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Biomedical subjects

L Hong

Publications and source records attributed to L Hong.

At least 91 records · Page 5Linked to original sources

Lysophosphatidic acid alters cerebrovascular reactivity in piglets.

Effects of the lipid mediator lysophosphatidic acid (LPA) were studied on the cerebral circulation of newborn pigs using closed cranial windows. Topical application of synthetic LPA caused dose-dependent vasoconstriction and inhibited vasodilation to hypercapnia and isoproterenol. These vasodilators elicited a rise in the adenosine 3',5'-cyclic monophosphate (cAMP) content of the cerebrospinal fluid, which was inhibited dose dependently by LPA. Pertussis toxin (1 microgram/ml) completely abolished LPA-induced vasoconstriction and the altered vascular reactivity, and LPA no longer decreased cAMP. Electrophysiological recording of currents evoked by LPA-like lipids in Xenopus oocytes showed that cerebrospinal fluid is normally devoid of LPA-like factors. In contrast, the amount of LPA-like factors generated 4 days after intrathecal injection of autologous blood was in the range of 1-10 microM LPA equivalents. The data indicate that LPA-like bioactive mediators were generated in an intracranial hematoma model and that these phospholipids might play a role in the pathophysiology of altered vascular reactivity often found in posthemorrhagic conditions and could also contribute to the development of posthemorrhagic vasoconstriction.

Animals↗

Alternative splicing of the Euglena gracilis chloroplast roaA transcript.

A novel gene, roaA (ribosomal operon-associated gene), encoding a potential RNA-binding protein has been identified in the rpl23 ribosomal protein operon of the Euglena gracilis chloroplast genome. The roaA gene is interrupted by one group III and three group II introns. Introns 1 and 2 of roaA can be interpreted as a twintron, formed from the insertion of a group II intron into the 5'splice site of a group III intron. Alternative splicing of the group III intron results in two distinct transcripts encoding proteins of 516 and 514 amino acids. Group III introns may play a role in the generation of alternatively spliced transcripts.

Alternative Splicing↗

A group III intron is formed from domains of two individual group II introns.

A 1352-nucleotide intron within the Euglena gracilis chloroplast ycf8 gene has been characterized as a complex twintron with overlapping internal introns and alternative splicing pathways. Partially spliced pre-mRNAs were characterized by a combination of cDNA cloning and sequencing, Northern hybridization, and S1 nuclease protection analyses. In the predominant pathway, two internal group II introns (601 and 392 nucleotides) are spliced from subdomain ID of an external group II intron (359 nucleotides). In an alternative pathway, following excision of the 601-nucleotide intron, splicing of a group III intron occurs. This group III intron is recruited from sequences of the external intron and the 392-nucleotide intron. This is the first evidence that a group III intron can be derived from portions of existing group II introns. The mechanism of group III intron formation may also be relevant to the evolution of nuclear introns from putative group II intron ancestors.

Alternative Splicing↗

Unusual structures of the tandem repetitive DNA sequences located at human centromeres.

The presence of the highly conserved repetitive DNA sequence d(AATGG)n.d(CCATT)n in human centromeres argues for a special role for this sequence in recognition, most probably through the formation of an unusual structure during mitosis. Quantitative one- and two-dimensional nuclear magnetic resonance (1D/2D NMR) spectroscopic studies reveal that the Watson-Crick duplex d(AATGG)n.d(CCATT)n adopts the usual B-DNA conformation as illustrated by taking d(AATGG)3.d(CCATT)3 as an example, whereas the d(CCATT)n strand is essentially a random coil. In contrast, the d(AATGG)n strand adopts an unusual stem-loop motif for repeat lengths n = 2, 3, 4, and 6. In addition to normal Watson-Crick A.T pairs, the stem-loop structures are stabilized by mismatched A.G and G.G pairs in the stem and G-G-A stacking in the loop. Stem-loop structures of d(AATGG)n are independently verified by gel electrophoresis and nuclease digestion studies and were also previously shown to be as stable as the corresponding Watson-Crick duplex d(AATGG)n.d(CCATT)n [Grady et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1695-1699]. Therefore, the sequence d(AATGG)n can, indeed, nucleate a stem-loop structure at little free energy cost, and if, during mitosis, it is located on the chromosome surface, it can provide specific recognition sites for kinetochore function.

Base Sequence↗

Toxicokinetics of oxazepam in rats and mice.

The comparative toxicokinetics of oxazepam were studied in F344 rats, B6C3F1 mice, and Swiss-Webster mice of both sexes after an i.v. dose of 20 mg/kg and oral gavage doses of 50, 200, and 400 mg/kg. In addition, the toxicokinetics of oxazepam in a 3-week dosed-feed study of male B6C3F1 mice at 125 and 2500 ppm were also investigated. Results indicated that the elimination of oxazepam from plasma after i.v. injection in both rats and mice were first-order and could be best described by a two-compartment model with a terminal elimination half-life of 4-5 h for rats and 5-7 h for mice. After oral gavage dosing the peak oxazepam plasma concentrations in most rodents were reached within 2-3.5 h. At all doses studied, female rodents had significantly higher plasma concentrations than males. Absorption of oxazepam was significantly extended at higher oral doses of 200 and 400 mg/kg. At 50 mg/kg, the bioavailability of oxazepam in rats (< 50%) was lower than in Swiss-Webster mice (> 80%). The bioavailability of oxazepam in both B6C3F1 and Swiss-Webster mice decreased with increasing dose. A dose proportionality of Cmax was not observed in rats and mice after gavage doses of 50, 200, and 400 mg/kg. Plasma concentrations of oxazepam in the dosed-feed study increased with the concentration of oxazepam in the feed, a quasi-steady-state of plasma concentrations of oxazepam was reached after approximately 4 days ad libitum exposure. In B6C3F1 mice, the estimated relative bioavailability of oxazepam from dosed feed (relative to gavage study at 50 mg/kg) was about 43%.

Animals↗

Gene structure and expression of a novel Euglena gracilis chloroplast operon encoding cytochrome b6 and the beta and epsilon subunits of the H(+)-ATP synthase complex.

The genes encoding cytochrome b6 of the chloroplast cytochrome b6/f complex (petB) and the ATP synthase CF1-beta subunit (atpB) and epsilon-subunit (atpE) were identified on the EcoD fragment of the Euglena gracilis chloroplast genome. The complete nucleotide sequence of these three genes was determined. The petB-atpB-atpE genes are cotranscribed as a tricistronic operon. This gene organization differs from that of land plants in which atpB-atpE form a discistronic operon, and petB is within the psbB-ycf8-psbH-petB-petD operon. Euglena cytochrome b6 and the beta-subunit of the chloroplast ATP synthase are very similar in derived amino acid sequence to the corresponding gene products from other organisms. The epsilon-subunit of the chloroplast ATP synthase complex is more divergent. In Euglena, the petB-atpB-atpE genes contain introns, including two twintrons, at eight different positions. All of the intron positions were confirmed by analysis of cDNAs. Two independent intercistronic RNA processing events and 11 splicing reactions lead to the accumulation of the mature petB, atpB and atpE monocistronic mRNAs.

Amino Acid Sequence↗

Histological observations of coated and prestressed dental implants under flexible or rigid loading conditions.

Hydroxyapatite-coated and hydroxyapatite-prestressed titanium implants were implanted under load-free, flexible-loaded and rigid-loaded conditions in dog mandibula in vivo. The bone response, bone contact and the quality of the coating were evaluated histologically and histometrically. Hydroxyapatite coating on titanium implants enhanced the bone apposition. The bone contact can still be present while the coating is resorbed. Gingiva contact with the coating led to a faster degradation of the coating. There was no significant difference in bone contact between hydroxyapatite-coated and -prestressed implants with regard to the loading systems: rigid, flexible and load-free. All three implants showed the highest amount of bone contact apically. Only at the mesial side of the implant did the flexible element show rather high amounts of bone contact compared to the rigid and load-free implants. The hydroxyapatite coating more or less degraded during the implantation period. The degree of degradation varied in the implant itself, between the implants and between the animals.

Analysis of Variance↗

Complete nucleotide sequence and tissue-specific expression of the rat phenylethanolamine N-methyltransferase gene.

The rat phenylethanolamine N-methyltransferase (PNMT) gene was isolated from a genomic library by cross-hybridization with a bovine PNMT cDNA probe. Complete nucleotide sequence analysis of a genomic clone showed that this gene contained three exons and spanned about 2.8 kb in length. There were the acute-phase response element, TATA, SP1, and GRE sequences. The physicochemical properties of rat adrenal PNMT were different from those of the brainstem PNMT. However, northern blot and reverse transcription-polymerase chain reaction analysis showed that the rat PNMT gene may not express the multiple forms of mRNA. These results suggest that the rat PNMT gene might produce a single enzyme protein, whose activity may be differentially modulated by tissue-specific environment in the central and peripheral systems.

Adrenal Glands↗

Variant mitochondrial plasmids of broad bean arose by recombination and are controlled by the nuclear genome.

Various cytoplasms of broad bean contain three mitochondrial plasmids (mtp1, 2 and 3), previously described. In cytoplasm 350 we have observed several additional mitochondrial plasmids, varying in number and in identity according to the nuclear background. Replacement of the nucleus by backcrossing led to the appearance or disappearance of additional plasmids, indicating that the nuclear genome controls either the creation or the copy level of mitochondrial plasmids. Analysis of eight variant additional plasmids (mtp4-11) suggests that they all result from a double recombination event between mtp1 and mtp2. In all cases, one recombination point was located within a 276-bp sequence, identical in both plasmids. For 7 plasmids, the region in which the second recombination event occurred could be narrowed down to a short stretch containing imperfect tandem repeats of a 31-bp motif. The largest sequence shared by the recombination regions was hexanucleotide GCGACG.

Base Sequence↗

Complete sequence of Euglena gracilis chloroplast DNA.

We report the complete DNA sequence of the Euglena gracilis, Pringsheim strain Z chloroplast genome. This circular DNA is 143,170 bp, counting only one copy of a 54 bp tandem repeat sequence that is present in variable copy number within a single culture. The overall organization of the genome involves a tandem array of three complete and one partial ribosomal RNA operons, and a large single copy region. There are genes for the 16S, 5S, and 23S rRNAs of the 70S chloroplast ribosomes, 27 different tRNA species, 21 ribosomal proteins plus the gene for elongation factor EF-Tu, three RNA polymerase subunits, and 27 known photosynthesis-related polypeptides. Several putative genes of unknown function have also been identified, including five within large introns, and five with amino acid sequence similarity to genes in other organisms. This genome contains at least 149 introns. There are 72 individual group II introns, 46 individual group III introns, 10 group II introns and 18 group III introns that are components of twintrons (introns-within-introns), and three additional introns suspected to be twintrons composed of multiple group II and/or group III introns, but not yet characterized. At least 54,804 bp, or 38.3% of the total DNA content is represented by introns.

Animals↗

Studies of the DNA binding properties of histone H4 amino terminus. Thermal denaturation studies reveal that acetylation markedly reduces the binding constant of the H4 "tail" to DNA.

The effect of acetylation on the DNA binding properties of the rigidly conserved histone H4 amino-terminal tail has been studied in detail using the technique of thermal denaturation. The quantitative DNA-binding parameters for both the non- and fully acetylated H4 amino terminus have been determined from thermal denaturation data for complexes of the peptides bound to mixed sequence 146-base pair DNA. We find that under dilute buffer conditions (5 mM Tris-HCl) the binding constant for the non-acetylated peptide to double-stranded DNA is 5 x 10(11) M-1 and that acetylation of lysine residues in the peptide reduces the binding constant to 1 x 10(5) M-1. The dramatic differences observed in the binding constants for the non- and fully acetylated peptides are probably due to the effect of acetylation on the even distribution of positively charged residues in the H4 amino terminus. In other experiments, the binding of both peptides to a 30-base pair oligonucleotide has been studied in solution with varying concentrations of sodium, magnesium, and phosphate ions. These experiments demonstrate that both magnesium and phosphate ions have strong effects on the binding of the H4 tail to DNA, especially weakening the binding of the acetylated peptide. For instance, the dissociation of the non-acetylated peptide from DNA requires 6 mM magnesium, yet the binding of the acetylated peptide is abolished in only 30 microM magnesium. The modulation of the DNA binding interactions of the H4 amino terminus by physiologically relevant ionic conditions, in addition to the effect of acetylation, can be important in the regulation of chromatin structure and function.

Acetylation↗

Intercellular adhesion molecule-1 expression induced by interleukin (IL)-1 beta or an IL-1 beta fragment is blocked by an IL-1 receptor antagonist and a soluble IL-1 receptor.

The effects of a recombinant human interleukin-1 (IL-1) receptor antagonist (IL-1ra) and a recombinant human soluble IL-1 receptor (sIL-1R) on cytokine-induced intercellular adhesion molecule-1 (ICAM-1) expression in a human glioblastoma cell line and a neuroblastoma cell line were determined. Cells were incubated with IL-1 beta, tumor necrosis factor (TNF) alpha and interferon (IFN) gamma. Cells were also tested under identical conditions with an IL-1 beta synthetic peptide fragment (IL-1 beta 208-240) previously shown to possess biological activity. IL-1 beta, TNF alpha and IFN gamma potentiated ICAM-1 expression in both cell lines in a dose-related manner. The IL-1 beta 208-240 fragments, corresponding to the rabbit, rat and human sequences, enhanced ICAM-1 expression in glioblastoma cells at high doses. ICAM-1 expression induced by IL-1 beta, rabbit IL-1 beta 208-240 and human IL-1 beta 208-240 was blocked by the IL-1ra, while TNF alpha- and IFN gamma-induced ICAM-1 expression were not. ICAM-1 expression induced by IL-1 beta and human IL-1 beta 208-240 was also blocked by the sIL-1R. Our findings suggest that IL1 beta 208-240 acts as an IL-1 beta agonist in enhancing ICAM-1 expression in vitro and that this effect is receptor-mediated.

Amino Acid Sequence↗

Expression of the human foamy virus bel-1 transactivator in insect cells.

The human foamy virus (HFV) bel-1 transactivator protein was expressed in insect cells by a recombinant baculovirus. For the generation of the recombinant baculovirus, Acbel-1, the bel-1 gene of an HFV mutant was used, that bears truncations in the bel-1 overlapping bel-2 open reading frame. Acbel-1 infected Sf9 cells produced high amounts of recombinant protein of the same electrophoretic mobility (36 kD) as bel-1 expressed in mammalian cells. The baculovirus expressed bel-1 protein was readily identified by a polyclonal rabbit serum directed against bel-1 in immunoblot assay. As in mammalian cells, bel-1 was predominantly localized to the nucleus of Acbel-1 infected insect cells. The baculovirus expressed bel-1 protein will be of use to determine the action of this novel viral transactivator more precisely.

Animals↗

Toxicokinetics of pentachloroanisole in F344 rats and B6C3F1 mice.

1. Toxicokinetics of pentachloroanisole (PCA) were studied in F344 rat and B6C3F1 mouse of both sexes by gavage at doses of 10, 20 and 40 mg/kg and by i.v. at 10 mg/kg. 2. PCA was rapidly demethylated to pentachlorophenol (PCP) in both rat and mouse and the resulting PCP plasma concentrations were much higher than that of parent PCA due to the much smaller apparent volume of distribution of PCP. 3. Peak plasma concentrations of PCA and PCP increased with dose in both rat and mouse. 4. Bioavailability of PCA was low in both rat and mouse and was sex independent. 5. The high plasma concentrations and relatively long biological half-life of PCP in both species after both i.v. and oral dosing with PCA indicate possible bioaccumulation of PCP upon multiple oral administrations of PCA.

Animals↗

[Analysis of the image of external pterygoid muscle during close and wide open of the jaw by condylo-pterygo-maxillo-oblique projection in magnetic resonance].

External pterygoid muscle is an important muscle. It is easy to be hurt by acute and chronic trauma which may become one of the chief cause of TMJDS. It is difficult to be studied, because it locates deeply. It is covered by thick soft and hard tissues. That is why the questions about its anatomy and physiological functions have been debated. In this paper, the changes of images form of external pterygoid muscle during the jaw closing and wide opening were observed and analysed in 14 cases, 25 sides, of adult men and women. By CPMOP in MR. The results can by concluded that (1) the upper head of this muscle inserts fully or mainly to the condyle pterygoid fossa; (2) the direction of the muscle fibers is not entirely horizontal but with some inclined and (3) the two heads of this muscle are relaxed in the jaw closing and are contracted in the jaw wide opening synchronously.

Adult↗