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Biomedical subjects

X Yang

Publications and source records attributed to X Yang.

At least 991 records · Page 55Linked to original sources

In vitro replication of cyanobacterial plasmids from Synechocystis PCC 6803.

Little knowledge of DNA replication in cyanobacteria is available. In this study, we report the development and characterization of an in vitro system for studies of replication of the endogenous plasmids from the unicellular cyanobacterium Synechocystis 6803. This system (fraction III) was isolated at high salt concentrations and partially purified on a heparin-agarose column. DNA polymerases in Synechocystis 6803 appeared to be associated with membranes and could be released by the addition of ammonium sulfate to 20% saturation. DNA synthesis in fraction III was dependent on the addition of cyanobacterial plasmids isolated from the same strain. The in vitro replication products consist mostly of the supercoiled form of the plasmids. Unlike replication of many Escherichia coli plasmids, replication of cyanobacterial plasmids did not require added ATP, was not inhibited by omission of the ribonucleotides, and was insensitive to the RNA polymerase inhibitor rifampicin and the gyrase inhibitor novobiocin, but was inhibited by ethidium bromide. These data suggest that RNA may not be involved in the initiation of replication of cyanobacterial plasmids from Synechocystis 6803. In addition, intermediates of replication have been detected by two-dimensional gel electrophoresis. Density labeling experiments also indicate that cyanobacterial plasmid synthesis in vitro occurs by a semiconservative replication.

Centrifugation, Density Gradient↗

Genetic and physical mapping of the GLUR5 glutamate receptor gene on human chromosome 21.

Glutamate receptors (GluRs) mediate excitatory neurotransmission and may have important roles in central nervous system disorders. To characterize the human GLUR5 gene, which is located on human chromosome 21q22.1, we isolated cDNAs, genomic phage lambda clones, and yeast artificial chromosomes (YACs) and developed sequence tagged sites (STSs) and simple sequence length polymorphisms (SSLPs) for GLUR5. Genetic mapping with a tetranucleotide AGAT repeat named GLUR5/AGAT (six alleles observed, 70% heterozygosity) placed GLUR5 5 cM telomeric to APP (D21S210) and 3 cM centromeric to SOD1 (D21S223). The human GLUR5 gene is located near the familial amyotrophic lateral sclerosis (FALS) locus; linkage analysis of GLUR5 SSLPs in FALS pedigrees yielded negative lod scores, consistent with the recent association of the FALS locus with the SOD1 gene. Physical mapping of GLUR5 using a YAC contig suggested that the GLUR5 gene spans approximately 400-500kb, and is within 280kb of D21S213. The large size of the GLUR5 gene raises questions regarding its functional significance. Our GLUR5 YAC contig includes clones found in the Genethon chromosome 21 YAC contig, and reference to the larger contig indicates the orientation centromere--D21S213-GLUR5 5' end-GLUR5/AGAT--GLUR5 3' end--SOD1. The development of GLUR5/AGAT should permit rapid determination of the status of the GLUR5 gene in individuals with partial trisomy or monosomy of chromosome 21. Such studies may provide insights concerning the possible role of GLUR5 in Down syndrome.

Base Sequence↗

A kalilo-like linear plasmid in Louisiana field isolates of the pseudohomothallic fungus Neurospora tetrasperma.

Two Louisiana strains of Neurospora tetrasperma contain a linear plasmid (LA-kalDNA) with a restriction map identical to the Hawaiian Neurospora intermedia senescence plasmid, kalDNA, but with termini 100 nucleotide pairs shorter. One of these strains also bore a circular plasmid similar to the Hawaiian circular plasmid Hanalei-2. One species probably acquired both plasmids from the other by horizontal transfer, at a time sufficiently distant for sequence divergence to take place. Many LA-kalDNA-bearing derivative strains senesced, but this plasmid does not guarantee senescence. Furthermore, LA-kalDNA does not insert into mtDNA. One senescent strain showed no LA-kalDNA. The plasmids are effectively transmitted via the pseudohomothallic sexual cycle. Single mating-type derivatives transmit plasmids maternally.

Biological Evolution↗

Vegetative incompatibility in Neurospora: its effect on horizontal transfer of mitochondrial plasmids and senescence in natural populations.

We have investigated the horizontal transfer of two mitochondrial plasmids and the Kalilo senescence phenotype in the fungus Neurospora without the use of heterokaryon-forcing markers. The Kalilo senescent state was only transferred between fully-compatible N. crassa strains, but not between strains differing at any of the loci het-c, het-d, het-e or mating-type. However, the linear plasmid kalDNA and the circular plasmid Han-2 were transferred following incompatible vegetative interactions. Our data suggest that vegetative incompatibility due to allelic differences at het-c is more effective in preventing transfer than that due to het-d, het-e or mating-type. Based on these observations we have developed a novel test for assessing vegetative incompatibility between Kalilo and non-Kalilo field isolates of N. intermedia. In this procedure combinations of Kalilo and non-Kalilo field isolates of N. intermedia were grown together and tested for senescence. Compatibility is inferred if the young non-Kalilo strain dies along with the senescent Kalilo strain, whereas incompatibility is inferred when the Kalilo strain dies without imposing its senescent state onto the non-Kalilo strain. Our results suggest that each of the nine Kalilo strains tested is incompatible with each of 20 non-Kalilo isolates from the same N. intermedia population of the Hawaiian island of Kauai. However, the observed incompatibility did not completely prevent cytoplasmic exchange, and in several cases plasmid transfer could be detected.

Crosses, Genetic↗

Developmental regulation of multiple nicotinic AChR channel subtypes in embryonic chick habenula neurons: contributions of both the alpha 2 and alpha 4 subunit genes.

Habenula neurons from both early and late stage embryonic chickens express multiple subtypes of nicotinic acetylcholine receptor channels (nAChRs). The channel subtypes expressed by habenula neurons are similar in functional properties, but apparently distinct in subunit composition, from their peripheral counterparts in autonomic ganglia. Early in development, nicotine activates four classes of neuronal bungarotoxin (nBGT)-sensitive channels (approx. conductance = 15, 30, 50, 60pS) that are intermingled on the surface of habenula neuronal somata. In neurons removed from older animals, nAChR channel activity has increased 4- to 40-fold and channel subtypes have become spatially segregated from one another. Analysis of the profile of nAChR subunit gene expression by polymerase chain reaction indicates that several of the alpha-type subunit genes, including alpha 2,3,4,5,7, and alpha 8, as well as both beta 2 and beta 4, are expressed. Treatment of the neurons with subunit specific antisense oligonucleotides reveals that the alpha 2 and alpha 4 (but not alpha 3) subunits contribute to the functional profile of native nAChRs expressed by habenula neurons. Consideration of the functional properties and apparent subunit composition of autonomic ganglion nAChRs in the chick suggests that habenula neurons may utilize a very distinct set of subunit combinations to produce an array of nAChR channel subtypes similar in both conductance and pharmacological profile to those expressed by sympathetic neurons.

Acetylcholine↗

Pharmacological evaluation of methylcarbamylcholine-induced drinking behavior in rats.

Methylcarbamylcholine (MCC), a structural analog of carbachol (an acetylcholine agonist), has been reported to be a specific nicotinic cholinergic receptor ligand. MCC produces a robust polydipsic response shortly following central administration. The purpose of the present study was to pharmacologically characterize this increase in drinking behavior. Male Wistar rats were implanted with intracerebroventricular (ICV) cannula guides directed at the left lateral ventricle. Following a recovery period, animals were injected ICV with saline or various doses of MCC (3-60 micrograms) and water consumption was quantified. MCC produced a dose-related, transient increase in water consumption that peaked at a dose of 30 micrograms. In contrast, nicotine, a potent nicotinic cholinergic receptor agonist, did not produce changes in drinking following ICV administration. MCC-induced increases in drinking were not blocked by pretreatment with several selective nicotinic receptor antagonists including dihydro-beta-erythriodine (DHBE), hexamethonium, and mecamylamine. However, pretreatment with the muscarinic antagonist atropine (0.01 or 1.0 microgram) completely abolished MCC-induced polydipsia. Following a chronic treatment regimen (MCC injected ICV twice daily for 10 days), no tolerance to MCC-induced changes in water consumption was observed. Previous studies have demonstrated that tolerance develops to nicotinic-receptor mediated responses following the identical chronic treatment paradigm. These results suggest that MCC-induced polydipsia is mediated through stimulation of muscarinic rather than nicotinic receptors.

Animals↗

Synergistic effect of ethanol and cycloheximide on activation of freshly matured bovine oocytes.

Bovine follicular oocytes were collected from ovarian antral follicles (2 to 7 mm in diameter) from slaughtered cattle. They were matured in vitro (IVM) for 23 to 24 h and then activated. In Experiment 1, 4 concentrations of ethanol were compared. The activation rates of oocytes were 4, 12, 36 and 27%, respectively, following exposure for 7 min to 0, 5, 7 and 10% ethanol. In Experiment 2, 7% ethanol was tested with exposure times of 0, 5, 7.5 and 10 min, and 6, 32, 27 and 33% of the oocytes were activated, respectively. In Experiment 3 the synergistic effect of ethanol and electric pulse was compared within 4 treatments: A) 7% ethanol alone, B) electric pulse alone, C) ethanol first and then electric pulse treatment, and D) electric pulse first followed by ethanol exposure. Of the oocytes activated, 37, 31, 28 and 51%, respectively, were from Treatments A through D. In Experiments 4 and 5 the possible synergistic effect of ethanol and a protein synthesis inhibitor, cycloheximide, was studied within 4 treatments: A) parthenogenetic control with no activation treatment, B) ethanol alone, C) cycloheximide alone, and D) ethanol treatment followed by cycloheximide. The oocyte activation rates in Experiment 4 in Treatments A through D, respectively, were 9, 44, 43 and 84%. Corresponding values for development of oocytes to the 2 to 8-cell stage after culture for 3 d (Experiment 5) were 9, 20, 14 and 45%, respectively (P<0.05). In conclusion, exposure to 7% ethanol for 5 min followed by incubation with cycloheximide was the best activation treatment for bovine IVM oocytes.

Journal Article↗

The toxic effects of cadmium on cell division and chromosomal morphology of Hordeum vulgare.

The effects of cadmium at different concentrations (0.5-20 ppm) on root growth, cell division and chromosomal morphology of Hordeum vulgare were studied. The rate of root growth and mitotic index decreased progressively with increasing cadmium concentration and treatment duration. Different concentrations of cadmium could cause mitotic irregularities comprising c-mitoses, anaphase bridges, breaks, stickiness, lagging and vagrant chromosomes and micronuclei. The intensity of the toxic effects is basically dependent on the cadmium concentration and duration of treatment.

Analysis of Variance↗

Tissue reactions to titanium implants containing bovine bone morphogenetic protein: a scanning electron microscopic investigation.

Cylindric titanium implants treated with bovine bone morphogenetic protein (bBMP) (Group I) and with bovine serum albumin (BSA) (Group II) were implanted in the edentulous mandibles of 15 adult dogs. They were examined 1, 2, 4, 8, and 12 weeks after insertion of the implants by scanning electron microscopy (SEM). In all specimens from Group I, active bone formation had occurred between the implant and host bone, with a well-adapted fit between the implant and the newly formed bone 4 weeks after implantation. The surrounding bone was of normal structure by week 8. However, 3 months after implantation, fibrous tissue was consistently seen surrounding parts of the implants in Group II. The results of this study suggest that the implants treated with bBMP are capable of forming new bone of normal structure at a faster rate than BSA controls and that this new bone is closely adapted to the implant surface.

Animals↗

Reproducibility of five anorectal morphologic measurements in defecography.

RATIONALE AND OBJECTIVES: We evaluated the inter- and intraobserver reproducibility of measuring five morphologic parameters of the anorectum in defecography (evacuation proctography). METHODS: Measurements from 42 defecographic studies were statistically analyzed. The parameters measured during resting, squeezing, and straining included two anorectal angles (posterior and axis), maximal width of the anal canal, maximal width of the rectal lumen, and size of the rectocele. RESULTS: The results demonstrated only fair interobserver agreement (kappa = 0.22-0.38) for almost all measurements of the five morphologic parameters. There were high correlations (kappa = 0.62-1.00) among most intraobserver measurements. CONCLUSION: For defecographic measurement, the five parameters we studied have relatively poor clinical value because of high inter- and intraobserver inconsistency.

Adult↗

Dietary alpha-linolenic and linoleic acids competitively affect metabolism of polyunsaturated fatty acids in Arctic charr (Salvelinus alpinus).

We investigated the effects of various levels of dietary 18:2(n-6) (linoleic acid) and 18: 3(n-3) (alpha-linolenic acid) on the growth, feeding, body composition and metabolism of polyunsaturated fatty acids in liver and muscle of Arctic charr. Fish were fed 8 or 12 casein-based semipurified diets in two separate experiments for 12 or 14 wk, respectively. Low levels of dietary 18:2(n-6) and 18:3(n-3) supplemented with pure methyl esters or linseed (flax) oil had an additive effect on fish growth. For each 100 g of diet, the addition of 1.6-1.7 g of 18:3(n-3) and 2.6 g of 18:2(n-6) led to significantly lower fish growth compared with the treatment diets with lower amounts of 18:2(n-6). Whereas 18:2(n-6) was converted to 20:4(n-6) and 22: 5(n-6) in the absence or presence of dietary 18: 3(n-3), the dominant product of 18:3(n-3) conversion was consistently 22:6(n-3). High levels of dietary 18: 3(n-3) markedly inhibited the conversion of 18:2(n-6), whereas the inhibition of dietary 18:2(n-6) on 18: 3(n-3) conversion was noted only when the ratio of dietary 18:2(n-6) to 18:3(n-3) changed from 1.0 to 1.5. Feeding diets rich in 18:3(n-3) led to a marked accumulation of 18:3(n-3) and 18:4(n-3) in fish muscle but a negligible change in 20:5(n-3) and 22:6(n-3), regardless of the level of 18:3(n-3) in the diets.

Animals↗

Zona-opening of hamster oocytes: a comparative study using macro- and micro-manipulation methods.

The present study compared a new macro-manipulation technique for zona-opening of hamster oocytes with existing micro-manipulation techniques. In experiment I the zona pellucida of hamster oocytes was partially opened with a 32 gauge steel needle (macro-manipulation) and these oocytes were then co-incubated with human spermatozoa at 36.5 degrees C in 5% CO2 in air for 3.5 h. Zona-free and zona-intact oocytes similarly inseminated served as controls. Of 113 oocytes, 30 (26%) lysed following zona-opening with the macro technique. The sperm penetration rates of zona-opened and zona-free oocytes were 37% (31/83) and 60% (49/81) respectively (P < 0.01), with 12 and 28% oocytes respectively showing polyspermia (P < 0.05). No sperm penetration occurred in zona-intact oocytes. In experiment II the zona pellucida of hamster oocytes was partially opened by macro- or micro-manipulation techniques before the oocytes were coincubated with human spermatozoa as in experiment I. Nine of 156 (6%) macro-manipulated and 11 of 133 (8%) micro-manipulated oocytes lysed following zona-opening. Of 147 remaining macro-manipulated and 122 remaining micro-manipulated oocytes, 80 (55%) and 79 (64%) respectively were penetrated (P > 0.05), with 30 and 37% respectively showing polyspermia (P > 0.05). These results demonstrate that macro-manipulation results in similar penetration and polyspermia rates as compared to conventional micro-manipulation.

Animals↗

Isolation and characterization of two protease inhibitors from bovine plasma.

Two protease inhibitors (Inh2 and Inh3) from bovine plasma have been isolated and characterized. The apparent molecular weights of the two proteins are 56 and 58 kDa, respectively. Although Inh2 and Inh3 both inhibit trypsin and human neutrophil elastase, only Inh3 is a good inhibitor of chymotrypsin and cathepsin G. Inh3 is much more sensitive to oxidation than Inh2. One murine monoclonal antibody recognizes Inh3 but not Inh2. Inh3 resembles human alpha 1-antitrypsin both structurally and functionally. Inh2, on the other hand, has some structural homology to human alpha 1-antichymotrypsin, but its specificity does not correspond to that of either human alpha 1-antitrypsin or human alpha 1-antichymotrypsin.

Amino Acid Sequence↗

Analysis of polarity of bovine and rabbit embryos by scanning electron microscopy.

Cellular polarization during preimplantation development of the embryo is believed to be a crucial event in the transition of a zygote to a blastocyst stage embryo with morphologically and functionally differentiated cell types. While extensive studies have been done on polarity development in mouse embryos, little information is available in other species, particularly in cattle. The objective of this study was to examine the initiation of polarity by microvilli distribution on blastomeres of cattle and rabbit embryos using scanning electron microscopy (SEM). Bovine embryos were obtained by in vitro fertilization of in vitro-matured follicular oocytes. Rabbit embryos of various stages were collected from superovulated rabbits. Intact embryos and isolated blastomeres were examined in both species. Blastomeres from 1- to 8-cell embryos in both cattle and rabbits showed no polarity. The onset of transitional polarization of microvillous distribution occurred in some blastomeres of cattle embryos at the 9- to 15-cell stage; but typical, distinct polarity was not manifested until after the 16-cell stage with approximately 40% polar cells per embryo. In the rabbit blastomere polarity occurred one cell cycle later, with 46% polar cells per embryo after the 32-cell stage. The difference in cell numbers at the time polarity is evident is probably related to the different cell stages for embryo compaction and blastocyst formation in the two species.

Animals↗

Injecting drug use and HIV infection in southwest China.

OBJECTIVES: To determine the prevalence of drug injection among drug users, the seroprevalence of HIV and risk factors for HIV infection among injecting drug users (IDU), and to determine heterosexual transmission of HIV among IDU and their spouses in southwest China. METHODS: Using a cross-sectional design, we conducted an HIV seroprevalence and behavioral survey in three rural counties of Yunnan province, Ruili, Longchuan and Luxi in southwest China, bordering Myanmar (Burma). A total of 860 drug users were recruited in randomly selected communities at the three study sites (response rate, 97%). In addition, a random sample of 62 wives of HIV-infected IDU were assembled from 460 known HIV-positive IDU in Ruili and Longchuan (response rate, 81%). RESULTS: In the sample of 860 drug users, 33% reported injecting drugs. Among the 282 subjects who injected drugs, 82% began intravenous drug use after 1988; 64% injected drugs at least once every day. All subjects shared needles but none cleaned the injection equipment with alcohol or bleach. Overall, 49% tested HIV-positive. HIV seropositivity was independently correlated with a longer history of drug injecting, daily injecting, frequent needle-sharing, being younger, and living in Ruili county. Among the 62 wives of HIV-positive IDU, none used condoms during sex and 10% tested HIV-positive. CONCLUSIONS: We conclude that the introduction of HIV into drug-using communities and the rapid increase in heroin injecting in this population appear to have triggered an explosive HIV epidemic among IDU in southwest China. We recommend that AIDS prevention efforts should begin immediately and focus on discouraging the shift from opium smoking to heroin injecting, needle-sharing, and unprotected sex among drug users and their partners.

Adult↗

Comparison of angiographic opacification of ioxaglate and iodixanol by digital region-of-interest processing technique in an experimental circulation model.

RATIONALE AND OBJECTIVES: By using digital ROI (region of interest) processing techniques, we measured and compared directly the angiographic opacification of ioxaglate and iodixanol in an experimental circulation model. METHODS: A pulsatile pump circulated water into a 2-mm plastic tube for digital subtraction angiography (DSA). Altogether, 90 digital imaging series were registered during injections of two contrast media with 320, 300, and 270 mg/mL iodine concentrations. By selecting a "vessel" ROI and a subtracted background ROI, a time-density curve (TDC) was created. From the TDC, the average density of contrast media and the appearance time of contrast media into the selected ROI were measured. RESULTS: The average density was statistically different (P < .01) among three various iodine concentrations of the contrast media, but no difference was found between the two contrast media. The appearance times of ioxaglate with 320 mg/mL iodine concentration were statistically shorter (P < .01) than those of iodixanol. In both contrast medium groups, the appearance times were statistically shorter (P < .05) with 270 mg/mL iodine concentration than with two higher iodine concentrations. CONCLUSION: The current experimental set-up facilitates reproducible measurement of angiographic opacification during the injection of contrast media. The average densities between ioxaglate and iodixanol are the same, but are significantly different among three iodine concentrations. The different appearance times of the two CM and the three iodine concentrations may be caused by the various viscosities of the contrast media.

Angiography, Digital Subtraction↗

Influence of carbon dioxide gas perfusion to thermal distribution of sapphire probe. A comparative study with saline.

RATIONALE AND OBJECTIVES: A previous study confirmed that the direct laser-thermal conduction of the sapphire probe with carbon dioxide gas perfusion increased the width of the laser-recanalized channel. This caused us to further investigate the thermal distribution characteristics of the sapphire probe when lasing with CO2 gas perfusion. METHODS: The surface temperature of a sapphire probe in a circulation model using 37 degrees C flowing whole blood was measured. Two hundred and sixteen measurements were obtained by directly contacting a flexible thermocouple wire onto the sapphire probe at different sites: 1) metal connector; 2) lateral side of the sapphire crystal; 3) top of the sapphire crystal; and 4) 3 mm in front of the sapphire probe. During lasing with a neodymium-yttrium-aluminum garnet (Nd-YAG) laser, the CO2 gas or saline was infused through the sapphire probe at different flow rates. RESULTS: The lateral side of the sapphire crystal was heated up to 75 degrees C when lasing without any perfusion, but up to 220 degrees C when lasing with CO2 gas perfusion. At all four sites, the mean temperature increases were statistically higher (P < .01) with CO2 gas than with saline perfusion. The mean peak temperatures increased with increasing flows of CO2 gas perfusion and decreased with increasing flows of saline perfusion. CONCLUSIONS: The thermal conduction from the sapphire probe can be significantly enhanced by increasing flows of CO2 gas perfusion. This may play an important role in creating a greater diameter of the recanalized channel and in better delaying the formation of restenosis or re-occlusion after laser recanalization of atheromatous arteries.

Aluminum Oxide↗

Adenosine deaminase inhibitor in cardioplegia enhanced function preservation of the hypothermically stored rat heart.

Adenosine (ADO) has been shown to be protective to the ischemic-reperfused myocardium. This study tested the hypothesis that inhibition of myocardial adenosine deaminase during cold storage will elevate tissue ADO content, improve the cardiac function, and preserve ATP. The isolated rat hearts (6-9 hearts/group) were flushed with a cardioplegic solution containing 0-75 microM erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and immersion-stored at 0 degree C for 9 hr. Function was assessed after 30 min working reperfusion. Function of the unstored hearts (n = 11, mean +/- SE) including heart rate (293 +/- 13 bpm), aortic flow (AF; 52.5 +/- 1.1 ml/min), coronary flow (CF; 23.5 +/- 1.3 ml/min), cardiac output (CO; 76.0 +/- 2.1 ml/min), systolic pressure (SP; 136 +/- 2 mmHg), diastolic pressure (DP; 63 +/- 1 mm Hg), work (90.5 +/- 3.4 g-m/min), and coronary vascular resistance (CVR; 2.77 +/- 0.14 mmHg-min/ml) served as controls. Heart rate in all stored hearts returned to normal after reperfusion. Recovery of other function in no-EHNA group was: AF, 52 +/- 7; CF, 55 +/- 5; CO, 53 +/- 6; SP, 79 +/- 4; DP, 93 +/- 3; work, 47 +/- 7; and CVR, 171 +/- 15% of control. EHNA improved functional recovery in a dose-dependent fashion. At the optimal concentration of 25 microM, the recovery was: AF, 83 +/- 6; CF, 68 +/- 4; CO, 78 +/- 5; SP, 90 +/- 3; DP, 105 +/- 5; work, 77 +/- 8; and CVR 151 +/- 9% of control. ADO A1 receptor antagonists, 8-phenyltheophylline (1 microM) and 1,3-dipropyl-8-cyclopentylxanthine (0.1 microM) blocked the effects of 25 microM EHNA; the recovery of CO was reduced to 65 +/- 3 and 50 +/- 2% of the control, respectively. Tissue ADO content in 25 microM EHNA hearts at the end of storage was 95 +/- 19 nmol/g dry wt, which was significantly elevated from 15 +/- 3 nmol/g dry wt in no-EHNA hearts. EHNA also caused a 45-fold increase in the release of ADO over no-EHNA group during the first 10 min of reperfusion. But EHNA treatment did not cause any change in either end-storage or end-reperfusion myocardial ATP levels. Thus EHNA in cardioplegic solution inhibited cardiac ADO catabolism during long-term hypothermic storage and improved function preservation partially via an ADO A1 receptor-mediated mechanism without invoking ATP conservation.

Adenine↗