EEG and behavioral effects of gamma-hydroxybutyrate in the rabbit

The effects of gamma-hydroxybutyrate (GHB) upon sleep wakefulness patterns and quantified nuchal muscle activity were examined in the rabbit in a dose-response paradigm (25–1,000 mg/kg). Relative to control (saline) values, there was no facilitation of sleep onset or epileptogenic activity at any of the dosages studied. However, at the higher GHB concentrations, slow wave sleep and tonic muscle activity were enhanced and a high amplitude, slow activity was superimposed on background EEG patterns. The highest concentration of GHB (1,000 mg/kg) was associated with depression of motor activity. An enhancement of paradoxical sleep observed at lower GHB levels in other species occured in attenuated form in the rabbit. The results indicate dose-related effects on both sleep and motor activation in the rabbit, but the absence of seizure activity for the concentrations of GHB studied.     

Phylogenetic diversification of immunoglobulin genes and the antibody repertoire

Immunoglobulins are encoded by a large multigene system that undergoes somatic rearrangement and additional genetic change during the development of immunoglobulin-producing cells. Inducible antibody and antibody-like responses are found in all vertebrates. However, immunoglobulin possessing disulfide-bonded heavy and light chains and domain-type organization has been described only in representatives of the jawed vertebrates. High degrees of nucleotide and predicted amino acid sequence identity are evident when the segmental elements that constitute the immunoglobulin gene loci in phylogenetically divergent vertebrates are compared. However, the organization of gene loci and the manner in which the independent elements recombine (and diversify) vary markedly among different taxa. One striking pattern of gene organization is the "cluster type" that appears to be restricted to the chondrichthyes (cartilaginous fishes) and limits segmental rearrangement to closely linked elements. This type of gene organization is associated with both heavy- and light-chain gene loci. In some cases, the clusters are "joined" or "partially joined" in the germ line, in effect predetermining or partially predetermining, respectively, the encoded specificities (the assumption being that these are expressed) of the individual loci. By relating the sequences of transcribed gene products to their respective germ-line genes, it is evident that, in some cases, joined-type genes are expressed. This raises a question about the existence and/or nature of allelic exclusion in these species. The extensive variation in gene organization found throughout the vertebrate species may relate directly to the role of intersegmental (V<==>D<==>J) distances in the commitment of the individual antibody-producing cell to a particular genetic specificity. Thus, the evolution of this locus, perhaps more so than that of others, may reflect the interrelationships between genetic organization and function.      

Characterization of a stable,anchorage-dependent clone obtained from a spontaneously transformed mouse cell line

Summary A variant nontransformed clone, I21, was selected from the spontaneously transformed mouse fibroblast line, IT22. Selection was done by plating IT22 in methylcellulose and picking single cells after 2 d. Cultures derived from these single cells were selected again and one clone, I21, derived from the second round of selection was characterized extensively. I21 and IT22 have the same plating efficiency (PE) on plastic, but in agarose they differ by 1000-fold. In comparison to IT22, I21 has a normal morphological appearance, a lower saturation density, a higher viability in stationary phase, an increased doubling time, an increased chromosome content, and is unable to form tumors in nude mice. I21 has remained remarkably stable in culture and has not reverted to the transformed phenotype for at least 300 generations in culture. Over 100 clones of I21, expanded to 10⁶ cells, failed to show an increased PE in agarose. Even expansion of the rare colonies of I21 that grow in agarose failed to produce clones similar to IT22. The research was supported by the Medical Research Council and the National Cancer Institute of Canada. R. Godbout was supported by a 1967 Science Scholarship and by an MRC Studentship. B. L. Gallie is a Research Associate of the Ontario Cancer Treatment and Research Foundation.     

Molecular Mechanism by Which a Potent Hepatitis C Virus NS3-NS4A Protease Inhibitor Overcomes Emergence of Resistance

Although optimizing the resistance profile of an inhibitor can be challenging, it is potentially important for improving the long term effectiveness of antiviral therapy. This work describes our rational approach toward the identification of a macrocyclic acylsulfonamide that is a potent inhibitor of the NS3-NS4A proteases of all hepatitis C virus genotypes and of a panel of genotype 1-resistant variants. The enhanced potency of this compound versus variants D168V and R155K facilitated x-ray determination of the inhibitor-variant complexes. In turn, these structural studies revealed a complex molecular basis of resistance and rationalized how such compounds are able to circumvent these mechanisms.     

Daily Changes of Plasma Corticosterone by an 8-h Daytime Feeding Occur Without Body Weight Loss or Severe Food Restriction in the Rat

Different studies have reported that daytime feeding entrains the circadian rhythm of corticosterone secretion in rats. However, it remained unclear whether calorie restriction or daytime feeding access have an effect. The aim of our study is to evaluate the effect of an 8-h daytime feeding access on the circadian rhythm of plasma corticosterone. Eleven adult male Wistar rats were assigned to two different conditions of access to food: ad lib feeding for one week and daytime feeding for the following two weeks. On the 7th, 14th and 21st day, blood samples were collected every 4 h from 08:00 to 04:00. Food intake and body weight were recorded daily. During daytime feeding, rats ingested 88% of the amount of food ingested over 24 h in the ad lib feeding period. However, body weight increased significantly from the first day to the end of experiment. Peak plasma corticosterone was 12 h shifted during daytime feeding period compared to the ad lib condition. This study showed that an 8-h daytime feeding entrained the circadian rhythm of plasma corticosterone without body weight loss or severe food restriction.     

Alternative splicing modulates Disabled-1 (Dab1) function in the developing chick retina

The Reelin-Disabled 1 (Dab1)-signaling pathway plays a critical role in neuronal cell positioning in the brain. We have isolated two alternatively spliced variants of Dab1 from chick retina, an early form (chDab1-E) expressed in undifferentiated cells and a late form (chDab1-L) expressed in amacrine and ganglion cells. A key difference between the two forms is the exclusion in chDab1-E of two Src-related tyrosine kinase recognition sites implicated in Reelin-mediated Dab1 tyrosine phosphorylation. Retinal cultures transfected with a chDab1-L expression construct undergo a dramatic change in morphology, accompanied by the formation of numerous thin elongated processes, increased tyrosine phosphorylation, activation of Src family kinase(s) and increased levels of the axonal outgrowth protein growth-associated protein-43. In contrast, chDab1-E transfectants retain an undifferentiated morphology. Mutational analysis implicates a specific tyrosine (tyr-198) in the morphological and biochemical alterations associated with chDab1-L expression. We propose that alternative splicing of chDab1 represents an effective and flexible way of regulating the Reelin-Dab1-signaling pathway in a mixed cell population, by ensuring that secreted Reelin activates the signaling cascade only in target neuronal cells.     

What can we learn from noncoding regions of similarity between genomes?

Background  

In addition to known protein-coding genes, large amounts of apparently non-coding sequence are conserved between the human and mouse genomes. It seems reasonable to assume that these conserved regions are more likely to contain functional elements than less-conserved portions of the genome.