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Biomedical subjects

D He

Publications and source records attributed to D He.

At least 91 records · Page 5Linked to original sources
PubMed1995

Nuclear matrix proteins as structural and functional components of the mitotic apparatus.

The eukaryotic nucleus is a membrane-enclosed compartment containing the genome and associated organelles supported by a complex matrix of nonhistone proteins. Identified as the nuclear matrix, this component maintains spatial order and provides the structural framework needed for DNA replication, RNA synthesis and processing, nuclear transport, and steroid hormone action. During mitosis, the nucleoskeleton and associated chromatin is efficiently dismantled, packaged, partitioned, and subsequently reassembled into daughter nuclei. The dramatic dissolution of the nucleus is accompanied by the assembly of a mitotic apparatus required to facilitate the complex events associated with nuclear division. Until recently, little was known about the fate or disposition of nuclear matrix proteins during mitosis. The availability of specific molecular probes and imaging techniques, including confocal microscopy and improved immunoelectron microscopy using resinless sections and related procedures, has enabled investigators to identify and map the distribution of nuclear matrix proteins throughout the cell cycle. This chapter will review the structure, function, and distribution of the protein NuMA (nuclear matrix mitotic apparatus) and other nuclear matrix proteins that depart the nucleus during the interphase/mitosis transition to become structural and functional components within specific domains of the mitotic apparatus.

Animals↗
PubMed1995

The effect of porphyrin and radiation on ferrochelatase and 5-aminolevulinic acid synthase in epidermal cells.

The effects of ultraviolet A (UVA) and blue light on ferrochelatase protein, and its mRNA level, in 5-aminolevulinic acid (ALA)-loaded A431 cells was evaluated. Western blot analysis of ferrochelatase protein showed a protein band of 43 kDa. There was a decrease in the protein concentration 24 h and 48 h after irradiation of these cells. In contrast, as judged by Northern blot analysis, there was no change in ferrochelatase mRNA level. Measurement of ALA synthase activity showed an ALA dose-dependent but radiation-independent decrease of enzyme activity, suggesting an end-product feedback inhibition. Since reactive oxygen species generated by porphyrin-induced photochemical reaction may be involved in the decrease in ferrochelatase protein, the effect of scavengers of reactive oxygen species was evaluated by measuring porphyrin accumulation in irradiated, ALA-loaded A431 cells. Porphyrin accumulation was significantly decreased in the presence of singlet oxygen scavenger sodium azide (0.05 mM, 40.6% suppression) or hydroxyl radical scavenger mannitol (5.0 mM, 45.0% suppression). These data suggest that the photochemical reaction induced by porphyrin and irradiation resulted in a decrease in ferrochelatase protein content, but had no effect on ferrochelatase mRNA level nor on ALA synthase activity. The decrease in protein was partly mediated by the reactive oxygen species.

5-Aminolevulinate Synthetase↗
PubMed1995

Muscle fibre type and dimension in genetically obese and lean Zucker rats.

Skeletal muscle structure and morphology may be altered in obesity. To study this further, muscles from six genetically obese (fa/fa) and six normal male rats were examined at 15 weeks of age. The gluteus medius, vastus lateralis and rectus abdominis muscles were dissected out and stained for histochemical fibre typing. In addition the fibre cross-sectional area was measured on a graphic tablet. The proportion of fast-twitch fibres was larger in the vastus lateralis and rectus abdominis muscles of the obese rats (P < 0.01); no difference was seen for the gluteus medius muscle. For the normal rats the cross-sectional area of the fast-twitch fibres was 2-3 times larger than the area of slow-twitch fibres in the same muscle. The cross-sectional area of the fast-twitch fibres in the obese rats was 40-47% less than in the control animals (P < 0.003), while no difference between the two groups was found for the slow-twitch fibre area. The data thus suggest that in the genetically obese rats the development of fast-twitch fibres was primarily affected. Moreover, in these animals some muscles may be more affected than others.

Animals↗
PubMed1995

The effect of ALA and radiation on porphyrin/heme biosynthesis in endothelial cells.

To study porphyrin biosynthesis in human microvascular endothelial cells, HMEC-1 cells, a transformed human microvascular endothelial cell line, were incubated with 5-aminolevulinic acid (ALA), the precursor of endogenous porphyrins, and porphyrin accumulation was measured spectro-fluorometrically. The HMEC-1 cells accumulated porphyrin in a concentration-related and a time-dependent fashion. Protoporphyrin was the predominant porphyrin accumulated in the cells. The effect of light on protoporphyrin accumulation was evaluated by exposing the ALA-loaded HMEC-1 cells to ultraviolet-A (UVA) and blue light, followed by another incubation with ALA for 2-24 h. Enhancement of protoporphyrin accumulation in irradiated HMEC-1 cells was observed 2-24 h after irradiation, which was associated with a decrease in ferrochelatase protein and activity. Porphyrin accumulation from ALA after irradiation was significantly decreased when catalase (750-3000 U/mL, 29.3-44.3% suppression) or superoxide dismutase (270 U/mL, 36.4% suppression) was present during irradiation. These data demonstrate that HMEC-1 cells were capable of porphyrin biosynthesis, and that exposure of protoporphyrin-containing HMEC-1 cells to UVA and blue light, which includes the Soret band spectrum, decreased the ferrochelatase activity and its protein. These changes were mediated, at least in part, by reactive oxygen species.

Aminolevulinic Acid↗
PubMed1994

Nuclear-mitotic apparatus protein: a structural protein interface between the nucleoskeleton and RNA splicing.

Vertebrate splicing factors are localized to discrete domains within the nuclei of somatic cells. The mechanism whereby such nuclear domains, identified as speckles by immunofluorescence microscopy, are generated is unclear. Recent studies suggest that the spatial order within the nucleus is maintained by nuclear matrix factors. Here we show that a protein in the nuclear matrix and mitotic apparatus [nuclear-mitotic apparatus protein, NuMA; Lydersen, B. & Pettijohn, D. (1980) Cell 22, 489-499] colocalizes with splicing factors in interphase nuclei and is associated with small nuclear ribonucleoproteins in a complex immunoprecipitated from HeLa extract with small nuclear ribonucleoprotein antibodies. Moreover, NuMA associates with splicing complexes that are reconstituted in vitro using wild-type pre-mRNA, but not with nonspecific RNA. Cumulatively, these observations suggest a function of NuMA or NuMA-like proteins in interphase cells in providing a bridge between RNA processing and the nucleoskeleton.

Antigens, Nuclear↗
PubMed1994

Localization of NuMA protein isoforms in the nuclear matrix of mammalian cells.

Using a monoclonal antibody 2D3 generated against a kinetochore-enriched human chromosome preparation, we identified a high molecular mass protein with nuclear staining in interphase and polar staining of the pericentriolar region in the mitotic spindle. Initially termed centrophilin, this protein associates with the minus-ends of spindle microtubules (MT) and appears to be important in spindle organization [Tousson et al., 1991: J. Cell Biol. 112:427-440]. Comparison of a partial cDNA sequence obtained for centrophilin with the full length cDNA sequence of nuclear mitotic apparatus protein (NuMA) [Compton et al., 1992: J. Cell Biol. 116:1395-1408; Yang et al., 1992: J. Cell Biol. 116:1303-1317] has indicated that NuMA and centrophilin are the same protein. Using a polyclonal NuMA antibody, we have provided further evidence that NuMA exists as isoforms as shown by peptide mapping and immunoblots. Sequential fractionation experiments along with immunofluorescence, immunoblotting, and EM immunogold labeling have demonstrated that NuMA isoforms are novel components of nuclear core filaments. Thus, NuMA, a long coiled-coil protein, appears to have dual functions in interphase and mitosis during the cell cycle. In interphase, NuMA likely plays a structural role in the nucleoskeleton that may be important in nuclear organization and functions, whereas in mitosis, NuMA appears to be associated with spindle MT organization and chromosome positioning.

Antigens, Nuclear↗
PubMed1994

The nature and origins of acid summer haze air pollution in metropolitan Toronto, Ontario.

During July and August of 1986, 1987, and 1988, a field study was conducted of ambient acidic aerosol levels in Toronto, Ontario. Fine particle mass (da < 2.5 microns) samples were collected twice daily at a central-city site for the determination of particulate-phase strong acidity (H+) and sulfate (SO4 =). Two additional H(+)-monitoring sites were concurrently operated during the summers of 1986 and 1987 to examine the spatial variability of H+ within the metropolitan area. During the summer of 1986, a quasi-continuous total sulfate/sulfuric acid analyzer was also deployed to allow a determination of the chemical form of H+. Results indicate that acid aerosol episodes (H+ > or = 100 nmole/m3) did occur in this city during the summer months, and that H+ peaks were well correlated with sulfate peaks. Virtually all of the H+ was found to be present as ammonium bisulfate (NH4HSO4). While H+ concentrations were highly correlated among the three monitoring sites (r = 0.9), the highest H+/SO4 = ratios prevailed during SO4 = episode periods and at the least urbanized site. This latter trend was apparently due to greater neutralization of H+ by local ammonia at the more urbanized sites. Comparisons of day vs night H+/SO4 = ratios, an examination of air mass back-trajectories, and contemporaneous H+ measurements at surrounding sites collectively indicated that transported regional haze air pollution from the United States is a major contributor to the H+ events recorded within Toronto.

Acids↗
PubMed1994

[Changes in vestibular function and furosemide test after experimental endolymphatic hydrops in guinea pigs].

The present study reports a successful anial model of endolymphatic hydrops (EH) in guinea pigs (n = 48) through a surgical obliteration of the endolymphatic sac and duct. Electronystagmography was calibrated by optokinetic nystagmus. During the formation of EH the vestibular functions (n = 16) were dynamically observed by ratation test. After eight weeks (n = 25) the furosemide tests with different doses were carried out by sinusoidal angular acceleration test. The results showed that the excitability of vestibular function in the operated ears was enhanced for the first week after operation, and then declined. At the eighth week it was significantly improved after intravenous injection of furosemide at the clinical equivalent dose level (1.25 mg/kg). The present results strongly support the usefulness of furosemide test in the diagnosis of EH. The possible mechanisms of its effect are discussed.

Animals↗
PubMed1994

Evaluation of defined antigen vaccines against Schistosoma bovis and S. japonicum in bovines.

Our objective is to contribute to the development of defined antigen vaccines for schistosomiasis by evaluating the protective efficacy of Schistosoma bovis and S. japonicum antigens in their natural bovine hosts. Antigens under evaluation include some already identified as vaccine candidates: glutathione S-transferases (GSTs); KLH, which shares protective epitopes with the protective antigen GP38 of S. mansoni; and Sj23, the analogue of the vaccine candidate Sm23 antigen. In another approach, since crude freeze/thaw schistosomular antigen plus BCG(F/T vaccine) has proved protective against S. japonicum in bovines, as it was against S. mansoni in mice, we are carrying out further evaluations both of this crude antigen and of recombinant-derived paramyosins. In a third line of work, novel vaccine candidate antigens identified by screening our cDNA libraries with various passively protective animal sera are being evaluated in animal experiments. In the Sudan we have shown that vaccination of calves with either native S. bovis GSTs or KLH induces high levels of fecundity-suppression without causing a significant reduction in adult worm recoveries. Therefore, recombinant-derived S. bovis 28kD GST is now being evaluated, as are the effects of combined GST/KLH vaccination. In China, sheep have been vaccinated with either S. japonicum GSTs, with KLH, or with the F/T vaccine, as a prelude to trials in bovines. As judged by adult worm recoveries, each type of vaccine induced significant protection, and there was also evidence, particularly with the GST and F/T vaccines, of fecundity-suppressive effects. As with the S. bovis/cattle system therefore, both GST and KLH showed protective effects against S. japonicum in sheep.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗