The preterm gut microbiota: changes associated with necrotizing enterocolitis and infection

Aim: To describe gut colonization in preterm infants using standard culture and 16S gene rRNA profiling, exploring differences in healthy infants and those who developed NEC/late onset sepsis (LOS). Methods: Ninety‐nine stools from 38 infants of median 27‐week gestation were cultured; 44 stools from 27 infants had their microbial profiles determined by 16S. Ordination analyses explored effects of patient variables on gut communities. Results: Standard microbiological culture identified a mean of two organisms (range 0–7), DGGE 12 (range 3–18) per patient. Enterococcus faecalis and coagulase negative staphylococci (CONS) were most common by culture (40% and 39% of specimens). Meconium was not sterile. No fungi were cultured. Bacterial community structures in infants with NEC and LOS differed from healthy infants. Infants who developed NEC carried more CONS (45% vs 30%) and less Enterococcus faecalis (31% vs 57%). 16S identified Enterobacter and Staphylococcus presence associated with NEC/LOS, respectively. Conclusions: Important differences were found in the gut microbiota of preterm infants who develop NEC/LOS. The relationship of these changes to current practices in neonatal intensive care requires further exploration.     

Gamma-carboxylated isoforms of recombinant human protein S with different biologic properties

Human protein S (HPS), a regulator of hemostasis, is a vitamin K- dependent plasma protein with potential clinical utility. We have obtained high-level expression of the cDNA for HPS in two mammalian cell lines. Both cell lines secreted single chain recombinant HPS (rHPS) in serum-free medium as determined by Western blot analysis. The ability of the rHPS from both cell lines to act as a cofactor for human protein C (HPC) was determined; the rHPS secreted from the human 293 cell line had an activity six times that of the rHPS from the AV12-664 Syrian hamster cell line. Furthermore, the relative specific cofactor activity of rHPS from the 293 cell line was actually 2.5-fold higher than that of single-chain human plasma-derived HPS. Essentially all of the rHPS secreted from the 293 cell line exhibited a calcium-dependent elution profile on anion exchange chromatography, whereas only 25% to 35% of the hamster cell-derived rHPS exhibited this profile. However, the calcium-eluted rHPS from the AV12 cell line had a high specific cofactor activity, equivalent to that of the 293-derived rHPS. A NaCl- elutable rHPS fraction (calcium nondependent) was isolated from the recombinant AV12-664 cell line, further purified, and found to have reduced activity, only 40% that of the calcium-dependent rHPS. The only observable difference in the calcium-dependent and nondependent rHPS molecules was in the content of gamma-carboxyglutamic acid (Gla); the calcium-dependent material contained approximately 10 mol Gla/mol protein whereas the calcium-nondependent material contained only approximately 8 mol Gla/mol of protein. In addition, the calcium- nondependent rHPS had reduced ability to interact with phospholipid vesicles as evidenced by an eightfold increase in the apparent kd. Our data demonstrate the isolation of rHPS with high specific activity, and show that a reduction in as few as two Gla residues dramatically decreases its functional cofactor activity for HPC, due to a reduction in ability to interact with the phospholipid bilayer.     

Prognosis in medically stabilized unstable angina: early Holter ST-segment monitoring compared with predischarge exercise thallium tomography

OBJECTIVE: To assess the relative value of invasive and noninvasive predictors of outcome in patients after unstable angina. DESIGN: Cohort of 54 patients with unstable angina who had 6-month follow-up after stabilization on medical therapy. SETTING: University-based hospital, tertiary referral center. PATIENTS: Consecutive patients with unstable angina whose symptoms resolved while receiving medical therapy. MEASUREMENTS AND MAIN RESULTS: We prospectively compared 24-hour Holter ST-segment monitoring at admission, quantitative exercise thallium tomography, and cardiac catheterization 5 +/- 2 days after admission and analyzed their value for predicting a cardiac event in patients with unstable angina within 6 months. When patients with a favorable outcome (n = 40) were compared with patients with an unfavorable outcome (n = 11) no statistical difference was found in duration of ST shift of 1 mm or more on Holter monitoring (51 +/- 119 min compared with 37 +/- 43 min), exercise duration by the standard Bruce protocol (8.0 +/- 3.6 min compared with 7.9 +/- 3.1 min), exercise-induced ST depression (0.6 +/- 0.9 mm compared with 1.0 +/- 1.0 mm), and contrast left ventricular ejection fraction (70% +/- 10% compared with 69% +/- 15%). Patients with a favorable outcome were distinguished from those with an unfavorable outcome by a higher maximum rate-pressure product (24 x 10(3) +/- 6 x 10(3) compared with 18 x 10(3) +/- 7 x 10(3), P = 0.0025), smaller size of the reversible scintigraphic perfusion defect expressed as a percentage of total myocardium imaged (6% +/- 11% compared with 17% +/- 18%, P = 0.05) and a smaller number of vessels with stenosis of 50% or more (1.1 +/- 1.2 compared with 2.1 +/- 1.0, P = 0.01). On multiple logistic regression analysis, a history of previous myocardial infarction was the most powerful predictor of outcome. In patients without myocardial infarction, reversible exercise thallium perfusion defect size was the only predictor. CONCLUSION: After stabilization of an episode of unstable angina, quantitative tomographic exercise thallium scintigraphy has greater value for risk stratification than Holter ST-segment monitoring, particularly in patients who have not had a previous infarction.     

Development of a multi-layered psychosocial care system for children in areas of political violence

Few psychosocial and mental health care systems have been reported for children affected by political violence in low- and middle income settings and there is a paucity of research-supported recommendations. This paper describes a field tested multi-layered psychosocial care system for children (focus age between 8-14 years), aiming to translate common principles and guidelines into a comprehensive support package. This community-based approach includes different overlapping levels of interventions to address varying needs for support. These levels provide assessment and management of problems that range from the social-pedagogic domain to the psychosocial, the psychological and the psychiatric domains. Specific intervention methodologies and their rationale are described within the context of a four-country program (Burundi, Sri Lanka, Indonesia and Sudan). The paper aims to contribute to bridge the divide in the literature between guidelines, consensus & research and clinical practice in the field of psychosocial and mental health care in low- and middle-income countries.     

The effect of PGI2 and theophylline on the response of platelets subjected to shear stress

A specially designed rotational viscometer was used to investigate the effects of the antiplatelet agent PGI2 in combination with theophylline on the response of human platelets subjected to shear stress. Samples of citrated platelet-rich plasma (PRP) were exposed to shear stress in the viscometer for a period of 5 min at 23 degrees C. The levels of stress studied ranged from 50 to 300 dynes/sq cm. Pretreatment of the platelets with 0.01 microM PGI2 and 500 microM theophylline before exposure to shear stress caused a large reduction in shear-induced platelet aggregation. However, it was also observed that the PGI2-- theophylline pretreatment concomitantly caused a large increase in shear-induced platelet lysis and serotonin release at stress levels equal to or greater than 150 dynes/sq cm. This observed increase in platelet fragility may have important implications for clinical applications of PGI2. The results are discussed and compared to those obtained in prior work in which platelets were pretreated with acetylsalicylic acid or with PGE1.