Hippocampal ceRNA networks from chronic intermittent ethanol vapor-exposed male mice and functional analysis of top-ranked lncRNA genes for ethanol drinking phenotypes.

The molecular mechanisms regulating the development and progression of alcohol use disorder (AUD) are largely unknown. While noncoding RNAs have previously been implicated as playing key roles in AUD, long-noncoding RNA (lncRNA) remains understudied in relation to AUD. In this study, we first identified ethanol-responsive lncRNAs in the mouse hippocampus that are transcriptional network hub genes. Microarray analysis of lncRNA, miRNA, circular RNA, and protein coding gene expression in the hippocampus from chronic intermittent ethanol vapor- or air- (control) exposed mice was used to identify ethanol-responsive competing endogenous RNA (ceRNA) networks. Highly interconnected lncRNAs (genes that had the strongest overall correlation to all other dysregulated genes identified) were ranked. The top four lncRNAs were novel, previously uncharacterized genes named Gm42575, 4930413E15Rik, Gm15767, and Gm33447, hereafter referred to as Pitt1, Pitt2, Pitt3, and Pitt4, respectively. We subsequently tested the hypothesis that CRISPR/Cas9 mutagenesis of the putative promoter and first exon of these lncRNAs in C57BL/6J mice would alter ethanol drinking behavior. The Drinking in the Dark (DID) assay was used to examine binge-like drinking behavior, and the Every-Other-Day Two-Bottle Choice (EOD-2BC) assay was used to examine intermittent ethanol consumption and preference. No significant differences between control and mutant mice were observed in the DID assay. Female-specific reductions in ethanol consumption were observed in the EOD-2BC assay for Pitt1, Pitt3, and Pitt4 mutant mice compared to controls. Male-specific alterations in ethanol preference were observed for Pitt1 and Pitt2. Female-specific increases in ethanol preference were observed for Pitt3 and Pitt4. Total fluid consumption was reduced in Pitt1 and Pitt2 mutants at 15% v/v ethanol and in Pitt3 and Pitt4 at 20% v/v ethanol in females only. We conclude that all lncRNAs targeted altered ethanol drinking behavior, and that lncRNAs Pitt1, Pitt3, and Pitt4 influenced ethanol consumption in a sex-specific manner. Further research is necessary to elucidate the biological mechanisms for these effects. These findings add to the literature implicating noncoding RNAs in AUD and suggest lncRNAs also play an important regulatory role in the disease.

Factors influencing choice between tracheostomy and prolonged translaryngeal intubation in acute respiratory failure: a prospective study.

One of the problems of prolonged ventilatory therapy in acute respiratory failure (ARF) is the need to choose between tracheostomy after 48 to 72 hours of translaryngeal (TL) tracheal intubation or the continuous use of the TL tube for a period of 10 days. Too often the choice has been based on retrospective studies or personal preference. To investigate this problem prospectively, 52 adults in ARF were divided sequentially into 2 groups on their 3rd day of TL intubation. Patients in group I (G-I) retained the TL tube for a total of 11 days; those in group II (G-II) were tracheostomized on the 3rd day. The following factors ere used to evaluate the efficiency and complications in each group: patient's epidemiologic variables, daily pulmonary functions, severity of respiratory infections, and scores of post-intubation airway lesions. No consistent statistically significant differences between the two procedures were seen in the pulmonary functions or the range of individual patient variables. However, with an early tracheostomy, there was an eightfold greater incidence of contamination of the airway by new organisms, airway lesions were more frequent and severe, and the need for the tracheal tube was extended. To identify the epidemiologic variables and the pulmonary functions that discriminate between patients with serious airway lesions and those with mild lesions, and to evaluate the ability of these variables to differentiate the patients who died from those who survived, the distribution of all factors was compared in the two categories. The epidemiologic variables separated the patients according to their airway lesions only, while the difference in pulmonary functions was statistically significant only between the patients who died and those who survived.

Cardiac arrest related to anesthesia. Contributing factors in infants and children.

A collaborative retrospective study undertaken to investigate cardiac arrest related to pediatric anesthesia in seven institutions between 1960 and 1972 showed 73 instances in which anesthesia was thought to have been either directly responsible or had played an important contributing role. About two thirds of these patients were successfully resuscitated. Cases were found to fit into one of two major categories: cardiovascular and respiratory. Among cardiovascular factors, blood loss, preoperative anemia, inappropriate administration of succinylcholine, and accidental administration of potassium were important contributing causes. Respiratory factors included failure to maintain a patent airway and ventilatory problems. In retrospect, most of these accidents were preventable. Such information should indicate where research emphasis needs to be placed and that our current methods of teaching and training need to be reevaluated.