Increasing the Effectiveness of Targeted Temperature Management.

TOPIC: Targeted temperature management and therapeutic hypothermia are essential components of the multimodal approach to caring for compromised patients after cardiac arrest and severe traumatic brain injury. CLINICAL RELEVANCE: The continuously evolving science necessitates summation of individual facets and concepts to enhance knowledge and application for optimally delivering care. Targeted temperature management is a complex therapy that requires fine-tuning the most effective interventions to maintain high-quality targeted temperature management and maximize patient outcomes. PURPOSE: To describe the underlying pathophysiology of fever and the importance of manipulating water temperature and of preventing and treating shivering during that process. CONTENT COVERED: This article discusses nursing considerations regarding the care of patients requiring targeted temperature management that are necessary to improve patient outcomes.

Prevalence of maternal cell contamination in amniotic fluid samples.

PURPOSE: The purpose of this study is to evaluate the incidence of maternal cell contamination (MCC) in the first few milliliters of amniotic fluid withdrawn during amniocentesis. METHODS: A prospective observational study was performed. The initial 2-3 ml of amniotic fluid withdrawn during amniocentesis was divided into direct analysis (uncultured) and cultured samples. A matching maternal buccal swab was obtained for MCC testing. MCC was determined by short-tandem repeat analysis. The primary outcome was measurement of clinically significant contamination (MCC >5%). Secondary outcomes included the determination of risk factors associated with MCC >5%. Outcomes were assessed by fisher's exact, independent t-test, binary logistic regression, and ANOVA. RESULTS: Direct analysis measured clinically significant contamination (MCC > 5%) in 26% of specimens, while any amount of MCC was present in 68% of specimens. Cultured specimens had MCC > 5% in 2%, and any amount of MCC in 24%. Only blood-tinged fluid was associated with an increased risk for MCC > 5%. Larger volumes of the discard sample were not associated with increased incidence of MCC greater than 5%. CONCLUSION: A significant amount of MCC is present with direct analysis of the initial few milliliters of amniotic fluid withdrawn and is not influenced by the volume of the discard sample. Our results suggest that the first few milliliters of amniotic fluid be removed and discarded when direct analysis is utilized for prenatal genetic testing.

The C. elegans DSB-2 protein reveals a regulatory network that controls competence for meiotic DSB formation and promotes crossover assurance.

For most organisms, chromosome segregation during meiosis relies on deliberate induction of DNA double-strand breaks (DSBs) and repair of a subset of these DSBs as inter-homolog crossovers (COs). However, timing and levels of DSB formation must be tightly controlled to avoid jeopardizing genome integrity. Here we identify the DSB-2 protein, which is required for efficient DSB formation during C. elegans meiosis but is dispensable for later steps of meiotic recombination. DSB-2 localizes to chromatin during the time of DSB formation, and its disappearance coincides with a decline in RAD-51 foci marking early recombination intermediates and precedes appearance of COSA-1 foci marking CO-designated sites. These and other data suggest that DSB-2 and its paralog DSB-1 promote competence for DSB formation. Further, immunofluorescence analyses of wild-type gonads and various meiotic mutants reveal that association of DSB-2 with chromatin is coordinated with multiple distinct aspects of the meiotic program, including the phosphorylation state of nuclear envelope protein SUN-1 and dependence on RAD-50 to load the RAD-51 recombinase at DSB sites. Moreover, association of DSB-2 with chromatin is prolonged in mutants impaired for either DSB formation or formation of downstream CO intermediates. These and other data suggest that association of DSB-2 with chromatin is an indicator of competence for DSB formation, and that cells respond to a deficit of CO-competent recombination intermediates by prolonging the DSB-competent state. In the context of this model, we propose that formation of sufficient CO-competent intermediates engages a negative feedback response that leads to cessation of DSB formation as part of a major coordinated transition in meiotic prophase progression. The proposed negative feedback regulation of DSB formation simultaneously (1) ensures that sufficient DSBs are made to guarantee CO formation and (2) prevents excessive DSB levels that could have deleterious effects.

Positive cell-free fetal DNA testing for trisomy 13 reveals confined placental mosaicism.

PURPOSE: We report on a case in which cell-free fetal DNA was positive for trisomy 13 most likely due to confined placental mosaicism. Cell-free fetal DNA testing analyzes DNA derived from placental trophoblast cells and can lead to incorrect results that are not representative of the fetus. METHODS: We sought to confirm commercial cell-free fetal DNA testing results by chorionic villus sampling and amniocentesis. These results were followed up by postnatal chromosome analysis of cord blood and placental tissue. RESULTS: First-trimester cell-free fetal DNA test results were positive for trisomy 13. Cytogenetic analysis of chorionic villus sampling yielded a mosaic karyotype of 47,XY,+13[10]/46,XY[12]. G-banded analysis of amniotic fluid was normal, 46,XY. Postnatal cytogenetic analysis of cord blood was normal. Karyotyping of tissues from four quadrants of the placenta demonstrated mosaicism for trisomy 13 in two of the quadrants and a normal karyotype in the other two. CONCLUSION: Our case illustrates several important aspects of this new testing methodology: that cell-free fetal DNA may not be representative of the fetal karyotype; that follow-up with diagnostic testing of chorionic villus sampling and/or amniotic fluid for abnormal test results should be performed; and that pretest counseling regarding the full benefits, limitations, and possible testing outcomes of cell-free fetal DNA screening is important.

Prevalence of hypertension and diabetes among Ethiopian adults.

OBJECTIVE: To determine the prevalence of hypertension and diabetes among members of an Ethiopian occupational cohort; and to examine the proportion of adults who were aware of their conditions. METHODS: A total of 2153 of subjects were included in this cross-sectional study. The World Health Organization STEPwise approach for non-communicable diseases was used to collect socio-demographic data, blood pressure measures and blood samples from participants. Prevalence estimates for hypertension and diabetes were determined separately. The 95% confidence intervals for prevalence estimates were also determined. RESULTS: The overall prevalence of hypertension was 19.1% (95%CI: 17.1-20.8) and 22% (95%CI: 20.2-23.8) and 14.9% (95%CI: 13.4-16.4) among men and women respectively. The overall prevalence of diabetes was 6.5% (95%CI: 5.4-7.6) and 6.4% (95%CI: 5.0-7.8) and 6.6% (95%CI: 4.8-8.4) among men and women correspondingly. Notably, 15% of hypertensives reported never having had their blood pressure checked prior to the present study examination. Approximately 45% of participants who had their blood pressure checked were never diagnosed with hypertension, but were found to be hypertensive in our study. Approximately 27% of newly diagnosed diabetics (during this study) reported never having a previous blood glucose test. Among those who had their blood glucose assessed prior to this study, 17.4% were found to have diabetes but were never diagnosed. CONCLUSION: The high prevalence of hypertension and diabetes reported in our study confirms findings from other Sub-Saharan Africa countries, and extends the literature to urban dwelling Ethiopians where non-communicable diseases are emerging as a major public health concern.

Interphase FISH demonstrates that human adipose stromal cells maintain a high level of genomic stability in long-term culture.

Human adipose stromal cells (ASCs) reside within the stromal-vascular fraction (SVF) in fat tissue, can be readily isolated, and include stem-like cells that may be useful for therapy. An important consideration for clinical application and functional studies of stem/progenitor cells is their capacity to maintain chromosome stability in culture. In this study, cultured ASC populations and ASC clones were evaluated at intervals for maintenance of chromosome stability. Uncultured SVF (uSVF) cells were included for comparison. G-banded chromosome analysis demonstrated that ASCs are diploid and have a normal karyotype. However since only approximately 20 cells are examined, low levels of chromosome instability would not be detected. To increase detection sensitivity, fluorescence in situ hybridization was employed, to permit chromosome enumeration in larger numbers of interphase cells. Seven cultured ASC populations, two ASC clones and four uSVF samples were examined. Chromosome X and 17 probes identified diploid, tetraploid, and aneuploid interphase cells. Both cultured ASC populations [up to approximately 35 Population Doublings (PDs)] and uSVF cells exhibited a similar level of diploidy (97.8% n = 6,355 and 98.83% n = 1,197, respectively) and numerical abnormalities, suggesting that cultured ASCs are genomically stable and supporting their suitability for transplantation applications. In comparison, cultured primary human chorionic villus cells exhibited marked genomic instability resulting in an 11.6% tetraploidy rate after 8-10 PD. Thus effects of culture on genomic stability may be cell type dependent and should be tested by appropriately scaled interphase fluorescence in situ hybridization analysis in any ex vivo expanded cell population destined for transplantation.

Semen analysis: the test techs love to hate.

At the heart of better semen analysis is professionalism. The walls of many labs are covered with slogans like, "at the end of every test is a worried patient who needs an answer". Semen analysis is no different. At its end is a couple desperate to have a child or start a family. In spite of the importance of semen analysis in fertility diagnosis and treatment, it remains in most clinical laboratories "the neglected laboratory test". The tips and recommendations in this article should help any lab improve the quality of semen analysis while reducing the effort required to produce better results. Knowledge and simple, repeatable procedures, combined with QC and competency benchmarks, can put the interest and satisfaction back into a test that is the gateway for fertility treatment. After all, what is not to love about a cell that swims and comes in so many interesting shapes?

Comparative pharmacodynamics of garenoxacin, gemifloxacin, and moxifloxacin in community-acquired pneumonia caused by Streptococcus pneumoniae: a Monte Carlo simulation analysis.

OBJECTIVE: This study used Monte Carlo simulations to assess the potential for attainment of pharmacodynamic targets with the fluoroquinolones garenoxacin, gemifloxacin, and moxifloxacin against Streptococcus pneumoniae in serum and epithelial lining fluid (ELF) from hospitalized patients with community-acquired pneumonia (CAP). METHODS: Data on the free AUC over 24 hours (fAUC(0-24)), a measure of drug exposure, were derived from previously described population pharmacokinetic models for therapeutic doses of the 3 fluoroquinolones. MIC distribution data for S pneumoniae were obtained from the Canadian Respiratory Organism Susceptibility Study. These data were used to produce the ratio of fAUC(0-24) to the MIC(90) (fAUC(0-24)/MIC(90)), a pharmacodynamic predictor of bacterial eradication. Monte Carlo simulations were used to analyze the potential for garenoxacin 400 mg QD, gemifloxacin 320 mg QD, and moxifloxacin 400 mg QD to achieve target fAUC(0-24)/MIC(90) ratios of 30, 40, 100, and 120 against S pneumoniae in serum and ELF from hospitalized patients with CAP. Target ratios of 30 and 40 were used to assess the probability of bacterial eradication, while ratios of 100 and 120 were used to assess the probability of preventing development of resistance. RESULTS: Monte Carlo simulations indicated that all 3 fluoroquinolones had a high probability (>90%) of attaining target fAUC(0-24)/MIC(90) ratios of 30 and 40 against S pneumoniae in both serum and ELF. Garenoxacin 400 mg QD was associated with a >95% probability of achieving target fAUC(0-24)/MIC(90) ratios of 100 and 120 in both serum and ELF. Both gemifloxacin 320 mg QD and moxifloxacin 400 mg QD were associated with high probabilities of attaining fAUC(0-24)/MIC(90) ratios of 100 and 120 in ELF (>95%); the probability of gemifloxacin and moxifloxacin attaining these targets in serum ranged from 78.3% to 88.0%. CONCLUSION: Based on these simulations, garenoxacin 400 mg QD, gemifloxacin 320 mg QD, and moxifloxacin 400 mg QD appeared likely to achieve target serum and ELF concentrations against S pneumoniae in hospitalized patients with CAP, with a low potential to select for resistance.