Biomedical subjects

W Xu

Publications and source records attributed to W Xu.

At least 415 records · Page 23Linked to original sources

PubMed1996

Mouse uroporphyrinogen decarboxylase: cDNA cloning, expression, and mapping.

Uroporphyrinogen decarboxylase (URO-decarboxylase; EC 4.1.1.37), the heme biosynthetic enzyme responsible for the conversion of uroporphyrinogen III to coproporphyrinogen III, is the enzymatic defect in porphyria cutanea tarda, the most common porphyria. The mouse URO-decarboxylase cDNA was isolated from a mouse adult liver cDNA library. The longest clone of 1.5 kb, designated pmUROD-1, had 5' and 3' untranslated sequences of 281 and 97 bp, respectively, and an open reading frame of 1104 bp encoding a 367-amino acid polypeptide with a predicted molecular mass of 40,595 Da. The mouse and human coding sequences had 87.8% and 90.0% nucleotide and amino acid identity, respectively. The authenticity of the mouse cDNA was established by expression of the active enzyme in Escherichia coli. In addition, the analysis of two sets of multilocus genetic crosses localized the mouse gene, Urod, on Chromosome (Chr) 4, consistent with the map location of the human gene to a position of conserved synteny on Chr 1. The availability of the mouse URO-decarboxylase should facilitate studies of the structure and organization of the mouse genomic sequence and the development of a mouse model of this inherited porphyria.

Amino Acid Sequence↗

PubMed1996

Cultured epidermal sheet grafting with Hemaseel HMN fibrin sealant on nude mice.

Grafting of cultured epidermal sheets is a promising technique for skin restoration in extensive burns, but the technique has some limitations, resulting in variable graft takes. These experiments were designed to evaluate the innocuity of Hemaseel HMN fibrin sealant in the grafting process and in vivo evolution of cultured epidermis. A total of 30 mice were grafted, 15 were controls, 15 received tissue sealant application before the deposition of the cultured human epidermal sheets. Seven days after transplantation, compared to controls, the percentage of graft take over the total surface area grafted was greater in animals that had received the tissue sealant application. No difference was found 14 and 21 days postgrafting. In contrast, the percentage of graft take over the bony area (spinal) was significantly increased in animals grafted with previous application of sealant compared to controls at 7, 14 and 21 days postgrafting. Immunohistological and ultrastructural analysis showed that the evolution of the cultured human epidermis after transplantation was similar in both groups. The basement membrane was well structured 21 days after transplantation. The sealant was present at 4 days but not at 21 days postgrafting. Therefore, we conclude that the application of fibrin sealant before cultured epidermal sheet deposition on nude mouse graft bed is innocuous and enhances their mechanical stability. Since in this nude mouse system Hemaseel HMN fibrin sealant increased the percentage of graft take over areas difficult to engraft, we think that it may be advantageous in cultured epidermal sheet grafting on burn patients.

Animals↗

PubMed1996

Spontaneous rates of sex chromosomal aneuploidies in sperm and offspring of mice: a validation of the detection of aneuploid sperm by fluorescence in situ hybridization.

This study was designed to evaluate the frequency of aneuploid sperm in young adult mice of the genotype (102/E1 x C3H/E1)F1 determined by the fluorescence in situ hybridization (FISH) procedure and to evaluate the frequencies of aneuploid sperm observed by FISH compared with the frequencies of aneuploid offspring. Three-chromosome FISH was applied to determine the fractions of hyperhaploid and diploid sperm with DNA probes specific for chromosomes X, Y and 8. The animals were treated with three common solvents. Sperm smears were prepared for FISH by two similar protocols and were scored by different persons and in two different laboratories. There were no significant differences between scorers or laboratories. The frequencies of the sex chromosome aneuploidies in sperm (Y-Y and X-Y) were compared to the frequencies of mice carrying sex chromosome aneuploidy among controls of the heritable translocation assay in studies conducted from 1975-1995. To identify aneuploid individuals, untreated males and females of the genotype (102/E1 x C3H/E1)F1 were mated to assess their fertility by observing three consecutive litters. Semisterile and sterile animals were further analysed by meiotic cytogenetics and by karyotyping to determine the incidence of reciprocal translocations and sex chromosome aneuploidies (XXY and XYY). Based on the analysis of 175247 sperm and 9840 progeny, the frequency of Y-Y sperm was 0.01% while 0.03% of the offspring were XYY. The frequency of X-Y sperm was 0.005% while 0.02% of the offspring were XXY. The frequencies of aneuploid sex chromosomes were not significantly different between sperm and offspring. This allows two conclusions. First, there was no detectable prenatal selection against these sex-chromosomal aneuploid offspring, and second, germ cell aneuploidy can be reliably determined in mice by sperm FISH analyses.

Aneuploidy↗

PubMed1996

Anatomy of vasculature of 850 spleen specimens and its application in partial splenectomy.

BACKGROUND: Anatomic knowledge of vasculature of the splenic lobe and segment is of great clinical significance in partial resections and transplantation of the spleen. METHODS: The methods of two-colored corrosion casting, roentgenography, and anatomic dissection were used to evaluate the vasculature of the splenic hilum and intraspleen. On this basis splenic lobectomy and segmentectomy were performed on 42 patients with traumatized spleens. RESULTS: In the observation of 850 spleen specimens the spleen showed a single lobar artery in 7 cases (0.8%), two lobar arteries in 730 cases (86%), three lobar arteries in 104 cases (12.2%), and multiple lobar arteries (i.e., more than three lobar arteries) in 9 cases (1%). In a subgroup of 276 specimens 17%, 53%, 24%, 4%, 1%, and 1% of spleen specimens had three, four, five, six, seven, and eight segmental arteries, respectively. The result from a subgroup of 280 specimens indicated that mean percentages of the existence of the superior and inferior polar arteries and of the coexistence of both polar arteries were 31.3%, 38.8%, and 13.3%, respectively. Relative avascular planes between segments or lobes were seen. Basic steps of splenic lobectomy or segmentectomy include mobilization of the injured spleen, ligation of vessels in the lobe or segment, transection of the splenic parenchyma, and sutures of the cut surface of the remaining spleen. The postoperative courses of all 42 patients undergoing partial splenectomy were uneventful. No postoperative bleeding and necrosis of the remaining spleen or infectious complications were registered. CONCLUSIONS: Anatomically the spleen is defined with two primary lobes (the superior lobe and inferior lobe), one accessory lobe, and three to five segments. This new classification facilitates surgeons to perform partial resections of the spleen and allotransplantation of the hemispleen from a living related donor in human beings.

Adolescent↗

PubMed1996

A murine model of prenatal cocaine exposure: effects on the mother and the fetus.

To develop and characterize a murine model for investigating the long-term effects of prenatal cocaine exposure, the present study established the route of drug administration and the doses to be used for pregnant C57BL/6 mice. Comparison of the effects of a high dose of cocaine (60 mg/kg) when gavaged or injected subcutaneously (SC) established patterns of pathology characteristic of administration route but no dominating logic for selecting one over the other route for prenatal studies; however, because of the fourfold greater brain levels, with no evidence of greater pathology, the SC route was selected. When injected daily during gestation days 12-18, the period of prenatal development of dopamine systems, cocaine at doses producing plasma concentrations consistent with its stimulatory effects reduced food ingestion and weight gains during pregnancy and fetal body and brain weights at term. The extent of these reductions was comparable to reports on babies exposed to cocaine prenatally. Furthermore, the present study suggests that maternal undernutrition is not a likely mediator of these perinatal effects and that differences in the amount of cocaine exposure may cause the contrasting effects of maternal cocaine noted in the human literature.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1996

A docking and modelling strategy for peptide-RNA complexes: applications to BIV Tat-TAR and HIV Rev-RBE.

BACKGROUND: In spite of the great interest in the interaction between RNAs and proteins, no general protocol for modelling these complexes is presently available. This methodological vacuum is particularly acute because the structure of few such complexes is known. RESULTS: A general strategy for docking and modelling RNA-protein complexes has been developed. The docking procedure involves minimizing electrostatic and van der Waals' interaction energies of conformationally rigid structures during docking. After docking, libraries of amino acid sidechain conformations are searched to obtain the best interactions between the peptide and the RNA. Using this method, we have reproduced the structure of a bovine immunodeficiency virus (BIV) Tat peptide bound to BIV TAR RNA and have developed a model for the structure of the arginine-rich HIV-1 Rev peptide (Rev34-50) interacting with the Rev-binding element (RBE). CONCLUSIONS: The resulting model of the Rev34-50-RBE complex predicts that although no single arginine sidechain is responsible for complex formation, residues Arg2, Arg5 and Arg11 are more important for binding than the other arginine residues in the peptide. One model is supported by binding measurements performed on wild-type and mutant RBE molecules with the peptide.

Animals↗

PubMed1996

The Arabidopsis XET-related gene family: environmental and hormonal regulation of expression.

Enzymes that modify cell wall components most likely play critical roles in altering size, shape, and physical properties of plant cells. Regulation of such modifying activity is expected to be important during morphogenesis and in eliciting developmental and physiological alterations that arise in response to environmental conditions. Previous work has shown that the Arabidopsis TCH4 gene encodes a xyloglucan endotransglycosylase (XET) which acts on the major hemicellulose of the plant cell wall. The expression of TCH4 is dramatically upregulated in response to several environmental stimuli (including touch, wind, darkness, heat shock, and cold shock) as well as the growth-enhancing hormones, auxin and brassinosteroids. This paper reports the presence of an extensive XET-related (XTR) gene family in Arabidopsis. In addition to TCH4, this family includes two previously identified genes, EXT and Meri-5, and at least five additional genes. The cDNAs of the XTR family share between 46 and 79% sequence identity and the predicted XTR proteins share from 37 to 84% identity. All eight proteins include potential N-terminal signal sequences and most have a conserved motif (DEIDFEFLG) that is also found in Bacillus beta-glucanase and may be important for enzyme activity. The members of the XTR gene family are differentially sensitive to environmental and hormonal stimuli. Magnitude and kinetics of regulation are distinct for the different genes. Differential regulation of expression of this complex gene family suggests a recruitment of related, yet distinct, cell wall-modifying enzymes that may control the properties of cell walls and tissues during development and in response to environmental cues.

Amino Acid Sequence↗

PubMed1996

A simple and rapid competitive enzyme-linked immunosorbent assay to identify HPA-1a (PlA1)-negative donor platelet units.

BACKGROUND: Alloantibodies to HPA-1a (PlA1) are the major cause of neonatal alloimmune thrombocytopenia and posttransfusion purpura and have been implicated in refractoriness to random-donor platelet transfusions. However, most assays used to phenotype platelets are cumbersome or time-consuming for large numbers of samples. STUDY DESIGN AND METHODS: A simple, competitive (inhibition) enzyme-linked immunosorbent assay for HPA-1a phenotyping of donor platelets was developed. A segment from the donor platelet unit transfer line was sealed to obtain a small aliquot of platelets. These platelets were washed once and added to a predetermined dilution of serum containing alloantibodies to HPA-1a. Residual anti-HPA-1a binding to the glycoprotein IIb/IIIa purified by lectin and high-performance liquid chromatography and coated on microtiter wells was detected with a conjugated antihuman IgG. A lack of inhibition equivalent to control (no platelets) was used to determine that the platelets were HPA-1b/b. RESULTS: Of the 557 platelet units tested, 14 (2.5%) were found to be HPA-1a negative, and they were confirmed to be HPA-1b/b by DNA genotyping. Two of the 14 HPA-1b/b units were also HPA-3b/b (approx. 0.35% of the random population). Use of the microtiter format allows 100 to 200 samples to be processed per day. CONCLUSION: This simple and inexpensive assay is useful for identifying HPA-1b/b units for platelet-compatible transfusions or for platelet antibody investigations.

Antigens, Human Platelet↗

PubMed1996

Biochemical characteristics of a human myeloid leukemia cell differentiation factor.

A human lymphocyte-derived maturation-inducing activity, which mediates the differentiation of human myeloid leukemia cells HL-60 to monocytic cells, was purified and characterized. The maturation inducer is a single polypeptide with a molecular mass of 54,300 Daltons. A Pl range of 8.0-8.5 was determined along with the amino acid composition of the inducer. In cell cultures, the purified inducer mediated a growth reduction and a differentiation of leukemia cells to mature monocytes-macrophages, possessing complement receptors and phagocytic capacity.

Amino Acids↗

PubMed1996

Ultrasonic assessment of human and bovine trabecular bone: a comparison study.

A comparison study is reported on the ultrasonic assessment of human trabecular and bovine trabecular bone samples. Both ultrasonic velocity and ultrasonic attenuation were evaluated through a transmission insertion technique and correlated with bone mineral density as determined with single photon absorptiometry. For a 1-MHz ultrasonic transducer pair and the human cancellous bone samples the correlations were 0.91 and 0.89 between density and velocity and attenuation, respectively. For a 500-kHz ultrasonic transducer pair the correlations were 0.89 and 0.81 between density and velocity and attenuation, respectively. For the bovine bone samples, the correlations were 0.90 and -0.31 for the velocity and attenuation, respectively, for the 1 MHz transducer pair. For the 500-kHz transducers, the correlations were 0.85 and -0.17 for the velocity and attenuation, respectively. By combining both velocity and attenuation in a multivariate regression, an improvement was achieved in the estimation of bone density in the human samples for both the 500-kHz and 1-MHz transducer pairs. No significant improvement was achieved in the multivariate regressions for the bovine bone samples. In conclusion, the results indicate that ultrasonic measurements are in general highly correlated with bone mineral density in trabecular bone samples. This correlation is more consistent and strong in relatively low density human samples compared with the higher density bovine samples.

Adult↗

PubMed1996

Modeling and measuring the elastic properties of an archaeal surface, the sheath of Methanospirillum hungatei, and the implication of methane production.

We describe a technique for probing the elastic properties of biological membranes by using an atomic force microscope (AFM) tip to press the biological material into a groove in a solid surface. A simple model is developed to relate the applied force and observed depression distance to the elastic modulus of the material. A measurement on the proteinaceous sheath of the archaebacterium Methanospirillum hungatei GP1 gave a Young's modulus of 2 x 10(10) to 4 x 10(10) N/m2. The measurements suggested that the maximum sustainable tension in the sheath was 3.5 to 5 N/m. This finding implied a maximum possible internal pressure for the bacterium of between 300 and 400 atm. Since the cell membrane and S-layer (wall) which surround each cell should be freely permeable to methane and since we demonstrate that the sheath undergoes creep (expansion) with pressure increase, it is possible that the sheath acts as a pressure regulator by stretching, allowing the gas to escape only after a certain pressure is reached. This creep would increase the permeability of the sheath to diffusible substances.

Archaea↗

PubMed1996

Positive charge modifications alter the ability of XIP to inhibit the plasma membrane calcium pump.

Exchange inhibitory peptide (XIP; RRLLFYKYVYKRYRAGKQRG) is the shortest peptide that inhibits the plasma membrane Ca pump at high Ca (A. Enyedi, T. Vorherr, P. James, D. J. McCormick, A. G. Filoteo, E. Carafoli, and J. T. Penniston, J. Biol. Chem. 264: 12313-12321, 1989). Sulfosuccinimidyl acetate (SNA)-modified XIP does not inhibit the Ca pump; SNA neutralizes the positive charge on Lys at positions 7, 11, and 17. Peptide 2CK-XIP (RRLLFYRYVYRCYCAGRQKG) inhibits the pump, but the iodoacetamido-modified peptide does not inhibit. Three peptide analogues, in which 7, 11, and 17 were Ala, Cys, or Lys, inhibited about as well as XIP. SNA modification of these analogues (each with 1 Lys) did not inhibit. SNA modification of 2CK-XIP results in a peptide that does not inhibit; thus position 19 is important. Our results suggest that it is critical that position 19 be positively charged, that positions 7, 11, and 17 are important contact points between XIP and the Ca pump (with at least one positively charged), and that, whereas it is not essential that residues 12 and 14 be positive, they cannot be negative.

Amino Acid Sequence↗

PubMed1996

Localization of thyrotropin-releasing hormone mRNA expression in the rat heart by in situ hybridization histochemistry.

Our laboratory has recently detected mRNA of thyrotropin-releasing hormone (TRH) in the rat heart. The density of mRNA for TRH is five-fold higher in the atria than in the left and right ventricle. We also found TRH receptor mRNA and 3H-TRH-binding sites in both ventricles. Cardiac contractility was stimulated after intracoronary administration of TRH. This study was performed to investigate the localization of TRH in the heart. We utilized in situ hybridization histochemistry (ISHH) to localize TRH mRNA expression in the rat heart. ISHH was performed on fresh frozen heart tissue sections which were hybridized with a specific 35S-TRH oligo probe and subsequently processed by autoradiography. The autoradiographic signals corresponding to TRH mRNA were analyzed with an image program. For positive controls TRH mRNA was identified in the hypothalamic paraventricular nucleus. This test confirms the specificity of the TRH oligo probe. Cardiac hybridization signals were observed predominantly in the atria and localized preferentially in atrial connective tissues, vascular adventitia and atrial cardiomyocytes. No hybridization signals were found in ventricular cardiomyocytes. These observations suggest that TRH is synthesized in atrial myocytes and atrial vascular structures. Based on studies which show synthesis of the TRH receptors in ventricular cardiomyocytes, we hypothesize that atrial TRH is an endocrine source for the stimulation of ventricular contractility and that endothelial and adventitial TRH may play a role(s) in the regulation of the growth and/or vasomotor tome of the cardiac vascular system.

Animals↗

PubMed1996

Morphologic and functional properties of bronchial cells isolated from normal and asthmatic subjects.

Recent advances in biomedical sciences have led to the development of various methods for the evaluation of the physiopathology of respiratory diseases. This study reports morphologic and functional features of cells isolated by a new method from bronchial biopsies of normal and asthmatic subjects. Both epithelial and fibroblastic cells were isolated from the same biopsies using collagenase. The cells were cultured for several passages and stored frozen. Two selective culture media were used in order to obtain pure epithelial and fibroblastic cell populations. Immunofluorescence analysis of intermediate filaments, keratins, and vimentin confirmed the type of the isolated cells. The proportions of alpha-actin-expressing cells varied among the fibroblastic cell populations isolated from normal and asthmatic subjects. Interestingly, the population containing high numbers of alpha-actin-expressing cells and presenting the fastest collagen contraction kinetic was isolated from bronchial biopsies of an asthmatic subject. Moreover, the fibroblastic cells that showed the best contractile properties 24 h after their seeding in floating collagen gels were isolated from bronchial biopsies of asthmatic patients having PC20 values below 1 mg/ml. On the basis of these data, we propose a new approach to isolate, culture and characterize human bronchial cells in vitro.

Adult↗

PubMed1996

Extreme obesity may be linked to markers flanking the human OB gene.

Mice with mutations of the ob gene are extremely obese, and the human homologue (OB) has been cloned and physically mapped. The protein product of the ob gene (leptin) reduces body fat in mice when given exogenously, and leptin has been proposed to provide a lipostatic signal that regulates adiposity. Variation in the OB gene may be one genetically determined cause of obesity in human populations. To test this hypothesis, we genotyped siblings from 78 families at markers flanking the human OB gene. Pairs of siblings with extreme obesity (BMI > or = 40; n = 59) shared haplotypes identical-by-descent for the region containing the OB gene at greater than chance levels (corrected P = 0.04). Furthermore, one haplotype containing the OB gene was transmitted by heterozygous parents to extremely obese (BMI > or = 40) offspring more frequently than expected by chance, indicting significant allelic disequilibrium (corrected P = 0.027). One explanation for these linkage findings is that some individuals with extreme obesity have an allelic variant of the OB gene, although other nearby genes could contribute to obesity in these families.

Adipose Tissue↗

PubMed1996

Cocaine/sex type effects on T lymphocytes: a preliminary report.

The influence of gender on the effects of cocaine (5 or 40 mg/kg) on immune function of sexually immature (35-day-old) C57BL/6J mice was assessed. The proliferation of T cells in thymus and spleen stimulated with phytohemagglutinin (PHA) and concanavalin-A(Con-A) was determined by 3H thymidine incorporation 24 hours after cocaine injections. The drug produced a dose-dependent reduction (25-51%) of Con-A and PHA-stimulated T cell proliferation in thymus from both male and female mice. Additionally, the impairment occurred at a lower cocaine dose for female than for male mice when the T cells were challenged with PHA. Under the experimental conditions, cocaine did not alter weights or cell numbers of thymus and spleen or T cell proliferation in spleen.

Analysis of Variance↗

PubMed1996

Life in a changing world: TCH gene regulation of expression and responses to environmental signals.

The Arabidopsis TCH genes were discovered as a consequence of their marked upregulation of expression in response to seemingly innocuous stimuli such as touch. Further analyses have indicated that these genes are upregulated by a variety of diverse stimuli. Understanding the mechanism(s) and factors that control TCH gene regulation will shed light on the signaling pathways that enable plants to respond to changing environmental conditions. The TCH proteins include calmodulin, calmodulin-related proteins and a xyloglucan endotransglycosylase. Expression analyses and localization of protein accumulation indicate that the potential sites of TCH protein function include expanding cells and tissues under mechanical strain. We hypothesize that the TCH proteins may collaborate in cell wall biogenesis.

Arabidopsis↗

PubMed1996

Genetic evidence for the neuronal nitric oxide synthase gene (NOS1) as a susceptibility locus for infantile pyloric stenosis.

The etiological role of the gene for neuronal nitric oxide synthase (NOS1) in infantile pyloric stenosis (PS) was investigated by analysis of two intragenic polymorphisms (NOS1a and NOS1b) in 27 families. There was significant overall transmission disequilibrium between PS and NOS1a (P = .006). Consideration of each allele independently revealed a highly significant tendency for allele 7 (210 bp) to be preferentially transmitted to the affected offspring (P = .0006). These observations suggest that NOS1 is a susceptibility locus for PS.

Alleles↗