1 to the RII promoter. Ectopic expression of Sp1 cDNA in MIA PaCa-2 cells led to an increase in RII promoter-chloramphenicol acetyltransferase activity and RII expression. Expression of Sp1 cDNA also caused a reduction in both growth and clonogenicity that was associated with restoration of responsiveness to TGF-beta. Conversely, cells that express RII (BxPC-3 and MIA PaCa-2 Sp1 transfectants) when treated with mithramycin, an inhibitor of Sp1 binding, showed a reduction in RII mRNA expression. The reduction of RII mRNA was attributed to a decrease in RII promoter-chloramphenicol acetyltransferase activity that was associated with a decrease in Sp1 binding to the RII promoter. These data indicate that transcriptional repression of the Sp1 gene in MIA PaCa-2 cells plays a role in the transcriptional inactivation of the RII gene and thus lack of responsiveness to TGF-beta.


Patterns of adult cross-racial friendships: A context for understanding contemporary race relations.
This study examined patterns, characteristics, and predictors of cross-racial friendships as the context for understanding contemporary race relations. A national survey included 1,055 respondents, of whom 55% were white, 32% were black, and 74% were female; ages ranged from 18 to ≥65 years. Focus groups were conducted to assess societal and personal benefits. Participants (n = 31) were racially diverse and aged 20 to 66 years. After accounting for multiple covariates, regression analysis revealed that Asians, Hispanics, and multiracial individuals are more likely than their white and black counterparts to have cross-racial friends. Females were less likely than males to have 8 or more cross-racial friends. Regression analysis revealed that the depth of cross-racial friendships was greater for women than men and for those who shared more life experiences. Increasing age was associated with lower cross-racial friendship depth. Qualitative analysis of open-ended questions and focus group data established the social context as directly relevant to the number and depth of friendships. Despite the level of depth in cross-racial friendships, respondents described a general reluctance to discuss any racially charged societal events, such as police shootings of unarmed black men. This study identified salient characteristics of individuals associated with cross-racial friendships and highlighted the influence of the social, historical, and political context in shaping such friendships. Our findings suggest that contemporary race relations reflect progress as well as polarization. (PsycINFO Database Record


Incidence of type II diabetes in Mexican Americans predicted by fasting insulin and glucose levels, obesity, and body-fat distribution.
Few data exist on predictors of non-insulin-dependent (type II) diabetes mellitus. We examined body mass index (BMI), ratio of subscapular-to-triceps skin fold (centrality index), and fasting glucose and insulin concentrations as predictors of decompensation to type II diabetes in Mexican Americans, a population at high risk for this disorder. Twenty-eight of 474 initially nondiabetic Mexican Americans developed type II diabetes after 8 yr of follow-up. Converters to diabetes were older and had higher BMIs, centrality indices, and fasting glucose and insulin concentrations than nonconverters. Subjects in the highest quartile of the insulin distribution had 6.6 times the risk of developing type II diabetes as subjects in the remaining three quartiles combined (95% confidence interval [CI] = 3.14-13.7). In multivariate analysis, fasting glucose (odds ratio [OR] = 5.80, 95% CI = 2.57-13.1) and insulin (OR = 3.12, 95% CI = 1.36-7.14) remained significantly related to conversion to diabetes. However, BMI and centrality index, which were significantly related to conversion in the univariate analysis, were no longer significant in the multivariate analysis once glucose and insulin concentrations were taken into consideration, suggesting that the effect of these variables may be mediated by insulin resistance. Nearly half of the incident cases developed in a subset of the population who were simultaneously in the highest quartile of both fasting insulin and glucose concentrations (population-attributable risk 44.2%). Our results support the insulin resistance/pancreatic exhaustion theory of type II diabetes.


CHRND mutation causes a congenital myasthenic syndrome by impairing co-clustering of the acetylcholine receptor with rapsyn.
The objective of this study was to analyse the mutations of the acetylcholine receptor (AChR) delta subunit gene (CHRND) in a patient with sporadic congenital myasthenic syndrome (CMS). Mutations in various genes encoding proteins expressed at the neuromuscular junction may cause CMS. Mutations of AChR subunit genes lead to end-plate AChR deficiency or to altered kinetic properties of the receptor. Mutations in the alpha, beta and delta subunits of the AChR are less frequent than mutations of the epsilon subunit; mutations in these subunits leading to AChR deficiency are often associated with a severe phenotype. A sporadic patient from Germany was studied, who presented with an early onset CMS associated with feeding difficulties, ptosis, a moderate general weakness responsive to anticholinesterase treatment and recurrent episodes of respiratory insufficiency provoked by infections. The CHRND gene was screened for mutations by RFLP, long-range PCR and sequence analysis. Subsequently, we conducted functional studies of AChR mutants co-transfected with rapsyn in HEK 293 cells. Heterozygously to a 2.2 kb microdeletion disrupting the CHRND gene, we identified a novel point mutation in the long cytoplasmic loop, CHRND E381K. The cytoplasmic loop of the AChR subunits is known to be essential for AChR-rapsyn co-clustering. We therefore studied the interaction of AChR containing the CHRND E381K mutation with rapsyn by evaluating expression and co-localization of rapsyn and mutated AChR subunits in co-transfected HEK 293 cells. Interestingly, the mutated receptor showed severely reduced cluster formation compared with the wild-type receptor. In contrast, the corresponding amino acid substitution in the cytoplasmic loop of the AChR epsilon (CHRNE E376K) as well as a recently reported CMS mutation affecting this domain (CHRNE N436del) had no impact on cluster formation. CHRND mutations are a rare cause for CMS but should be considered in patients with a severe, early onset disease form, clinically resembling a rapsyn phenotype with recurrent episodic apnoeas. Our results suggest that impairment of AChR-rapsyn co-clustering--a well-known molecular mechanism for rapsyn mutations--could also result from mutations in the delta subunit. Introduction of the same mutation in the epsilon subunit had no effect on AChR clustering indicating a special role of the delta subunit in AChR-rapsyn interactions.


Mathematic models for a ray tracing method and its applications in wireless optical communications.
This paper presents a new ray tracing method, which contains a whole set of mathematic models, and its validity is verified by simulations. In addition, both theoretical analysis and simulation results show that the computational complexity of the method is much lower than that of previous ones. Therefore, the method can be used to rapidly calculate the impulse response of wireless optical channels for complicated systems.


A recent increase in asthma mortality in the northwestern United States.
Epidemiologic studies have suggested a recent increase in asthma mortality in New Zealand and the United States. Vital statistics were used to calculate asthma mortality rates for Washington and Oregon from 1972 through 1983, adjusting for the nosologic redefinition of asthma in 1979. Between 1977 and 1983, age-adjusted mortality from asthma increased 82% in these two states. The combined age-specific asthma mortality rates for Washington and Oregon showed that the increase was concentrated in the greater than or equal to 75 age group, in which mortality increased from 4.7 to 12.0 per 100,000 between 1977 and 1983. A significant increasing trend in the proportion of death certificates listing a cardiac dysrhythmia along with asthma as a cause of death was observed overall, in the State of Washington alone, and in decedents 75 or more years of age. The same trend was not statistically significant in Oregon or among younger decedents. This increase in asthma mortality may be due to a change in therapy such as an increased reliance on beta-agonists and theophylline preparations in combination, a change in the clinical definition of asthma, or other as yet unrecognized factors. Further investigation of these findings is warranted.


Insights into the pores of microwave-assisted metal-imidazolate frameworks showing enhanced gas sorption.
Microwave heating (MW)-assisted synthesis has been widely applied as an alternative method for the chemical synthesis of organic and inorganic materials. In this work, we report MW-assisted synthesis of three isostructural 3D frameworks with a flexible linker arm of the chelating linker 2-substituted imidazolate-4-amide-5-imidate, named IFP-7-MW (M = Zn, R = OMe), IFP-8-MW (M = Co; R = OMe) and IFP-10-MW (M = Co; R = OEt) (IFP = Imidazolate Framework Potsdam). These chelating ligands were generated in situ by partial hydrolysis of 2-substituted 4,5-dicyanoimidazoles under MW- and also conventional electrical heating (CE)-assisted conditions in DMF. The structure of these materials was determined by IR spectroscopy and powder X-ray diffraction (PXRD) and the identity of the materials synthesized under CE-conditions was established. Materials obtained from MW-heating show many fold enhancement of CO2 and H2 uptake capacities, compared to the analogous CE-heating method based materials. To understand the inner pore-sizes of IFP structures and variations of gas sorptions, we performed positron annihilation lifetime spectroscopy (PALS), which shows that MW-assisted materials have smaller pore sizes than materials synthesized under CE-conditions. The "kinetically controlled" MW-synthesized material has an inherent ability to trap extra linkers, thereby reducing the pore sizes of CE-materials to ultra/micropores. These ultramicropores are responsible for high gas sorption.


Dual role of the Drosophila pattern gene tailless in embryonic termini.
One of the first zygotically active genes required for formation of the terminal domains of the Drosophila embryo is tailless (tll). Expression of the tll gene is activated ectopically in gain-of-function mutants of the maternal terminal gene torso (tor); this suggests that tor normally activates the tll gene in the termini. Ectopic expression of tll under the control of an inducible promoter results in differentiation of ectopic terminal-specific structures, the Filzkörper, and leads to the activation of at least one gene, hunchback, that is required to form these structures. Ectopic expression of the tll gene also represses segmentation by repressing the gap genes Krüppel and knirps and probably also pair rule genes.


Biomarkers of coagulation, fibrinolysis, endothelial function, and inflammation in arterialized venous blood.
Effects of venous blood arterialization on cardiovascular risk markers are still unknown. We evaluated biomarkers of inflammation, coagulation, fibrinolysis, and endothelial function in arterialized compared with regular venous blood. Cubital venipunctures were obtained from 10 healthy volunteers. Arterialization was generated by 10 min heating of the contralateral hand. Concentrations of albumin, C-reactive protein (CRP), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor type 1 (PAI-1), and von Willebrand factor (vWF) were measured by validated assays. Concentrations of albumin, CRP, and vWF were significantly lower in arterialized than in venous blood (albumin: 43.8 g/l and 44.8 g/l, P = 0.02). Differences in CRP and vWF became insignificant after adjusting for albumin. The endogenous thrombin potential (ETP) was significantly higher in arterialized than in venous blood (1929 nmol/l*min vs. 1872 nmol/l*min, P = 0.02). Addition of the FXIIa inhibitor Corn Trypsin Inhibitor (CTI) prior to the thrombin generation test eliminated the ETP difference suggesting that hand heating activates the FXII-dependent coagulation pathway.


Effect of Oral Insulin on the Severity and Recovery of Methotrexate-Induced Gastrointestinal Mucositis in the Rat.
Gastrointestinal (GI) mucositis is an adverse-effect of chemo- and radiotherapy. Oral insulin has been suggested as possible intestinal growth factor and possible intervention for GI mucositis. We aimed to determine the effect of oral insulin on the severity and recovery of mucositis in a methotrexate (MTX)-induced GI mucositis rat model. Male Wistar rats (n = 24) received a single injection of 60 mg/kg MTX intravenously at day 0. From day -3 oral insulin was added to the drinking water. Group MTX received normal drinking water, group MTX+INS0.5 received 0.5 U/mL insulin, and group MTX+INS1 received 1 U/mL insulin in drinking water. The severity of mucositis was determined by intake, bodyweight, illness, and plasma citrulline. In the recovery phase, the function of the gut was tested with an oral glucose tolerance test, and villus and crypt length of the small intestine were measured. MTX-induced mucositis in all 3 groups and oral insulin did not cause a change in the severity of mucositis, with comparable bodyweight, food intake, and water intake. Oral insulin did not alter the enterocyte mass, determined with plasma citrulline. The glucose level after bolus was higher in the MTX group than the MTX+INS1 group (P < 0.05). Histology was not significant different between all groups. Oral insulin does not alter the severity or the acceleration of recovery of mucositis. Therefore, we conclude that it is not useful to further study oral insulin as possible intervention to prevent or treat chemotherapy-induced GI mucositis.


Assessment of a small-scale fishery: Lane Snapper (Lutjanus synagris) using a length metric method.
Small-scale fisheries are hard to assess because of the limited availability of data. Therefore, a method requiring easy-to-obtain catch-data is important for the assessment and management of small-scale fisheries. The objective of this study was to assess the effect of fishing gear selectivity on a length-based metric method proposed by Froese by estimating three indicators using catch-data from Lane Snapper (Lutjanus synagris) collected in Honduras. These indicators are (1) the percentage of mature individuals in the catch, (2) the percentage of fish within the range of estimated optimal lengths to be captured, and (3) the percentage of fish larger than the optimal length. These indicators determine the level of overfishing. The indicators were estimated separately for catch-data corresponding to gill nets, and each indicator was estimated with and without selectivity correction. Selectivity and mesh sizes of the fishing gear had a major impact on the estimation of indicators 1 and 2. As for indicator 3, it consistently showed a high level of exploitation. The three estimated indicators suggested that the Lane Snapper fishery in Honduras is experiencing overfishing. Overall, the method appears to be promising for the assessment of small-scale fisheries, but it should be used cautiously.


The relationship between child protection contact and mental health outcomes among Canadian adults with a child abuse history.
Despite being a primary response to child abuse, it is currently unknown whether contact with child protection services (CPS) does more good than harm. The aim of the current study was to examine whether contact with CPS is associated with improved mental health outcomes among adult respondents who reported experiencing child abuse, after adjusting for sociodemographic factors and abuse severity. The data were drawn from the 2012 Canadian Community Health Survey-Mental Health (CCHS-2012), which used a multistage stratified cluster design (household-level response rate = 79.8%). Included in this study were individuals aged 18 years and older living in the 10 Canadian provinces (N = 23,395). Child abuse included physical abuse, sexual abuse, and exposure to intimate partner violence (IPV). Mental health outcomes included lifetime mental disorders, lifetime and past year suicidal ideation, plans, and attempts, and current psychological well-being and functioning and distress. All models were adjusted for sociodemographic factors and severity of child abuse. For the majority of outcomes, there were no statistically significant differences between adults with a child abuse history who had CPS contact compared to those without CPS contact. However, those with CPS contact were more likely to report lifetime suicide attempts. These findings suggest that CPS contact is not associated with improved mental health outcomes. Implications are discussed.


Epidemiology of cardiovascular diseases in Europe.
Within Europe large differences exist in mortality from coronary heart disease and stroke. These diseases show a clear West-East gradient with high rates in Eastern Europe. In spite the decreasing trend in age-adjusted cardiovascular disease mortality in Western European countries an increase in the number of cardiovascular patients is expected because of the ageing of the population. Consequently the health care cost for these diseases will increase. Total and HDL cholesterol are major determinants of coronary heart disease. Saturated and trans fatty acids have a total and LDL cholesterol elevating effect and unsaturated fatty acids a lowering effect. N-3 polyunsaturated fatty acids seem to have a protective effect on coronary heart disease occurrence independent of their effect on cholesterol. Dietary antioxidants could be of importance because they may prevent oxidation of the atherogenic cholesterol rich LDL lipoproteins. There is however no convincing evidence that either vitamin E, carotenoids or vitamin C protect against coronary heart disease. Observational research has shown that flavonols, polyphenols with strong antioxidant properties present in plant foods, may protect against coronary heart disease. Blood pressure is a major determinant of coronary heart disease and stroke. Historically salt is viewed as the most important dietary determinant of blood pressure. Recent research shows that also a low-fat diet rich in potassium, calcium and magnesium lowers blood pressure substantially. This suggests a multifactorial influence of different nutrients on blood pressure. It can be concluded that a diet low in saturated and trans fatty acids and rich in plant foods in combination with regular fish consumption is associated with a low risk of cardiovascular mortality.


Defens-IN! Human α-Defensin 5 Acts as an Unwitting Double Agent to Promote Shigella Infection.
Shigella pathogenesis has confounded researchers for years because of its narrow host selectivity and extraordinary infectious capability. In this issue of Immunity, Xu et al. (2018) identify a cunning mechanism whereby Shigella hijacks human α-defensin 5 to enhance its adhesion and subsequent invasion.


Striving to Make Massage Therapy Accessible to Older Adults: an Interview with Niki Munk, PhD, LMT.
First she became a licensed massage therapist in Kentucky. Then, in a quest to improve the care she provided to her elderly clinical population, Niki Munk continued her journey by furthering her education and receiving her doctorate in gerontology. Dr. Munk, a massage therapy researcher at Indiana University's IUPUI campus, shares in this interview her optimism on the future of the massage therapy profession and massage therapy research. Dr. Munk also provides insight into the way she makes decisions, and from whom she would like to receive advice.


Halothane inhibits two components of calcium current in clonal (GH3) pituitary cells.
The effect of halothane on isolated calcium (Ca2+) current of clonal (GH3) pituitary cells was investigated using standard whole-cell clamp techniques at room temperature. Halothane (0.1-5.0 mM) reversibly reduced both the low-threshold, transient [low-voltage-activated (LVA)] component and the high-threshold [high-voltage-activated (HVA)] component of Ca2+ current. Halothane had little effect on the voltage dependence of activation or inactivation of either component of Ca2+ current. Inhibition of the peak high-threshold Ca2+ current was half-maximal at about 0.8 mM halothane, with maximal inhibition (100%) occurring with 5 mM halothane. When measured at the end of a 190-msec command step, half-maximal reduction of high-threshold current occurred at less than 0.5 mM halothane. The low-threshold transient current was less sensitive to halothane, with half-maximal inhibition of peak transient current activated at -30 mV occurring at approximately 1.3 mM. The effect of halothane on the HVA current was apparently not mediated by changes in intracellular Ca2+ concentration. The ability of halothane to inhibit Ca2+ current was unaffected by either the inclusion of the rapid Ca2+ buffer 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid (BAPTA) in the recording pipette or exposure of the cell to 10 mM caffeine. To assess the selectivity of the effect of halothane, the actions of halothane on two components of voltage-activated potassium (K+) current observed in the absence of extracellular Ca2+ and on voltage-dependent sodium (Na+) current were also examined. Halothane had no effect on the voltage-dependent, inactivating K+ current of GH3 cells at concentrations up to 1.2 mM. In contrast, the non-inactivating K+ current, though less sensitive to halothane than either Ca2+ current, was reduced by about 40% by 1.2 mM halothane at +20 mV. Peak Na+ current was also blocked by halothane, but 50% block required around 2.6 mM halothane with little effect at 1.6 mM. Reduction of Na+ current was associated with a substantial negative shift in the steady-state inactivation curve. Although the results indicate that a number of voltage-dependent ionic currents are sensitive to halothane, both components of Ca2+ current exhibit a greater sensitivity to halothane than any of three other voltage-dependent currents in GH3 cells. These results show that GH3 cell Ca2+ currents are selectively inhibited by clinically appropriate concentrations of halothane and that the reduction of Ca2+ current can account for the inhibition by halothane of TRH- or KCl-induced prolactin secretion in GH3 cells.


Elemental mapping of human nervous tissue using the scanning proton microprobe.
The scanning proton microprobe (SPM) is a powerful multi-elemental analytical instrument capable of elemental mapping at the parts per million level of sensitivity. In this report we demonstrate that the SPM is sufficiently sensitive and versatile to distinguish different regions of the human central nervous system on the basis of differences in their normal endogenous elemental composition.


Continuous dopamine-receptor treatment of Parkinson's disease: scientific rationale and clinical implications.
Levodopa-induced motor complications are a common source of disability for patients with Parkinson's disease. Evidence suggests that motor complications are associated with non-physiological, pulsatile stimulation of dopamine receptors. In healthy brains, dopamine neurons fire continuously, striatal dopamine concentrations are relatively constant, and there is continuous activation of dopamine receptors. In the dopamine-depleted state, standard levodopa therapy does not normalise the basal ganglia. Rather, levodopa or other short-acting dopaminergic drugs induce molecular changes and altered neuronal firing patterns in basal ganglia neurons leading to motor complications. The concept of continuous dopaminergic stimulation proposes that continuous delivery of a dopaminergic drug will prevent pulsatile stimulation and avoid motor complications. In monkeys treated with MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) and patients with Parkinson's disease, long-acting or continuous infusion of a dopaminergic drug reduces the risk of motor complications. The current challenge is to develop a long-acting oral formulation of levodopa that provides clinical benefits but avoids motor complications.


Atg9 cycles between mitochondria and the pre-autophagosomal structure in yeasts.
Autophagy is a degradative process conserved among eukaryotic cells. It allows the elimination of cytoplasm including aberrant protein aggregates and damaged organelles. Accordingly, it is implicated in normal developmental processes and also serves a protective role in tumor suppression and elimination of invading pathogens, whereas defects in autophagy are associated with various human diseases including cancer and neurodegeneration. Atg proteins mediate the sequestration event that occurs at the preautophagosomal structure (PAS) by catalyzing the formation of double-membrane vesicles, termed autophagosomes. In Saccharomyces cerevisiae, the integral membrane protein Atg9 that is required for autophagy cycles through the PAS. Here, we demonstrate that Atg9 shuttles between this location and mitochondria. These data support a new model where mitochondria may provide at least part of the autophagosomal lipids and suggest a novel cellular function for this well-studied organelle.


Saving life and brain with extracorporeal cardiopulmonary resuscitation: A single-center analysis of in-hospital cardiac arrests.
Despite advances in medical care, survival to discharge and full neurologic recovery after cardiac arrest remains less than 20% after cardiopulmonary resuscitation. An alternate approach to traditional cardiopulmonary resuscitation is extracorporeal cardiopulmonary resuscitation, which places patients on extracorporeal membrane oxygenation during cardiopulmonary resuscitation and provides immediate cardiopulmonary support when traditional resuscitation has been unsuccessful. We report the results from extracorporeal cardiopulmonary resuscitation at the Thomas Jefferson University. Between 2010 and June 2014, 107 adult extracorporeal membrane oxygenation procedures were performed at the Thomas Jefferson University. Patient demographics, survival to discharge, and neurologic recovery of patients who underwent extracorporeal cardiopulmonary resuscitation were retrospectively analyzed with institutional review board approval. A total of 23 patients (15 male and 8 female; mean age, 46 ± 12 years) underwent extracorporeal cardiopulmonary resuscitation. All patients who met criteria were placed on 24-hour hypothermia protocol (target temperature 33°C) with initiation of extracorporeal membrane oxygenation. The mean duration of extracorporeal membrane oxygenation support was 6.2 ± 5.5 days. Nine patients died while on extracorporeal membrane oxygenation from the following causes: anoxic brain injury (4), stroke (4), and bowel necrosis (1). Two patients with anoxic brain injury on extracorporeal cardiopulmonary resuscitation donated multiple organs for transplant. The survival to discharge was 30% (7/23 patients) with approximately 100% full neurologic recovery. The extracorporeal cardiopulmonary resuscitation procedure provided reasonable patient recovery. Extracorporeal cardiopulmonary resuscitation also allowed for neurologic recovery and made multiorgan procurement possible. On the basis of the survival, extracorporeal cardiopulmonary resuscitation should be considered when determining the optimal treatment path for patients who need cardiopulmonary resuscitation. The proper use of extracorporeal cardiopulmonary resuscitation improved the hospital outcomes for patients with in-hospital cardiac arrest.


The frantic play of the concealed HIV envelope cytoplasmic tail.
Lentiviruses have unusually long envelope (Env) cytoplasmic tails, longer than those of other retroviruses. Whereas the Env ectodomain has received much attention, the gp41 cytoplasmic tail (gp41-CT) is one of the least studied parts of the virus. It displays relatively high conservation compared to the rest of Env. It has been long established that the gp41-CT interacts with the Gag precursor protein to ensure Env incorporation into the virion. The gp41-CT contains distinct motifs and domains that mediate both intensive Env intracellular trafficking and interactions with numerous cellular and viral proteins, optimizing viral infectivity. Although they are not fully understood, a multiplicity of interactions between the gp41-CT and cellular factors have been described over the last decade; these interactions illustrate how Env expression and incorporation into virions is a finely tuned process that has evolved to best exploit the host system with minimized genetic information. This review addresses the structure and topology of the gp41-CT of lentiviruses (mainly HIV and SIV), their domains and believed functions. It also considers the cellular and viral proteins that have been described to interact with the gp41-CT, with a particular focus on subtype-related polymorphisms.