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Biomedical subjects

L Wu

Publications and source records attributed to L Wu.

At least 595 records · Page 33Linked to original sources
PubMed1995

Immunoglobulin depositions in peripheral nerves in polymyositis.

An immunocytochemical study was performed in 6 peripheral nerve specimens from 6 cases of polymyositis. The results revealed that depositions of IgG, IgM, IgA and C3 were found in the epineurium, perineurium and the walls of capillaries. These findings demonstrated that depositions of immunoglobulins and the complement-mediated immunoreaction may play an important role in pathogenesis of polymyositis with peripheral nerve involvements.

Adult↗
PubMed1995

[The analysis of orthodontic force for distal movement of maxillary canine].

In this study, the finite element method combining with CAD technique was used to establish the three-dimensional mathematical model of the right maxillary canine and its periodontical tissue. We imitated the different kinds of force which were used during the orthodontic treatment, and analyzed the ways of the teeth moving and the initial stress on the teeth and periodontal tissue. Results showed that is was best to exert couple M and force P simultaneously, especially when M/P = 10 approximately 13 mm, the teeth could be moved fast and wholely without pathological destruction on the periodontical tissue. The results of this study provide some theoretical and experimental basis for clinical practice.

Biomechanical Phenomena↗
PubMed1994

The Cys(X)5Arg catalytic motif in phosphoester hydrolysis.

The Yersinia protein tyrosine phosphatase (PTPase) was identified in the genus of bacteria responsible for the plague or the Black Death and was shown to be essential for pathogenesis. The three-dimensional structure of the catalytic domain of the Yersinia PTPase has been solved, and this information along with a detailed kinetic analysis has led to a better understanding of the catalytic mechanism of the PTPase. Mutational and chemical modification experiments have established that an invariant Cys residue (Cys403) is directly involved in formation of a covalent phosphoenzyme intermediate. We have shown that Arg409 plays a critical role in PTPase action and that the Cys(X)5Arg active site motif forms a phosphate-binding loop which appears to represent the essential features necessary for catalysis by the PTPases, the dual specific phosphatases, and the low molecular weight acid phosphatases.

Amino Acid Sequence↗
PubMed1994

The catalytic role of Cys124 in the dual specificity phosphatase VHR.

The recombinant human Vaccinia virus H1-related protein tyrosine phosphatase, (VHR PTPase) possesses intrinsic Tyr and Thr/Ser phosphatase activities. Both activities were abolished by a single amino acid substitution, C124S. When VHR was incubated with a 32P-labeled phosphotyrosine-containing substrate and then rapidly denatured, enzyme-associated 32P was evident following SDS-polyacrylamide gel electrophoresis. The formation of 32P-labeled protein could be blocked in the presence of an unlabeled substrate. VHR-associated 32P was sensitive to iodine but insensitive to pyridine and hydroxylamine. The catalytically inactive C124S mutant would not form a 32P-labeled enzyme. Furthermore, VHR phosphatase could be selectively inactivated by the alkylating agent iodoacetate. The inactivation resulted from the specific covalent modification of Cys124. Collectively these results suggest that a thiol-phosphate enzyme intermediate is formed when Cys124 of VHR accepts a phosphate from the substrate. Our results also demonstrate that the dual specificity phosphatases and the tyrosine-specific PTPases employ similar catalytic mechanisms.

Amino Acid Sequence↗
PubMed1994

Thymic stem cells in mouse bone marrow.

There is still controversy concerning the nature of the stem cells from bone marrow that colonize the thymus during embryogenesis and continually throughout life. To identify the bone marrow stem cells that home to and populate the thymus, we screened murine bone marrow cells for the presence of a population of surface phenotype similar to the earliest known intrathymic precursor. We have identified a population characterized by expression of an intermediate level of heat stable antigen, a very low level of Thy-1, and high levels of CD44 and class I major histocompatibility complex antigens. It is negative for B-cell, granulocyte, macrophage, and erythrocyte markers (B220, Gr-1, Mac-1, and TER 119). All these markers are common to the intrathymic precursors and bone marrow stem cells. However, this new bone marrow population is Sca-2+, similar to the intrathymic precursor, which makes a clear distinction from the Sca-2- bone marrow hematopoietic stem cells previously characterized. The frequency of the new population in the normal mouse bone marrow is about 0.25%. When transferred intrathymically or intravenously to lethally irradiated mice, it has a higher expansion potential (2 x 10(5)) than has been found for the intrathymic precursors (10(3)), but less than was found for the Sca-2- multipotent stem cell (10(7)). These transfer studies also showed that it was pluripotent, in that its precursor activity was not restricted to the production of T or B lymphocytes. However, it gave a reduced spleen colony number and smaller colonies (day-12 colony-forming unit spleen) when compared with multipotent stem cells. Thus, the cell we have identified appears to be the latest pluripotent cells so far identified in bone marrow and is therefore a good candidate for a bone marrow prothymocyte, but it appears not to be T-cell-committed.

Animals↗
PubMed1994

Adenovirus-mediated gene transfer in the transplant setting. II. Successful expression of transferred cDNA in syngeneic liver grafts.

These experiments establish a model for gene transfer to transplanted liver grafts ex vivo using adenoviral vectors. Rat liver grafts (n = 8) were harvested, and preserved in UW or lactated Ringer's. The grafts were infected ex-vivo via portal vein perfusion with replication-defective Ad vectors encoding the beta-galactosidase (beta-gal) gene diluted in UW solution. The infected grafts were stored at 4 degrees C for 1 hr, then transplanted into syngeneic hosts. Liver biopsies were taken at 1, 7, and 15 days after transplantation. Infection rate was assessed by histochemical staining for beta-gal. Liver DNA and RNA were assayed for the beta-gal sequences, and recombinant protein production measured at 24 hr and 7 days after transplantation. Under conditions mimicking liver graft cold storage, efficient gene transfer was achieved with an infection rate of 10-15%, as assessed by X-gal staining. Viral DNA and RNA presence in the graft was confirmed; gene expression with protein production were verified by western blots and a functional protein assay. All studies were negative in control livers. Gene expression persisted for at least 2 weeks after transplantation. We conclude that efficient adenovirus-mediated gene insertion and expression of gene products can be accomplished in whole-liver grafts under hypothermic preservation conditions currently used in clinical transplantation.

Adenoviridae↗
PubMed1994

Assessment of CD4 expression by early T precursor cells and by dendritic cells in the human thymus.

The adult mouse thymus contains a minute population of early lymphoid precursor cells that express moderate levels of CD4. We searched for a corresponding population of early T precursors in the infant human thymus, by first depleting the majority of more mature thymocytes, then using immunofluorescence and flow cytometry to analyze cells bearing a range of early T lineage markers. No discrete population of early T precursors expressing CD4 was observed, in contrast to the murine thymus. Most putative very early human thymocytes were CD4-8-3-1-2lo44+34+7hi class I MHChi class II MHC-. However, a distinct population of human thymic dendritic cells expressing high levels of CD4 was isolated. These were CD4hi8-3-1-2-44+34-7- class I MHChi class II MHChi, and lacked markers of B cells, NK cells, or myeloid cells. They were large cells that exhibited dendritic morphology after brief periods of culture, and they were efficient stimulators of allogeneic T cells. The biologic implications of CD4 expression by thymic dendritic cells are discussed.

Adolescent↗
PubMed1994

Identification, assay, and purification of a Cdc2-activating threonine-161 protein kinase from human cells.

The biological activities of cyclin-dependent, proline-directed protein kinases (PDPKs) are highly regulated by a complex series of protein phosphorylation/dephosphorylation reactions involving both catalytic and regulatory subunits. In this paper we report on the enzymatic activation of p34cdc2/p58Cyclin A PDPK by a protein kinase present in human cells that targets threonine-161 of Cdc2. An assay for this Cdc2 kinase-kinase (PK161) was developed and specific enzyme activity was detected in a variety of mammalian cells and tissues. PK161 activity was rapidly stimulated by epidermal growth factor in human A431 epidermoid carcinoma cells. The development of an assay selective for PK161 phosphotransferase activity afforded the partial purification of the enzyme from human Wilms' tumors. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining of highly purified enzyme preparations revealed the presence of phosphoproteins migrating at approximately 42-44 and approximately 95 kDa, respectively, which correlated with enzyme activity upon fast-protein liquid chromatography gel permeation chromatography. Further purification was accomplished by immobilized peptide substrate affinity chromatography. The ability of PK161 to phosphorylate and activate p34cdc2/p58Cyclin A PDPK was confirmed by the use of purified recombinant subunits. Polyclonal antibodies directed against the Xenopus MO15 gene product (a putative Cdc2-activating kinase) cross-reacted with the purified 42- to 44-kDa phosphoprotein, thus identifying the catalytic subunit of human PK161 as a human homologue of Xenopus p40MO15. Subsequent immunoprecipitation experiments with metabolically labeled A431 cells identified a approximately 95-kDa phosphoprotein that coprecipitated with the approximately 42-kDa catalytic subunit. Taken together, these findings identify a human Cdc2-activating kinase as a growth factor-responsive enzyme system that may participate in the acute activation of cyclin-dependent protein kinases observed in mammalian somatic cells.

Animals↗
PubMed1994

Effects of elevated selenium concentration on selenium accumulation and nitrogen fixation symbiotic activity of Melilotus indica L.

Biological and soil factors which contribute to the successful colonization of an annual legume species. Melilotus indica L., in soils with elevated selenium (Se) were studied. This species was introduced into the Kesterson Reservoir in the fresh top soil that was brought in under the Kesterson Cleanup Action to fill lowering pond sites and prevent the formation of ephemeral pools containing hazardous levels of Se. In 4 years since its introduction, it has expanded its range of colonization from the fresh soil fill sites to the native soil sites and contributed 10 to 50% of biomass to the grassland communities. The plant and nodule tissue Se concentrations of the field grown plants were found to be negatively correlated with the soil sulfate concentration. Nutrient solution culture studies discovered that M. indica was able to accumulate 500 micrograms Se g-1 dry weight without a reduction of growth rate. Plants without nodulation were found to accumulate a greater amount of Se and more vulnerable to Se toxicity. Acetylene reduction rate measurements indicate that the nitrogen fixation symbiotic activity appears to be more susceptible to an elevated Se concentration than its host plant. M. indica is a winter weed, and it occurs naturally in the Se-rich soils. It grows actively over the winter and spring and complete its life cycle in May. If the root nodules and root tissues are incorporated into the soil, the rate of soil Se volatilization may be accelerated over the warm summer months. For disposal of the Se-rich plant materials the plant shoot tissues may be harvested for Se-deficient forage supplementation. Therefore, this species may be useful for field management and reclamation of Se-contaminated soils.

Biomass↗
PubMed1994

Herpes simplex virus type 1 replication and IL-1 beta gene expression in mouse peritoneal macrophages activated in vivo by an attenuated Salmonella typhimurium vaccine or Corynebacterium parvum.

Activated macrophages (M phi) from mice given Salmonella typhimurium or Corynebacterium parvum were compared with resident peritoneal macrophages at the molecular level for permissiveness for herpes simplex virus type 1 (HSV-1) replication and for expression of interleukin-1 beta (IL-1 beta). Peritoneal macrophages were harvested from mice injected 7 days previously with live, avirulent S. typhimurium (Sal-PM phi) or heat-killed C. parvum (CP-PM phi) and infected with HSV-1 in vitro. Both Sal-PM phi and CP-PM phi were activated as evidenced by characteristic changes in an ectoenzyme, by increased permissiveness for infectious virus production and viral cytopathic effect, and by induction of IL-1 beta mRNA. Analysis at the molecular level revealed that both types of activated M phi demonstrated increased patterns of HSV-1 immediate-early gene expression and viral DNA replication as compared with resident cells. A novel finding was that viral infection reduced IL-1 beta mRNA in both types of activated M beta. This observation has implications for the efficacy of Salmonella vaccines given in proximity to HSV-1 infection and for potential deleterious effects of HSV-1 infection in immunosuppressed patients receiving immunotherapy.

Animals↗
PubMed1994

The inhibition of silica-induced lung inflammation by dexamethasone as measured by bronchoalveolar lavage fluid parameters and peroxynitrite-dependent chemiluminescence.

The inhalation of silica has been shown to produce a dramatic inflammatory and toxic response within the lungs of humans and laboratory animals. Currently, no effective treatment exists for workers who may have been exposed to the inhalation of silica. The objective of this study was to develop an animal model in which we could evaluate the effect that anti-inflammatory steroids have on the acute silica-induced pulmonary inflammatory response. Male Fischer 344 rats were pretreated with either dexamethasone (2 mg/kg) or saline vehicle (i.p.) on days 1, 3, and 5. On day 6, the animals from the two groups were then intratracheally instilled with either silica (20 mg/0.5 ml saline vehicle) or saline vehicle (0.5 ml). Twenty-four hours after the instillations in the non-steroid group, significant increases occurred in total protein, total number of cells, neutrophils, and lymphocytes recovered from the lungs of animals treated with silica compared to saline controls. Silica also caused dramatic increases in the luminol-dependent chemiluminescence (LDCL) of lung tissue and bronchoalveolar lavage (BAL) cells. The LDCL reaction was markedly decreased by either superoxide dismutase (SOD) or N-nitro-L-arginine methyl ester hydrochloride (L-NAME). SOD is involved in the enzymatic breakdown of superoxide anion, while L-NAME, a nitric oxide (NO) synthase inhibitor, prevents the formation of NO. When the superoxide anion and NO react, they form the highly oxidizing substance peroxynitrite. This study then implicates peroxynitrite as an agent which may be involved in the silica-induced oxidant lung injury.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗
PubMed1994

Inositol 1,4,5-trisphosphate-, GTP-, arachidonic acid- and thapsigargin-mediated intracellular calcium movement in PANC-1 microsomes.

Inositol 1,4,5-trisphosphate (IP3)-, GTP-, arachidonic acid- and thapsigargin-mediated Ca2+ release from endoplasmic reticulum (ER)-enriched microsomes was studied in a PANC-1 cell line. IP3 maximally caused an approximately 20% release of actively accumulated Ca2+. This effect was completely blocked by heparin. In the presence of 3% polyethylene glycol (PEG), GTP maximally discharged about 60% of Ca2+ from the microsomes. This effect involved a GTP hydrolytic process, not the IP3-activated Ca2+ channel. Arachidonic acid maximally released approximately 80% of Ca2+ from PANC-1 microsomes. Metabolites of arachidonic acid did not appear to be involved in arachidonic acid-mediated Ca2+ release. However, other fatty acids also induced similar releasing effects suggesting that arachidonic acid-induced Ca2+ release appeared to be non-specific. Thapsigargin was shown to inhibit Ca2+ accumulation into and induce Ca2+ release from PANC-1 microsomes. The thapsigargin-releasable Ca2+ pool included the IP3- or arachidonic acid-sensitive pool. Studies on liposomes suggested that both arachidonic acid and thapsigargin did not exert either a Ca2+ ionophore-like or a membrane detergent-like effect. The present results have provided evidence for the existence of multiple non-mitochondrial Ca2+ pools in PANC-1 cells. These Ca2+ pools could be released by various Ca2+ mediators via different mechanisms.

Arachidonic Acid↗
PubMed1994

Enhanced calcium influx by parathyroid hormone in identified Helisoma trivolvis snail neurons.

The modulation of [Ca2+]i by parathyroid hormone (PTH) has been extensively studied in vertebrates. The present study examined the effects of PTH on [Ca2+]i in isolated invertebrate neurons B5 from buccal ganglia of the pond snail, Helisoma trivolvis, utilizing the Fura-2 fluorescence technique. Bovine PTH, bPTH-(1-84), induced a slow and sustained increase in [Ca2+]i in neurons B5. In contrast, the elevation of extracellular K+ concentration from 1.7 mM to 15 mM induced a rapid and transient increase in [Ca2+]i. Simultaneous application of 15 mM KCl and bPTH-(1-84), or application of 15 mM KCl in the presence of bPTH-(1-84) additively increased [Ca2+]i in neurons B5. An increase in [Ca2+]i in neurons B5 was also induced by a PTH agonist [bPTH-(1-34)], but not by a PTH antagonist [bPTH-(3-34)]. The absence of calcium, or the presence of lanthanum (2 mM) or omega-conotoxin (10 microM), in the bath solution abolished the effect of bPTH-(1-84) on [Ca2+]i. Furthermore, our results demonstrated that the effect of PTH on [Ca2+]i in neurons B5 was not due to the hormonal modulation of voltage-dependent Na+ or K+ channels or a Na+/Ca2+ exchanger. The results from this study show that PTH can modulate [Ca2+]i in an identified invertebrate neuron mainly by promoting extracellular calcium influx via the N-like voltage-dependent calcium channel.

Animals↗
PubMed1994

Effects of copper concentration on mineral nutrient uptake and copper accumulation in protein of copper-tolerant and nontolerant Lotus purshianus L.

One copper-tolerant and one copper-sensitive inbred line of Lotus purshianus L. derived from a copper mine waste site in Northern California and one inbred line of the same species derived from a pasture next to the mine waste were examined for the effects of excessive copper concentrations on mineral nutrient uptake and accumulation of copper in protein fractions. Plants were grown from seeds for a period of 24 days in a modified Hoagland nutrient solution culture supplemented with 3, 6, and 10 microM copper as copper sulfate. The basal nutrient solution without copper amendment was used as the control treatment. The uptake of Cu found in the roots was 100 times or more than that in the leaves. The root tissue copper concentrations reached a plateau under 6 microM copper treatment. The leaf tissue copper concentrations increased with the increase of copper concentration in the solution culture. No difference in pattern of copper uptake was detected between the copper-tolerant and nontolerant plants. The effects of excessive copper concentrations caused reduction of Ca uptake in the leaf tissue and P uptake in both the root and leaf tissues, and no difference was found between the copper-tolerant and nontolerant plants. Increased tissue copper concentration caused greater reduction of Fe, Mn, and Zn uptake in the nontolerant plants than in the tolerant plants; this difference may be important for the growth of the tolerant plants under conditions of excessive copper concentrations. Protein extracted from the roots and leaves of both the copper tolerant and nontolerant plants was subjected to Sephadex G-75 column separation. Two major peaks of protein fractions were detected. Under low (normal level) copper concentration treatment, the copper-tolerant and nontolerant plants had similar Cu/protein ratios. However, under high copper concentration challenged conditions the copper-tolerant plant had a considerably greater Cu/protein ratio (peak II protein) than the nontolerant plants. The amino acid composition of the copper-rich protein fraction (peak II) extracted from both the tolerant and nontolerant plants demonstrated a high asparate (about 25%) content. The contents of glutamate, cystine, and glycine were about 11, 2.5, and 10%, respectively, and the rest of the amino acids were in a range of 2 to 6%. This pattern of amino acid composition is different from the amino acid composition of the phytochelatin metallothionein-like proteins found in copper-tolerant plants which are very high in cysteine.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗
PubMed1994

Functional analysis of Mel-14+ and Mel-14- early precursor cells in the adult mouse thymus.

The earliest T-cell precursor population in the adult mouse thymus (low CD4 precursors) may be divided into 85% of cells expressing surface Mel-14 (LECAM-1, the lymphocyte homing receptor) and 15% of cells which are Mel-14. To date, this is the only surface marker for which we have found this population to be heterogeneous. The precursor activity of the Mel-14+ and Mel-14- subpopulation was assessed by both intrathymic and intravenous transfer of sorted cells into Ly5 congenic irradiated recipient mice. On both a cell-for-cell and a total activity basis, almost all precursor activity was associated with the Mel-14+ cells. No segregation was seen between T-cell, B-cell and dendritic cell precursor activity of the low CD4 population, all activities being concentrated in the Mel-14+ fraction. This strengthens the hypothesis that one precursor cell has the potential to form all three lineages.

Animals↗
PubMed1994

Male pseudohermaphroditism caused by mutations of testicular 17 beta-hydroxysteroid dehydrogenase 3.

Defects in the conversion of androstenedione to testosterone in the fetal testes by the enzyme 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) give rise to genetic males with female external genitalia. We have used expression cloning to isolate cDNAs encoding a microsomal 17 beta-HSD type 3 isozyme that shares 23% sequence identity with other 17 beta-HSD enzymes, uses NADPh as a cofactor, and is expressed predominantly in the testes. The 17 beta HSD3 gene on chromosome 9q22 contains 11 exons. Four substitution and two splice junction mutations were identified in the 17 beta HSD3 genes of five unrelated male pseudohermaphrodites. The substitution mutations severely compromised the activity of the 17 beta-HSD type 3 isozyme.

17-Hydroxysteroid Dehydrogenases↗
PubMed1994

Cloning and targeted mutations of G alpha 7 and G alpha 8, two developmentally regulated G protein alpha-subunit genes in Dictyostelium.

GTP-binding protein (G protein)-mediated signal transduction pathways play essential roles during the aggregation and differentiation process of Dictyostelium. In addition to the five known G protein alpha-subunit genes, we recently identified three novel alpha-subunit genes, G alpha 6, G alpha 7, and G alpha 8, using the polymerase chain reaction technique. We present here a more complete analysis of G alpha 7 and G alpha 8. The cDNAs of these two genes were cloned, and their complete nucleotide sequences were determined. Sequence analyses indicate that G alpha 8 possesses some unusual features. It lacks the "TCATDT" motif, a sequence of amino acids highly conserved among G alpha subunits, and has an additional 50 amino acids at its C-terminus consisting of long stretches of asparagine. Moreover, G alpha 8 is unusually resistant to protease digestion, which may indicate a slow GTP hydrolysis rate. The possible functions of these alpha-subunits were assessed by generating mutants lacking G alpha 7 or G alpha 8 by gene targeting through homologous recombination and by overexpressing G alpha 7 or G alpha 8 protein. Overexpression of G alpha 7 resulted in abnormal morphogenesis starting at the slug stage, whereas analysis of the other strains failed to reveal any obvious growth or developmental defects under either normal or stressful conditions. The implications of these results are discussed.

Amino Acid Sequence↗