Biomedical subjects

Y Xia

Publications and source records attributed to Y Xia.

At least 199 records · Page 11Linked to original sources

PubMed1997

Monoclonal antibodies to the extracellular domain of prostate-specific membrane antigen also react with tumor vascular endothelium.

Prostate-specific membrane antigen (PSMA), initially defined by monoclonal antibody (mAb) 7E11, is a now well-characterized type 2 integral membrane glycoprotein expressed in a highly restricted manner by prostate epithelial cells. 7E11 has been shown to bind an intracellular epitope of PSMA that, in viable cells, is not available for binding. Herein, we report the initial characterization of the first four reported IgG mAbs that bind the external domain of PSMA. Competitive binding studies indicate these antibodies define two distinct, noncompeting epitopes on the extracellular domain of PSMA. In contrast to 7E11, these mAbs bind to viable LNCaP cells in vitro. In addition, they show strong immunohistochemical reactivity to tissue sections of prostate epithelia, including prostate cancer. These mAbs were also strongly reactive with vascular endothelium within a wide variety of carcinomas (including lung, colon, breast, and others) but not with normal vascular endothelium. These antibodies should prove useful for in vivo targeting to prostate cancer, as well as to the vascular compartment of a wide variety of carcinomas.

Animals↗

PubMed1997

Decreased nitric-oxide synthase activity causes impaired endothelium-dependent relaxation in the postischemic heart.

Endothelial nitric-oxide synthase (eNOS) is an important regulator of endothelial function and vascular tone in biological tissues. While endothelial dysfunction occurs following ischemia and has been attributed to altered NO. formation, the biochemical basis for this dysfunction is unknown. Therefore, studies were performed to determine the effects of myocardial ischemia and reperfusion on eNOS in isolated rat hearts subjected to periods of global ischemia or ischemia followed by reperfusion. eNOS activity was assayed by L-[14C]arginine to L-[14C]citrulline conversion and alterations in the amount and distribution of eNOS determined by Western blotting and immunohistochemistry. While activity was preserved after 30 min of ischemia with a value of 1.1 +/- 0.1 pmol x min-1 x mg of protein-1, it decreased by 77% after 60 min and became nearly undetectable after 120 min. Reperfusion resulted in only a partial restoration of activity. The decline in activity with ischemia was due, in part, to a loss of eNOS protein. Hemodynamic studies showed that the onset of impaired vascular reactivity paralleled the loss of functional eNOS. Subjecting isolated eNOS to conditions of acidosis, which occur during ischemia, followed by restoration of pH as occurs on reperfusion, caused a combination of reversible and irreversible loss of activity similar to that seen in ischemic and reperfused hearts. Thus, loss of endothelial function following ischemia is paralleled by a loss of eNOS activity due to a combination of pH-dependent denaturation and proteolysis.

Animals↗

PubMed1997

Inhibition of constitutive nitric oxide synthase (NOS) by nitric oxide generated by inducible NOS after lipopolysaccharide administration provokes renal dysfunction in rats.

Excess NO generation plays a major role in the hypotension and systemic vasodilatation characteristic of sepsis. Yet the kidney response to sepsis is characterized by vasoconstriction resulting in renal dysfunction. We have examined the roles of inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) on the renal effects of lipopolysaccharide administration by comparing the effects of specific iNOS inhibition, -N6-(1-iminoethyl)lysine (L-NIL), and 2,4-diamino6-hydroxy-pyrimidine vs. nonspecific NOS inhibitors (nitro- -arginine-methylester). cGMP responses to carbamylcholine (CCh) (stimulated, basal) and sodium nitroprusside in isolated glomeruli were used as indices of eNOS and guanylate cyclase (GC) activity, respectively. LPS significantly decreased blood pressure and GFR (112+/-4 vs. 83+/-4 mmHg; 2.66+/-0.29 vs. 0. 96+/-0.22 ml/min, P < 0.05) and inhibited the cGMP response to CCh. GC activity was reciprocally increased. L-NIL and 2, 4-diamino-6-hydroxy-pyrimidine administration prevented the decrease in GFR (2.71+/-0.28 and 3.16+/-0.18 ml/min, respectively), restored the normal response to CCh, and GC activity was normalized. In vitro application of L-NIL also restored CCh responses in LPS glomeruli. Neuronal NOS inhibitors verified that CCh responses reflected eNOS activity. L-NAME, a nonspecific inhibitor, worsened GFR (0.41+/-0.15 ml/min), a reduction that was functional and not related to glomerular thrombosis, and eliminated the CCh response. No differences were observed in eNOS mRNA expression among the experimental groups. Selective iNOS inhibition prevents reductions in GFR, whereas nonselective inhibition of NOS further decreases GFR. These findings suggest that the decrease in GFR after LPS is due to local inhibition of eNOS by iNOS, possibly via NO autoinhibition.

Animals↗

PubMed1997

Superoxide and peroxynitrite generation from inducible nitric oxide synthase in macrophages.

Superoxide (O-2) and nitric oxide (NO) act to kill invading microbes in phagocytes. In macrophages NO is synthesized by inducible nitric oxide synthase (iNOS, NOS 2) from L-arginine (L-Arg) and oxygen; however, O-2 was thought to be produced mainly by NADPH oxidase. Electron paramagnetic resonance (EPR) spin trapping experiments performed in murine macrophages demonstrate a novel pathway of O-2 generation. It was observed that depletion of cytosolic L-Arg triggers O-2 generation from iNOS. This iNOS-mediated O-2 generation was blocked by the NOS inhibitor N-nitro-L-arginine methyl ester or by L-Arg, but not by the noninhibitory enantiomer N-nitro-D-arginine methyl ester. In L-Arg-depleted macrophages iNOS generates both O-2 and NO that interact to form the potent oxidant peroxynitrite (ONOO-), which was detected by luminol luminescence and whose formation was blocked by superoxide dismutase, urate, or L-Arg. This iNOS-derived ONOO- resulted in nitrotyrosine formation, and this was inhibited by iNOS blockade. iNOS-mediated O-2 and ONOO- increased the antibacterial activity of macrophages. Thus, with reduced L-Arg availability iNOS produces O-2 and ONOO- that modulate macrophage function. Due to the existence of L-Arg depletion in inflammation, iNOS-mediated O-2 and ONOO- may occur and contribute to cytostatic/cytotoxic actions of macrophages.

Animals↗

PubMed1997

Mitogenic signaling mediated by oxidants in Ras-transformed fibroblasts.

NIH 3T3 fibroblasts stably transformed with a constitutively active isoform of p21(Ras), H-RasV12 (v-H-Ras or EJ-Ras), produced large amounts of the reactive oxygen species superoxide (.O2-). .O2- production was suppressed by the expression of dominant negative isoforms of Ras or Rac1, as well as by treatment with a farnesyltransferase inhibitor or with diphenylene iodonium, a flavoprotein inhibitor. The mitogenic activity of cells expressing H-RasV12 was inhibited by treatment with the chemical antioxidant N-acetyl-L-cysteine. Mitogen-activated protein kinase (MAPK) activity was decreased and c-Jun N-terminal kinase (JNK) was not activated in H-RasV12-transformed cells. Thus, H-RasV12-induced transformation can lead to the production of .O2- through one or more pathways involving a flavoprotein and Rac1. The implication of a reactive oxygen species, probably .O2-, as a mediator of Ras-induced cell cycle progression independent of MAPK and JNK suggests a possible mechanism for the effects of antioxidants against Ras-induced cellular transformation.

3T3 Cells↗

PubMed1997

Suppression of cAMP formation by adenosine in myenteric ganglia from guinea-pig small intestine.

Effects of the adenosine receptor agonist 2-chloro-N6-cyclopentyl-adenosine (CCPA) on stimulation of cAMP formation by histamine, 5-hydroxytryptamine, substance P and forskolin were determined for enzymatically dissociated ganglia from the myenteric plexus of guinea-pig small intestine. Each of the 4 substances stimulated cAMP production. CCPA blocked the stimulation of cAMP by histamine, but not by 5-hydroxytryptamine or substance P. CCPA marginally suppressed stimulation by forskolin. CCPA alone suppressed basal levels of cAMP. The adenosine receptor antagonist 8-cyclopentyl-1,3-dimethylxanthine (CPT) reversed the inhibitory action of CCPA on stimulation of cAMP formation by histamine. Exposure to adenosine deaminase or CPT increased cAMP in the ganglia. The results are consistent with a hypothesis that stimulation of adenylate cyclase and elevation of intraneuronal cAMP in enteric neurons are steps in the signal transduction cascade for the excitatory actions of 5-hydroxytryptamine, substance P and histamine. They are consistent also with an original hypothesis from electrophysiologic studies which states that stimulation of adenosine A1 receptors suppresses cAMP formation and thereby slow synaptic excitation in response to histamine, but not to 5-hydroxytryptamine or substance P. The results support evidence from intracellular microelectrode studies which suggested that endogenous adenosine accumulates to levels sufficient for tonic suppression of cAMP formation in myenteric ganglia in vitro.

Adenosine↗

PubMed1997

Measurement of myeloperoxidase in leukocyte-containing tissues.

An improved method is reported for measurement of myeloperoxidase (MPO) activity in tissues. While spectrophotometric methods based on oxidation of O-dianisidine or other dyes have been reported for MPO measurement in pure polymorphonuclear leukocytes (PMNs), these methods often fail to accurately assay MPO activity in tissues. We observe that tissue myoglobin or vascular hemoglobin markedly effects the spectrophotometric assay for MPO. Under optimal conditions of 0.53 mM O-dianisidine, 0.15 mM H2O2, pH 6.0, either myoglobin or hemoglobin produced absorbance at 460 nm in a concentration-dependent manner similar to that of MPO. In perfused heart tissue, myoglobin caused a major problem with the assay resulting in an inability to obtain accurate linear results as a function of MPO concentration and PMN number. To eliminate the effect of tissue myoglobin or vascular hemoglobin on the assay, one-step gel filtration chromatography of tissue extracts was introduced. MPO, myoglobin, and hemoglobin were easily separated using a Sephadex G-75 column according to the difference in their molecular weights. A linear relationship between the MPO activity and PMN number was observed only after processing the tissue extracts through the Sephadex G-75 column. Thus, MPO activity in PMN-containing tissues can be precisely quantitated after one-step purification on a molecular exclusion column. Enzyme purification with removal of myoglobin is essential for obtaining accurate measurement of MPO activity and quantitation of PMNs in muscle tissue.

Animals↗

PubMed1997

Genetic evidence for the involvement of tau in progressive supranuclear palsy.

A dinucleotide repeat polymorphism in a tau intron was identified and used in a case-control study to analyze the genetic association of tau with several neurodegenerative diseases with tau pathology. Subjects with the homozygous tau AO alleles were excessively represented in the progressive supranuclear palsy (PSP) group, compared with the age-matched healthy control group. Consequently, this allele is more frequently found in PSP than in a group of healthy subjects. This trend was not found in Alzheimer's disease or parkinsonism-dementia complex of Guam, both of which are accompanied by major tau pathology. The result suggests a possible involvement of tau in the pathogenesis of PSP.

Base Sequence↗

PubMed1997

Locus of control and problem-solving interaction in families with adolescents.

The focus of this study is on further development and refinement of a classification system of family problem-solving interaction in families with adolescents. A rigorous qualitative analysis was conducted on the video-taped problem-solving sessions of 38 families which consisted of a father, mother and their adolescent son or daughter. The analysis concentrated on how families engaged the problem situations, how solutions were generated and evaluated, communication patterns, and how affect was managed. It was discovered through the analysis that family problem-solving interaction could be classified based on a concept the researchers call family locus of control (F-LOC). Four types of F-LOC were described: individualistic, collaborative, authoritarian and external. Internal classification reliability was assessed by three independent raters who were able to accurately classify 95% of the problem-solving sessions by F-LOC. Discussion focuses on comparing the classification system with previously developed classification systems.

Adolescent↗

PubMed1997

Postnatal expression of melanocortin-3 receptor in rat diencephalon and mesencephalon.

In situ hybridization was applied to examine the postnatal expression of melanocortin-3 (MC-3) receptor mRNA in the rat brain. Very weak and limited signals were seen in the hypothalamus on postnatal day 0 (P0) and in the dorsal lateral thalamus on P4. A marked increase was noted in several regions of the diencephalon and mesencephalon on P7. The highest levels were reached on P21, which was the time when an adult-like pattern was established. On P21, intense signals were seen in the ventromedial nucleus and the arcuate nucleus of the tuberal hypothalamus, the habenular nucleus of the epithalamus and the ventral tegmental area. [125I] Nle4, D-Phe7-alpha-MSH showed overlapping, but wider labelling of melanocortin receptors, that followed a similar developmental course. alpha-MSH-like immunoreactivity was seen widely in the forebrain and midbrain from P14. In contrast to the staining of alpha-MSH in neurons and their process, gamma 2-MSH-like immunoreactivity was detected strongly in the blood vessels. The neuronal localization of MC-3 receptor mRNA suggests that this receptor may mediate the neurotropic actions of melanocortin peptides in the developing brain.

Animals↗

PubMed1997

Disruption of monolayer integrity enables activation of a cystic fibrosis "bypass" channel in human airway epithelia.

Cystic fibrosis (CF) is a genetic disease characterized by marked reduction in Cl- conductance across many epithelia. Two kinds of Cl- channels have been associated with CF. One channel, termed the cystic fibrosis transmembrane conductance regulator (CFTR), is directly coded by the CF gene. The other channel is an outwardly rectifying depolarization induced Cl- channel (ORDIC) that is distinguished from other outwardly rectifying chloride channels (ORCCs) because its activity is induced most reliably by patch excision and depolarization. An issue in current CF research is whether ORDIC channels are indirectly activated by CFTR to contribute a significant portion of apical membrane Cl- conductance in airway cells. We now show that ORDIC channels are readily activated in patches excised and depolarized from isolated cells, but are rarer or refractory to activation in patches from the apical membranes of confluent human airway epithelia. These findings have important implications for proposed therapies that would bypass the CFTR conductance by activating ORDIC channels.

Calcium↗

PubMed1997

Age and gender differences in the self-esteem of Chinese children.

A Chinese version of the Self-Description Questionnaire 1 (SDQ-1; Marsh, 1988) was used to investigate age and gender differences in a sample of 303 male and 296 female 10-year-old children and 116 male and 116 female 13-year-old children attending typical Beijing public schools. Significant Age x Gender interaction effects were found on all 8 SDQ-1 scales. Main effects for age were found on the Physical Abilities, Reading, and School subscales and for gender on the same three subscales plus Peer Relations. Further analysis indicated that the older girls tended to report significantly lower self-esteem than both the younger girls and older boys in the areas of physical abilities, reading, mathematics, and general self-concept. The boys reported more positive self-perceptions on most nonacademic self-scales, but both the older boys and older girls reported less favorable self-esteem than their younger peers on the scales for reading and school in general.

Adolescent↗

PubMed1997

The glossy1 locus of maize and an epidermis-specific cDNA from Kleinia odora define a class of receptor-like proteins required for the normal accumulation of cuticular waxes.

Mutations at the glossy1 (gl1) locus of maize (Zea mays L.) quantitatively and qualitatively affect the deposition of cuticular waxes on the surface of seedling leaves. The gl1 locus has been molecularly cloned by transposon tagging with the Mutator transposon system. The epi23 cDNA was isolated by subtractive hybridization as an epidermis-specific mRNA from Senecio odora (Kleinia odora). The deduced amino acid sequence of the GL1 and EPI23 proteins are very similar to each other and to two other plant proteins in which the sequences were deduced from their respective mRNAs. These are the Arabidopsis CER1 protein, which is involved in cuticular wax deposition on siliques, stems, and leaves of that plant, and the protein coded by the rice expressed sequence tag RICS2751A. All four proteins are predicted to be localized in a membrane via a common NH2-terminal domain, which consists of either five or seven membrane-spanning helices. The COOH-terminal portion of each of these proteins, although less conserved, is predicted to be a water-soluble, globular domain. These sequence similarities indicate that these plant orthologs may belong to a superfamily of membrane-bound receptors that have been extensively characterized from animals, including the HIV co-receptor fusin (also termed CXCR4).

Algorithms↗

PubMed1997

Sequence analysis of the cloned glossy8 gene of maize suggests that it may code for a beta-ketoacyl reductase required for the biosynthesis of cuticular waxes.

The gl8 locus of maize (Zea mays L.) was previously defined by a mutation that reduces the amount and alters the composition of seedling cuticular waxes. Sixty independently derived gl8 mutant alleles were isolated from stocks that carried the Mutator transposon system. A DNA fragment that contains a Mu8 transposon and that co-segregates with one of these alleles, gl8-Mu3142, was identified and cloned. DNA flanking the Mu8 transposon was shown via allelic cross-referencing experiments to represent the gl8 locus. The gl8 probe revealed a 1.4-kb transcript present in wild-type seedling leaves and, in lesser amounts, in other organs and at other developmental stages. The amino acid sequence deduced from an apparently full-length gl8 cDNA exhibits highly significant sequence similarity to a group of enzymes from plants, eubacteria, and mammals that catalyzes the reduction of ketones. This finding suggests that the GL8 protein probably functions as a reductase during fatty acid elongation in the cuticular wax biosynthetic pathway.

3-Oxoacyl-(Acyl-Carrier-Protein) Reductase↗

PubMed1997

Developmental and hormonal regulation of the arabidopsis CER2 gene that codes for a nuclear-localized protein required for the normal accumulation of cuticular waxes.

The previously cloned CER2 gene is required for the normal accumulation of cuticular waxes and encodes a novel protein. Earlier reports suggested that the CER2 protein is either a membrane-bound component of the fatty acid elongase complex or a regulatory protein. Cell fractionation and immunoblot analyses using polyclonal antibodies raised against a chemically synthesized peptide with a sequence based on the predicted CER2 protein sequence have demonstrated that the 47-kD CER2 protein is soluble and nuclear localized. These results are consistent with CER2 being a regulatory protein. Detailed studies of plants harboring a CER2 promoter/GUS transgene (CER2-GUS), in combination with immunoblot analyses, revealed that CER2 is expressed and the CER2 protein accumulates in a variety of organs and cell types. Expression is highest early in the development of these organs and is epidermis specific in most tissues. In agreement with the activity of the CER2 promoter in hypocotyls, cuticular wax accumulates on this organ in a CER2-dependent fashion. In leaves CER2 expression is confined to the guard cells, trichomes, and petioles. However, application of the cytokinin 6-benzylaminopurine induces ectopic expression of CER2-GUS in all cell types of leaves that emerge following treatment.

Amino Acid Sequence↗

PubMed1997

Effect of chronic hypoxia on glucose transporters in heart and skeletal muscle of immature and adult rats.

Glucose transporter (GLUT) modulation can be an important mechanism that contributes to adaptation to hypoxic stress, but little is known about GLUT modulation in heart and skeletal muscle with prolonged hypoxia. In this work, the effect of chronic hypoxia on GLUT-4 and GLUT-1 mRNA and protein was studied in these two tissues in the adult and during development. Hypoxia (fractional inspired O2 = 9 +/- 0.5%) was administered to two groups, i.e., an immature group exposed from 3 to 30 days of age and an adult group exposed from 90 to 120 days of age. Rats were then killed and their heart and skeletal muscles were sampled for measurements of GLUT mRNA and protein with Northern and Western blots. In the adult, chronic hypoxia significantly decreased cardiac GLUT mRNA level by > 25% of control (P < 0.05), but had little effect on GLUT protein. A very different hypoxic effect was seen in the immature rat heart with a major increase in protein and no appreciable change in mRNA density. Adult skeletal muscle had no change in GLUT mRNA level but GLUT protein increased (15-20%, P < 0.05) while both GLUT mRNA and protein were significantly increased in the immature skeletal muscles (60-90% over control). We conclude that during chronic O2 deprivation, GLUT-1 and GLUT-4 expressions show a similar pattern but greatly depend on tissue type and age. These differences in GLUT regulation may be due to different strategies for coping with prolonged O2 deprivation in both immature and adult animals.

Aging↗

PubMed1997

Genetic association of the low-density lipoprotein receptor-related protein gene (LRP), an apolipoprotein E receptor, with late-onset Alzheimer's disease.

The presence of the APOE epsilon 4 allele encoding apolipoprotein E4 (apoE4) is the major genetic risk factor for late-onset Alzheimer's disease (AD). However, the molecular and cellular mechanisms by which APOE epsilon 4 renders AD risk are unclear. In this report, we present genetic evidence that an apoE receptor, LRP, may be associated with the expression of late-onset AD. Using a biallelic genetic marker in exon 3 of LRP, late-onset AD cases markedly differed from the control subjects in the distribution of LRP genotypes, and this difference was highly accentuated among AD cases with positive family history of senile dementia. Furthermore, the numbers of neutritic plaques were significantly altered as a consequence of different LRP genotypes in postmortem AD cases. Taken together, our results implicate the pathophysiology of LRP in the expression of late-onset AD.

Age of Onset↗

PubMed1997

RelB regulation of chemokine expression modulates local inflammation.

The resolution of acute inflammation is incompletely understood but presumably requires the elimination of both inflammatory cells and production of inflammatory cytokines. In the case of recruited bone-marrow-derived inflammatory cells such as granulocytes and macrophages, their short life span helps eliminate these cells and the cytokines they produce. By contrast, resident permanent cells such as fibroblasts require other mechanisms to stop the production of chemokines generated in response to inflammatory triggers such as lipopolysaccharide. Here we demonstrate that RelB is an important regulator of chemokine expression in fibroblasts, thereby playing a key role in the resolution of acute inflammation. Activation of normal fibroblasts by lipopolysaccharide induced a transient production of chemokines, closely followed by induction of RelB expression. However, stimulated RelB-/- fibroblasts exhibited dramatic persistent induction of seven chemokines (RANTES, MIP-1 alpha, MIP-1 beta, MIP-2, IP-10, JE/MCP-1, and KC/CINC). The persistent overexpression of chemokines correlated with increased NF- kappa B binding as well as with increased p50, p65/RelA, and I kappa B alpha expression. Transfection of RelB cDNA into RelB-deficient fibroblasts reversed the lipopolysaccharide-induced chemokine overexpression. In vivo, activated RelB-/- fibroblasts dramatically increased recruitment of granulocytes into tissues. In view of the apparent role of RelB in the resolution of acute inflammation in tissues and previous work showing a requirement for RelB in the initiation of immune responses through the differentiation of antigen-presenting cells, RelB may be an important factor regulating the transition from innate to adaptive immunity.

Animals↗