Staphylococcus aureus induces eosinophil cell death mediated by α-hemolysin

Staphylococcus aureus, a major human pathogen, exacerbates allergic disorders, including atopic dermatitis, nasal polyps and asthma, which are characterized by tissue eosinophilia. Eosinophils, via their destructive granule contents, can cause significant tissue damage, resulting in inflammation and further recruitment of inflammatory cells. We hypothesised that the relationship between S. aureus and eosinophils may contribute to disease pathology. We found that supernatants from S. aureus (SH1000 strain) cultures cause rapid and profound eosinophil necrosis, resulting in dramatic cell loss within 2 hours. This is in marked contrast to neutrophil granulocytes where no significant cell death was observed (at equivalent dilutions). Supernatants prepared from a strain deficient in the accessory gene regulator (agr) that produces reduced levels of many important virulence factors, including the abundantly produced α-hemolysin (Hla), failed to induce eosinophil death. The role of Hla in mediating eosinophil death was investigated using both an Hla deficient SH1000-modified strain, which did not induce eosinophil death, and purified Hla, which induced concentration-dependent eosinophil death via both apoptosis and necrosis. We conclude that S. aureus Hla induces aberrant eosinophil cell death in vitro and that this may increase tissue injury in allergic disease.     

Pregnancy and Infant Outcomes among HIV-Infected Women Taking Long-Term ART with and without Tenofovir in the DART Trial

Background

Few data have described long-term outcomes for infants born to HIV-infected African women taking antiretroviral therapy (ART) in pregnancy. This is particularly true for World Health Organization (WHO)–recommended tenofovir-containing first-line regimens, which are increasingly used and known to cause renal and bone toxicities; concerns have been raised about potential toxicity in babies due to in utero tenofovir exposure.

Methods and Findings

Pregnancy outcome and maternal/infant ART were collected in Ugandan/Zimbabwean HIV-infected women initiating ART during The Development of AntiRetroviral Therapy in Africa (DART) trial, which compared routine laboratory monitoring (CD4; toxicity) versus clinically driven monitoring. Women were followed 15 January 2003 to 28 September 2009. Infant feeding, clinical status, and biochemistry/haematology results were collected in a separate infant study. Effect of in utero ART exposure on infant growth was analysed using random effects models.382 pregnancies occurred in 302/1,867 (16%) women (4.4/100 woman-years [95% CI 4.0–4.9]). 226/390 (58%) outcomes were live-births, 27 (7%) stillbirths (≥22 wk), and 137 (35%) terminations/miscarriages (<22 wk). Of 226 live-births, seven (3%) infants died <2 wk from perinatal causes and there were seven (3%) congenital abnormalities, with no effect of in utero tenofovir exposure (p>0.4). Of 219 surviving infants, 182 (83%) enrolled in the follow-up study; median (interquartile range [IQR]) age at last visit was 25 (12–38) months. From mothers'' ART, 62/9/111 infants had no/20%–89%/≥90% in utero tenofovir exposure; most were also zidovudine/lamivudine exposed. All 172 infants tested were HIV-negative (ten untested). Only 73/182(40%) infants were breast-fed for median 94 (IQR 75–212) days. Overall, 14 infants died at median (IQR) age 9 (3–23) months, giving 5% 12-month mortality; six of 14 were HIV-uninfected; eight untested infants died of respiratory infection (three), sepsis (two), burns (one), measles (one), unknown (one). During follow-up, no bone fractures were reported to have occurred; 12/368 creatinines and seven out of 305 phosphates were grade one (16) or two (three) in 14 children with no effect of in utero tenofovir (p>0.1). There was no evidence that in utero tenofovir affected growth after 2 years (p = 0.38). Attained height- and weight for age were similar to general (HIV-uninfected) Ugandan populations. Study limitations included relatively small size and lack of randomisation to maternal ART regimens.

Conclusions

Overall 1-year 5% infant mortality was similar to the 2%–4% post-neonatal mortality observed in this region. No increase in congenital, renal, or growth abnormalities was observed with in utero tenofovir exposure. Although some infants died untested, absence of recorded HIV infection with combination ART in pregnancy is encouraging. Detailed safety of tenofovir for pre-exposure prophylaxis will need confirmation from longer term follow-up of larger numbers of exposed children.

Trial registration

www.controlled-trials.com ISRCTN13968779 Please see later in the article for the Editors'' Summary     

KATNAL1 regulation of sertoli cell microtubule dynamics is essential for spermiogenesis and male fertility

Spermatogenesis is a complex process reliant upon interactions between germ cells (GC) and supporting somatic cells. Testicular Sertoli cells (SC) support GCs during maturation through physical attachment, the provision of nutrients, and protection from immunological attack. This role is facilitated by an active cytoskeleton of parallel microtubule arrays that permit transport of nutrients to GCs, as well as translocation of spermatids through the seminiferous epithelium during maturation. It is well established that chemical perturbation of SC microtubule remodelling leads to premature GC exfoliation demonstrating that microtubule remodelling is an essential component of male fertility, yet the genes responsible for this process remain unknown. Using a random ENU mutagenesis approach, we have identified a novel mouse line displaying male-specific infertility, due to a point mutation in the highly conserved ATPase domain of the novel KATANIN p60-related microtubule severing protein Katanin p60 subunit A-like1 (KATNAL1). We demonstrate that Katnal1 is expressed in testicular Sertoli cells (SC) from 15.5 days post-coitum (dpc) and that, consistent with chemical disruption models, loss of function of KATNAL1 leads to male-specific infertility through disruption of SC microtubule dynamics and premature exfoliation of spermatids from the seminiferous epithelium. The identification of KATNAL1 as an essential regulator of male fertility provides a significant novel entry point into advancing our understanding of how SC microtubule dynamics promotes male fertility. Such information will have resonance both for future treatment of male fertility and the development of non-hormonal male contraceptives.     

A‐FABP—A Biomarker Associated With the Metabolic Syndrome and/or an Indicator of Weight Change?

Objective: Adipocyte fatty acid‐binding protein (A‐FABP) is a plasma biomarker recently associated with the metabolic syndrome. The aim of these studies was to investigate changes of A‐FABP during profound weight loss induced by laparoscopic adjustable gastric banding (LAGB). Methods and Procedures: In study one, 29 severely obese female subjects were examined before and 1 year after surgical treatment. A subgroup of 10 patients was investigated in 3‐month intervals. Metabolic parameters were determined using standard methods, and A‐FABP was detected using a commercially available enzyme‐linked immunosorbent assay. Results: Mean weight loss after 1 year was 24.9 kg (P < 0.001), mainly due to a decrease in fat mass. Metabolic parameters improved substantially. However, serum A‐FABP remained stable. In study two, a subgroup of 10 patients was examined quarterly to determine the time course of A‐FABP changes. Quarterly measurements of serum A‐FABP were significantly higher than baseline levels with the highest A‐FABP value after the first 3 months, where patients had highest weight loss. Discussion: Our results in study one show that A‐FABP serum levels are positively associated with body weight and fat mass. However, 1 year after pronounced weight loss A‐FABP levels remained unchanged. In study two, time course analyses revealed maximum increase of serum A‐FABP in parallel to highest weight loss, which allows to suppose that A‐FABP is not only a biomarker of the metabolic syndrome in the steady state, but also a marker of weight changes in dynamic situations.     

Randomised controlled trial assessing the impact of a nurse delivered,flow monitored protocol for optimisation of circulatory status after cardiac surgery

Objective To assess whether a nurse led, flow monitored protocol for optimising circulatory status in patients after cardiac surgery reduces complications and shortens stay in intensive care and hospital.Design Randomised controlled trial.Setting Intensive care unit and cardiothoracic unit of a university teaching hospital.Participants 174 patients who underwent cardiac surgery between April 2000 and January 2003.Interventions Patients were allocated to conventional haemodynamic management or to an algorithm guided by oesophageal Doppler flowmetry to maintain a stroke index above 35 ml/m².Results 26 control patients had postoperative complications (two deaths) compared with 17 (four deaths) protocol patients (P = 0.08). Duration of hospital stay in the protocol group was significantly reduced from a median of nine (interquartile range 7-12) days to seven (7-10) days (P = 0.02). The mean duration of hospital stay was reduced from 13.9 to 11.4 days, a saving in hospital bed days of 18% (95% confidence interval -12% to 47%). Usage of intensive care beds was reduced by 23% (-8% to 59%).Conclusion A nurse delivered protocol for optimising circulatory status in the early postoperative period after cardiac surgery may significantly shorten hospital stay.     

Monitoring populations of Western Sandpipers and Pacific Dunlins during northward migration on the Fraser River Delta,British Columbia, 1991–2013

The Fraser River Delta in British Columbia, Canada, is a globally significant stopover site for shorebirds, but the population status and trends of many species that use the site remain uncertain. We describe an ongoing program to monitor population trends of the two most abundant species, Western Sandpipers (Calidris mauri) and Dunlins (Calidris alpina), during northward migration. Counts of these species were conducted at a mudflat where large flocks assembled at mid‐tide from 15 April to 15 May, 1991–2013, and we estimated species‐specific counts as the product of daily total flock counts and species proportions obtained during supplementary sampling. The median peak count of both species combined was 177,000 birds, and occurred between 24 April and 3 May. Ratios (proportions) of the two species followed a predictable pattern during the migration period, with a low proportion of Western Sandpipers (3%–20%) in flocks before 20 April, followed by a rapid increase to 80%–100% between 25 April and 10 May and a variable decrease to 30%–80% by 15 May. Mean counts of Western Sandpipers showed no significant trend over the study period. Mean counts of Dunlins showed a non‐linear trend, decreasing until 2001 and then increasing to 2013. Bias and random error in field counts were quantified by comparing field counts to counts made from photographs taken during surveys, and analysis revealed that field counts had a downward, but predictable, bias, accounting for >90% of birds present, with a stochastic error rate of 28.0%. Uncertainty in total population estimates was high after accounting for the effect of length of stay and sampling error. Population estimates suggested that 600,000 Western Sandpipers and 200,000 Dunlins typically passed through the site during northward migration. Our estimates indicate the usefulness of daily counts at major stopover sites during northward migration as an effective tool for monitoring shorebird populations, and underscore the need for conserving such sites.