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Biomedical subjects

L Tang

Publications and source records attributed to L Tang.

At least 217 records · Page 12Linked to original sources
PubMed1996

The Intermittent Claudication Research Study: vascular outcomes research using home health nurses.

This article describes an ongoing, multicenter outcomes research study on intermittent claudication being conducted at 16 Chicago-area vascular surgery offices and clinics. The study relies on gathering data through a series of home health follow-up visits by visiting nurses who use portable Doppler equipment. A total of 624 patients (506 men and 118 women) have been enrolled to date. Data on study patients' baseline demographic characteristics, leg symptoms, peripheral blood flow, comorbidities, and walking impairments are presented. Initial physical functioning and walking-distance scores from patient self-report questionnaires are also presented, along with attrition and lower-extremity revascularization rates. The contribution of nursing to this type of patient outcomes research is discussed. The changing climate in health care delivery provides nursing with a great opportunity to influence the U.S. health care system. In particular, the participation of nurses can make a major contribution to research on patient outcomes and medical effectiveness, and especially to new methods of comparing treatment effects on patients' functional status. Interpretation and measurement of patient self-reported health status and quality of life are particularly relevant to nursing, which is based on interventions related to compromised function. Improvement of patients' physical functioning is the heart of nursing practice, and the focus of much of what is done in vascular medicine and surgery. This article describes the contribution of nursing skills and knowledge to an ongoing, government-funded, multicenter research study on intermittent claudication. The ability of nurses to make a unique contribution to outcomes research is demonstrated.

Activities of Daily Living↗
PubMed1996

Purification and characterization of the DNA-binding protein DnrI, a transcriptional factor of daunorubicin biosynthesis in Streptomyces peucetius.

The DnrI protein, essential for the biosynthesis of daunorubicin in Streptomyces peucetius, was purified almost to homogeneity from dnrI expression strains of Escherichia coll and S. peucetius through several steps of chromatography. The proteins purified from both organisms had identical chromatographic and electrophoretic behaviour. Purified Histagged or native DnrI was used to conduct DNA-binding assays by gel mobility-shift analysis, and the results showed no significant difference in the DNA-binding activity of native or His-tagged proteins. DnrI binds specifically to DNA segments containing the intergenic regions separating the putative dnrG-dpsABCD and dpsEF operons, and the dnrC gene and dnrDKPSQ operon. DNase I footprinting assays indicated that the DNA-binding sites for DnrI extended from upstream of the -10 to -35 regions of the dnrG or dpsE promoters to include about 65 bp of the dnrG-dpsE intergenic region and about 80 bp of the dnrC-dnrD intergenic region. Both binding sites contain imperfect inverted repeat sequences of 6-10 bp with a 5'-TCGAG-3' consensus sequence that was present in 4 out of 10 other promoter regions in the cluster of daunorubicin biosynthesis genes.

Base Sequence↗
PubMed1996

Role of an S4-S5 linker in sodium channel inactivation probed by mutagenesis and a peptide blocker.

A pair of conserved methionine residues, located on the cytoplasmic linker between segments S4 and S5 in the fourth domain of human heart Na channels (hH1), plays a role in the kinetics and voltage dependence of inactivation. Substitution of these residues by either glutamine (M1651M1652/QQ) or alanine (MM/AA) increases the inactivation time constant (tau) at depolarized voltages, shifts steady-state inactivation (h infinity) in a depolarized direction, and decreases the time constant for recovery from inactivation. The data indicate that the mutations affect the rate constants for both binding and unbinding of a hypothetical inactivation particle from its binding site. Cytoplasmic application of the pentapeptide KIFMK in Na channels mutated to remove inactivation produces current decays resembling inactivation (Eaholtz, G., T. Scheuer, and W.A. Catterall. 1994. Neuron. 12: 1041-1048.). KIFMK produces a concentration-dependent, voltage-independent increase in the decay rate of MM/QQ and MM/AA currents at positive membrane potentials (Ki approximately 30 microM), while producing only a small increase in the decay rate of wild-type currents at a concentration of 200 microM. Although MM/QQ inactivates approximately 2.5-fold faster than MM/AA in the absence of peptide, the estimated rate constants for peptide block and unblock do not differ in these mutants. External Na+ ions antagonize the block by cytoplasmic KIFMK of MM/AA channels, but not the inactivation kinetics of this mutant in the absence of peptide. The effect of external [Na+] is interpreted as a voltage-dependent knock-off mechanism. The data provide evidence that KIFMK can only block channels when they are open and that peptide block does not mimic the inactivation process.

Base Sequence↗
PubMed1996

Monoclonal antibody MIB1 in the assessment of cervical squamous intraepithelial lesions.

The aims of the study were to assess patterns of cell proliferation, using the monoclonal antibody MIB1, in a variety of non-neoplastic and neoplastic cervical squamous intraepithelial lesions. We also wished to determine whether this antibody is of value in the discrimination of different grades of cervical intraepithelial neoplasia (CIN) and in the distinction of CIN from non-neoplastic lesions that may histologically mimic CIN. Tissue sections of formalin-fixed, paraffin-embedded cervical colposcopic biopsies were stained immunohistochemically with MIB1 after microwave heating. Normal ectocervix and cervical specimens exhibiting immature squamous metaplasia, koilocytosis, CIN grades I, II, and III, and atrophic squamous epithelium were studied. In normal ectocervix, MIB1-positive cells were confined almost exclusively to the parabasal cell layer. Increased expression was seen in cases of koilocytosis and CIN, positive staining being present in progressively higher layers of epithelium in accordance with the degree of CIN. In immature squamous metaplasia, positive staining was generally confined to the parabasal layer with occasional scattered positive cells in higher layers of epithelium. There was little or no staining of atrophic squamous epithelium. The study confirms that proliferating cells, as demonstrated by MIB1 immunostaining, are found at progressively higher levels in cervical squamous epithelia from normal to koilocytosis to CIN grades I, II and III. Immunostaining with MIB1, which can be easily performed on routinely processed tissues, is of potential value as an adjunct to histology in the grading of CIN and the distinction of CIN from non-neoplastic lesions that may mimic CIN.

Antibodies, Monoclonal↗
PubMed1996

Altered contractile function in isoproterenol-induced hypertrophied rat heart.

OBJECTIVE: To study the calcium-dependent mechanisms contributing to altered contractile function in isoproterenol-induced ventricular hypertrophy of the rat heart. DESIGN: The force-interval relationship, systemic evaluation of changed contractile force and calcium sensitivity of the myofilaments were investigated using small trabecular muscle from hearts with isoproterenol-induced cardiac hypertrophy. METHODS: Cardiac hypertrophy was induced by daily subcutaneous injection of isoproterenol for 12 days. The calcium sensitivity of myofilaments was studied using non-ionic detergent (Triton-X-100)-skinned trabeculae. The contractile force was evaluated at various concentrations of extracellular calcium and muscle lengths. The force-interval relationship was used to reflect altered intracellular calcium handling. RESULTS: The isoproterenol-induced cardiac hypertrophy was associated with significantly enhanced contractile force at various concentrations of extracellular calcium and muscle lengths. Also, an amplified force-interval relationship in hypertrophied muscle at long rest intervals was found. However, this study revealed no change in calcium sensitivity of myofilaments. CONCLUSION: Altered intracellullar calcium handling contributes to enhanced contractile force in isoproterenol-induced cardiac hypertrophy.

Actin Cytoskeleton↗
PubMed1996

Cloning and characterization of a gene (msdA) encoding methylmalonic acid semialdehyde dehydrogenase from Streptomyces coelicolor.

A homolog of the mmsA gene of Pseudomonas aeruginosa, which encodes methylmalonic acid semialdehyde dehydrogenase (MSDH) and is involved in valine catabolism in pseudomonads and mammals, was cloned and sequenced from Streptomyces coelicolor. Of the two open reading frames (ORFs) found, which are convergently transcribed and separated by a 62-nucleotide noncoding region, the deduced amino acid sequence of the msdA ORF (homologous to mmsA) is similar to a variety of prokaryotic and eukaryotic aldehyde dehydrogenases that utilize NAD+, particularly to the MmsA protein from P. aeruginosa. No significant similarity was found between the deduced product of ORF1 and known proteins in the databases. An S. coelicolor msdA mutant, constructed by insertion of a hygromycin resistance gene (hyg) into the msdA coding region, lost the MSDH activity and the ability to grow in a minimal medium with valine or isobutyrate as the sole carbon source but grew on propionate. The msdA::hyg mutation was complemented by introduction of the msdA gene on a plasmid. When the S. coelicolor msdA gene was overexpressed in Escherichia coli under the control of the T7 promoter, a protein of 51-kDa, corresponding to the approximate mass of the predicted S. coelicolor msdA product (52.6 kDa), and specific MSDH activity were detected. These results strongly suggest that msdA indeed encodes the MSDH that is involved in valine catabolism in S. coelicolor.

Aldehyde Oxidoreductases↗
PubMed1996

Dopamine D2L receptor couples to G alpha i2 and G alpha i3 but not G alpha i1, leading to the inhibition of adenylate cyclase in transfected cell lines.

Previously, we showed that both D2 and D4 dopamine receptors inhibited adenylate cyclase in a pertussis toxin (Ptx)-sensitive manner in the dopamine-producing MN9D cell line, whereas only D2 receptors did so in a fibroblast cell line, CCL1.3. Of the known Ptx-sensitive G proteins, MN9D cells expressed G alpha i2, G alpha oA and G alpha oB, whereas CCL1.3 cells expressed only G alpha i2. Here we cotransfected MN9D and CCL1.3 cells with either the long form of the D2 receptor (D2L) or the D4 receptor and a mutant Ptx-resistant G protein alpha-subunit. When cotransfected CCL1.3 cell lines were tested for the ability of Ptx to block receptor-mediated inhibition of cyclic AMP accumulation, D2 receptors were found to couple to mutant G alpha i2 and G alpha i3 but not G alpha i1 or G alpha oA. D2 also coupled to mutant G alpha i2 but not G alpha oA in MN9D cells. In contrast, D4 receptors did not couple to either mutant G alpha i2 or G alpha oA subunits in MN9D cells. These data suggest that D4 receptor-mediated inhibition of adenylate cyclase is not coupled via the same mechanisms used by D2 receptors. D2L receptors are capable of coupling to more than one G protein in the modulation of cyclic AMP.

Adenylyl Cyclases↗
PubMed1996

Differentiation-associated overexpression of the cyclin-dependent kinase inhibitor p21waf-1 in human cutaneous squamous cell carcinoma.

p21waf-1 negatively regulates the cell cycle by inhibiting the activity of cyclin-dependent kinases. As p21waf-1 is a probable tumor suppressor, we sought to determine whether this cyclin-dependent kinase inhibitor is abnormally regulated in human cutaneous squamous cell carcinoma (SCC). An immunohistochemical technique was employed to assay p21waf-1 protein in SCCs chosen from sun-exposed and anogenital sites. We observed that p21waf-1 was greatly overexpressed in SCC versus adjacent benign epithelium. Furthermore, expression of p21waf-1 was consistently elevated in the superficial, differentiated cells versus basal keratinocytes. p21waf-1 expression correlated with the proliferative state of the cancers as measured by MIB-1 immunostaining. In vitro, keratinocytes grown in supplemented media upregulated p21waf-1 during differentiation, supporting our in vivo observations. We conclude that p21waf-1 overexpression is associated with differentiation in proliferating SCC but is not sufficient to suppress cancer development.

Aged↗
PubMed1996

[Studies on the time domain and power spectrum of high frequency ECG in normal mice].

The features of time domain and power spectrum of high frequency electrocardiogram (HF-ECG) were studied in normal Kunming mice using a microprocessor ECG system. The results were as follows (mean +/- SD): (1) P-R interval was 34.9 +/- 4.7 ms (n = 58), about one third of the cardiac cycle. (2) The duration and peak-to-peak amplitude of QRS complex were 9.2 +/- 1.2 ms and 1.456 +/- 0.480 mV (n = 74) respectively. (3) The duration and amplitude of T wave were 10.2 +/- 3.2 ms and 0.336 +/- 0.115 mV, respectively (n = 58). (4) Q-T interval was 19.4 +/- 3.2 ms (n = 58), about one fifth of the cardiac cycle. (5) The total number of notches and slurs of leads II of 73 mice were 3 and 26 respectively. (6) The relative power content of each frequency range was: 0-80 Hz: 45.48 +/- 15.32%; 80-200 Hz: 43.97 +/- 9.95%; 200-300 Hz: 8.89 +/- 7.83%; 300-1000 Hz: 1.66 +/- 2.74%; 80-1000 Hz: 54.52 +/- 15.32%.

Animals↗
PubMed1995

Detection of hepatitis C virus RNA in hepatocellular carcinoma by in situ hybridization.

BACKGROUND: Hepatocellular carcinoma frequently is associated with chronic hepatitis C virus (HCV) infection. The presence of HCV in hepatocellular carcinoma has been detected by reverse-transcription polymerase chain reaction of antigenomic HCV RNA, a tissue-specific replicative form of the virus. Now, however, this method of detecting the presence of HCV has been invalidated by reports of antigenomic RNA in the blood or in peripheral blood mononuclear cells. METHODS: In situ hybridization of HCV RNA was conducted with digoxigenin-labeled cDNA from the core region on surgical specimens of noncancerous and cancerous areas from 12 patients with chronic hepatitis C with or without cirrhosis associated with hepatocellular carcinoma. Several control experiments were also performed, including RNase digestion before hybridization, hybridization with the use of a negative control, and immunohistochemical staining of HCV-core protein. RESULTS: The in situ hybridization showed positive signals both in noncancerous and cancerous areas of the liver tissue in eight cases. Positive signals were confined to neoplastic cells and nonneoplastic hepatocytes. There were fewer HCV-positive cells in the cancerous area than in the surrounding noncancerous area. CONCLUSIONS: In situ detection of HCV presents direct evidence of HCV infection in the neoplastic cells of hepatocellular carcinoma and suggests that neoplastic cells may lose their affinity for HCV in the course of malignant transformation.

Carcinoma, Hepatocellular↗
PubMed1995

Regulation of expression of the valine (branched-chain amino acid) dehydrogenase-encoding gene from Streptomyces coelicolor.

Expression of the Streptomyces coelicolor (Sc) valine (branched-chain amino acid) dehydrogenase-encoding gene (vdh) is regulated by valine, glucose and NH+4 at the transcriptional level. The results of assays for the level of accumulated vdh mRNA in the Sc J802 strain by primer extension experiments and for the level of catechol dioxygenase (XylE) activity in Sc J802 (vdh::xylE) transformants show that transcription of the vdh gene is induced approx. 2.5-fold by valine, as compared to asparagine as the sole nitrogen source in the presence of glucose as carbon source. Valine induction is repressed by glucose, as compared to glycerol as the carbon source, and by NH+4. Glucose catabolite repression is relieved in Sc M480, a glucose kinase (glkA) deletion mutant. This suggests that glucose repression of vdh and carbohydrate metabolism are due to the same mechanism in Sc, which involves glucose kinase.

Amino Acid Oxidoreductases↗
PubMed1995

Channel gating governed symmetrically by conserved leucine residues in the M2 domain of nicotinic receptors.

In nicotinic acetylcholine receptors (nAChR), as well as glycine, GABAA (gamma-aminobutyric acid), serotonin (5-HT3), and GluCl glutamate receptors, a leucine residue at the approximate midpoint of the M2 transmembrane domain (the 9' position) is conserved across most known subunits. Structural data for the nAChR suggest that the Leu 9' residues occupy a 'kink' in each of the five M2 helices and point into the closed channel; in the opening step, the M2 helices rotate so that Leu 9' side chains no longer occlude the conduction pathway. Mutation of Leu 9' to one of several other residues slows desensitization and increases sensitivity to agonist. We have exploited the alpha 2 beta gamma delta stoichiometry of muscle nAChR to express receptors with ms* = 0 to 5 Leu 9'Ser mutated subunits. Strikingly, each Leu 9'Ser mutation shifts the dose-response relation for ACh to the left by approximately 10-fold; a nAChR with ms* = 4 is 10(4)-fold more sensitive than the wild type. The results suggest that each of the five Leu 9' residues participates independently and symmetrically in a key step in the structural transition between the closed and open states.

Acetylcholine↗
PubMed1995

Heterologous expression and enzymatic properties of a selenium-independent glutathione peroxidase from the parasitic nematode Brugia pahangi.

A full-length cDNA from the parasitic nematode Brugia pahangi encoding a secreted homolog of glutathione peroxidase in which the codon for the active site selenocysteine is substituted naturally by a cysteine codon has been expressed in Spodoptera frugiperda (insect) cells via Autographa californica nuclear polyhedrosis virus (baculovirus). The recombinant protein was glycosylated and secreted from the cells in tetrameric form. The purified protein showed glutathione peroxidase activity with a range of organic hydroperoxides, including L-alpha-phosphatidylcholine hydroperoxide, but no significant activity against hydrogen peroxide. Glutathione was the only thiol tested that served as a substrate for the enzyme, which showed no activity with the thioredoxin system (thioredoxin, thioredoxin reductase, and NADPH). No glutathione-conjugating activity was detected against a range of electrophilic compounds that are common substrates for glutathione S-transferases. The apparent (pseudo)m for glutathione was determined as 4.9 mM at a fixed concentration of linolenic acid hydroperoxide (3 microM). The enzyme showed low affinity for hydroperoxide substrates (apparent Km for linolenic acid hydroperoxide and L-alpha-phosphatidylcholine hydroperoxide of 3.8 and 9.7 mM, respectively at a fixed glutathione concentration of 3 mM).

Animals↗
PubMed1995

Isolation of the cell cycle control gene cdc2 from Paramecium tetraurelia.

The polymerase chain reaction was used to amplify a cdc2 homologous sequence from Paramecium tetraurelia. The corresponding amino acid sequences exhibits about 50% similarity to the cdc2 proteins of other species. The Paramecium cdc2 encoded protein is 11 amino acids longer than S. pombe p34cdc2. There is one amino acid change in the conserved PSTAIRE region. Southern blot analysis indicates that Paramecium has multiple cdc2 genes. Northern blotting results shows that Paramecium cdc2 is much more expressed in actively dividing cells. It is almost undetectable in starved stationary cells. The mRNA level of cdc2 gene does not change during the vegetative cell cycle.

Amino Acid Sequence↗
PubMed1995

Brugia malayi: localisation and differential expression of extracellular and cytoplasmic CuZn superoxide dismutases in adults and microfilariae.

We have determined the levels of superoxide dismutase (SOD) in different stages of the lymphatic filarial nematode parasite of man, Brugia malayi. Adult male worm extracts showed the highest levels of enzyme activity at 34.5 U mg-1, and there was no significant difference in the overall levels of SOD in extracts of adult female worms and microfilariae (27.1 and 26.7 U mg-1, respectively). SOD activity was detected in the culture medium of parasites maintained in vitro, with particularly high levels of specific activity in media in which males and females were maintained (357 and 339 U mg-1, respectively), indicative of active secretion. In all cases, this was accounted for predominantly by CuZn SOD, assessed by potassium cyanide inhibition. Northern blots with cDNA probes specific for cytoplasmic and extracellular CuZn SODs indicated that levels of mRNA for the cytoplasmic form were similar between adults and microfilariae, whereas expression of the extracellular form was 10x higher in adult worms. Western blots with an antibody to recombinant CuZn SOD demonstrated that higher levels of the extracellular protein were present in adult male worms, whereas the cytoplasmic form was present in roughly equivalent amounts in males, females, and microfilaria. Iodination and immunoprecipitation experiments indicated that the extracellular enzyme was accessible to surface labeling of both male and female adult worms, but not microfilaria. Immuno-electron microscopy showed that CuZn SOD was localised predominantly in the hypodermis of adult parasites, with an asymmetric distribution in the intercordal regions suggestive of compartmentalisation into several distinct syncytia. No labeling was evident in the cuticle, and thus the accessibility of the extracellular enzyme to extrinsic iodination in adult worms remains unclear. No binding of antibody was demonstrable in the glandular region of the oesophagus or the uterus of females, presumed to be major sites of synthesis for secreted proteins. Dense labeling was observed in the seminal fluid surrounding spermatazoa in the vas deferens of male parasites. These data also suggest that, as observed in mammals, nematode spermatazoa are particularly susceptible to oxidative damage and are protected during storage by secreted anti-oxidant enzymes.

Animals↗
PubMed1995

Brugia malayi: differential susceptibility to and metabolism of hydrogen peroxide in adults and microfilariae.

The sensitivity of microfilariae and adult Brugia malayi to hydrogen peroxide (H2O2) was determined in vitro, and parasite viability assessed by incorporation of 2-deoxy-D-[1-3H]glucose. Both stages were surprisingly resistant to peroxide stress. Microfilariae tolerated the direct addition of H2O2 to medium in which they were incubated at concentrations up to 50 microM, whereas adult worms survived the addition of 100 microM H2O2 and showed slightly impaired viability at 150 microM H2O2. Higher concentrations were lethal in both cases. This observation of differential susceptibility was reproducible when parasites were subjected to continuous generation of H2O2 via glucose/glucose oxidase. Microfilariae remained viable over a 4-hr period when challenged with concentrations which generated 20 microM H2O2 in the absence of parasites. Adults survived higher concentrations of glucose oxidase, which generated 200 microM H2O2 over the same time period. Under these conditions the parasites effectively countered the rate of peroxide generation by metabolising the product. Protein carbonyl formation was detectable at sublethal concentrations of glucose/glucose oxidase, but malonaldehyde formation was only detectable coincident with parasite death. The rate of H2O2 consumption by parasites was determined and showed that adult worms metabolised it at a rate 23x faster than microfilariae, expressed as activity per wet weight. Assessment of enzyme activities in parasite extracts demonstrated that H2O2 metabolism was effected principally by catalase activity, which was elevated in adult worms relative to microfilariae. Cytochrome c peroxidase activity was also detected and was roughly equivalent in both stages. Glutathione peroxidase and NADH/NADPH-dependent consumption of H2O2 were absent, and the rate of nonenzymic reduction of H2O2 coupled to glutathione oxidation did not contribute significantly to metabolism. Glutathione reductase activity and total glutathione content were equivalent in adults and microfilariae. This study illustrates that Brugia malayi are much more resistant to H2O2 than other filarial species examined to date and can effectively metabolise levels in excess of those potentially generated by activated leucocytes.

Animals↗
PubMed1995

Social aspects of malaria in Heping, Hainan.

This paper presents findings from a study conducted in Heping Town, Qiongzhong County, Hainan Province, China. The study, conducted in 1992, used qualitative as well as quantitative methods to gather social, cultural and behavioural data associated with the acquisition, transmission and prevention of malaria, and the diagnosis and treatment of disease. These methods included focus groups, key informant and other in-depth interviews, and observations, a household survey and tests of school children of knowledge of malaria. The study is among the first to our knowledge that has utilized this broad mix of methods for tropical disease research in China.

Adolescent↗