Spontaneous Sleep-Like Brain State Alternations and Breathing Characteristics in Urethane Anesthetized Mice

Brain state alternations resembling those of sleep spontaneously occur in rats under urethane anesthesia and they are closely linked with sleep-like respiratory changes. Although rats are a common model for both sleep and respiratory physiology, we sought to determine if similar brain state and respiratory changes occur in mice under urethane. We made local field potential recordings from the hippocampus and measured respiratory activity by means of EMG recordings in intercostal, genioglossus, and abdominal muscles. Similar to results in adult rats, urethane anesthetized mice displayed quasi-periodic spontaneous forebrain state alternations between deactivated patterns resembling slow wave sleep (SWS) and activated patterns resembling rapid eye movement (REM) sleep. These alternations were associated with an increase in breathing rate, respiratory variability, a depression of inspiratory related activity in genioglossus muscle and an increase in expiratory-related abdominal muscle activity when comparing deactivated (SWS-like) to activated (REM-like) states. These results demonstrate that urethane anesthesia consistently induces sleep-like brain state alternations and correlated changes in respiratory activity across different rodent species. They open up the powerful possibility of utilizing transgenic mouse technology for the advancement and translation of knowledge regarding sleep cycle alternations and their impact on respiration.     

Myocardial preconditioning factors evoke mesenteric ischemic tolerance via opioid receptors and K(ATP) channels

We have shown that a reverse-phase concentrate generated from the effluent of preconditioned (PC) rabbit hearts evokes a cardioprotective effect in virgin acceptor hearts. With the use of a model of sustained (1 h) simulated ischemia in isolated, spontaneously contracting rabbit jejunum, our current aims were to 1) determine whether protective factor(s) released from PC hearts can improve ischemic tolerance in noncardiac tissue; and 2) obtain preliminary insight into the mediator(s) involved in triggering and eliciting this remote protection. Recovery of contractile force following reoxygenation (our index of ischemic tolerance) was enhanced in jejunal segments pretreated with concentrate generated from PC hearts (33 +/- 3% of baseline, P < 0.01) versus segments that received no concentrate (21 +/- 2%) and segments treated with concentrate from normoxic hearts (16 +/- 3%; P < 0.01). Protection achieved with PC concentrate was attenuated by coadministration of naloxone or glibenclamide, thereby implicating the involvement of opioids and ATP-sensitive potassium channels. Moreover, evaluation of purified subfractions of the crude PC concentrate identified a specific bioactive fraction that may participate in triggering the improved jejunal ischemic tolerance.     

Substituted indanylacetic acids as PPAR-alpha-gamma activators

A series of oxazole-substituted indanylacetic acids were prepared which show a spectrum of activity as ligands for PPAR nuclear receptor subtypes.     

Cloning and expression of human nebulin cDNAs and assignment of the gene to chromosome 2q31-q32

We have isolated two nonoverlapping cDNAs encoding human nebulin, a muscle-specific protein. Northern hybridization analysis shows that nebulin is encoded by a huge message at least 25 kb in length. By hybridizing two nonoverlapping cDNAs to DNA isolated from rodent X human cell hybrids, we assign this presumably single-copy gene to human chromosome 2; sublocalization studies indicate that the nebulin gene is on the long arm of the chromosome, in the region 2q31-q32.     

Microbial growth associated with granular activated carbon in a pilot water treatment facility

The microbial dynamics associated with granular activated carbon (GAC) in a pilot water treatment plant were investigated over a period of 16 months. Microbial populations were monitored in the influent and effluent waters and on the GAC particles by means of total plate counts and ATP assays. Microbial populations between the influent and effluent waters of the GAC columns generally increased, indicating microbial growth. The dominant genera of microorganisms isolated from interstitial waters and GAC particles were Achromobacter, Acinetobacter, Aeromonas, Alcaligenes, Bacillus, Chromobacterium, Corynebacterium, Micrococcus, Microcyclus, Paracoccus, and Pseudomonas. Coliform bacteria were found in small numbers in the effluents from some of the GAC columns in the later months of the study. Oxidation of influent waters with ozone and maintenance of aerobic conditions on the GAC columns failed to appreciably enhance the microbial growth on GAC.     

Approaches to sequence analysis of 125I-labeled RNA.

A method is described for the initial steps of sequence analysis of RNase T1-and pancreatic RN-ase-resistant oligonucleotides of RNA containing cytidylate residues labeled in vitro with 125I. In many cases an oligonucleotide sequence can be deduced from a consideration of (i) its relative position in the two-dimensional fingerprint (with DEAE thin layer homochromatographic second dimension), (ii) its electrophoretic mobility on DEAE paper at pH 1.9, and (iii) identification of its products of further enzymatic digestion by comparison with a set of marker oligonucleotides. Additional methods including analysis of oligonucleotides following chemical blocking of uridylate residues with CMCT and analysis of products of incomplete enzymatic digestion are also discussed.     

Pasteuria sp. Parasitizing Trophonema okamotoi in Florida

Two populations of Trophonema okamotoi parasitized by Pasteuria sp. were found on Liquidambar styraciflua (sweetgum) and on an unidentified tropical grass in north-central Florida. Endospores of this Pasteuria sp. attached to motile vermiform second-stage juveniles (J2) and males of T. okamotoi, but not to other developmental stages. Sporangia and new endospores were produced only inside the bodies of swollen and sedentary third- and fourth-stage juveniles and females that developed in the host roots. No egg masses were produced by infected T. okamotoi females. The endospore diameter from the tropical grass population was 4.93 μm and the central core diameter was 1.97 μm; measurements of endospores from the sweetgum populations were similar. Endospores that were collected from T. okamotoi and added to uninfected T. okamotoi and other plant-parasitic nematodes attached/to J2 of T. okamotoi but did not attach to juveniles and adults of Helicotylenchus pseudorotrustus, Pratylenchus brachyurus, or to J2 of either Meloidogyne arenaria race 1, M. incognita race 1, M. javanica, or Tylenchulus semipenetrans. Pasteuria sp. from T. okamotoi differed from the described Pasteuria species in endospore size, host preference, and rate of attachment.     

The deubiquitinating enzyme USP37 enhances CHK1 activity to promote the cellular response to replication stress

The deubiquitinating enzyme USP37 is known to contribute to timely onset of S phase and progression of mitosis. However, it is not clear if USP37 is required beyond S-phase entry despite expression and activity of USP37 peaking within S phase. We have utilized flow cytometry and microscopy to analyze populations of replicating cells labeled with thymidine analogs and monitored mitotic entry in synchronized cells to determine that USP37-depleted cells exhibited altered S-phase kinetics. Further analysis revealed that cells depleted of USP37 harbored increased levels of the replication stress and DNA damage markers γH2AX and 53BP1 in response to perturbed replication. Depletion of USP37 also reduced cellular proliferation and led to increased sensitivity to agents that induce replication stress. Underlying the increased sensitivity, we found that the checkpoint kinase 1 is destabilized in the absence of USP37, attenuating its function. We further demonstrated that USP37 deubiquitinates checkpoint kinase 1, promoting its stability. Together, our results establish that USP37 is required beyond S-phase entry to promote the efficiency and fidelity of replication. These data further define the role of USP37 in the regulation of cell proliferation and contribute to an evolving understanding of USP37 as a multifaceted regulator of genome stability.