Biomedical subjects

X Jiang

Publications and source records attributed to X Jiang.

At least 289 records · Page 16Linked to original sources

PubMed1997

Effect of vasoactive intestinal peptide (VIP) on cytokine production and expression of VIP receptors in thymocyte subsets.

Intrathymic T cell precursors undergo a programmed sequence of developmental changes resulting in the production of mature, self-MHC restricted, single positive T lymphocytes which migrate to the periphery. The intrathymic T cell development is controlled by various factors, including cytokines and possibly neuroendocrine hormones. Our previous studies indicate that vasoactive intestinal peptide (VIP) inhibits IL-2 and IL-4 production in thymocytes through different molecular mechanisms. Thymocytes acquire the competence to express IL-2 and IL-2R during thymic development in a maturation-dependent manner. In this study we investigate the effect of VIP on IL-2 production, and the expression of VIP-R1 and VIP-R2 mRNA in different thymocyte subsets in comparison to T cell lines. All thymocyte subsets and T cell lines tested express VIP-R2. In contrast, only single positive, CD4+8- and CD4-8+ thymocytes express VIP-R1. VIP inhibits IL-2 production in CD4+8+ and single positive CD4+8- and CD4-8+ thymocytes and in TH1 cells stimulated through the TCR. No inhibition is observed in CD3-4-8- and single positive CD4+8- and CD4-8+ thymocytes, or in TH1 cells stimulated by a combination of calcium ionophores and phorbol esters. These findings suggest that VIP inhibits IL-2 production through VIP-R2, and that it interferes with a TCR-connected transduction pathway. We also investigate the expression of VIP mRNA in thymocyte subsets and T cell lines, and conclude that thymocytes as well as antigen-specific T cells may function as VIP sources within the lymphoid organs.

Animals↗

PubMed1997

Bridging of neutrophils to target cells by opsonized zymosan enhances the cytotoxicity of neutrophil-produced H2O2.

Hydrogen peroxide (H2O2) is a well-established cytotoxic agent released by activated neutrophils into the extracellular environment. However, a maximum of only 5 microM H2O2 was detected in the medium when 10(6) neutrophils/ml were activated with opsonized zymosan (OZ), more than 50-fold lower than the concentration of exogenous H2O2 required to produce equivalent killing of a cell line. In addition PMA-activated neutrophils were noncytotoxic, despite the capacity of PMA to generate two- to fourfold as much H2O2 for five times longer. The basis for this discrepancy was explored. NaN3 increased cytotoxicity to >90% only when neutrophils were activated with OZ due in part to inhibition of myeloperoxidase-mediated hydrolysis of H2O2, while catalase completely prevented cytotoxicity of OZ-activated neutrophils. These results indicate that H2O2 was solely responsible for the observed cytotoxicity. OZ-mediated cytotoxicity was prevented by intermittent agitation of the cultures or by the addition of soluble complement receptor type 1, suggesting that a physical association between neutrophils and target cells mediated by OZ was required to generate a cytotoxic environment. Significant numbers of neutrophil-target cell aggregates were observed by microscopic examination only under low hydrodynamic shear conditions. We conclude that the cytotoxic potency of H2O2 produced by neutrophils activated with OZ was due to a localized high concentration of H2O2 to which the target cells were exposed as a result of their labile adherence to OZ. This phenomenon may reflect a mechanism that neutrophils have acquired for maximizing the antimicrobial power of extracellular oxidants toward microbes that escape phagocytotosis.

Catalase↗

PubMed1997

Ligand-specific opening of a gated-porin channel in the outer membrane of living bacteria.

Ligand-gated membrane channels selectively facilitate the entry of iron into prokaryotic cells. The essential role of iron in metabolism makes its acquisition a determinant of bacterial pathogenesis and a target for therapeutic strategies. In Gram-negative bacteria, TonB-dependent outer membrane proteins form energized, gated pores that bind iron chelates (siderophores) and internalize them. The time-resolved operation of the Escherichia coli ferric enterobactin receptor FepA was observed in vivo with electron spin resonance spectroscopy by monitoring the mobility of covalently bound nitroxide spin labels. A ligand-binding surface loop of FepA, which normally closes its transmembrane channel, exhibited energy-dependent structural changes during iron and toxin (colicin) transport. These changes were not merely associated with ligand binding, but occurred during ligand uptake through the outer membrane bilayer. The results demonstrate by a physical method that gated-porin channels open and close during membrane transport in vivo.

Bacterial Outer Membrane Proteins↗

PubMed1997

Double mutagenesis of a positive charge cluster in the ligand-binding site of the ferric enterobactin receptor, FepA.

Siderophores and colicins enter bacterial cells through TonB-dependent outer membrane proteins. Using site-directed substitution mutagenesis, we studied ligand recognition by a prototypic Escherichia coli siderophore receptor, FepA, that binds the iron chelate ferric enterobactin and colicins B and D. These genetic experiments identified a common binding site for two of the three ligands, containing multiple positive charges, within cell surface residues of FepA. Elimination of single residues in this region did not impair the adsorption or transport of ferric enterobactin, but double mutagenesis in the charge cluster identified amino acids (Arg-286 and Arg-316) that participate in siderophore binding and function in FepA-mediated killing by colicins B and D. Ferric enterobactin binding, furthermore, prevented covalent modification of FepA within this domain by either a fluorescent probe or an arginine-specific reagent, corroborating the involvement of this site in ligand recognition. These results identify, for the first time, residues in a TonB-dependent outer membrane protein that participate in ligand binding. They also explain the competition between ferric enterobactin and the colicins on the bacterial cell surface: all three ligands interact with the same arginine residues within FepA during their penetration through the outer membrane.

Amino Acid Sequence↗

PubMed1997

Immune CD8+ T lymphocytes lyse Listeria monocytogenes-infected hepatocytes by a classical MHC class I-restricted mechanism.

Hepatocytes constitute the principal site of listerial replication in the livers of mice infected i.v. CD8+ T lymphocytes play a predominant role in the host defenses to Listeria monocytogenes. In vitro experiments by others undertaken to delineate the functions of CD8+ T lymphocytes have focused primarily on their interaction with Listeria-infected macrophages. Such experiments do not address directly the role of CD8+ T lymphocytes in eliminating the bulk of Listeria replicating within the liver. Here, we report that immune CD8+ T cells at an E:T cell ratio > or = 10:1 lysed Listeria-infected hepatocytes as judged by the following two criteria. Aspartate aminotransferase activity in the culture supernatants, indicative of hepatocyte damage, increased significantly. Conversely, infected hepatocytes cocultured with immune CD8+ T cells exhibited a marked reduction in viable intracellular Listeria assessed by CFUs. Neither immune CD4+ T cells nor nonimmune CD8+ T cells caused a similar increase in aspartate aminotransferase activity released or a decrease in intracellular bacteria. Immune CD8+ T cell-mediated lysis of infected hepatocytes was restricted by classical MHC class I (H-2Kb) molecules and was inhibited by the presence of either brefeldin A or mAb specific for CD8. These results suggest that the predominant role of CD8+ T lymphocytes in host resistance to listerial infections of the liver may be due to their capacity to lyse infected hepatocytes.

Animals↗

PubMed1997

Incidence of human calicivirus and rotavirus infection in patients with gastroenteritis in South Africa.

Human caliciviruses (HuCVs) are reportedly responsible for 2.5-4% of nonbacterial sporadic gastroenteritis. The incidence of HuCV infection in South Africa is unknown. Stool specimens from 1,296 South African patients with sporadic gastroenteritis were screened for the presence of HuCVs using electron microscopy, recombinant enzyme immunoassays for Norwalk (NV) and Mexican (MX) viruses, and the reverse transcriptase-polymerase chain reaction (RT-PCR). RT-PCR products were sequenced to ascertain which HuCV genogroups were present. HuCVs were detected in 43/1,296 (3.3%) specimens examined, with RT-PCR proving to be the most sensitive detection method. Genetic analysis of the isolates indicated that 81% were Snow Mountain Agent, or MX-like; 8% were NV-like; and 11% were HuCV/Sapporo-like. This study indicates that a combination of assays is needed for the accurate detection of HuCVs. Comparative data on hospitalised patients showed that the incidence of rotavirus infection was approximately ten times greater than that of HuCV infection.

Adolescent↗

PubMed1997

Phylogenetic analysis of the Caliciviruses.

A phylogenetic portrait of the genus Calicivirus in the family Caliciviridae was developed based upon published sequences and newly characterized calicivirus (CV) strains, including additional Sapporo-like HuCV strains in pediatric diarrhea stool specimens from South Africa, the United Kingdom, and the United States. Distance and parsimony methods were applied to nucleotide and amino acid sequences of human and animal calicivirus 3D RNA-dependent RNA polymerase (approximately 470nt) and capsid hypervariable regions (approximately 1,200nt) to generate phylogenetic trees. Pairwise amino acid identity in the 3D region among the Sapporo-like strains ranged from 61% to 100%. Human and animal caliciviruses (HuCVs and AnCVs) separated into five genogroups: small round-structured viruses (SRSV), Sapporo-like, and hepatitis E virus (HEV)-like HuCVs and rabbit-, and vesicular exanthema of swine virus (VESV)-like AnCVs, each with a distinct genome organization. Each genogroup, including the Sapporo-like HuCVs, subdivided further into subgenogroups. The capsid region trees had higher levels of confidence than the 3D region trees and limited conclusions about genogroups could be drawn from the 3D region analyses. This analysis suggested that CVs include five potential virus subfamilies.

Amino Acid Sequence↗

PubMed1997

Sapporo-like human caliciviruses are genetically and antigenically diverse.

The Sapporo-like human caliciviruses (HuCVs) comprise one of three genogroups of HuCVs associated with acute gastroenteritis. Phylogenetic analysis has shown that Sapporo-like HuCVs are related more closely to animal caliciviruses than to other known HuCVs. We produced 3.2 kb cDNA fragments from the 3' end to three Sapporo-like HuCVs that were associated with acute gastroenteritis in children (Houston/86, Houston/90, and London/92). Sequence analysis of the 3.2 kb cDNAs showed that two of the three viruses had a genomic organization similar to that of other Sapporo-like strains and the third strain (London/92) lacked an open reading frame overlapping the 5' end of the capsid gene. Alignment of the capsid sequences of these three strains showed 44-78% amino acid identity among the three strains. Phylogenetic analysis of the aligned sequences indicated the three strains are related but each belongs to a distinct genetic cluster. The genetic differences are associated with antigenic differences in that an enzyme immune assay (EIA) specific for the prototype Sapporo/82 strain detected the Houston/86 strain, but not the Houston/90 and London/92 strains. In vitro transcription and translation of viral cDNA containing the predicted capsid gene of Houston/90 resulted in a protein of 63 K, which is immunoprecipitated by sera from children infected with the strain. Genetically and antigenically distinct strains in the Sapporo-like HuCVs have not been described previously and the occurrence of such diverse strains in the same community likely increases the importance of these strains as a cause of illness in children.

Amino Acid Sequence↗

PubMed1997

Thymotrophic effect of ether lipid 1-O-octadecyl-2-O-methoxy-rac-glicero-3-phosphocholine in the mouse.

1-O-octadecyl-2-O-methoxy-rac-glicero-3-phosphocholine (ET-18-OCH3) is a synthetic derivate of 2-lysophosphatidyl-choline, endowed with some immunomodulatory and anticancer effects. In the present work we report that a chronic (1 microgram/g b.w. for 2 weeks) or acute single dose injection of ET-18-OCH3 produced a recovery of thymus weight and thymocytes cellularity in two different strains of mice, C57BL6 and Swiss mice, undergoing thymus age-dependent involution. This effect was significant when the thymus weight was reduced at 50% and it was without effect on thymus lacking age dependent involution, such as young mice. The ability of ET-18-OCH3 to produce thymus weight and thymocyte cellularity recovery was also demonstrated in adult mice showing hypotrophy of thymus induced by chronic corticosterone treatment, suggesting that this compound could be effective against thymus hypotrophy induced by external stimuli. This thymotrophic effect of ET-18-OCH3 was not dependent on direct action on thymocyte proliferation, but probably it was dependent on its action on thymic epithelial cells to produce hormone thymulin, which level was found significantly increased in the plasma. These results provide further immunomodulatory propriety of ET-18-OCH3 and open the possibility to use this compound to counteract thymus hypotrophy.

Aging↗

PubMed1997

Postanesthetic hypotropia: a unique syndrome in left eyes.

PURPOSE: To investigate a recurring syndrome of postoperative hypotropia after cataract surgery performed under local anesthesia. SETTING: Hawaiian Eye Center, Wahiawa, Hawaii, USA. METHODS: Thirty-one left eyes in 2143 cataract surgeries developed postoperative hypotropia. Phacoemulsification and intraocular lens implantation were performed by one surgeon; the anesthetic (containing bupivacaine) was administered by one anesthesiologist who had treated 4200 patients without incident. We conducted epidemiologic, outcomes, and prevention studies to identify the etiology, evaluate hypotropia treatment, and test specific anesthetic techniques, respectively. RESULTS: Changes in preoperative and intraoperative management and surgical methods were unsuccessful in preventing the hypotropia. In 55% of the cases, strabismus surgery was required and in 13%, prism glasses. Early recession of the inferior rectus and conjunctiva was successful. After repeated observations of the anesthetic technique, it was noted that the right-handed anesthesiologist misdirected the retrobulbar needle into the left orbit. This resulted in a change in the anesthetic technique, and there has been no incidence of postoperative hypotropia in 3000 surgeries. CONCLUSION: We believe this syndrome resulted from mytoxicity or perimuscular inflammation, producing contracture hypotropia and restricted elevation of the globe. We propose the term postbupivacaine hypotropia for this chiefly left-sided syndrome.

Anesthesia, Local↗

PubMed1997

Plasma insulin-like growth factor 1 distribution and its relation to blood pressure in adolescents: the Bogalusa Heart Study.

The distribution of circulating insulin-like growth factor-1 (IGF-1) and its relationship to blood pressure was examined in a community study of 1073 biracial (black-white) adolescents aged 11 to 18 years. Girls of both races displayed higher levels of plasma IGF-1 than did their male counterparts (P < .01), independent of age and sexual maturation. In boys, IGF-1 was correlated positively with height (r = 0.37 P < .001), weight (r = 0.26, P < .001), Tanner stage (r = 0.31, P < .001), and age (r = 0.11, P < .05). Girls, on the other hand, showed an inverse association with age (r = -0.38, P < .001) and Tanner stage (r = -0.10, P < .05). Plasma IGF-1 was correlated positively with systolic blood pressure in boys of both races (r = 0.21 to 0.25, P < .01) and with diastolic blood pressure in white boys (r = 0.18, P < .05), but not in girls of either race. Boys with elevated levels of IGF-1 (>80th percentile) showed significantly higher blood pressure levels, especially during early to middle stages of puberty. Multivariate analysis revealed that IGF-1 was associated with systolic and diastolic blood pressure, independent of age, race, sexual maturation, height, weight, and insulin in boys. These results suggest that plasma IGF-1 may contribute to the regulation of blood pressure only in males during puberty.

Adolescent↗

PubMed1997

Expression of soluble, active human macrophage colony stimulating factor in Escherichia coli.

Human macrophage colony stimulating factor (M-CSF) has been successfully overexpressed in Escherichia coli AD494 (DE3) with an expression level of approximate 26% of the total cellular proteins. The truncated human M-CSF gene encoding the amino-terminal 149 amino acids was subcloned into the prokaryotic expression vector pET11d under the control of the inducible T7 promoter. Nearly 40% of the recombinant protein was in the soluble fraction which showed obvious stimulating effects on mouse macrophage colony formation and had an M-CSF specific activity of approximately 1 x 10(6) units/mg soluble protein.

Animals↗

PubMed1997

Epidemiology of Mexico virus, a small round-structured virus in Yorkshire, United Kingdom, between January 1992 and March 1995.

The epidemiology of the small round-structured virus, Mexico virus (MxV), was investigated in North and West Yorkshire, United Kingdom, between January 1992 and March 1995 using a type-specific antigen ELISA. The results indicate that an epidemic of MxV occurred during the winter of 1993-1994, when this strain was associated with 45 of 99 outbreaks and sporadic childhood cases of gastroenteritis. Only 4 MxV-like isolates were found during the 1992-1993 winter season and none in the 1994-1995 season. This descriptive epidemiologic study suggests that MxV has an epidemic pattern of infection.

Caliciviridae↗

PubMed1997

Renaturation, purification, and characterization of human plasminogen activator inhibitor type 2 (PAI-2) accumulated at high level in Escherichia coli.

Plasminogen activator inhibitor 2 (PAI-2) is an important regulator of plasminogen activation, which inhibits both tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA). In this study we have developed a high-level expression system by inserting a modified PAI-2 gene downstream of the T7 promoter. The expression level of recombinant PAI-2 amounted to 55-60% of total microbial protein. By efficient renaturation and one-step purification, the recombinant protein was purified to homogeneity. The specific activity and yield of recombinant PAI-2 reached 33,000 IU/mg and 10 mg per gram wet weight of Escherichia coli cells, respectively. The second-order rate constant for uPA was 2.6-2.8 x 10(6) M(-1) x s(-1).

Cloning, Molecular↗

PubMed1997

Effect of bile on Vibrio parahaemolyticus.

Many enteric pathogens are thought to enter a viable but nonculturable state when deprived of nutrients. Virulent strains of the enteric pathogen Vibrio parahaemolyticus are rarely isolated from their low-nutrient aquatic environments, possibly due to their nonculturability. Host factors such as bile may trigger release from dormancy and increase virulence in these strains. In this study, the addition of bile or the bile acid deoxycholic acid to estuarine water-cultured bacteria led to an increase in the direct viable count and colony counts among the virulent strains. This effect was not demonstrated in the nonvirulent strains, and it was reversed by extraction of bile acids with cholestyramine. Bile-treated V. parahaemolyticus had lower levels of intracellular calcium than untreated cells, and this effect coincided with an increase in the number of metabolically active cells. Chelation of intracellular calcium with BAPTA/AM (R. Y. Tsien, Biochemistry 19:2396-2402, 1980) produced similar results. Addition of bile to V. parahaemolyticus cultures in laboratory medium enhanced factors associated with virulence such as Congo red binding, bacterial capsule size, and adherence to epithelial cells. These results suggest that a bile acid-containing environment such as that found in the human host favors growth of virulent strains of V. parahaemolyticus and that bile acids enhance the expression of virulence factors. These effects seem to be mediated by a decrease in intracellular calcium.

Bacterial Adhesion↗

PubMed1997

Study of Norwalk virus and Mexico virus infections at Ga-Rankuwa Hospital, Ga-Rankuwa, South Africa.

Small round-structured viruses (SRSVs) or Norwalk-like viruses have been implicated as an important causative agent of gastroenteritis outbreaks. We used the relatively newly developed recombinant enzyme immunoassays (EIAs) to determine the seroprevalence of Norwalk virus (NV) and Mexico virus (MxV) in a family-based cohort and an antenatal clinic cohort at Ga-Rankuwa Hospital, Ga-Rankuwa, South Africa. High prevalences (96 to 99%) of anti-NV and anti-MxV antibodies were detected in both cohorts. We also investigated the pattern of antibody acquisition in a cohort of infants and young children without gastroenteritis and found that by 48 months of age all children had acquired adult antibody levels to both these viruses. Lastly, we tested 276 stool specimens collected from infants and young children with gastroenteritis for the presence of NV or MxV antigen by recombinant EIAs to each virus, by electron microscopy (EM), and by reverse transcription (RT)-PCR. NV and MxV antigens were present in 1.8 and 4.3% of the stool specimens, respectively, by the recombinant EIAs; 9.2% were positive for SRSVs by EM, and 25% of these SRSVs gave a positive result by RT-PCR for primer pair 35-36 directed to a region of the RNA-dependent RNA polymerase gene. The seroprevalence studies indicate a high level of exposure to these viruses in both children and adults. Although the viral antigens are not highly prevalent in diarrheal stools, it was determined by the two assays for NV and MxV that children are, nevertheless, infected early in life.

Adolescent↗

PubMed1997

Glycosylation differences between a cystic fibrosis and rescued airway cell line are not CFTR dependent.

Altered glycosylation of mucus and membrane glycoconjugates could explain reported differences in binding of bacterial pathogens to cystic fibrosis (CF) versus normal tissue. However, because bacteria can alter cell surface glycoconjugates, it is not possible to assess the role of cystic fibrosis transmembrane conductance regulators (CFTR) in glycosylation in these studies. To address this issue, we have developed quantitative lectin binding assays to compare cell surface glycosylation in well-matched immortalized CF cells and rescued cell lines. The CF airway bronchial epithelial cell line IB3-1 consistently bound more peanut agglutinin (PNA) than its clonal derivative S9, which stably expresses functional wild-type CFTR. Pretreatment with neuraminidase increased PNA binding and abolished the difference between the two cell lines. However, infection of the IB3-1 cells with a replication-deficient recombinant adenovirus encoding CFTR restored CFTR function but did not alter PNA binding to cells. In contrast, treatment with the weak base ammonium chloride increased PNA binding to both cell lines as expected. Our data show that even clonally related CF and rescued cells can exhibit significant differences in carbohydrate processing. Although the differences that we found are consistent with the proposed role for CFTR in modulating intraorganellar pH, our data strongly suggest that they are CFTR independent. These studies add a cautionary note to the interpretation of differences in glycosylation between CF and normal primary tissues and immortalized cells.

Adenoviridae↗

PubMed1997

Serum Calmodulin Activity in Male Lead-exposed Workers.

Serum calmodulin (CaM) activity was studied in 75 lead-exposed and 21 non-exposed male workers. The lead-exposed workers were divided into groups with low blood lead (BPb < 50 µg/dL) and high blood lead (BPb>/=50 µg/dL). The concentrations of lead, calcium, magnesium, copper, zinc, and free erythrocytic protoporphyrin (FEP) in blood were determined. Serum samples were heated in a water bath (100 degrees C) for 3 minutes and centrifuged for 15 minutes at 4 degrees C (18,000 x g). The supernatants obtained were used to measure CaM activity. The results showed that: 1) Average blood lead concentrations in workers with both low and high levels of exposure were significantly higher than those in controls (p < 0.05). 2) Serum CaM activity in the high-exposure group (31.09 +/- 7.84 µg/dL) was significantly lower than that in controls (78.11 +/- 15.13 µg/dL, p < 0.05). The biological threshold of BPb inhibition of CaM activity was less than 50 µg/dL. 3) Multiple correlation analysis showed a negative dose-response relationship between BPb and CaM activity. The stepwise regression procedure indicated that lead had negative, and calcium and magnesium positive, effects on serum CaM activity. The regression equation was Y = 66.1383 - 1.0857 X&inf1; + 2.9676 X&inf2; + 5.2222 X&inf3; (Y:CaM; X&inf1;:Pb; X&inf2;: Ca; X&inf3;:Mg). These results of the first such study carried out in male lead-exposed workers suggest that lead can inhibit CaM activity in humans.

Journal Article↗