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Biomedical subjects

J Wang

Publications and source records attributed to J Wang.

At least 289 records · Page 16Linked to original sources

Use of myocardial tissue Doppler imaging for intraoperative monitoring of left ventricular function.

BACKGROUND: Detection of myocardial ischaemia during surgery is usually by assessment of regional wall motion using two-dimensional transoesophageal echocardiography (TOE). Tissue Doppler imaging (TDI) may assist this assessment and improve its accuracy. METHODS: We measured peak myocardial velocities in the anterior mid-wall of the left ventricle by TOE and pulsed-wave TDI in addition to transmitral flow velocity, two-dimensional echocardiography and cardiovascular variables. We studied 42 patients before and after coronary bypass surgery with left internal mammary artery grafts. RESULTS: Peak systolic and early and late diastolic velocity measurements of the anterior mid-wall were obtained in all patients. Variation between and within observers was small (<6%). Peak systolic thickening velocity correlated with visual assessment of anterior wall motion score, fractional area change of the left ventricle and left ventricular systolic wall stress. Because of the wide overlap of systolic velocity between the segments with normal and abnormal wall motion, it was not possible to separate normal from abnormal segments on the basis of TDI-derived velocity alone. The diastolic velocity in the anterior wall reflected the transmitral filling pattern. After surgery, the peak systolic and late diastolic anterior wall velocities increased (from 4.2 (95% confidence interval 4.0, 4.7) to 5.7 (4.8, 6.3) cm s(-1) and from 3.5 (3.2, 3.9) to 6.0 (5.1, 6.9) cm s(-1) respectively), while the ratio of early to late diastolic velocity decreased from 1.5 (1.2, 1.7) to 1.0 (0.8, 1.2). TDI changes characteristic of new myocardial ischaemia were not seen in any patient. CONCLUSION: Intraoperative measurement of TDI in the anterior wall of the left ventricle is feasible and provides additional quantitative information on both regional and global systolic and diastolic function. We found changes in myocardial velocities indicating improvement in the systolic and impairment in the diastolic function of the anterior wall of the left ventricle immediately after mammary artery grafting.

Aged↗

Antidepressant-like and anorectic effects of the cannabinoid CB1 receptor inverse agonist AM251 in mice.

Psychopathological disorders, and depression in particular, are strongly linked to eating attitude in obese patients. The identification of cannabinoid CB1 receptors (CB1Rs) in areas of the central nervous system (CNS) that have been implicated in regulation of mood and food intake suggests that these receptors may mediate such a behavioral link. The goal of this study was to evaluate CB1R modulation of antidepressant-like effects and food intake. For this purpose, 129/SVE and C57BL/6 male mice were acutely dosed intraperitoneally (i.p.) with the CB1R inverse agonist AM251 (3-30 mg/kg) and tested, respectively, in the tail-suspension test (TST) and in the forced-swim test (FST), which have been used widely as tests sensitive to antidepressant compounds. Like the antidepressant desipramine (DMI, 16 mg/kg), AM251 significantly reduced immobility at 10 mg/kg in the TST and at 1 and 10 mg/kg in the FST. Such a decrease of immobility was not accompanied by an increase in motor activity in the open field, suggesting that occupancy of CB1R by AM251 induced antidepressant-like effects. This was supported by two additional experiments. First, the co-administration of the CB1R agonist CP55940, at a dose that did not induce motor impairment or profound hypothermia (0.01 mg/kg), reversed effects of AM251 in the TST. Secondly, effects of AM251 in the FST were absent in CB1R knockout (KO) mice. In addition to an antidepressant-like effect, AM251 reduced fasting-induced hyperphagia over a comparable dose range. Taken together, these data suggest that regulation of mood and food intake might be obtained through inverse agonism of CB1R.

Affect↗

The vibrio pathogenicity island of epidemic Vibrio cholerae forms precise extrachromosomal circular excision products.

The Vibrio pathogenicity island (VPI) in epidemic Vibrio cholerae is an essential virulence gene cluster. Like many pathogenicity islands, the VPI has at its termini a phage-like integrase gene (int), a transposase-like gene (vpiT), and phage-like attachment (att) sites, and is inserted at a tRNA-like locus (ssrA). We report that the VPI precisely excises from the chromosome and that its left and right ends join to form an extrachromosomal circular excision product (pVPI). Two-stage nested PCR analysis and DNA sequencing confirmed the int-att-vpiT junction and that the core attP of pVPI is identical to the chromosomal VPI attR site. Excision was independent of toxR and toxT. Excision was independent of recA, suggesting that it is mediated by site-specific recombination. Interestingly, while excision was detected in int and vpiT mutants, excision was abolished in a double (int vpiT) mutant and was restored by plasmids containing genes for either recombinase. Excision results in deletion of A361 in the ssrA locus, which flanks the right junction of the VPI. Since A361 encodes U70 in the critical G. U base pair in the acceptor stem of the ssrA RNA that is the determinant for aminoacylation with alanine, this deletion might have deleterious effects on ssrA function. Also, vpiT may have undergone interchromosomal translocation or may represent an independent integration event, as it was found downstream of hutA in some isolates. Our results provide new insight into the molecular biology of the VPI, and we propose that the process of excision and circularization is important in the emergence, pathogenesis, and persistence of epidemic V. cholerae.

Chromosomes, Bacterial↗

A collagen-based scaffold for a tissue engineered human cornea: physical and physiological properties.

Stabilized collagen-glycosaminoglycan scaffolds for tissue engineered human corneas were characterized. Hydrated matrices were constructed by blending type I collagen with chondroitin sulphates (CS), with glutaraldehyde crosslinking. A corneal keratocyte cell line was added to the scaffolds with or without corneal epithelial and endothelial cells. Constructs were grown with or without ascorbic acid. Wound-healing was evaluated in chemical-treated constructs. Native, noncrosslinked gels were soft with limited longevity. Crosslinking strengthened the matrix yet permitted cell growth. CS addition increased transparency. Keratocytes grown within the matrix had higher frequencies of K+ channel expression than keratocytes grown on plastic. Ascorbic acid increased uncrosslinked matrix degradation in the presence of keratocytes, while it enhanced keratocyte growth and endogenous collagen synthesis in crosslinked matrices. Wounded constructs showed recovery from exposure to chemical irritants. In conclusion, this study demonstrates that our engineered, stabilized matrix is well-suited to function as an in vitro corneal stroma.

Animals↗

The PCR-based detection of Leishmania major infections in Meriones libycus (Rodentia: Muridae) from southern Iran.

Cutaneous leishmaniasis (CL) is an increasing public-health problem in the countries of the Mediterranean region. In southern Iran, for example, the incidence of CL has more than doubled over the last decade. As part of an investigation of this worrying trend, the leishmanial infections of the wild rodents that may act as reservoir hosts were investigated in the Iranian province of Fars. Overall, 82 rodents, 56 of them Libyan jirds (Meriones libycus), were collected, in live traps, from a new, hypo-endemic focus of zoonotic CL. When Giemsa-stained smears of ear tissue or any lesions on the rodents were prepared and checked under a light microscope, amastigotes were found in the smears from six of the rodents, all M. libycus. None of the other rodents investigated was found to be infected. The infected rodents were encountered in two of the three areas studied. In PCR-based assays, the amastigotes seen in the ear smears and those from two recent human cases of CL from the focus were all found to be identical to a reference strain of L. major. It therefore seems that M. libycus is the main active reservoir host for ZCL in the focus.

Animals↗

Expression of regulator of G-protein signalling protein 2 (RGS2) in the mouse uterus at implantation sites.

Establishment of an active dialogue between the maternal endometrium and the implanting blastocyst is essential for successful implantation. The aim of this study was to identify genes that are explicitly expressed at implantation sites of the mouse uterus by subtractive hybridization. One expressed sequence tag of the genes identified showed 92% identity to the regulator of G-protein signalling protein 2 (RGS2). The full cDNA sequence of this gene was amplified by PCR and subsequently registered in GenBank. The sequence of its open reading frame encoding 211 amino acids was the same as that of mouse RGS2, with the exception of four amino acids. Northern blot analysis showed that the expression of this gene was much higher at implantation sites than at inter-implantation sites on days 5-8 of pregnancy. In situ hybridization localized this mRNA predominantly to the stromal cells at the implantation sites. These results indicate that RGS2 has a role during implantation, possibly by regulating the intracellular Ca(2+) mobilization and T-cell proliferation at the maternal-fetal interface.

Animals↗

RNA amplification strategies for cDNA microarray experiments.

The biological materials available for cDNA microarray studies are often limiting. Thus, protocols have been developed to amplify RNAs isolated from limited amounts of tissues or cells. RNA amplification by in vitro transcription is the most widely used among the available amplification protocols. Two means of generating a dsDNA template for the RNA polymerase are a combination of reverse transcription with conventional second-strand cDNA synthesis and a combination of the switch mechanism at the 5' end of RNA templates (SMART) with reverse transcription, followed by PCR. To date, there has been no systematic comparison of the efficiency of the two amplification strategies. In this study, we performed and analyzed a set of six microarray experiments involving the use of a "regular" (unamplified) microarray experimental protocol and two different RNA amplification protocols. Based on their ability to identify differentially expressed genes and assuming that the results from the regular protocol are correct, our analyses demonstrated that both amplification protocols achieved reproducible and reliable results. From the same amount of starting material, our results also indicated that more amplified RNA can be obtained using conventional second-strand cDNA synthesis than from the combination of SMART and PCR. When the critical issue is the amount of starting RNA, we recommend the conventional second-strand cDNA synthesis as the preferred amplification method.

DNA, Complementary↗

Transgenic expression of CD36 in the spontaneously hypertensive rat is associated with amelioration of metabolic disturbances but has no effect on hypertension.

Spontaneously hypertensive rats (SHR/NIH strain) harbor a deletion variant in the Cd36 fatty acid transporter and display defective fatty acid metabolism, insulin resistance and hypertension. Transgenic rescue of Cd36 in SHR ameliorates insulin resistance and improves dyslipidemia. However, the role of Cd36 in blood pressure regulation remains controversial due to inconsistent blood pressure effects that were observed with transgenic expression of Cd36 on the SHR background. In the current studies, we developed two new SHR transgenic lines, which express wild type Cd36 under the control of the universal Ef-1 alpha promoter, and examined the effects of transgenic expression of wild type Cd36 on selected metabolic and cardiovascular phenotypes. Transgenic expression of Cd36 in the new lines was associated with significantly decreased serum fatty acids, amelioration of insulin resistance and glucose intolerance but failed to induce any consistent changes in blood pressure as measured by radiotelemetry. The current findings confirm the genetic association of defective Cd36 with disordered insulin action and fatty acid metabolism in the SHR/NIH strain and suggest that Cd36 is linked to other gene(s) on rat chromosome 4 that regulate blood pressure.

Adipose Tissue↗

An outbreak of Methicillin-resistant Staphylococcus aureus cutaneous infection in a saturation diving facility.

We present a molecular epidemiological investigation of an outbreak of cutaneous tissue infection, which involved six divers during a 45 day saturation exposure dive. The cutaneous infection manifested as boils, foliculitis and small abscesses involving different body sites, including nose, external ear canal, necks, back, extremities, and buttocks. Staphylococcus aureus was consistently isolated from the skin lesions of affected divers. A study of the antibiogram revealed that all Staphylococcus aureus isolates were uniformly resistant to penicillin, oxacillin and erythromycin, but sensitive to clindamycin, tetracycline, trimethoprim-sulfamethoxazole, rifampin and vancomycin. Molecular typing by pulse field gel electrophoresis (PFGE) demonstrated that all the Methicillin-resistant Staphylococcus aureus (MRSA) isolates had an indistinguishable pulsed field gel electrophoresis pattern. The source of outbreak was identified as a colonized diver (diver D). Personal contact was most likely the mode of transmission among the six divers. Infection with MRSA should be suspected in outbreaks of boils that are not responding to standard antibiotic therapy among healthy divers and their close contacts. To our knowledge, this is the first report of Methicillin-resistant Staphylococcus aureus (MRSA) outbreak in a saturation diving facility.

Adult↗

Purification, molecular characterization and reactivity with aromatic compounds of a laccase from basidiomycete Trametes sp. strain AH28-2.

A recently isolated basidiomycete, Trametes sp. strain AH28-2, can be induced to produce a high level of laccases when grown on a cellobiose-asparagine liquid medium. After induction by kraft lignin, two major isozymes were detected in the fermentation supernatant of the fungus. The principal component laccase A, which accounts for about 85% of the total activity, can be purified to electrophoretic homogeneity by three chromatographic steps: DEAE-Sepharose FF, Superdex-200 and Mono-Q. The solution containing purified laccase is blue in color, and the ratio of absorbance at 280 nm to that at 600 nm is 22. The molecular mass of laccase A is estimated to be 62 kDa by SDS-PAGE, 57 kDa by FPLC, and measured as 58522 Da by MALDI mass spectrum. Laccase A is a monomeric glycoprotein with a carbohydrate content of 11-12% and an isoelectric point of 4.2. The optimum pH and temperature for oxidizing guaiacol are 4.5 and 50 degrees C, respectively. The half-life of the enzyme at 75 degrees C is 27 min. The enzyme shows a good stability from pH 4.2 to pH 8.0. The K(m) values of the enzyme toward substrates 2,2'-azino-bis (3-ethylbenzothazoline-6-sulfonate) (ABTS), guaiacol and 2,6-dimethoxyphenol are 25, 420 and 25.5 microM, respectively, and the corresponding V(max) values are 670, 66.8, and 79 microM min(-1) x mg(-1), respectively. Laccase A activity is strongly inhibited by 0.1 mM NaN(3) or 0.1 mM cyanide. Two units of laccase A alone is able to completely oxidize 100 micromol 2,6-chlorophenol in 6 h. In the presence of 1 mM ABTS and 1-hydroxybenzotriazole, 15.0 U laccase A is able to oxidize 45% and 70% of 50 micromol fluorene in 12 and 18 h, respectively. The laccase A gene was cloned by a PCR method, and preliminary analysis of its sequence indicates 87.0% similarity to the corresponding segment in the phenoloxidase gene from Coriolus hirsutus.

Amino Acid Sequence↗

Binding affinities and geometries of various metal ligands in peptide deformylase inhibitors.

Removal of the N-terminal formyl group from newly synthesized proteins by the enzyme peptide deformylase (PDF) is essential for normal growth of bacteria but not higher organisms. Recently, PDF has been explored as a target for novel antibiotics. Screening a collection of natural products for antimicrobial activity identified actinonin and two matlystatin analogs as potent PDF inhibitors. A number of synthetic analogs of these natural products were prepared and their inhibitory potency determined. Previous work has shown that PDF is an iron metalloproteinase also containing a catalytic glutamic acid residue. Ligation of the ferrous cation is an essential feature of potent inhibitors. The structures of actinonin, a matlystatin analog and a synthetic inhibitor complexed with PDF were determined by crystallography. A quantum mechanics/molecular mechanics (QM/MM) method was used to reproduce the geometry of known complexes, to predict the protonation state in the active site and to predict the geometry of additional complexes. The requirement for protonation of the active site glutamate anion is an important factor in understanding the potency of inhibitors with acidic iron-ligating groups such as hydroxamate and carboxylate. Even though potent inhibitors of PDF have been discovered, their bacteriostatic mechanism of action and the rapid development of resistance in vitro may limit their potential as antibacterial drugs.

Amidohydrolases↗

Limiting temperatures and the equation of state of nuclear matter.

From experimental observations of limiting temperatures in heavy ion collisions we derive the critical temperature of infinite nuclear matter Tc=16.6+/-0.86. Theoretical model correlations between Tc, the compressibility modulus K, the effective mass m*, and the saturation density rho s are then exploited to derive the quantity (K/m*)1/2 rho -1/3 s. This quantity together with calculations employing Skyrme and Gogny interactions indicates a value of K in moderately excited nuclei that is in excellent agreement with the value determined from giant monopole resonance data.

Journal Article↗

Search for the KL-->pi 0 pi 0 e+e- decay in the KTeV experiment.

The recent discovery of a large CP violating asymmetry in KL-->pi+pi-e+e- mode has prompted us to seach for the associated KL-->pi 0 pi 0 e+e- decay mode in the KTeV-E799 experiment at Fermilab. In 2.7 x 10(11) K(L) decays, one candidate event has been observed with an expected background of 0.3 event, resulting in an upper limit for the KL-->pi 0 pi 0 e+e- branching ratio of 6.6 x 10(-9) at the 90% C.L.

Journal Article↗

Evidence for segregation of heterologous GPI-anchored proteins into separate lipid rafts within the plasma membrane.

Cholesterol and glycosphingolipid-rich membrane rafts, which are rich in GPI-anchored proteins and are distinct from caveolae, are believed to serve as platforms for signal transduction events and protein recycling. GPI-anchored proteins with diverse functions as well as caveolin may be recovered in a membrane fraction insoluble in cold non-ionic detergent. This study tests for possible heterogeneity in the protein composition of the lipid rafts and detergent-insoluble membrane complexes by examining the two GPI-anchored homologous human folate receptors (FR)-alpha and -beta, the GPI-anchored human placental alkaline phosphatase (PLAP), and caveolin (control) in transfected CHO cells. Both FR and PLAP showed the equal distribution of cell-surface vs. sequestered (recycling) protein typical of GPI-proteins. Quantitative affinity purification of detergent-insoluble complexes using biotinylated folate or specific antibodies demonstrated a strong association of the homologous FR-alpha and FR-beta in the same detergent-insoluble complex and separate complexes containing either PLAP or caveolin. Immunogold localization experiments using antibody crosslinking to produce larger aggregates of GPI-anchored proteins for visualization by electron microscopy also showed a clear separation between FR- and PLAP-rich membrane microdomains. Thus, even though functionally diverse and heterologous GPI-anchored proteins are known to share endocytic and recycling vesicles, they may be segregated in distinct lipid rafts on the basis of their ecto(protein) domains facilitating clustering, compartmentalization and homotypic protein interactions.

Animals↗

Radiative decay width measurements of neutral kaon excitations using the primakoff effect.

We use K(L)'s in the 100-200 GeV energy range to produce 147 candidate events of the axial vector pair K1(1270)-K1(1400) in the nuclear Coulomb field of a Pb target and determine the radiative widths Gamma(K1(1400)-->K0+gamma)=280.8+/-23.2(stat)+/-40.4(syst) keV and Gamma(K1(1270)-->K0+gamma)=73.2+/-6.1(stat)+/-28.3(syst) keV. These first measurements appear to be lower than the quark-model predictions. We also place upper limits on the radiative widths for K(*)(1410) and K(*)(2)(1430) and find that the latter is vanishingly small in accord with SU(3) invariance in the naive quark model.

Journal Article↗

Hypoxia enhances the expression of autocrine motility factor and the motility of human pancreatic cancer cells.

The incidence of distant metastases is higher in the tumours with low oxygen pressure than in those with high oxygen pressure. It is well known that hypoxia induces the transcription of various genes involved in angiogenesis and anaerobic metabolism necessary for the growth of tumour cells in vivo, suggesting that hypoxia may also induce the transcription of metastasis-associated genes. We sought to identify the metastasis-associated genes differentially expressed in tumour cells under hypoxic conditions with the use of a DNA microarray system. We found that hypoxia enhanced the expression of autocrine motility factor mRNA in various cancer cells and also enhanced the random motility of pancreatic cancer cells. Autocrine motility factor inhibitors abrogated the increase of motility under hypoxic conditions. In order to explore the roles of hypoxia-inducible factor-1alpha, we established hypoxia-inducible factor-1alpha-transfectants and dominant negative hypoxia-inducible factor-1alpha-transfectants. Transfection with hypoxia-inducible factor-1alpha and dominant-negative hypoxia-inducible factor-1alpha enhanced and suppressed the expression of autocrine motility factor/phosphohexase isomerase/neuroleukin mRNA and the random motility, respectively. These results suggest that hypoxia may promote the metastatic potential of cancer cells through the enhanced autocrine motility factor/phosphohexase isomerase/neuroleukin mRNA expression and that the disruption of the hypoxia-inducible factor-1 pathway may be an effective treatment for metastasis.

Blotting, Northern↗

Mycobacterium tuberculosis and Mycobacterium fortuitum osteomyelitis of the foot and septic arthritis of the ankle in an immunocompetent patient.

Mycobacteria, both tuberculous and nontuberculous, are recognized as a cause of chronic bone and joint infection. However, the diagnosis of mycobacterial infection is easily missed because of the absence of systemic involvement. Moreover, specific microbiologic techniques are required to detect mycobacteria in clinical specimens. Infections due to uncommon pathogens such as mycobacteria are more likely to occur in the immunocompromised host. A case of septic arthritis of the ankle and osteomyelitis of the foot due to both tuberculous and nontuberculous mycobacteria in an immunocompetent host is reported here.

Adult↗

Measurement of the K(L) charge asymmetry.

We present a measurement of the charge asymmetry delta(L) in the mode K(L)-->pi(+/-)e(-/+)nu based on 298 x 10(6) analyzed decays. We measure a value of delta(L) = [3322+/-58(stat)+/-47(syst)]x10(-6), in good agreement with previous measurements and 2.4 times more precise than the current best published result. The result is used to place more stringent limits on CPT and DeltaS = DeltaQ violation in the neutral kaon system.

Journal Article↗