Aberrant patterns of cellular communication in diabetes-induced embryopathy in rats: II, apoptotic pathways

OBJECTIVE: The objective was to test the hypothesis that hyperglycemia-induced injury of yolk sac cell membranes is associated with disruption of cellular apoptotic signaling pathways. STUDY DESIGN: Pregnant rats were induced to become diabetic by injection of streptozotocin. Fourteen normal control and 24 diabetic rats were killed on day 12 of gestation. Yolk sac membranes in 3 conceptus groups (nondiabetic, diabetic with normal, or diabetic with malformed conceptus) were collected for study. DNA was extracted from yolk sac cells and assayed for fragmentation by using gel electrophoresis, which indicates apoptosis. Protein expression was evaluated by Western blot assays. Statistical analyses were performed with the Student t -test. RESULTS: The level of phosphorylated Akt was significantly decreased, whereas that of the proapoptotic protein Bax was increased. These changes were correlated with the presence of DNA fragmentation in yolk sac cells of the diabetic malformed conceptuses. CONCLUSION: Hyperglycemia-induced embryopathy involves apoptosis, during which the expression of proapoptotic protein Bax is upregulated and the activity of the cell-survival factor, Akt kinase, is decreased in yolk sac cells. These observations suggest that hyperglycemia of maternal diabetes triggers apoptotic signaling pathways and inhibits cell survival pathways, leading to embryonic malformations.     

The effect of processing and cryopreservation on nucleated umbilical cord blood cells

Recovery of nucleated cord blood cells after storage in liquid nitrogen was evaluated. Red cells were depleted using Ficoll-Paque or Puregene red cell lyses. Freeze Medium contained 10% dimethylsulfoxide and 20% serum for cryoprotection. Recovery of the original cell population remaining serviceable for fluorescence activated cell sorting (FACS) was 12 +/- 10% (average +/- standard deviation), with a range of 1% to 55%. Viability measured by FACS analysis after freezing was significantly lower than that of the same specimens prior to freezing, 62 +/- 20% compared to 91+/-11% (p<0.001). Percentage CD45+34+ cells were the same for fresh and frozen cells. Gestational age at which specimens were collected had no effect on the percent cells carrying the CD45+34+ markers. We conclude that better cryoprotective supplements are needed to insure consistent high recovery of viable nucleated umbilical cord blood cells after preservation in liquid nitrogen.     

Identification to the species level and differentiation between strains of Aspergillus clinical isolates by automated repetitive-sequence-based PCR

A commercially available repetitive-sequence-based PCR (rep-PCR) DNA fingerprinting assay adapted to an automated format, the DiversiLab system, enables rapid microbial identification and strain typing. We explored the performance of the DiversiLab system as a molecular typing tool for 69 Aspergillus isolates (38 A. fumigatus, 15 A. flavus, and 16 A. terreus isolates) had been previously characterized by morphological analysis. Initially, 27 Aspergillus isolates (10 A. fumigatus, 9 A. flavus, and 8 A. terreus isolates) were used as controls to create a rep-PCR-based DNA fingerprint library with the DiversiLab software. Then, 42 blinded Aspergillus isolates were typed using the system. The rep-PCR-based profile revealed 98% concordance with morphology-based identification. rep-PCR-based DNA fingerprints were reproducible and were consistent for DNA from both hyphae and conidia. DiversiLab dendrogram reports correctly identified all A. fumigatus (n = 28), A. terreus (n = 8), and A. flavus (n = 6) isolates in the 42 blinded Aspergillus isolates. rep-PCR-based identification of all isolates was 100% in agreement with the contiguous internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) sequence-based identification of the respective isolates. Additionally, the DiversiLab system could demonstrate strain-level differentiation of A. flavus and A. terreus. Automated rep-PCR may be a time-efficient, effective, easy-to-use, novel genotyping tool for identifying and determining the strain relatedness of fungi. This system may be useful for epidemiological studies, molecular typing, and surveillance of Aspergillus species.     

A Study in Sexual Health Applying the Principles of Community-Based Participatory Research

The principles of community-based participatory research were applied to an exploratory sexual health study that examined "cruising for sex" among men on a college campus. In the context of a study seeking a broad interpretation of the health implications of cruising, and when faced with methodological challenges, the researchers found these principles to provide invaluable guidance. A review of the research process is offered and the manner in which the principles of community-based participatory research were operationalized for this study is described.     

Prevalence of Mycoplasma bacteria in amniotic fluid at the time of genetic amniocentesis using the polymerase chain reaction

OBJECTIVE: To use the polymerase chain reaction (PCR) to detect the presence of bacteria and Mycoplasma in amniotic fluid at the time of genetic amniocentesis and to assess their relationship to amniotic fluid interleukin-6 (IL-6) concentration and preterm delivery. STUDY DESIGN: A prospective study was performed on 78 asymptomatic pregnancies presenting for genetic amniocentesis. Amniotic fluid samples were analyzed by PCR for the detection of bacterial 16S ribosomal RNA, by PCR/enzyme-linked immunosorbent assay (ELISA) for Mycoplasma and by ELISA analysis for IL-6 concentration. RESULTS: All 78 samples were analyzed, and bacterial RNA was detected in 18% of them. However, no sample tested positive for Mycoplasma. The mean IL-6 concentration was 435 pg/mL. There was no difference in IL-6 levels or gestational age between bacteria-positive and -negative groups. CONCLUSION: Amniotic fluid may not be sterile in the midtrimester, yet the presence of bacteria, as detected by PCR, does not always result in an inflammatory response or adverse perinatal outcome.     

Testing of the PHS-ES: a measure of Perimenopausal Health Self-Efficacy

This article summarizes the development and psychometric analysis of the Perimenopausal Health Self-Efficacy Scale (PHS-ES) designed to assess women's health promotion self-efficacy related to mid-life changes in health. Items were generated from a qualitative study of HRT decision-making and recommended health promotion activities. The PHS-ES was administered 2 weeks apart to 98 university-based women ages 45 to 64 along with the measures of functional health status, stress, and the self-concept. Internal consistency (alpha = .88 and .90) and test-retest reliabilities (.86) were acceptable. Four factors emerged during factor analysis with 21 of the items explaining 50% of the variance and which were consistent with the conceptual basis of the PHS-ES. The PHS-ES was significantly correlated with functional health status, self-concept, stress, age and body mass index (BMI). In conjunction with stress and BMI, the PHS-ES predicted 50% of the variance of functional health. Further reliability and validity assessments are recommended with more racially and socioeconomically heterogeneous groups of perimenopausal women. It was concluded that the PHS-ES adequately demonstrated reliability and validity in this study.