Methicillin-resistant Staphylococcus capitis with reduced vancomycin susceptibility causes late-onset sepsis in intensive care neonates

Background

Coagulase-negative staphylococci, mainly Staphylococcus epidermidis, are the most frequent cause of late-onset sepsis (LOS) in the neonatal intensive care unit (NICU) setting. However, recent reports indicate that methicillin-resistant, vancomycin-heteroresistant Staphylococcus capitis could emerge as a significant pathogen in the NICU. We investigated the prevalence, clonality and vancomycin susceptibility of S. capitis isolated from the blood of NICU infants and compared these data to adult patients.

Methodology/Principal Findings

We conducted a retrospective laboratory-based survey of positive blood cultures in NICU infants ≥3 days of age (n = 527) and in adult ICU patients ≥18 years of age (n = 1473) who were hospitalized from 2004 to 2009 in two hospital centers in Lyon, France. S. capitis was the most frequent pathogen in NICU infants, ahead of S. epidermidis (39.1% vs. 23.5% of positive blood cultures, respectively). Conversely, S. capitis was rarely found in adult ICU patients (1.0%) compared to S. epidermidis (15.3%). S. capitis bloodstream isolates were more frequently resistant to methicillin when collected from NICU infants than from adult patients (95.6% vs. 53.3%, respectively). Furthermore, we collected and characterized 53 S. capitis bloodstream isolates from NICU infants and adult patients from six distant cities. All methicillin-resistant S. capitis isolates from NICU infants were clonally related as determined by pulsed-field gel electrophoresis. These isolates harbored a type V-related staphylococcal chromosomal cassette mec element, and constantly showed either vancomycin resistance (37.5%) or heteroresistance (62.5%). Conversely, the isolates that were collected outside of the NICU were genetically diverse and displayed much lower rates of vancomycin resistance and heteroresistance (7.7% and 23.1%, respectively).

Conclusions/Significance

A clonal population of methicillin-resistant S. capitis strains has spread into several French NICUs. These isolates exhibit reduced susceptibility to vancomycin, which is the most widely used antimicrobial agent in the NICU setting.     

Assessment of the release of rehabilitated vervet monkeys into the Ntendeka Wilderness Area, KwaZulu-Natal, South Africa: a case study

In South Africa, vervet monkeys (Chlorocebus aethiops) are frequently persecuted, resulting in large numbers of injured and/or orphaned animals. Rehabilitation centres aim to care for these monkeys and ultimately return them to the wild whenever possible. However, it is unknown whether rehabilitation is successful in its goal of creating wild-living, independent, self-sustaining troops due to limited published research in this area. This study describes the release and subsequent fate of a troop of rehabilitated vervet monkeys over a 6-month period. A troop of 16 monkeys was released into the Ntendeka Wilderness Area, a protected part of Ngome Forest, by the WATCH (Wild Animal Trauma Centre and Haven) rehabilitation centre in KwaZulu-Natal, South Africa. Monitoring data were evaluated with regard to survival, mortality, suitability of the release site, breeding, condition, troop composition, behaviour, group dynamics, ranging patterns and the effectiveness of monitoring tools. The release was considered to be a partial success in that the troop exhibited behaviour, group dynamics and ranging patterns similar to wild conspecifics. However, the survival rate was low and the troop was judged to be non-self-sustaining. The main problems identified were the limited lifetimes of radio collars, which resulted in missing animals and caused monitoring to be cut short, illegal hunting activities, predation and a small troop size with few adults. The authors recommend improvements that may increase success, such as retaining troops in release enclosures for longer periods, releasing a larger troop with more adults that more closely matches wild troop composition, selecting a release site at least 3 km from the nearest human settlement and the use of GPS collars to allow for a longer monitoring period encompassing all seasonal conditions. Furthermore, all primates for release should be medically screened so as to avoid potential negative impacts on wild populations.     

Distribution and population estimate for the chacma baboon (Papio ursinus) in KwaZulu-Natal, South Africa

We report the current species distribution and population estimate for the chacma baboon (Papio ursinus) in KwaZulu-Natal Province (KZN), South Africa, based on an analysis of estimated area of occupancy and estimated home range size. This estimate suggests a total population size of approximately 11,000 individuals for KZN. Much of the province is uninhabited, with a density in occupied areas of approximately 1.8 animals per km(2). The current population size may be more than an order of magnitude smaller than historical population size. Chacma baboons now exhibit a highly fragmented and discontinuous distribution in KZN, with 58% of the population residing within protected areas, and more than half of these troops reside in areas >1,500?m above average sea level. The small population and highly fragmented distribution of chacma baboons in KZN, combined with rapidly increasing human population size and transformation of natural habitat, suggest this species requires greater conservation attention.     

Urocortin 3 Marks Mature Human Primary and Embryonic Stem Cell-Derived Pancreatic Alpha and Beta Cells

The peptide hormone Urocortin 3 (Ucn 3) is abundantly and exclusively expressed in mouse pancreatic beta cells where it regulates insulin secretion. Here we demonstrate that Ucn 3 first appears at embryonic day (E) 17.5 and, from approximately postnatal day (p) 7 and onwards throughout adult life, becomes a unifying and exclusive feature of mouse beta cells. These observations identify Ucn 3 as a potential beta cell maturation marker. To determine whether Ucn 3 is similarly restricted to beta cells in humans, we conducted comprehensive immunohistochemistry and gene expression experiments on macaque and human pancreas and sorted primary human islet cells. This revealed that Ucn 3 is not restricted to the beta cell lineage in primates, but is also expressed in alpha cells. To substantiate these findings, we analyzed human embryonic stem cell (hESC)-derived pancreatic endoderm that differentiates into mature endocrine cells upon engraftment in mice. Ucn 3 expression in hESC-derived grafts increased robustly upon differentiation into mature endocrine cells and localized to both alpha and beta cells. Collectively, these observations confirm that Ucn 3 is expressed in adult beta cells in both mouse and human and appears late in beta cell differentiation. Expression of Pdx1, Nkx6.1 and PC1/3 in hESC-derived Ucn 3⁺ beta cells supports this. However, the expression of Ucn 3 in primary and hESC-derived alpha cells demonstrates that human Ucn 3 is not exclusive to the beta cell lineage but is a general marker for both the alpha and beta cell lineages. Ucn 3⁺ hESC-derived alpha cells do not express Nkx6.1, Pdx1 or PC1/3 in agreement with the presence of a separate population of Ucn 3⁺ alpha cells. Our study highlights important species differences in Ucn 3 expression, which have implications for its utility as a marker to identify mature beta cells in (re)programming strategies.     

hERG1 Channels and Glut-1 as Independent Prognostic Indicators of Worse Outcome in Stage I and II Colorectal Cancer: A Pilot Study

BACKGROUND: There is a need to identify new markers to assess recurrence risk in early-stage colorectal cancer (CRC) patients. We explored the prognostic impact of ether-a-gò-gò-related gene 1 channels and some hypoxia markers, in patients with nonmetastatic (stage I, II, and III) CRC. METHODS: The expression of hERG1, vascular endothelial growth factor A (VEGF-A), glucose transporter 1, carbonic anhydrase IX (CA-IX), epidermal growth factor receptor (EGF-R), and p53 was tested by immunohistochemistry in 135 patients. The median follow-up was 35 months. Clinicopathologic parameters and overall survival were evaluated. RESULTS: hERG1 displayed a statistically significant association with Glut-1, VEGF-A, CA-IX, and EGF-R; p53 with VEGF-A and CA-IX; Glut-1 with the age of the patients; and EGF-R with TNM and mucin content. TNM and CA-IX were prognostic factors at the univariate analysis; TNM, hERG1, and Glut-1, at the multivariate analysis. Risk scores calculated from the final multivariate model allowed to stratify patients into four different risk groups: A) stage I-II, Glut-1 positivity, any hERG1; B) stage I-II, Glut-1 and hERG1 negativity; C) stage I-II, Glut-1 negativity, hERG1 positivity; D) stage III, any Glut-1 and any hERG1. CONCLUSIONS: hERG1 positivity with Glut-1 negativity identifies a patient group with poor prognosis within stage I-II CRC. The possibility that these patients might benefit from adjuvant therapy, independently from the TNM stage, is discussed. IMPACT: More robust prognostic and predictive markers, supplementing standard clinical and pathologic staging, are needed for node-negative patients.     

Nutrition and epigenetics: an interplay of dietary methyl donors, one-carbon metabolism and DNA methylation

DNA methylation is the most extensively studied mechanism of epigenetic gene regulation. Increasing evidence indicates that DNA methylation is labile in response to nutritional and environmental influences. Alterations in DNA methylation profiles can lead to changes in gene expression, resulting in diverse phenotypes with the potential for increased disease risk. The primary methyl donor for DNA methylation is S-adenosylmethionine (SAM), a species generated in the cyclical cellular process called one-carbon metabolism. One-carbon metabolism is catalyzed by several enzymes in the presence of dietary micronutrients, including folate, choline, betaine and other B vitamins. For this reason, nutrition status, particularly micronutrient intake, has been a focal point when investigating epigenetic mechanisms. Although animal evidence linking nutrition and DNA methylation is fairly extensive, epidemiological evidence is less comprehensive. This review serves to integrate studies of the animal in vivo with human epidemiological data pertaining to nutritional regulation of DNA methylation and to further identify areas in which current knowledge is limited.     

The Dac-tag, an affinity tag based on penicillin-binding protein 5

Penicillin-binding protein 5 (PBP5), a product of the Escherichia coli gene dacA, possesses some β-lactamase activity. On binding to penicillin or related antibiotics via an ester bond, it deacylates and destroys them functionally by opening the β-lactam ring. This process takes several minutes. We exploited this process and showed that a fragment of PBP5 can be used as a reversible and monomeric affinity tag. At ambient temperature (e.g., 22°C), a PBP5 fragment binds rapidly and specifically to ampicillin Sepharose. Release can be facilitated either by eluting with 10mM ampicillin or in a ligand-free manner by incubation in the cold (1-10°C) in the presence of 5% glycerol. The "Dac-tag", named with reference to the gene dacA, allows the isolation of remarkably pure fusion protein from a wide variety of expression systems, including (in particular) eukaryotic expression systems.     

Changes in Serum Zinc Levels Associated with Giardiasis and Dietary Zinc Intake in Mice

The association of giardiasis with the malabsorption of zinc remains controversial. This study investigated changes in serum zinc levels in Giardia-infected mice subjected to different dietary zinc regimens. Thirty-five mice (strain C₃H/H_eJ) were randomly categorized into two groups. The first group was inoculated with 5 × 10⁶ Giardia trophozoites (n = 18), and the second group remained Giardia free (n = 17). Each group (Giardia infected and Giardia free) was randomly classified into three subgroups and given low (9 mg Zn/kg), normal (33 mg Zn/kg), and high levels (288 mg Zn/kg) of dietary zinc over a 2-week period for acclimation. Fourteen days post-Giardia infection, all of the mice were euthanized and blood samples were collected. The number of trophozoites was quantified (hematocytometer), and serum zinc levels were determined via atomic absorption spectrophotometry. Significant increases in the median weights were only found in the Giardia-free mice (p < 0.05). A higher final median weight was found in the Giardia-free group when compared with that of the Giardia-infected group given low dietary zinc (p = 0.013). In the Giardia-infected group with low dietary zinc, the geometric mean of trophozoites was 3,498 ± 101 (SE) per milliliter. The Giardia-infected group had lower serum zinc levels than did the Giardia-free group with the high dietary zinc regimens (p < 0.05). Our results are consistent with studies among human populations, but further studies are required to elucidate the actual mechanism governing the zinc–giardiasis interaction.     

Deep-sea bacteria enriched by oil and dispersant from the Deepwater Horizon spill

The Deepwater Horizon oil spill resulted in a massive influx of hydrocarbons into the Gulf of Mexico (the Gulf). To better understand the fate of the oil, we enriched and isolated indigenous hydrocarbon-degrading bacteria from deep, uncontaminated waters from the Gulf with oil (Macondo MC252) and dispersant used during the spill (COREXIT 9500). During 20 days of incubation at 5°C, CO(2) evolution, hydrocarbon concentrations and the microbial community composition were determined. Approximately 60% to 25% of the dissolved oil with or without COREXIT, respectively, was degraded, in addition to some hydrocarbons in the COREXIT. FeCl(2) addition initially increased respiration rates, but not the total amount of hydrocarbons degraded. 16S rRNA gene sequencing revealed a succession in the microbial community over time, with an increase in abundance of Colwellia and Oceanospirillales during the incubations. Flocs formed during incubations with oil and/or COREXIT in the absence of FeCl(2) . Synchrotron radiation-based Fourier transform infrared (SR-FTIR) spectromicroscopy revealed that the flocs were comprised of oil, carbohydrates and biomass. Colwellia were the dominant bacteria in the flocs. Colwellia sp. strain RC25 was isolated from one of the enrichments and confirmed to rapidly degrade high amounts (approximately 75%) of the MC252 oil at 5°C. Together these data highlight several features that provide Colwellia with the capacity to degrade oil in cold, deep marine habitats, including aggregation together with oil droplets into flocs and hydrocarbon degradation ability.