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Biomedical subjects

G M Chen

Publications and source records attributed to G M Chen.

At least 19 recordsLinked to original sources

Analytical solution for the extremums of cell water volume and cell volume using a two-parameter model.

Based on a two-parameter model [Cryobiology 37 (1998) 271-289], the analytical solution for the extremums of cell water volume and cell volume for a two-solute system are obtained. Compared with the numerical solution, the analytical solution offers an accurate but simple choice. The approximate solution [Cryobiology 40 (2000) 64-83] for the extremum of cell water volume is also discussed, the reason for the deviation is presented.

Cell Membrane Permeability↗

Molecular and phylogenetic analyses of 16 novel simian T cell leukemia virus type 1 from Africa: close relationship of STLV-1 from Allenopithecus nigroviridis to HTLV-1 subtype B strains.

A serological survey searching for antibodies reacting with human T-cell leukemia virus type 1 (HTLV-1) antigens was performed on a series of 263 sera/plasma obtained from 34 monkey species or subspecies, originating from different parts of Africa. Among them, 34 samples exhibited a typical HTLV-1 Western blot pattern. Polymerase chain reaction was performed with three primer sets specific either to HTLV-1/STLV-1 or HTLV-2 and encompassing gag, pol, and tax sequences, on genomic DNA from peripheral blood mononuclear cells of 31 animals. The presence of HTLV-1/simian T-cell leukemia virus type 1 (STLV-1) related viruses was determined in the 21 HTLV-1 seropositive animals tested but not in the 10 HTLV-1 seronegative individuals. Proviral DNA sequences from the complete LTR (750 bp) and a portion of the env gene (522 bp) were determined for 16 new STLV-1 strains; some of them originating from species for which no STLV-1 molecular data were available as Allenopithecus nigroviridis and Cercopithecus nictitans. Comparative and phylogenetic analyses revealed that these 16 new sequences belong to five different molecular groups. The A. nigroviridis STLV-1 strains exhibited a very strong nucleotide similarity with HTLV-1 of the subtype B. Furthermore, four novel STLV-1, found in Cercocebus torquatus, C. m. mona, C. nictitans, and Chlorocebus aethipos, were identical to each other and to a previously described Papio anubis STLV-1 strain (PAN 503) originating from the same primate center in Cameroon. Our data extend the range of the African primates who could be permissive and/or harbor naturally STLV-1 and provide new evidences of cross-transmission of African STLV-1 between different monkey species living in the same environment and also of STLV-1 transmissions from some monkeys to humans in Central Africa.

Africa↗

Evolutionary inferences of novel simian T lymphotropic virus type 1 from wild-caught chacma (Papio ursinus) and olive baboons (Papio anubis).

A serological survey of 22 wild-caught South African (Transvaal) chacma baboons (Papio ursinus) and eight olive baboons (Papio anubis) from Kenya indicates that 13 P. ursinus and one P. anubis have antibodies reacting with human T cell leukemia/lymphoma virus type 1 (HTLV-1) antigens, whereas three P. ursinus had a indeterminate reactivity on Western blot analysis. With six primer sets specific to either HTLV-1-Simian T-cell leukemia virus type 1 (STLV-1) or HTLV-2 and encompassing long terminal repeat (LTR), gag, pol, env, and tax sequences, polymerase chain reaction was performed on genomic DNA from peripheral blood mononuclear cells of 18 animals, and the presence of HTLV-1-STLV-1-related viruses was determined in 13 seropositive and three seroindeterminate animals but not in the two HTLV seronegative individuals. Proviral DNA sequences from env (522 bp), pol (120 bp), and complete (755 bp) or partial (514 bp) LTR were determined for three STLV-1-infected P. ursinus and one P. anubis. Comparative and phylogenetic analyses revealed that P. anubis (Pan-486) sequence clusters with one (Pan-1621) of two previously described P. anubis STLV-1. Likewise, P. ursinus viruses (Pur-529, Pur-539, and Pur-543) form a distinct group, different from all known HTLV-1 but closely affiliated with two STLV-1 strains from South African vervets (Cercopithecus aethiops pygerythrus). This study, reporting the first STLV-1 sequences from wild-caught P. ursinus and P. anubis, corroborates the hypothesis of cross-species transmissions of STLV-1 in the wild. Further, phylogenetic analyses indicate that the known HTLV-1 strains do not share a common origin with nonhuman primates STLV in South Africa.

Animals↗

[Qualitative and quantitative studies of 1000 cases of isolated ventricular septal defect and follow-up using two-dimensional color Doppler echocardiography].

Color Doppler echocardiography (CDE) was used to evaluate 1,000 cases of isolated ventricular septal defect (IVSD). The types of IVSD included perimembrane (854), outflow tract (131) and muscular (15). Compared to surgical findings, CDE had 100% of sensitivity in the diagnosis of IVSD. The accuracy was 98.0% in determining the types of IVSD. CDE was significantly correlated with surgery in assessing sizes of defects (r = 0.893, P < 0.001). Continuous wave Doppler was significantly correlated with catheterization in measuring pulmonary systolic pressure (r = 0.838, P < 0.001). MPA/Ao value > 1.27 and enlargement of right ventricle were both considered good indices for reflecting pulmonary hypertension. A follow-up datum showed that 21 out of 80 cases of IVSD were closed spontaneously in their age of 7 months to 8 years, and the spontaneous closure was related to the types of defects. The complications of IVSD were also discussed.

Adolescent↗

Expression of the major inner capsid protein of the group B rotavirus ADRV: primary characterization of genome segment 5.

A complete cDNA copy of the fifth RNA segment of the human group B rotavirus, ADRV, has been cloned into plasmid AD512. Gene segment 5 contains 1269 bases and encodes one long open reading frame of 391 amino acids beginning at base 31 and terminating at base 1203. The gene 5 polypeptide, expressed in vitro in a rabbit reticulocyte lysate, comigrates with the 44-kDa major inner capsid protein present on EDTA treated ADRV virions. The gene 5 protein is immunoprecipitable by hyperimmune serum to ADRV, human ADRV convalescent serum and by a group B-specific monoclonal antibody. In addition, this protein shares amino acid identity and similarity with the VP6 proteins from group C and group A rotavirus strains. The ADRV VP6 equivalent protein appears to be more closely related to the group C VP6 than the Group A VP6 polypeptide and a common ancestral rotavirus VP6 precursor protein is suggested. As a result, the fifth RNA segment of ADRV defines the major inner capsid protein, or VP6 equivalent, in the group B rotavirus. Expression of the ADRV VP6 equivalent protein is potentially useful for evaluating the prevalence of serum antibodies to group B rotavirus in human and animal populations as well as for generating antibodies for the direct detection of group B rotavirus antigen.

Amino Acid Sequence↗

The role of mixed function oxidase (MFO) in the metabolism of the spin trapping agent alpha-phenyl-N-tert-butyl-nitrone (PBN) in rats.

It has been previously demonstrated that alpha-phenyl-N-tert-butyl-nitrone (PBN), one of the most widely used free radical trapping agents, is rapidly absorbed, evenly distributed among a wide range of tissues, and metabolized to form one metabolite. The objective of this study was to determine if the distribution and metabolism of PBN can be affected by inducers or inhibitors of the microsomal mixed function oxidase (MFO) system. Rats were pretreated with MFO inducers (phenobarbital and 3-methylcholanthrene) or MFO inhibitors (metyrapone and piperonyl butoxide) before 14C-PBN was injected (i.p.) The concentrations of 14C-PBN and its metabolite were measured in plasma, urine, liver, lung and kidney 2 hours after injection. The results indicated that when MFO was induced, the concentration of 14C-PBN metabolite was significantly reduced in all tissues measured. The maximum concentration of PBN parent compound in the tissues where MFO was induced was 50% of that found in saline controls. Manipulation of tissue MFO levels with inducers and inhibitors altered the ratio of 14C-PBN parent compound to the PBN-metabolite. When 14C-PBN was incubated with rat liver microsomes at 37 degrees C in the presence of NADPH, the rate of metabolism was 1,752 dpm of 14C-PBN-metabolite formed/nmole P-450/min. Inactivation of MFO by heat (80 degrees C for 1 min) or deletion of NADPH diminished the formation of PBN metabolite in vitro. It is concluded that the MFO system may be responsible for the metabolism of PBN. Tissue concentrations of PBN can be affected by drugs or toxins which are inducers or inhibitors of MFO.

Animals↗

Identification of the gene encoding the group B rotavirus VP7 equivalent: primary characterization of the ADRV segment 9 RNA.

Gene segment 9 of the adult diarrheal rotavirus, ADRV, has been cloned, and the full-length gene 9 cDNA has been sequenced. Sequences at the 5' and 3' termini of the gene 9 double-stranded RNA were confirmed by direct RNA sequencing. The ninth RNA segment is 814 bases in length with a single open reading frame beginning at base 10 and terminating at base 757. The encoded polypeptide contains 249 amino acids with a calculated molecular weight of 28.5 kDa. The protein contains three potential N-linked glycosylation sites and a hydrophobic signal-like sequence at its amino terminus. A search of the NBRF protein data base with the encoded polypeptide revealed extensive similarities with VP7 proteins from a number of group A rotaviruses. Direct comparisons of the ADRV gene 9 polypeptide and the group A rotavirus VP7 demonstrate that the two proteins share 78% amino acid similarity and 28% identity as well as predicted secondary structure similarities. These findings suggest that the ADRV gene segment 9 encodes the VP7 polypeptide equivalent of group A rotaviruses.

Amino Acid Sequence↗

cDNA cloning of each genomic segment of the group B rotavirus ADRV: molecular characterization of the 11th RNA segment.

The group B noncultivatable rotavirus, ADRV, was purified from infected stool specimens. Double-stranded RNA was extracted, polyadenylated, reverse transcribed into cDNA, and cloned into plasmid vector pAT153. Each cDNA clone hybridized to a single ADRV RNA segment and cDNA clones of each genomic RNA segment were identified. Sequencing of genomic RNA and a full-length cDNA clone (AD63) determined that ADRV RNA segment 11 is 631 bases in length and contains a single open reading frame of 170 amino acids with a calculated molecular weight of 19.9 kDa and a pl of 6.2. The RNA 5'- and 3'-termini contain untranslated regions of 58 and 63 bases, respectively, and are complementary to each other. A comparison of encoded ADRV gene 11 amino acids with the NS26 protein of group A rotaviruses demonstrates a distant similarity between the two proteins. Further analysis and use of these ADRV clones should aid in determining the gene coding assignments for group B proteins as well as for diagnostic detection of group B or ADRV-specific nucleic acids in specimens.

Amino Acid Sequence↗

Excretion, metabolism and tissue distribution of a spin trapping agent, alpha-phenyl-N-tert-butyl-nitrone (PBN) in rats.

The objective of this study is using radiolabelled PBN to determine the tissue distribution, excretion, and metabolism of PBN in rats in order to evaluate the effective time to trap free radical in appropriate tissue(s). Our results demonstrated that PBN is rapidly absorbed when it is injected intraperitoneally in the animal. PBN can be used as an effective spin trapping agent for a variety of tissues since it is evenly distributed among a wide range of tissues measured. Since there is no difference in the tissue concentrations and distribution pattern of PBN at 15, 30 and 60 min after injection of PBN, it is appropriate to choose any of these time intervals to terminate the experiment and extract the spin adduct. The excretion of PBN, however, is slow. The majority of the radioactivity (70%) was excreted by the first 3 days. Only 5.7% of radioactivity was collected from 3 to 14 days. The remaining 25% of the radioactivity may be in the form of expired 14CO2. Trace amounts of radioactivity were recovered in the feces. PBN has probably only one major form of metabolite excreted in the urine. A small amount of the parent compound, however, was also excreted in the urine. The chemical structure of the metabolite(s) is still unknown.

Animals↗

Seroepidemiology and molecular epidemiology of the Chinese rotavirus.

The Chinese rotavirus which causes epidemics of diarrhoea in adult humans was isolated in 1983. This virus, designated adult diarrhoea rotavirus (ADRV), resembles typical rotaviruses morphologically and has a genome made up of 11 discrete segments of double-stranded RNA. Because the Chinese rotavirus has a unique RNA pattern on polyacrylamide gel electrophoresis and is antigenically distinct from group A rotaviruses, it is tentatively included in group B. Infection with ADRV or ADRV-related viruses (as shown by serological study) is detected in human populations as widespread as mainland China, Hong Kong, Australia, the United States and Canada, and in some domestic animals. RNA co-electrophoresis has shown homology of isolates from 12 different outbreaks (with some minor variations at segments 10, 11, 3 and 5). cDNA probes and monoclonal antibodies have been prepared to improve the detection and further characterization of the virus.

Adult↗

Effects of two negative staining methods on the Chinese atypical rotavirus.

The Chinese atypical (Group B) rotavirus, previously called the adult diarrhoea rotavirus (ADRV), was examined by transmission electron microscopy using either uranyl acetate or potassium phosphotungstate (PTA) as negative stains. Complete rotavirus particles were seen using uranyl acetate which were indistinguishable morphologically from typical rotaviruses. In the same preparations virus particles with differing degrees of degradation were seen after staining with PTA. This effect was not related to pH of the PTA and could be prevented by fixation of the specimen by 0.1 per cent glutaraldehyde. It is concluded that the use of PTA can give rise to falsely negative results for specimens containing this virus.

Glutaral↗

Prevalence of antibody to group B (atypical) rotavirus in humans and animals.

Enzyme-linked immunosorbent assays were developed for the detection of group B rotavirus antigen and antibody. The specificities of both assays were evaluated for antigens and serum specific for rotavirus groups A to D. Serum collected in the United Kingdom from different animal species exhibited the following high prevalence of group B rotavirus-specific antibody: pigs, 97%; cattle, 71%; sheep, 91%; and goats, 91%. In human serum, a lower prevalence of group B-specific antibody was detected; serum from blood donors showed 10% prevalence, and serum from veterinarians showed 4% prevalence.

Animals↗