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J Xu

Publications and source records attributed to J Xu.

At least 1,027 records · Page 57Linked to original sources

Mouse Brca1: localization sequence analysis and identification of evolutionarily conserved domains.

The human genes BRCA1, conferring susceptibility to early-onset breast and ovarian cancer, has recently been isolated. Here we describe isolation of cDNAs, sequence analysis, and genomic localization of the murine homolog, Brac1. The mouse cDNA sequence predicts a protein of 1812 amino acids; a number of small gaps account for the 51 fewer residues in the mouse protein relative to human BRCA1. While the predicted mouse and human proteins display on the whole a high level of homology (58% identity, 73% similarity), the regions of greatest homology are at the respective amino and carboxyl termini. Most reported disease-associated missense mutations in human BCRA1 occurred within these more highly conserved terminal regions. A predicted zinc-building RING finger domain near the amino terminus lies within a 50 amino acid stretch that is perfectly conserved in both species. The strong conservation during mammalian evolution argues for the importance of this domain, perhaps mediating a role for BRCA1 in DNA and/or protein binding. We have also identified a conserved highly acidic domain in the carboxyl terminal half of the BCRA1 protein resembling acidic transactivation domains of certain transcription factors. Using an interspecific backcross panel, Brca1 was mapped to a region of mouse chromosome 11 that exhibits conserved linkage with 17q21. The sequence and isolated cDNAs will provide useful reagents for studying the expression of Brca1 in the mouse, and for testing the importance of the evolutionarily conserved domains.

Amino Acid Sequence↗

Comparison of a rapid nonisotopic polymerase chain reaction assay with four commonly used methods for the early diagnosis of human immunodeficiency virus type 1 infection in neonates and children.

To initiate antiretroviral therapy and prophylaxis for Pneumocystis carinii pneumonia, it is important to identify human immunodeficiency virus (HIV-1)-infected infants as soon after birth as possible. This study was undertaken to evaluate a novel 5-hour nonisotopic (NI) polymerase chain reaction (PCR) assay (Amplicor PCR; Roche Molecular Systems) and four other commonly used HIV-1 diagnostic tests including culture, oligonucleotide hybridization PCR, p24 antigen and immune complex-dissociated (ICD) p24 antigen tests and to determine the optimal age at which to perform these tests for the early and rapid diagnosis of HIV-1 in infants and children. We prospectively evaluated 225 infants and children, including 114 neonates, for HIV-1 infection. HIV-1 infection was defined as 2 positive HIV cultures. Of the 225 infants and children, 57 were infected, 138 were uninfected and 30 were of unknown (Centers for Disease Control and Prevention Classification P0) status. The sensitivity of NI PCR was 60% in cord blood, 40% at 0 to 2 days, 67 to 80% in the neonate (3 to 30 days) and 95 to 100% after 1 month of age. NI PCR was as sensitive as oligonucleotide hybridization PCR, culture, p24 antigen and ICD p24 antigen in the first 2 months of life and was more sensitive than p24 antigen or ICD p24 antigen thereafter. Specificity was 94% for cord blood and 99 to 100% for all age groups. The majority of HIV-1-infected newborns can be identified with NI PCR if testing is performed at birth and again between the third and fourth weeks of life. For older infants and children 2 NI PCR tests can correctly diagnose 98.5% of infected and uninfected infants and children. NI PCR of umbilical cord blood might also be useful as an initial screening test to identify infants who may be infected with HIV-1.

AIDS Serodiagnosis↗

Superior extension of the parascapular free flap for cervical burn scar contracture.

A newly designed superior extension of the parascapular free flap and its free transfer clinically for the repair of 11 cases of cervical burn scar contracture without any loss are presented. The flap sizes ranged from 16 x 7 cm to 32 x 11 cm. Based on the anatomic distribution of the circumflex scapular vessels, the flap in the scapular territory can be taken much longer and wider than before. There is less tension in this vertically oriented flap than in the horizontally oriented flap in obtaining primary closure. This method extends the application of the scapular territory as a donor site for free flaps.

Adolescent↗

aldB, an RpoS-dependent gene in Escherichia coli encoding an aldehyde dehydrogenase that is repressed by Fis and activated by Crp.

Escherichia coli aldB was identified as a gene that is negatively regulated by Fis but positively regulated by RpoS. The complete DNA sequence determined in this study indicates that aldB encodes a 56.3-kDa protein which shares a high degree of homology with an acetaldehyde dehydrogenase encoded by acoD of Alcaligenes eutrophus and an aldehyde dehydrogenase encoded by aldA of Vibrio cholerae and significant homology with a group of other aldehyde dehydrogenases from prokaryotes and eukaryotes. Expression of aldB is maximally induced during the transition from exponential phase to stationary phase. Its message levels are elevated three- to fourfold by a fis mutation and abolished by an rpoS mutation. In addition, the expression of an aldB-lacZ fusion was decreased about 20-fold in the absence of crp. DNase I footprinting analysis showed that five Fis binding sites and one Crp binding site are located within the aldB promoter region, suggesting that Fis and Crp are acting directly to control aldB transcription. AldB expression is induced by ethanol, but in contrast to that of most of the RpoS-dependent genes, the expression of aldB is not altered by an increase in medium osmolarity.

Aldehyde Dehydrogenase↗

Fis activates the RpoS-dependent stationary-phase expression of proP in Escherichia coli.

Fis is a general nucleoid-associated protein in Escherichia coli whose expression is highly regulated with respect to growth conditions. A random collection of transposon-induced lac fusions was screened for those which give increased expression in the presence of Fis in order to isolate a ProP-LacZ protein fusion. We find that proP, which encodes a low-affinity transporter of the important osmoprotectants proline and glycine betaine, is transcribed from two promoters. proP1 is transiently induced upon subculture and is upregulated by increases in medium osmolarity. As cells enter stationary phase, a second promoter, proP2, is strongly induced. This promoter can also be induced by high medium osmolarity in exponential phase. The activity of proP2 depends on Fis and the stationary-phase sigma factor sigmas. In the presence of Fis, proP2 expression is increased over 50-fold, as judged by the LacZ activity of cells carrying the proP-lacZ fusion as well as by direct RNA analysis, making this the most strongly activated promoter by Fis that has been described. Two Fis binding sites centered at positions -41 (site I) and -81 (site II) with respect to the transcription initiation site of P2 have been defined by DNase I footprinting. Mutations in site I largely abolish stationary-phase activation, while mutations at site II have a minor effect, suggesting that direct binding of Fis to site I is important for Fis-mediated activation of this promoter. In addition to Fis and sigmas, sequences located over 108 bp upstream of the proP2 transcription initiation site are required for efficient expression.

Amino Acid Sequence↗

Identification of genes negatively regulated by Fis: Fis and RpoS comodulate growth-phase-dependent gene expression in Escherichia coli.

Fis is a nucleoid-associated protein in Escherichia coli that has been shown to regulate recombination, replication, and transcription reactions. It is expressed in a transient manner under batch culturing conditions such that high levels are present during early exponential phase and low levels are present during late exponential phase and stationary phase. We have screened a random collection of transposon-induced lac fusions for those which give decreased expression in the presence of Fis. Thirteen different Fis-repressed genes were identified, including glnQ (glutamine high-affinity transport), mglA (methyl-galactoside transport), xylF (D-xylose-binding protein), sdhA (succinate dehydrogenase flavoprotein subunit), and a newly identified aldehyde dehydrogenase, aldB. The LacZ expression patterns revealed that many of the fusions were maximally expressed at different stages of growth, including early log phase, mid- to late log phase, and stationary phase. The expression of some of the late-exponential- and stationary-phase genes was dependent on the RpoS sigma factor, whereas that of others was affected negatively by RpoS. We conclude that Fis negatively regulates a diverse set of genes and that RpoS can function to both activate and inhibit the expression of specific genes.

Amino Acid Sequence↗

Structure and expression of histone H3.3 genes in Drosophila melanogaster and Drosophila hydei.

We demonstrate that in Drosophila melanogaster the histone H3.3 replacement variant is encoded by two genes, H3.3A and H3.3B. We have isolated cDNA clones for H3.3A and cDNA and genomic clones for H3.3B. The genes encode exactly the same protein but are widely divergent in their untranslated regions (UTR). Both genes are expressed in embryos and adults; they are expressed in the gonads as well as in somatic tissues of the flies. However, only one of them, H3.3A, shows strong testes expression. The 3' UTR of the H3.3A gene is relatively short (approximately 250 nucleotides (nt)). H3.3B transcripts can be processed at several polyadenylation sites, the longest with a 3' UTR of more than 1500 nt. The 3' processing sites, preferentially used in the gonads and somatic tissues, are different. We have also isolated the Drosophila hydei homologues of the two H3.3 genes. They are quite similar to the D. melanogaster genes in their expression patterns. However, in contrast to their vertebrate counterparts, which are highly conserved in their noncoding regions, the Drosophila genes display only limited sequence similarity in these regions.

Animals↗

Mechanical strain-enhanced fetal lung cell proliferation is mediated by phospholipase C and D and protein kinase C.

The signaling pathways by which intermittent strain (60 cycles/min, 15 min/h) regulates proliferation of mixed fetal rat lung cell in vitro have been investigated. Adenosine 3',5'-cyclic monophosphate (cAMP) content and cAMP-dependent protein kinase (PKA) activity were not affected by strain. The stimulatory effect of strain on DNA synthesis was also not influenced by the cyclic nucleotide-dependent protein kinase inhibitors H-8 or HA-1004, the adenylate cyclase inhibitor SQ-22536, or a PKA inhibitor and cAMP antagonist, adenosine 3',5'-cyclic monophosphothioate (Rp-cAMPS). In contrast, intracellular concentrations of two second messengers, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG), were dramatically increased after a short period of strain. This increase in second messengers was accompanied by an increased tyrosine phosphorylation of phospholipase C-gamma 1. Phospholipase D activity was also increased by strain. Mechanical strain elicited a shift in the subcellular distribution of PKC activity from cytosol to membranes shortly after the onset of strain. The specific activity of PKC in the membranes increased 6- to 10-fold within 5-15 min and remained increased throughout a 48-h period of intermittent strain. Strain-induced PKC activation and DNA synthesis were blocked by the PKC inhibitors H-7, staurosporine, and calphostin C, as well as by the phospholipase C inhibitor U-73,122. We conclude that mechanical strain of mixed fetal rat lung cells activates phospholipid turnover via phospholipases, followed by PKC activation, which then triggers the downstream events that lead to cell proliferation.

Animals↗

Segregation analysis of HDL3-C levels in families of patients undergoing coronary arteriography at an early age.

HDL cholesterol (HDL-C) level is a risk factor for coronary heart disease. Studies have shown a strong genetic influence on HDL-C levels in addition to environmental influences, but no definite major gene control has been demonstrated. Since HDL subfractions may better reflect the actions of distinct metabolic alterations than total HDL2 we tested the hypothesis that the amount of cholesterol in the denser HDL3 subfraction (HDL3-C) is under the control of a major gene. The study population included 676 family members of 116 probands who underwent coronary arteriography at an early age (men < or = 50 and women < or = 60 years). HDL3-C level was measured by using enzymatic methods after preparative ultracentrifugation at a density of 1.125 g/mL. HDL3-C was adjusted for age, gender, alcohol consumption, and smoking, which combined accounted for 3% of its variance. Segregation analysis was conducted on adjusted HDL3-C by using regressive models. The familial correlations for HDL3-C levels were spouse .03 +/- .08, parent-offspring .14 +/- .05, and sibling .24 +/- .05. The data strongly supported a codominant mendelian model, with the common allele coding for lower HDL3-C levels and the rarer allele (frequency, 25%) coding for higher HDL3-C levels. This major gene explained 34% of the variation in HDL3-C levels and 9% of the variation in total HDL-C levels. These results suggest that HDL3-C levels exhibit clearer genetic control than total HDL-C and may therefore be a useful target for further genetic studies.

Adult↗

Survival of pseudorabies virus on swabs maintained under standard field sample shipping conditions.

Pseudorabies virus survival was compared using three different types of applicator swabs in Eagle's minimum essential medium held under shipping conditions (packed with frozen gel packs) for up to 96 hours. Virus titer decay rates for dacron-tipped applicators were not statistically different from those of controls. Titer decay rates were statistically different from controls for cotton- and calcium alginate-tipped applicators. With the lowest input virus titer, virus was detectable up to 96, 72, or 24 hours after inoculation for dacron-, cotton-, and calcium alginate-tipped applicators, respectively. Dacron-tipped applicators were chosen to evaluate pseudorabies virus survival on tonsil swabs collected from experimentally challenged or contact control pigs to simulate field sampling and shipping conditions. Virus was still detectable in 20 of 24 swab samples after 72 hours in cell culture medium under shipping conditions.

Alginates↗

FGF-8 isoforms activate receptor splice forms that are expressed in mesenchymal regions of mouse development.

The Fgf8 gene is expressed in developing limb and craniofacial structures, regions known to be important for growth and patterning of the mouse embryo. Although Fgf8 is alternatively spliced to generate at least 7 secreted isoforms that differ only at their mature amino terminus, the biological significance of these multiple isoforms is not known. In this report, we demonstrate that multiple FGF-8 isoforms are present at sites of Fgf8 expression during mouse development. To address the possibility that the FGF-8 isoforms might interact with different fibroblast growth factor receptors, we prepared recombinant FGF-8 protein isoforms. We examined the ability of these proteins to activate alternatively spliced forms of fibroblast growth factor receptors 1-3, and fibroblast growth factor receptor 4. Recombinant FGF-8b and FGF-8c activate the 'c' splice form of FGFR3, and FGFR4, while FGF-8b also efficiently activates 'c' splice form of FGFR2. No activity could be detected for recombinant or cell expressed FGF-8a. Furthermore, none of the isoforms tested interact efficiently with 'b' splice forms of FGFR1-3, or the 'c' splice form of FGFR1. These results indicate that the FGF-8b and FGF-8c isoforms, produced by ectodermally derived epithelial cells, interact with mesenchymally expressed fibroblast growth factor receptors. FGF-8b and FGF-8c may therefore provide a mitogenic signal to the underlying mesenchyme during limb and craniofacial development.

Alternative Splicing↗

[Simplified enzyme-linked immunoadsorbent assays for myelin basic protein and antibodies to myelin basic protein].

Synchronous quantitative determination using simplified enzyme-linked immuno adsorbent assay has been developed for myelin basic protein (MBP) and antibodies to myelin basic protein (Anti-MBP). Antisera were prepared by immunizing rabbits with purified MBP from human white matter. Antibodies to MBP was conjugated with horseradish peroxidase (HRP) by using modified periodate oxidation method. Minimum detection amounts of MBP and Anti-MBP are 0.25 ng/ml and 0.20 ng/ml respectively. The activities of Anti-MBP-HRP conjugates are stable (stored at -30 degrees C) for at least 2 years. The detection procedure can be accomplished within 24 hours.

Animals↗

Modification of IFSSH system for evaluation of impairment of hand function.

We modified the examination and evaluation standard of impairment of hand function proposed by the International Federation of Societies for Surgery of the Hand (IFSSH). 1) The thumb adduction and opposition examination is changed from measuring distance alone to calculation of the ratio of thumb length to the distance of movement. 2) Sensation grading and two-point discrimination test are included in the sensory function evaluation. 3) In the estimation of anatomic loss of the hand, instead of measuring the length of the loss, calculation of the area of loss is proposed. 4) In the evaluation of joint movement, besides the angle of movement and position of the joint itself, influence of the defect distal to the joint thereon is also considered. 5) In the original evaluation standard, sensory disturb ances of the palmar and dorsal aspects of the hand as well as the anatomic loss of tissues through the palm are not taken into account. The present work pays special attention to this point. The software of the above mentioned work has been developed, approved by experts and adopted nation wide in China. The data of hand function examinations can be fed into the computer and the function loss in percentage can be displayed, or printed out. This software, using a graphic display, can also take care of the filling and retrieving of case history. At present the menu and instructions are in Chinese, which can be changed to suit other language.(ABSTRACT TRUNCATED AT 250 WORDS)

Anthropometry↗

[Uteroplacental blood flow monitoring by color Doppler flow imaging in pregnancy induced hypertension].

The uteroplacental blood flow was monitored using color Doppler flow imaging (CDFI) in 31 cases of pregnancy induced hypertension (PIH) and 71 cases of normal pregnancy. At the same time the values of serum estriol (E3), human placental lactogen (hPL), thromboxane (TXB2), 6-KP and TXB2/6-KP ratio were measured. The results showed that time average velocity (TAV) and volume of blood flow (Q) of uterine and umbilical arteries in PIH cases were much lower than those in normal pregnant women and fetuses (P < 0.05). After treatment with magnesium sulfate and qingxintong there was a marked improvement in placental blood flow. TAV and Q of uterine and umbilical arteries was also increased (P < 0.05). In PIH cases the E3 and hPL levels were much lower, while the ratio of TXB2/6-KP was significantly higher than that in normal pregnancy. Using CDFI to observe the velocity and volume of uteroplacental blood flow can monitor the blood supply of uterus and placenta. It is a valuable predictive index of fetal and placental function.

Acetophenones↗

[Expression of endothelin-1 mRNA, endothelin receptor-A and nitric oxide synthase mRNA in pulmonary artery and right ventriculus cordis of rats exposed to hypoxia].

Endothelins (ETs) are a family of novel regulatory peptides which can constrict the vascular and promote the proliferation of vascular smooth muscle cells. Serum ET-1 was elevated, and nitric oxide (NO) levels were reduced in hypoxic pulmonary hypertension. However, the exact mechanism remains unclear, and no study has elucidated if hypoxia could stimulate directly overgrowth of right ventricle in patients with chronic cor pulmonale. To investigate the role of ET-1 and NO in hypoxia-induced pulmonary hypertension and right ventricular hypertrophy, we measured the levels of ET-1 mRNA, ET receptor-A (ETR-A) mRNA and nitric oxide synthase (NOS) mRNA in the pulmonary artery and right ventricle of rats exposed to hypoxia (FiO = 0.1, 8 hours daily for 1, 2 and 3 weeks) by reverse transcription-polymerase chain reaction (RT-PCR). ET-1 mRNA level of pulmonary artery raised after 1 week's hypoxia (P < 0.05), and after 2 weeks' hypoxia, it returned to near normal, but elevated significantly again after 3 weeks' hypoxia. Pulmonary artery ETR-A mRNA in 1 and 2 weeks hypoxic groups showed no significant change, but it raised significantly in 3 weeks' hypoxic group. After exposure to hypoxia for 1, 2 and 3 weeks, NOS mRNA in the pulmonary artery all reduced significantly (P < 0.05). The right ventriculus cordis showed a significant increase in weight after 3 and 2 weeks' hypoxia. ET-1 mRNA showed no significant change but ETR-A mRNA increased significantly after 2 weeks' hypoxia; both ET-1 mRNA and ETR-A mRNA showed significant increase in weight after 3 and 2 weeks' hypoxia. In the ventriculus cordis of rats exposed to hypoxia for 2 and 3 weeks, NOS mRNA had no significant change.

Animals↗