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Biomedical subjects

J Xiao

Publications and source records attributed to J Xiao.

At least 19 recordsLinked to original sources
PubMed1999

Effects of glutamatergic, cholinergic and gabaergic antagonists on tectal cells in toads.

The present paper using microiontophoresis analysis describes transmitters and their receptor subtypes used in retinotectal and isthmotectal transmission, and suggests several modes converging retinotectal and isthmotectal afferents on tectal neurons in toads (Bufo bufo gargarizans). Neuronal responses of tectal cells were extracellularly recorded to both visual stimulation and electrical stimulation of the nucleus isthmi, and effects of glutamatergic, cholinergic, GABAergic and glycinergic antagonists on these responses examined. Visual responses in 80% of tectal cells were reversibly blocked by the N-methyl-D-aspartate antagonist 3-Rs-2-carboxypiperazin-4-yl-propyl-1-phosphonic acid, and those of the remaining 20% of cells by the muscarinic antagonist atropine, suggesting that there may be at least two kinds of retinotectal synapse that use glutamate and N-methyl-D-aspartate receptors, and acetylcholine and muscarinic receptors, respectively. Electrical stimulation of the nucleus isthmi elicited excitatory responses in 67% of tectal cells, excitatory-inhibitory responses in 16% of cells, and inhibitory responses in 17% of cells examined. The excitatory responses were reversibly abolished by atropine, but not affected by either 3-Rs-2-carboxypiperazin-4-yl-propyl-1-phosphonic acid or the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, whereas the inhibitory responses were released by the GABA receptor A antagonist bicuculline, but not influenced by the GABA receptor B antagonist 2-hydroxysaclofen and glycinergic antagonist strychnine. Excitatory and inhibitory components in the excitatory-inhibitory responses were blocked by atropine and bicuculline, respectively. It appears that glutamatergic and cholinergic afferents from the retina, and cholinergic and GABAergic afferents from the nucleus isthmi may converge on tectal neurons in at least five modes of synaptic connections, in agreement with the heterogeneous populations of tectal cells in amphibians.

Animals↗
PubMed1999

Dual role for nitric oxide in dynorphin spinal neurotoxicity.

The pharmacological effects of nitric oxide synthase (NOS) inhibitors, NO donor, and NOS substrate on dynorphin(Dyn) A(1-17) spinal neurotoxicity were studied. Intrathecal (i.t.) pretreatment with both 7-nitroindazole 1 micromol, a selective neuronal constitutive NOS (ncNOS) inhibitor, and aminoguanidine 1 micromol, a selective inducible NOS (iNOS) inhibitor, 10 min prior to i.t. Dyn A(1-17) 20 nmol significantly ameliorated Dyn-induced neurological outcome. Both 7-nitroindazole and aminoguanidine significantly antagonized the increases of cNOS and iNOS activities measured by conversion of 3H-L-arginine to 3H-L-citrulline in the ventral spinal cord, and blocked the Dyn-induced increases of ncNOS-immunoreactivity in the ventral horn cells 4 h after i.t. Dyn A(1-17) 20 nmol. Pretreatment with Nomega-nitro-L-arginine methyl ester (L-NAME) 1 micromol, a cNOS inhibitor nonselective to both ncNOS and endothelial NOS (ecNOS), did not antagonize Dyn A(1-17) 20 nmol-induced permanent paraplegia but aggravated Dyn A(1-17) 10 nmol-induced transient paralysis and caused permanent paraplegia. Pretreatment with L-NAME 1 micromol 10 min before i.t. Dyn A(1-17) 1.25 and 2.5 nmol, which produced no significant motor dysfunction alone, induced transient paralysis in seven out of 12 and five out of seven rats, respectively. L-NAME 1 micromol plus Dyn A(1-17) 10 nmol induced ncNOS-immunoreactivity expression in ventral horn cells. Both low and high doses of aminoguanidine (0.2-30 micromol) did not affect spinal motor function, but high doses of L-NAME (5-20 micromol) induced dose-dependent hindlimb and tail paralysis associated with spinal cord injury in normal rats. Pretreatment with low-dose Spermine NONOate, a controlled NO releaser, 0.1 and 0.5 micromol 10 min before i.t. Dyn A(1-17) 20 nmol, significantly prevented Dyn spinal neurotoxicity, and high-dose Spermine NONOate 2 micromol i.t. per se induced transient and incomplete paraplegia. But pretreatment with L-Arg 10 micromol 10 min before Dyn A(1-17) 20 nmol produced only partial blockade of Dyn-induced paraplegia. These results demonstrated that relatively specific inhibition of ncNOS and iNOS block Dyn-induced increases in cNOS and iNOS activities and ncNOS-immunoreactivity in ventral spinal cord, but nonspecific inhibition of ncNOS and ecNOS aggravated Dyn spinal neurotoxicity. It suggested that both ncNOS and iNOS play an important role, but ecNOS might be beneficial in Dyn spinal neurotoxicity. Moderate production of NO (at vascular level) has an apparently neuroprotective effect, and overproduction of NO (at cellular level) induces neurotoxicity.

Animals↗
PubMed1998

A discordancy test approach to identify outliers of length of hospital stay.

A discordancy test approach is proposed to identify outliers of inpatient length of stay. This has implications not only for benchmarking service delivery but also for linking budget allocation procedures to efficiency of health service provisions. The effects of shifting trim point thresholds on hospital payments are assessed in a case study of a group of obstetrical patients.

Adolescent↗
PubMed1998

The BRC repeats in BRCA2 are critical for RAD51 binding and resistance to methyl methanesulfonate treatment.

The BRCA2 gene was identified based on its involvement in familial breast cancer. The analysis of its sequence predicts that the gene encodes a protein with 3,418 amino acids but provides very few clues pointing to its biological function. In an attempt to address this question, specific antibodies were prepared that identified the gene product of BRCA2 as a 390-kDa nuclear protein. Furthermore, direct binding of human RAD51 to each of the four single 30-amino acid BRC repeats located at the 5' portion of exon 11 of BRCA2 was demonstrated. Such an interaction is significant, as BRCA2 and RAD51 can be reciprocally coimmunoprecipitated by each of the individual, specific antibodies and form complexes in vivo. Inferring from the function of RAD51 in DNA repair, human pancreatic cancer cells, Capan-1, expressing truncated BRCA2 were shown to be hypersensitive to methyl methanesulfonate (MMS) treatment. Exogenous expression of wild-type BRCA2, but not BRC-deleted mutants, in Capan-1 cells confers resistance to MMS treatment. These results suggest that the interaction between the BRC repeats of BRCA2 and RAD51 is critical for cellular response to DNA damage caused by MMS.

BRCA2 Protein↗
PubMed1998

A new slicing method for the lower vertebrate brain: brain mould and adjustable stage.

This paper describes a new method for slicing the lower vertebrate brain which is too small and soft to be sliced using conventional methods. The brain is sliced in a pre-prepared agar mould glued to a special stage placed in a conventional vibratome. The mould is constructed from a plaster model prepared by embedding a paraformaldehyde fixed brain in a paraffin and vaseline mixture. The brain to be sliced is placed within two prepared agar half cylinders which are in turn placed in a pre-prepared larger agar collar glued to a special stage. The study describes in detail the preparation of the interlocking collar and inner cylinders of agar. The plexiglass stage allows the agar block containing the brain, to be rotated and inclined to improve the angle of cut. By using this method, small and soft brain even with thin walls and large ventricles could be sliced coronally, sagittally, horizontally or obliquely. Brain slices obtained by this method have good viability showing spontaneous and evoked activity.

Animals↗
PubMed1998

Environmental occurrence and potential toxicity of planar, mono-, and di-ortho polychlorinated biphenyls in the biota.

Four non-ortho-, eight mono-ortho-, and two di-ortho-chlorinated congeners have been determined in fresh water and salt water mussels, fish, snapping turtles, mallard, seals, and in human milk and adipose tissue. The planar PCB congeners are separated from the remainder of PCBs by activated carbon chromatography or HPLC on porous graphitic carbon followed by gas chromatography with electron capture detection. PCB toxic equivalency factors (TEFs) recommended by WHO [1] for 3 non-ortho, 8 mono-ortho, and 2 di-ortho PCBs and a TEF for congener 81 suggested by Harris et al. [2] were used for calculation of the contribution to dioxin-like toxicity to each life form. In all the biota examined, PCB congener IUPAC number 126 was the major contributor to PCB toxic equivalents. Congeners IUPAC number 118, 114, 105, 156, 157, 77, 81, and 170 also contributed significantly to PCB toxic equivalents. The ability to separate out planar PCBs from the majority of PCBs has allowed the use of TCDD toxicity equivalence to compare the relative dioxin-like potency of PCB residues in various species from different locations.

Adipose Tissue↗
PubMed1998

Identification of trait-improving quantitative trait loci alleles from a wild rice relative, Oryza rufipogon.

Wild species are valued as a unique source of genetic variation, but they have rarely been used for the genetic improvement of quantitative traits. To identify trait-improving quantitative trait loci (QTL) alleles from exotic species, an accession of Oryza rufipogon, a relative of cultivated rice, was chosen on the basis of a genetic diversity study. An interspecific BC2 testcross population (V20A/O. rufipogon//V20B///V20B////Ce64) consisting of 300 families was evaluated for 12 agronomically important quantitative traits. The O. rufipogon accession was phenotypically inferior for all 12 traits. However, transgressive segregants that outperformed the original elite hybrid variety, V20A/Ce64, were observed for all traits examined. A set of 122 RFLP and microsatellite markers was used to identify QTL. A total of 68 significant QTL were identified, and of these, 35 (51%) had beneficial alleles derived from the phenotypically inferior O. rufipogon parent. Nineteen (54%) of these beneficial QTL alleles were free of deleterious effects on other characters. O. rufipogon alleles at two QTL on chromosomes 1 and 2 were associated with an 18 and 17% increase in grain yield per plant, respectively, without delaying maturity or increasing plant height. This discovery suggests that the innovative use of molecular maps and markers can alter the way geneticists utilize wild and exotic germplasm.

Alleles↗
PubMed1998

Conditional adherence of Enterococcus faecalis to extracellular matrix proteins.

The adherence of 44 clinical isolates of Enterococcus faecalis, a common cause of endocarditis, and 13 Enterococcus faecium to substrates of six extracellular matrix (ECM) proteins was examined using 35S-labeled bacteria. One E. faecalis strain, isolated from a patient with endocarditis, adhered to collagen types I and IV and another E. faecalis strain adhered to laminin and to collagen types I and IV. However, most isolates showed little adherence ( < 5% of added cells adhered) when grown at 37 degrees C regardless of their source (endocarditis, urine or fecal sample). When grown at 46 degrees C (but not when grown in CO2 or nutrient limited media), most isolates of E. faecalis increased their adherence to immobilized laminin, collagen types I and IV but not to fibronectin, fibrinogen or bovine serum albumin, whereas none of the E. faecium increased adherence when grown at 46 degrees C or 50 degrees C. The adherence of E. faecalis was eliminated by digestion with trypsin, suggesting that a protein is somehow important, directly or indirectly, for adherence to occur. Pre-incubation of bacteria with soluble collagen types I and IV inhibited the adherence to these ECM proteins. These results demonstrate that in E. faecalis, adherence to ECM proteins is produced during routine in vitro growth conditions by occasional isolates and can be produced during certain stressful growth conditions by others. Whether this adherence relates to the propensity of E. faecalis to cause endocarditis remains to be determined.

Animals↗
PubMed1998

In vivo protein binding and functional analysis of cis-acting elements in the U3 region of the bovine leukemia virus long terminal repeat.

Bovine leukemia virus (BLV) is a member of the human T-cell leukemia virus (HTLV)/BLV group of retroviruses. These viruses regulate their own transcription by producing Tax, a protein which activates the virus promoter region, the long terminal repeat (LTR). To explore the molecular mechanisms involved in the transactivation, we identified protein binding elements by in vivo footprinting and analyzed their function by site- directed mutagenesis. We used in vivo dimethyl sulfate footprinting by ligation-mediated PCR to detect constitutive in vivo protein-DNA interactions in a BLV-producing cell line, Bat2Cl6. The U3 region and part of the R region of the LTR were footprinted. In addition to the cis-acting elements (three cyclic AMP-responsive elements [CREs] and two AP4 sites) reported by others to be important for Tax-mediated activation of the BLV LTR, we found footprints in regions flanking these elements and in the core promoter region. The importance of these sites for transcriptional activation was studied by site-directed mutagenesis followed by promoter function analysis of the mutants with a chloramphenicol acetyltransferase reporter system. Our data corroborate those of others showing that the CREs are necessary for transactivation of the LTR, and they identify two new functional sites not previously reported by others. We show that the middle region of the BLV U3 contains multiple dual-functioning cis-acting elements which act as either positive or negative regulatory elements depending on the cell type tested. This is the first report of a functional mapping of the cis-acting elements of a virus of the HTLV/BLV group.

Animals↗
PubMed1998

Human cytomegalovirus glycoprotein B contains autonomous determinants for vectorial targeting to apical membranes of polarized epithelial cells.

We previously reported that human cytomegalovirus (CMV) glycoprotein B (gB) is vectorially transported to apical membranes of CMV-infected polarized human retinal pigment epithelial cells propagated on permeable filter supports and that virions egress predominantly from the apical membrane domain. In the present study, we investigated whether gB itself contains autonomous information for apical transport by expressing the molecule in stably transfected Madine-Darby canine kidney (MDCK) cells grown on permeable filter supports. Laser scanning confocal immunofluorescence microscopy and domain-selective biotinylation of surface membrane domains showed that CMV gB was transported to apical membranes independently of other envelope glycoproteins and that it colocalized with proteins in transport vesicles of the biosynthetic and endocytic pathways. Determinants for trafficking to apical membranes were located by evaluating the targeting of gB derivatives with deletions in the lumen, transmembrane (TM) anchor, and carboxyl terminus. Derivative gB(Delta717-747), with an internal deletion in the luminal juxtamembrane sequence that preserved the N- and O-glycosylation sites, retained vectorial transport to apical membranes. In contrast, derivatives that lacked the TM anchor and cytosolic domain (gBDelta646-906) or the TM anchor alone (gBDelta751-771) underwent considerable basolateral targeting. Likewise, derivatives lacking the entire cytosolic domain (gBDelta772-906) or the last 73 amino acids (gBDelta834-906) showed disrupted apical transport. Site-specific mutations that deleted or altered the cluster of acidic residues with a casein kinase II phosphorylation site at the extreme carboxyl terminus, which can serve as an internalization signal, caused partial missorting of gB to basolateral membranes. Our studies indicate that CMV gB contains autonomous information for apical targeting in luminal, TM anchor, and cytosolic domain sequences, forming distinct structural elements that cooperate in vectorial transport in polarized epithelial cells.

Amino Acid Sequence↗
PubMed1998

Synergetic protective effects of combined blockade by two kinds of autolesion mediator receptor on neurological function after cervical cord injury.

OBJECTIVE: To investigate the effects of combined blockade by platelet activating factor (PAF) receptor antagonist BN52021 in combination with opiate receptor antagonist naloxone on neurological function and neurological tissue damage after cervical cord injury. METHODS: Spinal cord contusion at C6 segment was made with Allen method in cats, which were randomly divided into four groups: saline control group; BN52021 group; naloxone group; and combined treatment group with BN52021 and naloxone. Alteration of cervical cord blood flow, blood barrier permeability of the spinal cord, cervical cord tissue pathology and neurological functional scores were studied after experimental cervical cord injury. RESULTS: The animals treated with BN52021 or naloxone had significantly better functional scores than saline controls 6 weeks after injury (P < 0.05). Moreover, the combined treatment showed significantly better neurologic recovery than either naloxone or BN52021 treated animals (P < 0.05). The other indexes in combined treatment animals were also superior to those in naloxone or BN52021 treated animals. CONCLUSIONS: Combined blockade by two kinds of autolesion mediator receptor can more effectively inhibit secondary damage production and development after cervical cord injury and improve neurologic function.

Animals↗
PubMed1998

[Study on the activity of fibroblast in hypertrophic scar].

To determine the state of fibroblast during the process of development of hypertrophic scar (HS), 40 specimens of HS in different periods were collected. The expressions of prolifrating cell nuclear antigen (PCNA) and Ag-protein in nucleolar organizer regions (Ag NORs) as well as the content of total amino acids in the tissues were examined. The hypertrophic scar of 1st and 3rd month old, the expression of PCND and Ag NORs were the highest. In the 9th and 12th month old, althrough PCNA was nearly negative, but the expression of Ag NORs was low. The content of total amino acid was increased gradually as HS developed but the increase of amount of hydroxyproline was markedly slowed down in 9 month old HS. It was suggested that: (1) in the developing process of HS the proecess of overproliferation of fibroblasts was short and limitted in 1-3 months period in the process of wound lealing; (2) the synthesis of collagen was nearly stopped at 6 months, but that of other extracellular matrix such as fibronectin and proteoglycan might be continued to aggregate after 12 months.

Adult↗
PubMed1998

Tegmental inhibition on isthmic neurons is mediated by the decussatio veli in amphibians.

This in vitro study on toads (Bufo bufo gargarizans) provides the first electrophysiological evidence that electrical stimulation of the decussatio veli in the midbrain tegmentum predominantly produced inhibition on neurons extracellularly recorded from the nucleus isthmi (NI). These cells were distributed throughout the nucleus. The present study lends strong support to the suggestion that the extratectal input to NI is mainly inhibitory and gives a good explanation for the previously reported inhibition of isthmic neurons by the contralateral optic nerve stimulation (Wu and Wang, 1995a).

Animals↗
PubMed1997

The eye-specification proteins So and Eya form a complex and regulate multiple steps in Drosophila eye development.

Sine oculis (so) and eyes absent (eya) are required for Drosophila eye development and are founding members of the mammalian Six and Eya gene families. These genes have been proposed to act with eyeless (Pax6) to regulate eye development in vertebrates and invertebrates. so encodes a highly diverged homeobox transcription factor and eya encodes a novel nuclear protein. We demonstrate that So and Eya (1) regulate common steps in eye development including cell proliferation, patterning, and neuronal development; (2) synergize in inducing ectopic eyes; and (3) interact in yeast and in vitro through evolutionarily conserved domains. We propose that an So/Eya complex regulates multiple steps in eye development and functions within the context of a network of genes to specify eye tissue identity.

Animals↗
PubMed1997

Cloning of the BssHII restriction-modification system in Escherichia coli : BssHII methyltransferase contains circularly permuted cytosine-5 methyltransferase motifs.

BssHII restriction endonuclease cleaves 5'-GCGCGC-3' on double-stranded DNA between the first and second bases to generate a four base 5'overhang. BssHII restriction endonuclease was purified from the native Bacillus stearothermophilus H3 cells and its N-terminal amino acid sequence was determined. Degenerate PCR primers were used to amplify the first 20 codons of the BssHII restriction endonuclease gene. The BssHII restriction endonuclease gene (bssHIIR) and the cognate BssHII methyltransferase gene (bssHIIM) were cloned in Escherichia coli by amplification of Bacillus stearothermophilus genomic DNA using PCR and inverse PCR. BssHII methyltransferase (M.BssHII) contains all 10 conserved cytosine-5 methyltransferase motifs, but motifs IX and X precede motifs I-VIII. Thus, the conserved motifs of M. BssHII are circularly permuted relative to the motif organizations of other cytosine-5 methyltransferases. M.BssHII and the non-cognate multi-specific phiBssHII methyltransferase, M.phiBss HII [Schumann,J. et al . (1995) Gene, 157, 103-104] share 34% identity in amino acid sequences from motifs I-VIII, and 40% identity in motifs IX-X. A conserved arginine is located upstream of a TV dipeptide in the N-terminus of M.BssHII that may be responsible for the recognition of the guanine 5' of the target cytosine. The BssHII restriction endonuclease gene was expressed in E.coli via a T7 expression vector.

Amino Acid Sequence↗
PubMed1997

Chromosomal regions associated with segregation distortion of molecular markers in F2, backcross, doubled haploid, and recombinant inbred populations in rice (Oryza sativa L.).

Chromosomal regions associated with marker segregation distortion in rice were compared based on six molecular linkage maps. Mapping populations were derived from one interspecific backcross and five inter-subspecific (indica/japonica) crosses, including two F2 populations, two doubled haploid (DH) populations, and one recombinant inbred (RI) population. Mapping data for each population consisted of 129-629 markers. Segregation distortion was determined based on chi-square analysis (P < 0.01) and was observed at 6.8-31.8%, of the mapped marker loci. Marker loci associated with skewed allele frequencies were distributed on all 12 chromosomes. Distortion in eight chromosomal regions bracketed previously identified gametophyte (ga) or sterility genes (S). Distortion in three other chromosomal regions was found only in DH populations, where japonica alleles were over-represented, suggesting that loci in these regions may be associated with preferential regeneration of japonica genotypes during anther culture. Three additional clusters of skewed markers were observed in more than one population in regions where no gametophytic or sterility loci have previously been reported. A total of 17 segregation distortion loci may be postulated based on this study and their locations in the rice genome were estimated.

Alleles↗
PubMed1997

Zn2+ promotes the self-association of human immunodeficiency virus type-1 integrase in vitro.

It has been recently demonstrated that the Mg(2+)-dependent 3'-processing activity of purified human immunodeficiency virus type-1 (HIV-1) integrase is stimulated by the addition of exogenous Zn2+ [Lee, S. P., & Han, M. K. (1996) Biochemistry 35, 3837-3844]. This activation was hypothesized to result from integrase self-association. In this report, we examine the Zn2+ content of purified HIV-1 integrase by atomic absorption spectroscopy and by application of a thiol modification reagent, p-(hydroxymercuri)benzenesulfonate, with a metallochromic indicator, 4-(2-pyridylazo)resorcinol. We find that the Zn2+ content of HIV-1 integrase varies from 0.1 to 0.92 equiv of Zn2+ per monomer depending on the conditions of protein purification. In vitro activity assays, time-resolved fluorescence emission anisotropy, and gel filtration chromatographic analyses all indicate that EDTA yields an apoprotein which is predominantly monomeric and less active with Mg2+. Further, sedimentation equilibrium studies reveal that reconstitution of the apoprotein with Zn2+ results in a monomer-tetramer-octamer transition. These results suggest that Zn2+ promotes a conformation with enhanced oligomerization and thereby stimulates Mg(2+)-dependent 3'-processing. This may also imply that multimers larger than dimers (tetramers and possibly octamers) are required for in vitro activity of integrase in the presence of Zn2+ and Mg2+. It should be noted, however, that the content of Zn2+ did not significantly affect the 3'-processing and strand transfer reactions with Mn2+ in vitro.

Chromatography, Gel↗