Temporal Control of Metabolic Amplitude by Nocturnin

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Cytochrome c activation of CPP32-like proteolysis plays a critical role in a Xenopus cell-free apoptosis system.

In a cell-free system based on Xenopus egg extracts, Bcl-2 blocks apoptotic activity by preventing cytochrome c release from mitochondria. We now describe in detail the crucial role of cytochrome c in this system. The mitochondrial fraction, when incubated with cytosol, releases cytochrome c. Cytochrome c in turn induces the activation of protease(s) resembling caspase-3 (CPP32), leading to downstream apoptotic events, including the cleavage of fodrin and lamin B1. CPP32-like protease activity plays an essential role in this system, as the caspase inhibitor, Ac-DEVD-CHO, strongly inhibited fodrin and lamin B1 cleavage, as well as nuclear morphology changes. Cytochrome c preparations from various vertebrate species, but not from Saccharomyces cerevisiae, were able to initiate all signs of apoptosis. Cytochrome c by itself was unable to process the precursor form of CPP32; the presence of cytosol was required. The electron transport activity of cytochrome c is not required for its pro-apoptotic function, as Cu- and Zn-substituted cytochrome c had strong pro-apoptotic activity, despite being redox-inactive. However, certain structural features of the molecule were required for this activity. Thus, in the Xenopus cell-free system, cytosol-dependent mitochondrial release of cytochrome c induces apoptosis by activating CPP32-like caspases, via unknown cytosolic factors.     

Physical mapping of the HOXA1 gene and the hnRPA2B1 gene in a YAC contig from human chromosome 7p14-p15

A cluster of homeobox-containing genes (HOXA) and a heterogeneous nuclear ribonucleoprotein (hnRPA2B1) have both previously been assigned to chromosome 7p15 by in situ hybridization. In this report, we constructed a YAC contig from chromosome 7p14-p15, between markers D7S2496 and D7S1838, and determined the position of the HOXA1 gene and the hnRPA2B1 gene in this YAC contig. Received: 19 November 1996 / Revised: 2 January 1997     

Localization and Characterization of the Human ADP-Ribosylation Factor 5 (ARF5) Gene

ADP-ribosylation factor 5 (ARF5) is a member of the ARF gene family. The ARF proteins stimulate thein vitroADP-ribosyltransferase activity of cholera toxin and appear to play a role in vesicular traffickingin vivo.We have mapped ARF5, one of the six known mammalian ARF genes, to a well-defined yeast artificial chromosome contig on human chromosome 7q31.3. In addition, we have isolated and sequenced an 3.2-kb genomic segment that contains the entire ARF5 coding region, revealing the complete intron–exon structure of the gene. With six coding exons and five introns, the genomic structure of ARF5 is unique among the mammalian ARF genes and provides insight about the evolutionary history of this ancient gene family.     

The X-linked gene G4.5 is responsible for different infantile dilated cardiomyopathies.

Barth syndrome (BTHS) is an X-linked disorder characterized clinically by the associated features of cardiac and skeletal myopathy, short stature, and neutropenia. The clinical manifestations of the disease are, in general, quite variable, but cardiac failure as a consequence of cardiac dilatation and hypertrophy is a constant finding and is the most common cause of death in the first months of life. X-linked cardiomyopathies with clinical manifestations similar to BTHS have been reported, and it has been proposed that they may be allelic. We have recently identified the gene responsible for BTHS, in one of the Xq28 genes, G4.5. In this paper we report the sequence analysis of 11 additional familial cases: 8 were diagnosed as possibly affected with BTHS, and 3 were affected with X-linked dilated cardiomyopathies. Mutations in the G4.5 gene were found in nine of the patients analyzed. The molecular studies have linked together what were formerly considered different conditions and have shown that the G4.5 gene is responsible for BTHS (OMIM 302060), X-linked endocardial fibroelastosis (OMIM 305300), and severe X-linked cardiomyopathy (OMIM 300069). Our results also suggest that very severe phenotypes may be associated with null mutations in the gene, whereas mutations in alternative portions or missense mutations may give a "less severe" phenotype.     

Do secondary substances in the thallus of a lichen promote CO2 diffusion and prevent depression of net photosynthesis at high water content?

Many lichens show seriously depressed net photosynthesis (NP) at high thallus water contents due to increased carbon dioxide diffusion resistance through blockage of diffusion pathways by water. The soil lichen Diploschistes muscorum, however, shows no depression and NP is close to maximal even at the highest thallus water content. We investigated whether lichen substances (lecanoric and diploschistesic acids) in the cortex and medulla contributed to this ability to maintain high NP. Dry thalli were extracted with water-free acetone and, after this treatment, were found to be fully viable to the extent of continued growth after replanting in the field. No differences were found in the response of NP to thallus water content between the normal and extracted thalli, in fact the response curves were often nearly identical. Thus, in this species it seems that lichen substances did not maintain the water-free diffusion pathways and some other explanation, possibly structural, needs to be sought. Received: 5 April 1997 / Accepted: 26 April 1997     

Serum concentrations of melatonin in prepubertal or postpubertal gilts exposed to artificial lighting or sunlight

To determine if the type of environmental lighting or reproductive status influences secretory patterns of serum melatonin, gilts were exposed to artificial light or full sunlight during the summer months. In Experiment 1, eight prepubertal and eight postpubertal gilts (Hampshire x Yorkshire x Duroc) were exposed to light intensity of 700 lux in an environmentally controlled room from 0730 to 1900 h daily. An additional eight prepubertal and eight postpubertal gilts were reared outdoors in an open modified-front gestation building and fed on a concrete apron outdoors where light intensity approached 50,000 lux in full sunlight. After 2 mo of acclimating to these environmental conditions, blood samples were drawn from each gilt at 2-h intervals from 1000 to 0200 h. Serum concentrations of melatonin were assayed utilizing Guildhay antisera. The experiment was repeated during the same months of the following year utilizing different gilts (Experiment 2). During both replications, neither light intensity nor reproductive status affected the secretory patterns of melatonin during the sampling period (P >0.05). In both prepubertal and postpubertal gilts, serum concentrations of melatonin were not reduced (P >0.05) by exposure to direct sunlight. Since baseline concentrations of serum melatonin were not reduced by sunlight during the day, the incidence of nocturnal rises of melatonin was not increased (P >0.05) in either prepubertal or postpubertal gilts.     

Influence of larval feeding history on the body condition of Amphiprion melanopus

The cellular condition of liver hepatocytes and the height of gut epithelium cells of larval Amphiprion melanopus were sensitive indicators of feeding condition. Muscle fibres of the trunk showed marked separation in fish fed every third day just prior to settlement. Low feeding regimes also caused reductions in growth, increases in larval duration and reductions in size at metamorphosis. Gut epithelium cell height was also influenced by fish standard length and age. This study suggests that gut epithelium cell height is a useful index for an examination of the importance of starvation of larvae in tropical waters; however, size and age standardization is required prior to comparisons of wild caught fish through time or with laboratory samples.