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Biomedical subjects

M M Lai

Publications and source records attributed to M M Lai.

At least 55 records · Page 3Linked to original sources
PubMed1999

Serum cholesterol levels and prevalence of hypercholesterolemia in school-aged Taiwanese children and adolescents: the Taichung Study.

BACKGROUND: Atherosclerosis has become one of the leading causes of death in Taiwan. Hypercholesterolemia is a major risk factor for coronary atherosclerosis. To evaluate the mean total cholesterol values and the prevalence of hypercholesterolemia in school-aged Taiwanese children and adolescents, an epidemiologic survey was conducted. METHODS: After two-stage sampling of 52 primary schools and 26 junior high schools in Taichung City, we randomly selected 3,924 children (2,070 boys and 1,854 girls). Blood total cholesterol (TC), triglyceride (TG) and high-density lipoprotein (HDL)-cholesterol were measured. Response rate was 86.8%. Subjects' ages ranged from seven to 14 years old. RESULTS: From this cross-sectional survey, the following characteristics (mean +/- SD) were documented for boys and girls, respectively: TC, 162 +/- 28 and 165 +/- 29 mg/dl; TG, 74 +/- 33 and 80 +/- 32 mg/dl; HDL-cholesterol, 61 +/- 13 and 59 +/- 12 mg/dl; and low-density lipoprotein (LDL)-cholesterol, 87 +/- 24 and 90 +/- 25 mg/dl. The mean TC across all age groups ranged from 149 to 172 mg/dl for boys and 157 to 170 mg/dl for girls. In boys, the TC concentration was highest at age 11 (172 mg/dl), and in girls was highest at age 7 (170 mg/dl). Borderline hypercholesterolemia (TC > 170 mg/dl) was found in 36.5% of boys (13.5-53.1%) and 39.7% of the girls (28.3-48.6%). Hypercholesterolemia (TC > 200 mg/dl) was found in 9.7% of males (3.1-16.6%) and 10.3% of the females (5.0-14.9%). CONCLUSIONS: The prevalence of borderline hypercholesterolemia was 38.0% for TC and 17.2% for LDL-C, and for hypercholesterolemia, it was 9.9% for TC and 5.8% for LDL-C. The higher TC levels in Taiwanese children were primarily due to higher HDL-C and TG levels.

Adolescent↗
PubMed1999

Association between serum uric acid and cardiovascular risk factors among elderly people in Taiwan.

In order to understand the distribution of serum uric acid and the relationship between serum uric acid and the cardiovascular risk factor among elderly people, a cross-sectional study was conducted in Chung-Shing-Shin-Tseun community in Taiwan in May 1998. All individuals aged 65 and over were collected. A total of 1123 persons, out of 1774 registered residents, were contacted by face-to-face interview. The response rate was 63.3%. However, only 586 respondents had blood tests and completed questionnaires. Analysis in this study was based on these 586 subjects. The mean uric acid values were 7.4 +/- 1.8 mg/dl in men and 6.3 +/- 1.6 mg/dl in women, respectively (p < 0.001). Multivariate linear regression showed that serum uric acid was significantly correlated with sex and body mass index. Simple correlation showed that serum uric acid was significantly correlated with diastolic pressure, total cholesterol, triglyceride and creatinine. Age, systolic pressure and fasting glucose were not related to serum uric acid. In our conclusion, the uric acid values are high among elderly people. The serum uric acid levels are significantly associated with the cardiovascular risk factors among elderly people.

Age Factors↗
PubMed1998

Characterization of mRNA for hepatitis delta antigen: exclusion of the full-length antigenomic RNA as an mRNA.

Hepatitis delta virus (HDV) encodes a single protein, the hepatitis delta antigen (HDAg), which is thought to be translated from a 0. 8-kb RNA of antigenomic sense. This subgenomic RNA species is present in very small amounts in HDV-infected liver tissues and in cultured cells infected or transfected with HDV, and in some cases it cannot be detected at all. In contrast, HDAg protein is present in large amounts in all natural and experimental models of HDV infection. This study addresses whether other HDV RNA species, such as the antigenomic-sense, genome-size HDV RNA can also serve as the mRNA for HDAg synthesis. Taking advantage of the ability of herpes simplex virus (HSV) to degrade only polyadenylated mRNAs, we examined the effect of HSV coinfection on HDAg synthesis. It was shown that HSV infection did degrade the subgenomic 0.8-kb HDV mRNA but not HDV genome-length RNA. Under such conditions, HDAg synthesis was completely inhibited. Furthermore, the genome-length HDV RNA was found not to be associated with polysomes. Finally, in vitro translation studies demonstrated that HDAg could not be translated directly from the genome-length antigenomic-sense HDV RNA. These results suggest that only the subgenomic RNA species of HDV possesses properties characteristic of the mRNA for HDAg and that the genome-length RNA cannot be used for translating HDAg. In addition, we found that HDV RNA replication did not depend on de novo HDAg synthesis.

5' Untranslated Regions↗
PubMed1998

Inhibition of Cellular RNA polymerase II transcription by delta antigen of hepatitis delta virus.

Hepatitis delta virus (HDV) contains a circular, viroid-like RNA and the hepatitis delta antigen (HDAg) protein. The viral RNA is replicated via RNA-dependent RNA synthesis, which is thought to be mediated by host DNA-dependent RNA polymerase II (pol II). The precise mechanism of HDV RNA replication using RNA as a template remains to be elucidated, though it is clear that HDAg is involved. We demonstrate here that both SP1-activated and basal pol II transcription are inhibited by HDAg. This inhibitory effect of HDAg was observed in vivo in transient cotransfection assays as well as in vitro in HeLa nuclear extracts with purified, recombinant HDAg. The in vitro inhibition of pol II transcription could be reversed with excess HeLa nuclear extracts. Furthermore, HDAg specifically inhibited pol II-mediated transcription but not pol I- or III-mediated transcription. These results provide support for the model in which HDAg participates in a complex with host cell pol II transcription factors to mediate pol II-dependent HDV RNA replication, concomitantly cellular pol II transcription.

Chloramphenicol O-Acetyltransferase↗
PubMed1998

Cain, a novel physiologic protein inhibitor of calcineurin.

Calcineurin is a widely distributed protein phosphatase regulated by calcium and calmodulin. It mediates the immunosuppressive actions of drugs such as cyclosporin and FK506, and has been implicated in a number of calcium-sensitive pathways in the nervous system, including regulation of neurotransmitter release and modulation of long-term changes in synaptic plasticity. Calcineurin associates physiologically with other proteins, including calmodulin, FKBP12 (FK506-binding protein), the ryanodine receptor, and the inositol 1,4,5-trisphosphate receptor. We now report the identification, molecular cloning, and functional characterization of a novel protein, cain (calcineurin inhibitor), that interacts with and inhibits calcineurin. The full-length cain cDNA predicts a 240-kDa protein with no significant homology to any known protein. Cain associates with calcineurin both in vitro and in vivo, leading to a non-competitive inhibition of calcineurin activity. The putative calcineurin-binding domain of cain, a 38-amino acid region defined by mutational analysis, is highly basic. Like calcineurin, cain has a prominent neuronal expression and a wide tissue distribution. Cain's expression pattern in the brain closely resembles that of calcineurin, indicating a physiologic association between the two proteins.

Amino Acid Sequence↗
PubMed1998

Hepatitis C virus core protein interacts with heterogeneous nuclear ribonucleoprotein K.

Hepatitis C virus (HCV) core protein, a component of viral nucleocapsid, has been shown to modulate cellular and viral promoter activities. To identify potential cellular targets for HCV core protein, a human liver cDNA library was screened for core-interacting proteins using the yeast two-hybrid system. Among the proteins identified was heterogeneous nuclear ribonucleoprotein K (hnRNP K), which has been demonstrated to be a transcriptional regulator. The interaction of HCV core protein with hnRNP K was confirmed by glutathione S-transferase fusion protein binding assay, protein-protein blotting assay, and coimmunoprecipitation in vitro and in vivo. Additionally, these two proteins were shown to be partially colocalized in the nucleus. The hnRNP K-binding site in HCV core protein was mapped to the region from amino acid residues 25-91, a hydrophilic area near the N terminus. The HCV core protein-binding domain was located within amino acid residues 250 to 392, which contain the three proline-rich domains, of hnRNP K. Furthermore, HCV core protein relieved the suppression effect of hnRNP K on the activity of the human thymidine kinase gene promoter. The specific binding of HCV core protein to hnRNP K suggests that multiple functions of hnRNP K may be disrupted by the core protein during HCV infection and thus explains, in part, the pathogenesis of HCV.

Amino Acid Sequence↗
PubMed1998

Neurabin is a synaptic protein linking p70 S6 kinase and the neuronal cytoskeleton.

p70 S6 kinase (p70(S6k)) is a mitogen-activated protein kinase that plays a central role in the control of mRNA translation. It physiologically phosphorylates the S6 protein of the 40s ribosomal subunit in response to mitogenic stimuli and is a downstream component of the rapamycin-sensitive pathway, which includes the 12-kDa FK506 binding protein and includes rapamycin and the 12-kDa FK506 binding protein target 1. Here, we report the identification of neurabin (neural tissue-specific F-actin binding protein), a neuronally enriched protein of 1,095 amino acids that contains a PDZ domain and binds p70(S6k). We demonstrate the neurabin-p70(S6k) interaction by yeast two-hybrid analysis and biochemical techniques. p70(S6k) and neurabin coimmunoprecipitate from transfected HEK293 cells. Site-directed mutagenesis of neurabin implicates its PDZ domain in the interaction with p70(S6k), and deletion of the carboxyl-terminal five amino acids of p70(S6k) abrogates the interaction. Cotransfection of neurabin in HEK293 cells activates p70(S6k) kinase activity. The mRNA of neurabin and p70(S6k) show striking colocalization in brain sections by in situ hybridization. Subcellular fractionation of rat brain demonstrates that neurabin and p70(S6k) both localize to the soluble fraction of synaptosomes. By way of its PDZ domain, the neuronal-specific neurabin may target p70(S6k) to nerve terminals.

Amino Acid Sequence↗
PubMed1998

Variability of persisting MHV RNA sequences constituting immune and replication-relevant domains.

Survivors of acute infection with the neurotropic JHM strain of mouse hepatitis virus develop a persistent infection of the central nervous system associated with chronic ongoing demyelination. Persistence is characterized by viral RNA in the absence of infectious virus. To associate persistence with possible immune evasion and/or replication defects, viral RNA from brains of acutely and persistently infected mice was examined for mutations by reverse transcriptase-PCR. Sequences analyzed included the encapsidation sequence (ECS), the transmembrane domains of the matrix (M) protein, and a cytotoxic T cell (CTL) epitope within the nucleocapsid (N) protein. The ECS, present only on genomic RNA, revealed minimal variability and was detected out to 120 days postinfection, suggesting low levels of replication. The M gene sequence also remained stable during persistence despite random mutations during the acute phase. Although the N gene sequence exhibited the greatest diversity, mutations were random and not selected for during persistence. A single exception was detected comprising a prominent Pro to Ser substitution in a region of N not associated with any known regulatory or immune function. Of the N gene mutations found within the CTL epitope in responder mice (H-2d), one resulted in reduced CTL recognition with no evidence of antagonist activity. However, this mutation was also detected in nonresponder mice (H-2b), suggesting that escape variants arising from CTL pressure play no role in establishing persistence in immunocompetent hosts infected as adults.

Animals↗
PubMed1998

Cellular factors in the transcription and replication of viral RNA genomes: a parallel to DNA-dependent RNA transcription.

Viral RNA replication and transcription involves not only viral RNA-dependent RNA polymerases, but also cellular proteins, the majority of which are subverted from the RNA-processing or translation machineries of host cells. These factors interact with viral RNA or polymerases to form transcription or replication ribonucleoprotein complexes and may provide template specificity for RNA-dependent RNA synthesis, suggesting a close parallel to the mechanism of DNA-dependent RNA synthesis. The types of cellular proteins involved and their modes of action are reviewed.

Animals↗
PubMed1998

Expression of hemagglutinin/esterase by a mouse hepatitis virus coronavirus defective-interfering RNA alters viral pathogenesis.

A defective-interfering (DI) RNA of mouse hepatitis virus (MHV) was developed as a vector for expressing MHV hemagglutinin/esterase (HE) protein. The virus containing an expressed HE protein (A59-DE-HE) was generated by infecting cells with MHV-A59, which does not express HE, and transfecting the in vitro-transcribed DI RNA containing the HE gene. A similar virus (A59-DE-CAT) expressing the chloramphenicol acetyltransferase (CAT) was used as a control. These viruses were inoculated intracerebrally into mice, and the role of the HE protein in viral pathogenesis was evaluated. Results showed that all mice infected with parental A59 or A59-DE-CAT succumbed to infection by 9 days postinfection (p.i.), demonstrating that inclusion of the DI did not by itself alter pathogenesis. In contrast, 60% of mice infected with A59-DE-HE survived infection. HE- or CAT-specific subgenomic mRNAs were detected in the brains at days 1 and 2 p.i. but not later, indicating that the genes in the DI vector were expressed only in the early stage of viral infection. No significant difference in virus titer or viral antigen expression in brains was observed between A59-DE-HE- and A59-DE-CAT-infected mice, suggesting that virus replication in brain was not affected by the expression of HE. However, at day 3 p.i. there was a slight increase in the extent of inflammatory cell infiltration in the brains of the A59-DE-HE-infected mice. Surprisingly, virus titers in the livers of A59-DE-HE-infected mice were 3 log10 lower than that of the A59-DE-CAT-infected mice at day 6 p.i. Also, substantially less necrosis and viral antigen were detected in the livers of the A59-DE-HE-infected mice. This may account for the reduced mortality of these mice. The possible contribution of the host immune system to this difference in pathogenesis was analyzed by comparing the expression of four cytokines. Results showed that both tumor necrosis factor-alpha and interleukin-6 mRNAs increased in the brains of the A59-DE-HE-infected mice at day 2 p.i., whereas interferon-gamma and interleukin-1 alpha mRNAs were similar between A59-DE-HE- and A59-DE-CAT-infected mice. These data suggest that the transient expression of HE protein enhances an early innate immune response, possibly contributing to the eventual clearance of virus from the liver. This study indicates the feasibility of the DI expression system for studying roles of viral proteins during MHV infection.

Animals↗
PubMed1998

Cellular protein hnRNP-A1 interacts with the 3'-end and the intergenic sequence of mouse hepatitis virus negative-strand RNA to form a ribonucleoprotein complex.

We previously showed that several cellular proteins specifically bind to the 3'-end and the intergenic sequences of the negative-strand RNA of mouse hepatitis virus (MHV), and proposed that these distant RNA sequences can be brought together by cellular and viral proteins (Furuya and Lai, 1993; Zhang et al., 1994; Zhang and Lai, 1995). The cellular protein p35 has been identified as a heterogeneous nuclear ribonucleoprotein (hnRNP) A1. We have now expressed hnRNP-A1 as a glutathione-S-transferase (GST) fusion protein and demonstrated that the amino terminal two-thirds of hnRNP-A1 interacted with the two MHV regulatory RNA sequences (3'-end and intergenic sequences) through protein-RNA interaction while its carboxy-terminal glycine-rich domain mediated homomeric (protein-protein) interactions. In a partially reconstituted reaction, in which the two MHV RNA fragments and the purified GST-hnRNP-A1 fusion protein were mixed, an RNP complex was formed. Depletion of either hnRNP-A1 or one of the RNA components abolished the complex formation. These results indicate that hnRNP-A1 can mediate the formation of an MHV RNP complex, which includes both the negative-strand leader and intergenic sequences. Site-directed mutagenesis revealed that mutations in the MHV intergenic sequences, which inhibited MHV RNA transcription, also inhibited the RNP complex formation. Deletion analysis showed that the amino terminal RNA-binding domains of hnRNP-A1 is essential for the RNP complex formation while the carboxy-terminal protein-binding domain enhanced the complex formation by 90-fold. These findings provide direct evidence demonstrating that the negative-strand leader RNA and intergenic sequences can form an RNP complex mediated by cellular protein hnRNP-A1.

Animals↗
PubMed1998

Using a defective-interfering RNA system to express the HE protein of mouse hepatitis virus for studying viral pathogenesis.

We have developed a defective-interfering (DI) RNA of mouse hepatitis virus (MHV) as a vector for expressing a variety of cellular and viral genes including the chloramphenicol acetyltransferase (CAT), hemagglutinin' esterase (HE), and gamma interferon. Here, we used the HE-expressing DI RNA for examining the role of HE protein in viral pathogenesis. The pseudorecombinant virus containing an expressed HE protein was generated by infecting cells with MHV-A59, which does not express, HE, and transfecting the in vitro-transcribed DI RNA containing the HE gene. These pseudorecombinant viruses (DE-HE A59) were then inoculated intracerebrally into mice. Viruses recovered from cells infected with A59 and transfected with DI RNA expressing the CAT gene (DE-CAT A59) were used as a control. At various time points after inoculation, mice were observed for clinical symptoms. Tissues (brains and livers) were obtained for determining the replication of DI RNA by RT-PCR, virus replication by plaque assay, antigen expression by immunohistochemistry, and pathological changes. Results showed that all mice infected with DE-CAT A59 succumbed to infection by 9 days postinfection (d p.i). These data are identical to the pathogenesis of the parental A59 virus, demonstrating that inclusion of the DI RNA did not by itself alter pathogenesis. In contrast, only 40% of mice infected with DE-HE A59 succumbed to infection. The subgenomic mRNAs transcribed from the DI vector were detected at 1 and 2 d p.i. but not at subsequent time points, indicating that the genes in the DI vector were expressed only at an early stage of viral infection. No significant difference in virus replication in the brains was detected between these two groups of mice, suggesting that virus replication in brains was not affected by the expression of the HE. Histopathological examination showed only a small increase in the extent of inflammatory cell infiltration and reduced viral antigen in the mice infected with DE-HE A59. There was no difference in virus replication in the livers at 2 and 4 d p.i., but a 3 log10 reduction was detected in the livers of mice infected with DE-HE A59 at 6 d p.i. Histological examination showed a significant reduction in viral antigen, inflammation and necrosis in mice infected with DE-HE A59. These results indicate that the expression of HE from the DI vector altered the viral pathogenesis. This study thus demonstrates the usefulness of this system in studying the role of viral or cellular genes expressed locally at the sites of viral infection in viral pathogenesis.

Animals↗
PubMed1998

Viral evolution and CTL epitope stability during JHMV infection in the central nervous system.

The JHM strain of mouse hepatitis virus (JHMV) establishes a persistent infection in the murine central nervous system (CNS) associated with chronic ongoing demyelination in the absence of detectable virus. To distinguish between immune and replication associated mechanisms of persistence, brains from acutely and persistently infected mice were analyzed for viral RNA mutations in the encapsidation sequence (ECS) and regions encoding either the transmembrane domains of the matrix (M) protein or a protective CTL epitope in the nucleocapsid (N) protein. Detection of the ECS to 120 days post infection (p.i.) indicated low levels of replication. The ECS remained stable whereas the fragment encoding the CTL epitope revealed extensive diversity with mutation frequencies in the order of 2.0 per 1000 nts. The M gene also remained stable despite random mutations during the acute phase. Mutations in the N gene were random and not selected for during persistence, with the exception of a single prominent Pro363 to Ser substitution in a region not associated with any known regulatory function or immune response. Mutations within the CTL epitope affecting CTL recognition were found early in responder BALB/c mice (H-2d), but also in non-responder C57BL/6 (H-2b) mice, suggesting that CTL escape variants play no significant role in establishing persistence.

Animals↗
PubMed1998

Neural actions of immunophilin ligands.

Immunophilins, protein receptors for immunosuppressant drugs such as cyclosporin A and FK506, are enriched far more in the brain than in the immune system. Drug-immunophilin complexes bind to calcineurin, inhibiting its phosphatase activity and leading to immunosuppressant effects. The immunophilin FKBP-12 (FK506 binding protein, 12 kDa) forms a complex with the ryanodine and inositol (1,4,5) trisphosphate (IP3) receptors to regulate their physiological release of intracellular Ca2+. Here, Solomon Snyder and colleagues describe how non-immunosuppressant as well as immunosuppressant immunophilin ligands are neurotrophic for numerous classes of damaged neurones, both in culture systems and intact animals. Their ability to stimulate functional regrowth of damaged sciatic, cortical cholinergic, dopamine and 5-HT neurones may have therapeutic relevance.

Animals↗
PubMed1998

Apoptosis and viral hepatitis.

Viral hepatitis is a diffuse inflammatory reaction of the liver caused by hepatotropic viruses. Pathomorphologic studies have shown acidophilic bodies and hepatocyte dropout, features that are compatible with apoptosis. Using in situ terminal transferase labeling, the number of hepatocytes showing features of apoptosis in patients with chronic hepatitis B and C was found to be small but higher than healthy subjects, indicating that apoptosis is involved in the pathogenesis of these diseases. There is evidence that the immune (cytotoxic T lymphocyte)-mediated pathways of apoptosis are activated, suggesting that the apoptosis of liver cells may at least in part be related to the host immune defense. Whether other cytokine-mediated and cellular constitutive apoptotic pathways are involved or not remains to be studied. There are recent data suggesting that hepatitis B and C viral proteins may modulate apoptosis. The exact role of these observations in relation to pathogenesis remains to be established.

Apoptosis↗