NOTCH1 Gain of Function in Germ Cells Causes Failure of Spermatogenesis in Male Mice

NOTCH1 is a member of the NOTCH receptor family, a group of single-pass trans-membrane receptors. NOTCH signaling is highly conserved in evolution and mediates communication between adjacent cells. NOTCH receptors have been implicated in cell fate determination, as well as maintenance and differentiation of stem cells. In the mammalian testis expression of NOTCH1 in somatic and germ cells has been demonstrated, however its role in spermatogenesis was not clear. To study the significance of NOTCH1 in germ cells, we applied a cre/loxP approach in mice to induce NOTCH1 gain- or loss-of function specifically in male germ cells. Using a Stra8-icretransgene we produced mice with conditional activation of the NOTCH1 intracellular domain (NICD) in germ cells. Spermatogenesis in these mutants was progressively affected with age, resulting in decreased testis weight and sperm count. Analysis of downstream target genes of NOTCH1 signaling showed an increased expression of Hes5, with a reduction of the spermatogonial differentiation marker, Neurog3 expression in the mutant testis. Apoptosis was significantly increased in mouse germ cells with the corresponding elevation of pro-apoptotic Trp53 and Trp63genes' expression. We also showed that the conditional germ cell-specific ablation of Notch1 had no effect on spermatogenesis or male fertility. Our data suggest the importance of NOTCH signaling regulation in male germ cells for their survival and differentiation.

Molecular Mechanisms Involved in Regulating the Mucoid-to-Nonmucoid Conversion of Pseudomonas aeruginosa Associated with Cystic Fibrosis Patients

Pseudomonas aeruginosa is an opportunistic pathogen that has received attention because of its close association with cystic fibrosis (CF). Chronic pulmonary infection with the mucoid P. aeruginosa is the leading cause of mortality in CF patients. This bacterium has the ability to sense and adapt to the harsh environment in the CF lung by converting from a nonmucoid to a mucoid state. The mucoid phenotype is caused by overproduction of a polysaccharide called alginate. Alginate production is regulated by the algT/U operon containing five genes, algT/U-mucA-mucB-mucC-mucD. Alginate overproduction in CF isolates has been partially attributed to a loss-of-function mutation in mucA that results in the overexpression of algT. This mucoid phenotype is unstable, reverting to the nonmucoid form when the isolates are cultured outside of the CF lung. This study was undertaken to determine the mechanisms involved in the conversion from the mucoid to the nonmucoid form. Thirty-six spontaneous nonmucoid variants of a known mucoid isolate with a mucA mutation were analyzed. Ten of these isolates were complemented in trans by plasmids containing the algT operon and the algT gene. Chromosomal DNA was extracted and the mucA and algT genes were amplified by the polymerase chain reaction. Sequence analysis of the genes showed that these mutants retained the original mucA mutation but acquired secondary mutations in the algT gene.

Loss of electricity and refrigerated foods: avoiding the danger zone

Recent events such as Winter Storm [Hurricane] Sandy and Hurricane Katrina have demonstrated that local food supplies must last as long as possible. Current recommendations are to dispose of all refrigerated food four hours after the power is lost. The purpose of this study was to determine if it is possible to safely hold food longer than four hours without power. The results indicate that the food can be held for up to six hours if the door is not opened. If ice is added to the refrigerator, then it will take the food approximately 10 hours to reach 5°C (41°F).

Does Obesity Affect Outcomes in Children Admitted from Trauma Centers?

Introduction and Research Objectives: Pediatric obesity has reached epidemic proportions in the United States. In the critical care setting, obesity has yet to be fully studied. We sought to evaluate the effects of obesity in children who are admitted to a hospital from trauma centers using Kid's Inpatient Database (KID) during 2009. Methods: The study examined inpatient admissions from pediatric trauma patients in 2009 using the Kids´ Inpatient Database (KID). Patients (n=27599) were selected from the KID based on Age (AGE>1) and Admission Type (ATYPE=5) and assessed on Race, Sex, Length of Stay (LOS), Number of Diagnoses and Procedures, Severity of Illness (SOI), Risk of Mortality (ROM), Co-morbidities, and Intubation by comparing obese and non-obese cohorts. Chi-square test and student t-test were used to analyze the data. All variables were weighted to get national estimates. Results: The overall prevalence of obesity (those coded as having obesity as co-morbidity) was 1.6% with significantly higher prevalence among Blacks (1.8%), Hispanics (2.3%), and Native Americans (4.1%; p<0.001). Obesity was more prevalent among females (2.4% vs 1.2%; p<.001). Overall mortality in the cohort was 4.8%. Obesity was significantly lower among children who died during hospitalization (0.5% vs 1.6%; p<0.002). However, obese children had significantly longer LOS, greater number of diagnoses, more procedures and greater than expected loss of function due to SOI when compared with nonobese cohort (p<.001). Deficiency anemia, diabetes, hypertension, liver disease, and fluid and electrolyte disorders are all strongly associated with the presence of obesity (p<.005). The rate of intubation is similar between obese and non-obese cohorts. Conclusion: Our study using KID national database found that obese children who are admitted from trauma centers have a higher morbidity and LOS but lower mortality. Racial and gender inequalities of obesity prevalence is consistent with previous reports.