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Biomedical subjects

Z Y Yuan

Publications and source records attributed to Z Y Yuan.

At least 19 recordsLinked to original sources
PubMed2001

PCR Based Cloning and Sequence Analysis of the Pichia pastoris Cystathionine beta-Synthase Gene.

The cystathionine beta-Synthase (CBS) gene of Pichia pastoris has been cloned by homology to the CBS gene of Saccharomyces cerevisiae. First, based on the homology alignment of CBS genes from different sources, a pair of degenerate PCR primers were designed to amplify a conservative fragment of Pichia pastoris CBS gene. After the sequence was revealed, 5' RACE and 3' RACE were performed separately. The whole sequence of Pichia pasotris CBS gene was assembled. This gene encodes a polypeptide of 501 residues. Comparison of deduced amino acid sequence of this new gene with that of S. cerevisiae CBS showed 54% identity. Disruption of CBS gene resulted in a Cys(-) auxotrophy in Pichia pastoris. Thesequence was submitted to GenBank/EMBL/DDBJ under Accession No.AF367364.

Journal Article↗
PubMed2000

[The influence on oxgen-carrying capacity of procine hemoglobin while attached PEG to increase its total molecular weight].

Increasing the total molecular weight of hemoglobin through PEG modification has been proved to be a better choice in prolonging vascular retention time. As a long, linear, hydrophilic molecular, PEG exerts significant influence on the oxygen-carrying properties of porcine hemoglobin (pHb) when attached. PEG-hemoglobins exhibit a wide range of differences in their properties depending on which molecular weight PEG is selected, how many PEGs are bound and whether the allosteric cofactors exist or not. Furthermore, DBBF intracross-linked pHbs are bound to various active PEG. As a result, with the combination of the three methods: DBBF intracross-linking, allosteric cofactors involving and PEG conjugating, a tetramer stable and high oxygen-carrying capacity pHb derivitive with large molecular weight is obtained.

Animals↗
PubMed1995

Defects in primer-template binding, processive DNA synthesis, and RNase H activity associated with chimeric reverse transcriptases having the murine leukemia virus polymerase domain joined to Escherichia coli RNase H.

The RNase H domain of murine leukemia virus (MuLV) reverse transcriptase (RT) was replaced with Escherichia coli RNase H, and the effect on RNase H activity and processive DNA synthesis was studied, using RNA-DNA hybrids containing sequences from the MuLV polypurine tract (PPT). Two chimeric RTs, having the entire polymerase domain or all but the last 19 amino acids, were expressed. In both cases, these RTs made multiple cuts in PPT-containing substrates, whereas wild-type cleavages occurred primarily at sites consistent with the distance between the polymerase and RNase H active sites. Primer extension assays performed with the chimeric RTs, an RNase H-minus RT, and wild-type showed that the presence of a wild-type viral RNase H domain facilitates processive DNA synthesis. When wild-type RT was bound to primer-template, two retarded bands could be detected in band-shift assays. In the absence of primer extension, a high proportion of enzyme-bound primer-template was associated with the faster-migrating band, whereas with DNA synthesis, more of the bound radioactivity was in the super-shifted complex. This suggests that the super-shifted complex contains the active form of RT. The mutant RTs were deficient in formation of this complex, but the chimeric RTs were somewhat less defective than the RNase H-minus mutant. Our results demonstrate that in the wild-type enzyme, the RNase H domain is required to stabilize the interaction between RT and primer-template.

Base Sequence↗
PubMed1995

Evidence that the L-arginine pathway plays a role in the regulation of pumping activity in bovine mesenteric lymphatic vessels.

The objective of this study was to investigate the role of the L-arginine pathway in the regulation of lymphatic pumping. Bovine mesenteric lymphatic vessels (8 to 12 cm in length containing four to six lymphangions) were immersed in an organ bath with input provided by a reservoir filled with Krebs solution. The vessels were stimulated to pump by applying a 6 cm H2O transmural pressure. The addition of 10(-7)-10(-4) M oxyhemoglobin, NG-monomethyl-L-arginine (L-NMMA), or methylene blue to the reservoir resulted in a reduction in lymphatic pumping. L-Arginine (10(-7)-10(-4) M) had no effect on spontaneous pumping activity. However, L-arginine reversed the inhibition caused by oxyhemoglobin and L-NMMA. When tested between 10(-7) and 10(-6) M, sodium nitroprusside (sNP) had variable effects on lymphatics. sNP depressed pumping in approximately 2/3 of the vessels and increased pumping in the remaining 1/3 of ducts. When the results were meaned, sNP caused a significant depression in activity. However, the lower concentration of sNP (10(-7) M) was able to reverse the inhibitory effects of oxyhemoglobin, L-NMMA, and methylene blue whereas the higher concentration (10(-6) M) caused further reductions in pumping activity. These results suggest that bovine lymphatic vessels produce nitric oxide or a related compound. L-Arginine metabolites appear to facilitate the pumping response by an as yet undefined mechanism.

Animals↗
PubMed1994

Effect of phosphatidylcholine on lymphatic drainage and fluid loss from the peritoneal cavity of sheep.

The purpose of this investigation was to test the hypothesis that phosphatidylcholine enhances net ultrafiltration by decreasing lymphatic drainage of the peritoneal cavity. Twelve sheep were used in this study. Six animals received 50 ml/kg intraperitoneal infusions of Dianeal 4.25% (490 mOsm/liter) and six received similar volumes of premixed phosphatidylcholine-Dianeal (510 mOsm/liter). Labeled albumin (25 microCi 125I-human serum albumin) was added to the dialysate as a lymph flow marker. Lymph drainage of the peritoneal cavity was estimated from the appearance of the intraperitoneally administered tracer in the blood. Net ultrafiltration was significantly enhanced by phosphatidylcholine at each hour up to 6 hours post-infusion, and over this period reached 30.3 +/- 3.8 ml/kg in the phosphatidylcholine animals compared to 12.2 +/- 2.1 ml/kg in the control group. Phosphatidylcholine treatment decreased the volume removed by lymphatics; by six hours 5.5 +/- 1.1 ml/kg in the animals receiving phosphatidylcholine, and 10.3 +/- 1.0 ml/kg in the control group was drained as lymph. Fluid loss (estimated from the tracer disappearance from the peritoneal cavity) was slightly less in the phosphatidylcholine-treated animals, averaging 15.8 +/- 1.6 in this group versus 16.8 +/- 1.7 ml/kg in the control sheep. However, these differences were not significant. Phosphatidylcholine significantly increased transcapillary ultrafiltration (estimate of volume movement into peritoneal cavity without fluid loss) from 27.6 +/- 1.5 ml/kg in the controls to 43.8 +/- 3.4 ml/kg in the animals receiving phosphatidylcholine.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

51Cr-RBCs and 125I-albumin as markers to estimate lymph drainage of the peritoneal cavity in sheep.

The purpose of this study was to compare the use of 125I-labeled human serum albumin (125I-HSA) and autologous 51Cr-labeled red blood cells (51Cr-RBCs) as lymph flow markers to estimate lymph drainage of the peritoneal cavity in conscious sheep. In one group, we assessed lymph drainage from the appearance of intraperitoneally administered tracer in the bloodstream. To determine distribution of drainage into discrete lymph compartments, in a second group of studies, lymph was collected from the caudal mediastinal lymph node and the thoracic duct, both of which are involved in lymphatic drainage of the ovine peritoneal cavity. Ringer lactate solution (50 ml/kg) containing 8-10 microCi each of 125I-HSA and 51Cr-RBCs was infused into the peritoneal cavity. Lymph drainage was calculated by dividing the change in mass of tracer in the blood or lymph compartments by the average intraperitoneal tracer concentration. In noncannulated animals, lymph drainage averaged over 6 h was higher with 125I-HSA as tracer (1.35 +/- 0.12 vs. 0.62 +/- 0.19 ml.h-1.kg-1 with 51Cr-RBCs). A similar pattern was noted in terms of drainage into the caudal lymphatic (0.89 +/- 0.23 and 0.52 +/- 0.19 ml.h-1.kg-1 with 125I-HSA and 51Cr-RBCs, respectively) and thoracic duct (0.16 +/- 0.06 and 0.05 +/- 0.02 ml.h-1.kg-1 with 125I-HSA and 51Cr-RBCs, respectively). Analysis of 125I-HSA and 51Cr-RBC concentrations in lymph and intraperitoneal fluid suggested sieving of RBCs at the diaphragmatic stomata or lymph nodes. Using 125I-HSA as tracer and combining data from noncannulated and cannulated sheep, we estimated peritoneal lymph drainage to be 1.35 ml.h-1.kg-1, with 66% of this flow drained by the caudal vessel, 22% by the parasternal pathway (right lymph duct), and 12% by the thoracic duct.

Animals↗
PubMed1994

Long-term continuous ambulatory peritoneal dialysis in diabetics.

Of 147 diabetic patients with end-stage renal disease who were treated in our CAPD program between 1978 and 1991, 6 men and 1 woman (5 had type II and 2 type I diabetes) with a mean age of 54 (range 21-70) years have been on CAPD for more than five years (mean: 76 mos, range: 65-109 mos) and on peritoneal dialysis (IPD+CAPD) for an average of 85 (range: 67-118) mos. They had a variety of comorbid conditions at the start of CAPD: Retinopathy (5/7), blindness (3/7), hypertension (5/7), peripheral neuropathy (7/7), peripheral vascular disease (3/7), congestive heart failure (3/7), myocardial infarction (1/7), ischemic heart disease (2/7). Two were smokers and five over the age of 65. Peritonitis rate was 1 episode/11.4 pt mos, exit-site infection 1/76.4 pt mos and average hospitalization rate 32.8 days/patient/year. Hypertension was well-controlled with discontinuation of all medications; after initiation of CAPD two of them remained without medications throughout the study but in the rest, medications had to be restarted. As assessed by HbA1c, blood glucose control improved with IP administration of insulin. Residual renal function progressively decreased. None of them developed severe hyperparathyroidism. Peripheral neuropathy remained stable in four and deteriorated in two. Total protein, albumin, cholesterol and triglycerides decreased during the last two years indicating a degree of malnutrition. Our experience with these seven patients suggests that diabetic patients, even the aged and those with many comorbid conditions and complications, can survive for long periods on CAPD.

Comorbidity↗
PubMed1993

Lymphatic drainage of hypertonic solution from peritoneal cavity of anesthetized and conscious sheep.

Lymphatic drainage of the peritoneal cavity may reduce ultrafiltration in continuous ambulatory peritoneal dialysis. We assessed lymphatic drainage of the peritoneal cavity in sheep under dialysis conditions by cannulation of the relevant lymphatic vessels and compared lymphatic drainage in anesthetized and conscious animals. Lymph was collected from the caudal mediastinal lymph node and the thoracic duct, both of which are involved in the lymphatic drainage of the ovine peritoneal cavity. Volumes of a hypertonic dialysis solution (50 ml/kg 4.25% Dianeal) containing 25 microCi 125I-human serum albumin were instilled into the peritoneal cavity, and lymph flows and the appearance of labeled protein in the lymphatic and vascular compartments were monitored for 6 h. Intraperitoneal pressures increased 4-5 cmH2O above resting levels after infusion of dialysate. On the basis of the appearance of tracer in the lymph, drainage of peritoneal fluid into the caudal lymphatic was calculated to be 3.09 +/- 0.69 and 14.14 +/- 2.86 ml/h in anesthetized and conscious sheep, respectively. Drainage of peritoneal fluid into the thoracic duct preparations was calculated to be 1.32 +/- 0.33 and 14.69 +/- 5.73 ml/h in anesthetized and conscious sheep, respectively. Significant radioactivity was found in the bloodstream, and at least a portion of this was likely contributed by the right lymph duct, which was not cannulated in our experiments.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthesia↗
PubMed1993

Clinical associations of glycemic control in diabetics on CAPD.

Diabetic control in 110 diabetics (39 type I and 71 type II), who had been on continuous ambulatory peritoneal dialysis (CAPD) for at least 3 months, was considered good (group G, n = 63) or poor (group P, n = 47) if > 50% or < or = 50% of glucose measurements, respectively, were within 3.3-11.1 mmol/L. Compared to group P, group G had more type I diabetics and fewer type II diabetics; higher serum cholesterol and lower serum creatinine; higher rates of blindness, autonomic neuropathy, congestive heart failure, myocardial infarction, cerebrovascular accidents and extremity gangrene; higher annual rates of peritonitis (1.47 +/- 1.31 vs 0.98 +/- 1.19 episodes/patient-year), exit-site/tunnel infection (0.83 +/- 1.14 vs 0.39 +/- 0.68 episodes/patient-year), and catheter loss (0.81 +/- 0.59 vs 0.39 +/- 0.52 episodes/patient-year); and longer hospitalization (38 +/- 31 vs 14 +/- 15 days yearly). All differences were significant at p = 0.05 or lower. According to life-table analysis, median patient survival was 25 +/- 3 months in group P and 85 +/- 17 months in group G (p < 0.0001). Technique survival was 14 +/- 2 months for group P and 28 +/- 4 months for group G (p < 0.0001). Good diabetic control in diabetics on CAPD is associated with better outcome and constitutes, therefore, a desirable therapeutic goal.

Blood Glucose↗
PubMed1992

[The relationship between plasma atrial natriuretic peptide (ANP) and glomerular ANP receptors in 5/6 nephrectomized rats].

Chronic renal failure was induced in male Wistar rats with 5/6 nephrectomy (group I) and sham-operation was carried in the controls (group II). The results showed that in group I, plasma ANP levels increased progressively as Scr elevated. The plasma levels of renin and angiotensin raised simultaneously as compared with the controls (P < 0.001). At the 20th week after operation, urine volume and Na decreased significantly (P < 0.05) and the number of glomerular ANP receptors decreased significantly at the 12th week (P < 0.05) and 20th week (P < 0.01). Our data suggest that in 5/6 nephrectomized rats: 1. The elevation of plasma ANP level might be partly caused by damage of glomerular receptors. 2. The elevated plasma ANP could not cause its diuretic, natriuretic, blood pressure depression and R-A inhibition effect due to the damage of kidney ANP receptors.

Aldosterone↗
PubMed1992

CAPD in end stage patients with renal disease due to diabetes mellitus--an update.

Large numbers of diabetics with renal failure have been treated by continuous ambulatory peritoneal dialysis (CAPD). Overall 1-year patient survival varies from 51% to 87%. Mortality is due to cardiovascular disease in more than 50% of the cases. Young diabetics with good blood pressure control and without cardiac disease have a chance at long survival on CAPD. In comparison to hemodialysis, CAPD yields better patient survival for young diabetics and worse for old diabetics, worse technique survival, probably greater overall morbidity, and similar rates of progression of retinopathy, neuropathy and peripheral vascular disease. Adequacy of peritoneal clearance and peritoneal ultrafiltration characteristics are similar between diabetics and non-diabetics on CAPD. CAPD is associated with better preservation of renal function than hemodialysis in diabetics. The rates of CAPD peritonitis do not differ substantially between diabetics and non-diabetics. However, diabetes appears to be associated with higher incidence of tunnel infection. Hyperlipidemia is generally less severe in diabetics than non-diabetics on CAPD, but malnutrition is more frequent in diabetics. CAPD has many attractive features and several drawbacks for the management of diabetics with end stage renal failure (ESRF). Its ultimate success will depend on the outcome of efforts to improve cardiovascular mortality, malnutrition, hyperlipidemia and catheter-related infections.

Adolescent↗
PubMed1992

CAPD and pancreatitis: no connection.

Autopsy studies have shown that approximately 56% of patients on long-term continuous ambulatory peritoneal dialysis (CAPD) develop various pancreatic abnormalities, such as acute and chronic pancreatitis, fibrosis, and acinar dilatation. This prevalence of anatomical abnormalities is similar to that observed in patients on hemodialysis and higher than that in those with normal renal function. However, clinical acute pancreatitis is an uncommon complication of CAPD (0.9%), and this prevalence is similar to that (1.7%) of patients on hemodialysis. We can attribute acute pancreatitis in CAPD patients to no single factor. Perhaps preexisting anatomical abnormalities of the pancreas make the CAPD patient susceptible to acute pancreatitis when exposed to a variety of physiological and nonphysiological influences. The diagnosis of acute pancreatitis in CAPD patients is difficult, because symptoms and signs are similar to those of dialysis-associated peritonitis. Serum amylase values three times greater than the upper limit of normal and effluent amylase greater than 100 U/L suggest the diagnosis of acute pancreatitis. Serum lipase, isoamylase, and pancreatic secretory trypsin inhibitor are not helpful. In confirming the diagnosis, a computed tomography (CT) scan is more helpful than ultrasound, although it is positive in only 50-60% of cases. One should harbor a high index of suspicion concerning acute pancreatitis if a CAPD patient presenting with suspected peritonitis has either a negative effluent culture or does not respond to antibiotic therapy.

Acute Disease↗
PubMed1991

A continuous fluorescent assay for measuring protease activity using natural protein substrate.

A continuous caseinolytic activity assay has been developed and characterized with trypsin, a serine protease, and transin, a metalloproteinase. Beta-casein labeled with both N-(7-dimethylamino-4-methylcoumarinyl)-maleimide (DACM) and fluorescein isothiocyanate (FITC) is used as the substrate in this assay. The effect of proteolysis of the substrate is a reduction of the intermolecular energy transfer from DACM to FITC. The caseinolytic activity is then monitored by the fluorescence increase. The activities of both proteases obey Michaelis-Menten kinetics with Km = 1.6 +/- 0.2 microM for trypsin and Km = 13.2 +/- 1.9 microM for transin. Protease concentrations as low as 10 ng/mL can be utilized. The pH dependence of the caseinolytic activity has been determined for both enzymes.

Caseins↗
PubMed1991

Mutational analysis of the transin (rat stromelysin) autoinhibitor region demonstrates a role for residues surrounding the "cysteine switch".

The family of mammalian extracellular matrix metalloproteases (MMPs) are secreted by cells in an inactive (latent) proenzyme form. A highly conserved amino acid sequence, PRCGVPDV, is found near the COOH-terminal end of the pro-domain of these MMPs and believed to act as an "autoinhibitor." Recent studies (Springman, E. B., Angleton, E. L., Birkedal-Hansen, H., and Wart, H. E. V. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 364-368) indicate the Cys of this sequence ligands to the active-site zinc keeping the proenzyme in an inactive state, and mutational analysis (Sanchez-Lopez, R., Nicholson, R., Gesnel, M. C., Matrisian, L. M., and Breathnach, R. (1988) J. Biol. Chem. 263, 11892-11899) suggests that the conserved residues surrounding this Cys are required for latency. We have constructed 16 new site-directed mutations of the PRCGVPDV autoinhibitor region of the MMP transin (rat stromelysin) and tested whether these mutant enzymes are produced in a latent or activated form. We find that the conserved Arg as well as the Cys are essential for maintaining latency. The Cys cannot be replaced by other zinc-liganding amino acids, and the Arg cannot be replaced by Lys. Residues immediately surrounding the Cys are sensitive to even conservative amino acid substitutions. We show that a synthetic peptide PRCGVPDV is capable of acting as a weak inhibitor of transin and that replacement of the Cys with a Ser abolishes inhibition by the peptide. A review of the current knowledge of MMP substrate specificity in combination with these new results suggests that the PRCGVPDV sequence does not inhibit activity by mimicking the known substrates of the protease.

Amino Acid Sequence↗
PubMed1990

Relation between plasma atrial natriuretic peptide (ANP) and glomerular ANP receptors in 5/6 nephrectomized rats.

Chronic renal failure (CRF) was induced in male wistar rats (Group I) by 5/6 nephrectomy and the sham-operated ones served as control (Group II). The results showed that in Group I, plasma atrial natriuretic peptide (ANP) levels increased progressively as the Scr was elevated. Plasma R-A rose simultaneously compared to the normal (P less than 0.001). At the 20th week after operation, urine volume and Na decreased significantly (P less than 0.05). The number of glomerular receptors decreased markedly at the 12th week (P less than 0.05) and 20th week (P less than 0.01). Our data suggest that in 5/6 nephrectomized rats, the elevation of plasma ANP level might be partly caused by the damage of glomerular ANP receptors, and the elevated plasma ANP could not play its role in diuresis, natriuresis, blood pressure depression and R-A inhibition as a result of the damage of kidney ANP receptors.

Animals↗
PubMed1988

Molecular cloning and sequencing of DNA complementary to chicken liver fatty acid synthase mRNA.

The cDNA corresponding to 4.18 kilobases (kb) of the mRNA of chicken liver fatty acid synthase has been cloned and sequenced. The cDNA corresponds to the 3' end of the mRNA and consists of a 1.87-kb noncoding tail and a 2.31-kb region encoding 769 amino acids of the C terminus of the enzyme. The thioesterase at the C terminus, preceded by the acyl carrier protein, can be identified from known amino acid sequences. However, the identity of the enzymes N terminal to the acyl carrier protein could not be ascertained. The partial amino acid sequence of the chicken liver fatty acid synthase shows greater than 70% similarity with the rat mammary gland enzyme.

Amino Acid Sequence↗
PubMed1986

Fluorescence studies of chicken liver fatty acid synthase. Segmental flexibility and distance measurements.

The 4'-phosphopantetheine of chicken liver fatty acid synthase was specifically labeled with the fluorescent substrate analog coenzyme A 6-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]aminohexanoate at low salt concentrations. A serine at the active site of the thioesterase was specifically labeled with the fluorescent compounds 6-[7-nitrobenz-2-oxa-1,3-diazol-4-yl]aminopentylmethylphosphono fluoridate and/or pyrenebutyl methylphosphonofluoridate. Dynamic anisotropy measurements indicate the thioesterase has considerable segmental flexibility, whereas the fluorescent labeled 4'-phosphopantetheine does not display detectable local or segmental flexibility. Fluorescence resonance energy transfer measurements indicate that the distance between the fluorescent label at the end of the 4'-phosphopantetheine and NADPH bound to the beta-ketoacyl reductase or enoyl reductase site on the same polypeptide chain is essentially the same, approximately 38 A. The two types of reductases were distinguished by specifically blocking enoyl reductase with pyridoxal 5'-phosphate. No significant energy transfer occurs between sites on different polypeptide chains so that the distances must be greater than 55 A. The distance between the serine on the thioesterase and the 4'-phosphopantetheine on the same polypeptide is 48 A; again no interpolypeptide chain energy transfer was observed. The distance between the serines of the two thioesterases within a fatty acid synthase molecule is greater than 56 A. The monomeric enzyme obtained at 1 degree C does not have beta-ketoacyl synthase and reductase activities. Also fluorescent titrations indicate NADPH is not bound to beta-ketoacyl reductase in monomeric enzyme. The addition of potassium phosphate to the monomers at 1 degree C rapidly dimerizes the enzyme and restores the beta-ketoacyl reductase activity. The beta-ketoacyl synthase activity is slowly restored when the dimer is raised to room temperature. The results obtained suggest that relatively large conformational changes may be part of the catalytic cycle.

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