Local atomic order and individual pair displacements of Fe46.5Ni53.5 and Fe22.5Ni77.5 from diffuse x-ray scattering studies.
Explore the source record for details and available documents.
Journal Article↗
Biomedical subjects
X Jiang
Publications and source records attributed to X Jiang.
Microsatellite analysis of endometriosis reveals loss of heterozygosity at candidate ovarian tumor suppressor gene loci.
Endometriosis is a very common gynecological condition in which tissue similar to endometrium proliferates at sites outside the uterine cavity, most commonly the ovary. Although it generally remains a benign condition, malignant transformation has been documented. and it is commonly found in association with endometrioid subtype ovarian cancer. Tumor suppressor genes are commonly altered in ovarian cancers, and the development of endometriosis may involve mutations in the same class of genes. We have investigated this possibility by examining DNA from 40 cases of endometriosis for clonal status, alterations in TP53 and RASK, and allelic losses at candidate ovarian tumor suppressor loci on chromosome arms 6q, 9p, l1q, 17p, l7q, and 22q. The majority of endometriotic cysts were monoclonal, but interestingly, 8 of 10 normal endometrial glands were also monoclonal, demonstrating that both are able to develop from a single progenitor cell. No mutations were detected in TP53 or RASK. Loss of heterozygosity (LOH) was detected on chromosomes 9p (18%), 1lq (18%), and 22q (15%). In total, 11 of 40 (28%) cases demonstrated LOH at one or more of these loci. This study, which is the first to report LOH in endometriosis, supports the notion that tumor suppressor gene inactivation may play a role in the development of at least a subset of cases.
Base Sequence↗
Expression of Norwalk virus capsid protein in transgenic tobacco and potato and its oral immunogenicity in mice.
Alternatives to cell culture systems for production of recombinant proteins could make very safe vaccines at a lower cost. We have used genetically engineered plants for expression of candidate vaccine antigens with the goal of using the edible plant organs for economical delivery of oral vaccines. Transgenic tobacco and potato plants were created that express the capsid protein of Norwalk virus, a calicivirus that causes epidemic acute gastroenteritis in humans. The capsid protein could be extracted from tobacco leaves in the form of 38-nm Norwalk virus-like particles. Recombinant Norwalk virus-like particle (rNV) was previously recovered when the same gene was expressed in recombinant baculovirus-infected insect cells. The capsid protein expressed in tobacco leaves and potato tubers cosedimented in sucrose gradients with insect cell-derived rNV and appeared identical to insect cell-derived rNV on immunoblots of SDS/polyacrylamide gels. The plant-expressed rNV was orally immunogenic in mice. Extracts of tobacco leaf expressing rNV were given to CD1 mice by gavage, and the treated mice developed both serum IgG and secretory IgA specific for rNV. Furthermore, when potato tubers expressing rNV were fed directly to mice, they developed serum IgG specific for rNV. These results indicate the potential usefulness of plants for production and delivery of edible vaccines. This is an appropriate technology for developing countries where vaccines are urgently needed.
Administration, Oral↗
Regional and laminar differences in synaptic localization of NMDA receptor subunit NR1 splice variants in rat visual cortex and hippocampus.
Changes in N-methyl-D-aspartate (NMDA) receptor expression may represent a molecular substrate for differences in synaptic plasticity between early postnatal and adult brains (Fox and Zahs [1994] Curr. Opinion Neurobiol. 4:112-119). We have, therefore, examined the regional and laminar distribution of NR1, the essential subunit of the NMDA receptor, in two regions in which synaptic plasticity has been most thoroughly studied: primary visual cortex and hippocampus. To study NR1 expression at the light and electron microscopic levels we have used a new antiserum (NR1-C1; Sheng et al. [1994] Nature 368:144-147) directed against a differentially spliced C-terminal exon ("C1"). The most striking result was that the pattern of NR1-C1 labeling in the adult was more restricted than that of previously published NR1-specific antibodies. Specifically, NR1-C1 did not label cells in the CA3, dentate gyrus or subicular regions of the hippocampus or in layer 4 of the visual cortex. Quantitative ultrastructural analysis revealed that these differences were paralleled by differential expression of NR1-C1 at synapses. In sharp contrast to the pattern in the adult, NR1-C1 immunoreactivity was distributed more widely in the developing brain. At postnatal day 11, NR1-C1 splice variants were expressed in all layers of the visual cortex and in all regions of the hippocampus. The transient expression of NR1-C1 splice variants in layer 4 of visual cortex suggests that NR1-C1 may play a role in determining the critical period for binocular plasticity. Continued expression of NR1-C1 in upper and lower layers of the adult cortex and in CA1 of the hippocampus may provide a substrate for plasticity in corticocortical connections and Schaffer collateral synapses beyond the critical period. In addition to abundant postsynaptic staining, NR1-C1 immunoreactivity was found in a large number of axon terminals in the dorsal subiculum, but in very few terminals in visual cortex. This strongly suggests that presynaptic NMDA receptors play a major role in neuronal processing of hippocampal output through the subiculum, but play a relatively minor role in visual processing.
Alternative Splicing↗
Effects of ether lipid 1-O-octadecyl-2-methoxy-rac-glycero- 3-phosphocholine and its analogs PAF and CPAF on the release of nitric oxide in primary cultures of rat astrocytes.
Ether lipid 1-O-octadecyl-2-O-methoxy-rac-glicero-3-phosphocholine (ET-18-OCH3) is an immunomodulator with antineoplastic activity. Its analog compounds PAF and CPAF share some of its biological effects. In our experiments, even very small amounts of ET-18-OCH3 released a remarkable quantity of nitric oxide (NO) from rat astrocytes cultured in vitro. The NO biosynthesis was inhibited by pretreatment with the antagonist BN 50730. The effect of ET-18-OCH3 was greater than that of the LPS inducer. PAF did not produce NO, even at high doses, while the nonmetabolizable CPAF only induced a significant release of NO from 12 micrograms/ml onwards. These results demonstrate that ET-18-OCH3 is functionally active also in astrocyte cultures. Stimulation of NO biosynthesis is of a great value on account of its the known effect as a neurotransmitter, potentiator of immune defences and possible modulator of cerebral circulation.
Animals↗
The clonal progression in the neoplastic process of nasopharyngeal carcinoma.
The clonality of a total of 70 human nasopharyngeal carcinomas (NPC) was analyzed using the structure of the terminal fragment of episomal Epstein-Barr virus (EBV). Thirty female samples heterozygous for the BstXI polymorphism of the phosphoglycerokinase (PGK) gene were analyzed using polymerase chain reaction (PCR) amplification of X-chromosome linked PGK gene for restriction fragment length polymorphism (RFLP). All NPC samples analyzed were shown to be monoclonal, with two exceptions that were polyclonal. Clonal determination was also performed for non-cancerous cell populations: normal, and simple hyperplastic, grade I (mild) and grade II-III (severe) atypical hyperplastic epithelia. It was found that the normal and simple hyperplastic and 3 grade I (mild) atypical hyperplastic epithelia were polyclonal, whereas the grade II-III (severe) atypical hyperplastic samples were monoclonal. The analysis of the clonality of various stages in the neoplastic process suggested that NPC might originate from several cells, after clonal selection; finally a large majority of NPC has been demonstrated to be monoclonal, also indicating that the alteration of clonal nature might have occurred at a very early stage.
Base Sequence↗
Proinflammatory agents, IL-8 and IL-10, upregulate inducible nitric oxide synthase expression and nitric oxide production in avian osteoclast-like cells.
Nitric oxide synthase (NOS) isoenzymes generate nitric oxide (NO), a sensitive multifunctional intercellular signal molecule. High NO levels are produced by an inducible NOS (iNOS) in activated macrophages in response to proinflammatory agents, many of which also regulate local bone metabolism. NO is a potent inhibitor of osteoclast bone resorption, whereas inhibitors of NOS promote bone resorption both in vitro and in vivo. The possibility that osteoclasts, like macrophages, express a regulated iNOS and produce NO as a potential autocrine signal following inflammatory stimulation was investigated in well-characterized avian marrow-derived osteoclast-like cells. NO production (reflected by medium nitrite levels) was markedly elevated in these cells by the proinflammatory agents lipopolysaccharide (LPS) and the synergistic action of IL-1 alpha, TNF alpha, and IFN gama. inhibitors of NOS activity (aminoguanidine, L-NAME) or iNOS induction (dexamethasone, TGF beta) reduced LPS-stimulated nitrite production. LPS also increased the NOS-associated diaphorase activity of these cells and their reactivity with anti-iNOS antibodies. RT-PCR cloning, using avian osteoclast-like cell RNA and human iNOS primers, yielded a novel 900 bp cDNA with high sequence homology (76%) to human, rat, and mouse iNOS genes. In probing osteoclast-like cell RNA with the PCR-derived iNOS cDNA, a 4.8 kb mRNA species was detected whose levels were greatly increased by LPS. Induction of iNOS mRNA by LPS, or by proinflammatory cytokines, occurred prior to the rise of medium nitrite in time course studies and was diminished by dexamethasone. Moreover, osteoclast-like cells demonstrated an upregulation of NO production and iNOS mRNA by IL-8 and IL-10, regulatory mechanism's not previously described. It is concluded that osteoclast-like cells express a novel iNOS that is upregulated by inflammatory mediators, leading to NO production. Therefore, NO may serve as both a paracrine and autocrine signal for modulating osteoclast bone resorption.
Animals↗
Antigenic mapping of the recombinant Norwalk virus capsid protein using monoclonal antibodies.
Norwalk virus (NV) is the prototype strain of a group of noncultivatable caliciviruses that infect humans and cause outbreaks of epidemic acute nonbacterial gastroenteritis. The NV virion is composed of 180 copies of a single structural protein that, when expressed in insect cells infected with a recombinant baculovirus, assembles into empty recombinant Norwalk virus-like particles (rNV VLPs) which are morphologically and antigenically similar to native NV. We have begun to dissect the antigenic structure of the rNV particles using monoclonal antibodies made to the rNV VLPs. Ten MAbs made to rNV particles were characterized for their reactivity as detector antibodies by ELISA, as capture antibodies in an ELISA to detect NV in stools, by Western blot, and by immunoprecipitation. Seven of the MAbs recognize discontinuous epitopes, requiring the rNV capsid protein to remain at least partially folded, while the other three recognize continuous epitopes. Eight of the MAbs map to the C-terminal half of the capsid protein as they react by Western blot and by immunoprecipitation with a 32K trypsin cleavage product of the full-length 58K capsid protein, suggesting that the C-terminal half of the capsid protein may contain the immunodominant epitopes. The three MAbs that recognize continuous epitopes map to the extreme C terminus of the capsid protein, between amino acids 457 and 530, in a region that is relatively conserved among different human calicivirus capsid proteins. These MAbs which were assigned into three antigenic groups will be useful as tools to further dissect the structural and antigenic topography of the NV virion, and as unlimited reagents to detect NV in diagnostic assays.
Animals↗
Regulation of phospholipase D by protein kinase C is synergistic with ADP-ribosylation factor and independent of protein kinase activity.
Phospholipase D (PLD) which was partially purified from membranes of porcine brain could be stimulated by multiple cytosolic components; these included ADP-ribosylation factor (Arf) and RhoA, which required guanine nucleotides for activity, and an unidentified factor which activated the enzyme in a nucleotide-independent manner (Singer, W. D., Brown, H. A., Bokoch, G. M., and Sternweis, P. C. (1995) J. Biol. Chem. 270, 14944-14950). Here, we report purification of the latter factor, its identification as the alpha isoform of protein kinase C (PKCalpha), and characterization of its regulation of PLD activity. Stimulation of PLD by purified PKCalpha or recombinant PKCalpha (rPKCalpha) occurred in the absence of any nucleotide and required activators such as Ca2+ or phorbol ester. This action was synergistic with stimulation of PLD evoked by either Arf or RhoA. Dephosphorylation of rPKC alpha with protein phosphatase 1 or 2A resulted in a loss of its kinase activity, but had little effect on its ability to stimulate PLD either alone or in conjunction with Arf. Staurosporine inhibited the kinase activity of PKCalpha without affecting activation of PLD. Finally, gel filtration of PKCalpha that had been cleaved with trypsin demonstrated that stimulatory activity for PLD coeluted with the regulatory domain of the enzyme. These data indicate that PKC may regulate signaling events through direct molecular interaction with downstream effectors as well as through its well characterized catalytic modification of proteins by phosphorylation.
ADP-Ribosylation Factors↗
Study on occurrence of human calicivirus (Mexico strain) as cause of sporadic cases and outbreaks of calicivirus-associated diarrhoea in the United Kingdom, 1983-1995.
The use of a recently developed EIA using antisera raised against purified baculovirus expressed recombinant Mexico virus (rMx) capsid protein is described for screening for human calicivirus in stools. The results show that MX-like viruses have been circulating in the UK periodically since 1983 and were an occasional cause of sporadic cases of diarrhoea in infants and outbreaks of infection among elderly patients in hospitals and old people's homes. Further evidence is presented that some strains of caliciviruses with characteristic surface morphology (HuCVs) and some with an indistinct appearance, small round structured viruses (SRSVs) are antigenically related to MxV. Tests on SRSVs from four unrelated outbreaks typed as UK3 failed to react in the Mx EIA or recombinant Norwalk virus (rNV) EIA. A 2-month survey of 206 children treated in two London hospitals for diarrhoea showed that only one was positive for MxV, a child known to be infected with HIV-1. None of the samples reacted in the rNV EIA.
Adult↗
Outbreaks of gastroenteritis in elderly nursing homes and retirement facilities associated with human caliciviruses.
Eleven outbreaks of acute gastroenteritis, eight of which were in nursing homes or retirement facilities, were reported in virginia during the winter of 1993-1994. Serum samples (four outbreaks) and stool samples (two outbreaks) from involved people were tested for human calicivirus (HuCV) infection by enzyme immune assays (EIAs) using recombinant Norwalk virus (rNV) and Mexico virus (rMX) capsid antigens and reverse transcription-polymerase chain reaction (RT-PCR). Of the 31 pairs of acute and convalescent serum specimens tested, 24 had a fourfold or more titer increase to rMX and 4 responded to rNV. In all four outbreaks, the geometric mean titers (GMTs) against rMX were significantly higher than those against rNV in the convalescent, but not in the acute phase of illness. The antibody response to rMX among these patients was also higher than to rNV (summary mean 32-fold increase vs. 0.7-fold increase, respectively, P < .001). Antigen was detected in 5 of 21 stool specimens tested by the rMX EIA, RNA in 12 of 17 stool specimens tested by RT-PCR, and small round structured virus (SRSV) particles in 12 of 21 by electron microscopy (EM); none were positive by the rNV EIA. Sequence analysis of the RT-PCR-amplified products from the viral RNA polymerase region revealed 92-93% amino acid identity with Snow Mountain agent (SMA), 86% with MX, 58-59% with NV, and 31-32% with Sapporo HuCV, suggesting that these viruses belong to the SMA HuCV genogroup.
Aged↗
The effect of ether lipid 1-O-octadecyl-2-O-methoxy-rac-glycero-3-phosphocholine and its analogues platelet activating factor and carbamyl-platelet activating factor on the biosynthesis of interleukin-6 in human thymic epithelial cells cultivated in vitro.
The effect of ether lipid 1-O-Octadecyl-2-O-methoxy-rac-glycero-3-phosphocholine (ET-18-OCH3), platelet activating factor (PAF) and carbamyl activating factor (CPAF) on the release of IL-6 in human thymic epithelium cultivated in vitro was studied. ET-18-OCH3 significantly increased the release of IL-6, giving the greatest effect at the dose of 2 micrograms/ml. PAF had no effect. The non-metabolizable CPAF was less effective than ET-18-OCH3 and showed the greatest release of IL-6 at the dose of 1 microgram/ml. Ether lipid ET-18-OCH3 has a thymotrophic effect as has also been demonstrated by other results obtained in our previous work.
Azepines↗
Structure of Norwalk virus.
Norwalk virus is the major cause of epidemic viral gastroenteritis of humans. Attempts to grow this virus in laboratory cell lines have been unsuccessful. However, the Norwalk virus capsid protein, when expressed in insect cells infected with a recombinant baculovirus, spontaneously assembles into virus-like particles. We have determined the 3-dimensional structure of baculovirus-expressed Norwalk virus using electron cryomicroscopy and computer image reconstruction to a resolution of approximately 22 A. These particles, having a diameter of 380 A exhibit T = 3 icosahedral symmetry. The 3-dimensional structure is composed of 90 dimers of the 58000 molecular weight (58 K) capsid protein, each of which forms an arch-like capsomere. The structure of the protein subunit is modular with three distinct domains. The distal globular domain that appears bilobed is connected to the lower shell domain by a central stem-like domain. We also have been able to grow crystals of the baculovirus-expressed Norwalk virus particles suitable for high resolution X-ray crystallography.
Capsid↗
Genetic and antigenic diversity of human caliciviruses (HuCVs) using RT-PCR and new EIAs.
RT-PCR using primers from conserved regions of calicivirus genomes, followed by sequencing, permits characterization of genetic variation within the family. EIAs based on baculovirus-expressed viral capsid proteins and hyperimmune antisera against the capsid proteins were developed to detect HuCV antigens and antibodies. Serologic surveys using recombinant Norwalk virus (rNV) and recombinant Mexico virus (rMX, a SMA-like virus) EIAs showed that infections by HuCVs are common and that children acquire antibodies to HuCVs at an early age in both developed and developing countries. Three HuCV genogroups have been described that are represented by Norwalk virus (NV), Snow Mountain agent (SMA), and Sapporo virus, although recently accumulated sequences of HuCV strains indicate these genogroups can be further divided. These genogroups also correspond to distinct antigenic groups based on the results of the recombinant EIAs. The three genogroups co-circulate and have a worldwide distribution, although the SMA genogroup seems to be predominant currently. Application of these new assays for further characterization of the genetic and antigenic properties of HuCVs remains an important task for HuCV research.
Antibodies, Viral↗
Partial characterization of the genome of nine animal caliciviruses.
Caliciviruses (CVs) include at least 42 distinct serotypes. Seventeen CV serotypes have been isolated from marine sources and are called San Miguel sea lion caliciviruses (SMSVs). CVs also have been isolated from reptiles, primates, and other terrestrial animals. Nucleotide sequences from portions of genome of prototype strains for six SMSV serotypes, the reptile CV, Cro-1, the cetacean CV, Tur-1, and the primate CV, Pan-1, are presented. cDNA products of the polymerase (all strains characterized) and capsid (SMSV-17) regions were produced by reverse transcription-polymerase chain reaction using Pan-1 primers. Comparisons of nucleotide and amino acid identity among these and published CV sequences indicated that the nine characterized CVs fall into a phylogenetic group that includes SMSV-1 and SMSV-4 and that is more closely related to other characterized animal CVs than to most human CVs. The phylogenetic analysis also indicated that distinct genera exist among the Caliciviridae. SMSV-17 and SMSV-4 are predicted to be closer to each other than other caliciviruses of known serotype; 574 (82%) of the 704 amino acids in the SMSV-17 and SMSV-4 capsid genes were identical.
Amino Acid Sequence↗
Urinary N-acetyl-beta-D-glucosaminidase changes in relation to age, sex, race, and diastolic and systolic blood pressure in a young adult biracial population. The Bogalusa Heart Study.
Increased urinary activity of N-acetyl-beta-D-glucosaminidase (NAG) has been reported in many clinical conditions, including essential hypertension. Since hypertension is increasingly recognized as beginning in childhood, we hypothesized that urinary NAG changes with increasing blood pressure may start early in life and may also be the evidence of the existence of early hypertensive disease. We analyzed the urinary NAG changes in 980 young adults, ages between 18 to 32, in relation to age, race, sex, and systolic and diastolic blood pressure. We observed that black women had the highest level of NAG, with or without adjustment for creatinine. With aging, urinary NAG significantly increased in men. As blood pressure increased, urinary NAG excretion appeared to increase, and this was more apparent in black women (P < .05). Significant correlations between NAG excretion and systolic (r = 0.12, P = .04) and diastolic (r = 0.18, P = .003) blood pressures existed in the oldest age group, 28 to 32 years old. These findings show that a significant association between urinary NAG and blood pressure exists in normal young adults and changes in urinary NAG may be evidence of early hypertensive disease.
Acetylglucosaminidase↗
Evaluation of an antigen capture ELISA based on recombinant mexico virus capsid protein.
BACKGROUND: The diagnosis of gastrointestinal infections caused by small round structured viruses (SRSV) has relied upon electron microscopy and antigen/antibody assays based on Norwalk virus. We investigated cases of gastroenteritis associated with SRSVs employing a new sandwich enzyme-linked immunosorbent assay (ELISA) using hyperimmune animal anti-sera against recombinant Mexico virus capsid protein (rMXV). STUDY DESIGN: One hundred and thirty-five specimens from 86 episodes of gastroenteritis associated with SRSVs, collected in the UK between October 1993 and September 1994, were tested in the rMXV assay. RESULTS: Forty-seven (35%) specimens from 35 of 86 (41%) episodes were positive in the rMXV ELISA and these could further be divided into high and low reactors. Sequencing of a 266-base region of the RNA polymerase gene revealed that strains highly reactive in the rMXV assay demonstrated a high degree of similarity to MXV (97-99% at the nucleotide level), whereas low-reactive strains consist of Mexico-like strains and a heterogeneous group of viruses exhibiting 70-75% similarity to MXV. CONCLUSION: Our results indicate that the rMXV ELISA is predominantly a type specific assay, although some cross reactivity with other genogroup 2 SRSVs was observed. MXV was responsible for 26% of SRSV-associated gastrointestinal infections investigated in the UK during one year's surveillance.
Journal Article↗
Loss of heterozygosity on chromosome 5q in ovarian cancer is frequently accompanied by TP53 mutation and identifies a tumour suppressor gene locus at 5q13.1-21.
Forty-nine ovarian tumours were examined for loss of heterozygosity (LOH) on chromosome 5 using eight microsatellite markers spanning both arms, including one at the APC locus. LOH on 5q was a frequent event, detectable in 23 of 49 (47%) tumours, whereas 5p LOH was detected in only 1 of 22 tumours (5%). Six tumours showed partial LOH on 5q, enabling the candidate region to be localised to a 22 cM region proximal to APC, flanked by D5S424 and D5S644. An association was found between 5q LOH and TP53 mutation, with 18 of 23 (78%) tumours with LOH on 5q also harbouring a TP53 mutation. LOH on 5q was observed in 6 of 18 (33%) stage I tumours, suggesting that it may be an early event in the molecular pathogenesis of certain ovarian carcinomas.
Base Sequence↗