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Biomedical subjects

E Wang

Publications and source records attributed to E Wang.

At least 163 records · Page 9Linked to original sources
PubMed1999

[Traverse connectivity of innervation in the vestibular sensory epithelium].

OBJECTIVE: To further understand the traverse connections of nerve components in the vestibular sensory epithelium. METHODS: The nerve innervations in the vestibular sensory epithelia and infraepithelia of ten healthy guinea pigs (16 ears) were observed by transmission electron microscopy. RESULTS: Type I sensory cells were intricately connected with type II cells. Afferent and efferent fibers as well as their endings had also complicated connections with nerve trunks and branches. The traverse connections appeared among nerve components in the vestibular sensory epithelium were extremely complicated, forming multilevel connections. They stemmed from and end in the crista ampullaris and the maculae. CONCLUSION: These complicated and multilevel connections are the basis of nerve activity in the vestibular apparatus.

Animals↗
PubMed1998

Acylation of naturally occurring and synthetic 1-deoxysphinganines by ceramide synthase. Formation of N-palmitoyl-aminopentol produces a toxic metabolite of hydrolyzed fumonisin, AP1, and a new category of ceramide synthase inhibitor.

Fumonisin B1 (FB1) is the predominant member of a family of mycotoxins produced by Fusarium moniliforme (Sheldon) and related fungi. Certain foods also contain the aminopentol backbone (AP1) that is formed upon base hydrolysis of the ester-linked tricarballylic acids of FB1. Both FB1 and, to a lesser extent, AP1 inhibit ceramide synthase due to structural similarities between fumonisins (as 1-deoxy-analogs of sphinganine) and sphingoid bases. To explore these structure-function relationships further, erythro- and threo-2-amino, 3-hydroxy- (and 3, 5-dihydroxy-) octadecanes were prepared by highly stereoselective syntheses. All of these analogs inhibit the acylation of sphingoid bases by ceramide synthase, and are themselves acylated with Vmax/Km of 40-125 for the erythro-isomers (compared with approximately 250 for D-erythro-sphinganine) and 4-6 for the threo-isomers. Ceramide synthase also acylates AP1 (but not FB1, under the conditions tested) to N-palmitoyl-AP1 (PAP1) with a Vmax/Km of approximately 1. The toxicity of PAP1 was evaluated using HT29 cells, a human colonic cell line. PAP1 was at least 10 times more toxic than FB1 or AP1 and caused sphinganine accumulation as an inhibitor of ceramide synthase. These studies demonstrate that: the 1-hydroxyl group is not required for sphingoid bases to be acylated; both erythro- and threo-isomers are acylated with the highest apparent Vmax/Km for the erythro-analogs; and AP1 is acylated to PAP1, a new category of ceramide synthase inhibitor as well as a toxic metabolite that may play a role in the diseases caused by fumonisins.

Acylation↗
PubMed1998

The Epstein-Barr virus nuclear protein SM is both a post-transcriptional inhibitor and activator of gene expression.

The Epstein-Barr virus (EBV) nuclear protein BS-MLF1 (SM) is expressed early after entry of EBV into the lytic cycle. SM transactivates reporter gene constructs driven by a wide variety of promoters, but the mechanism of SM action is poorly understood. In this study, we demonstrate that the SM protein inhibits expression of intron-containing genes and activates expression of intron-less genes. We demonstrate that SM has the predicted inhibitory effect on expression of a spliced EBV gene but activates an unspliced early EBV gene. SM inhibited gene expression at the post-transcriptional level by preventing the accumulation of nuclear and cytoplasmic RNA transcripts. Conversely, SM led to increased accumulation of nuclear mRNA from intron-less genes without affecting the rate of transcription, indicating that SM enhances nuclear RNA stability. The ratio of cytoplasmic to nuclear polyadenylated mRNA was increased in the presence of SM, suggesting that SM also enhances nucleo-cytoplasmic mRNA transport. The degree of transactivation by SM was dependent on the sequence of the 3'-untranslated region of the target mRNA. Finally, we demonstrate that the amino-terminal portion of SM fused to glutathione-S-transferase binds radioactively labeled RNA in vitro, indicating that SM is a single-stranded RNA binding protein. Importantly, the latent and immediate-early genes of EBV contain introns whereas many early and late genes do not. Thus, SM may down-regulate synthesis of host cell proteins and latent EBV proteins while simultaneously enhancing expression of specific lytic EBV genes by binding to mRNA and modulating its stability and transport.

Cell Line↗
PubMed1998

Genomic organization and cloning of the human homologue of murine Sipa-1.

Murine Sipa-1 (signal-induced proliferation associated protein) is a mitogen induced GTPase activating protein (GAP). While mapping candidate genes for multiple endocrine neoplasia type 1 (MEN1) at 11q13, we cloned the human homologue of Sipa-1. Herein, we report the complete cDNA sequence, expression, and genomic organization of SIPA-1. SIPA-1 consists of 16 exons with highly conserved exon-intron boundaries. The predicted SIPA-1 protein is highly homologous to the mouse protein, particularly in the region of the GAP-related domain at the amino terminus and the leucine zipper at the carboxy terminus. It is widely expressed, including in fetal tissues, but is most highly expressed in lymphoid organs. During the course of cloning SIPA-1, the MEN1 gene was identified, thus excluding human SIPA-1 as a candidate for this disease.

Amino Acid Sequence↗
PubMed1998

Synthesis and evaluation of compounds that facilitate the gastrointestinal absorption of heparin.

A family of novel compounds (delivery agents) that promote the gastrointestinal absorption of USP heparin in rats and primates has been discovered. The delivery agents in combination with heparin were administered either orally or intracolonically in an aqueous propylene glycol solution and caused dramatic increases in both plasma heparin concentrations (anti-Factor Xa) and clotting times (APTT). Using one of the most effective delivery agents in this series, an estimated relative bioavailability of 8% can be achieved following oral administration to cynomolgus monkeys. To establish a correlation between the in vivo data and an in vitro parameter, immobilized artificial membrane (IAM) chromatography was performed. Log relative k' values were correlated to the efficiency of oral heparin delivery.

Amino Acids↗
PubMed1998

Apoptosis rate can be accelerated or decelerated by overexpression or reduction of the level of elongation factor-1 alpha.

Peptide chain elongation factor-1 alpha (EF-1 alpha) is required for the binding of aminoacyl-tRNAs to acceptor sites of ribosomes during protein synthesis. More recently, EF-1 alpha has been shown to be involved in cytoskeletal organization. The elongation factor functions in actin bundling and microtubule severing. Moreover, it can activate the phosphatidylinositol-4 kinase whose substrates are involved in regulation of actin polymerization. The expression level of EF-1 alpha is regulated in many situations such as growth arrest, transformation, and aging. Because of this regulation of EF-1 alpha in various states of cell life, and its key position in protein synthesis as well as cytoskeletal organization, we chose to investigate the effect of its expression levels on apoptosis. Apoptosis is a complex event regulated through numerous activators and inhibitors. In some situations, protein synthesis is required for apoptosis to be triggered. Investigation of the effect of altered levels of elongation factor-1 alpha on apoptosis is of particular interest since it may affect both protein synthesis and cytoskeletal organization. For example, reduction of EF-1 alpha leads to a reduced protein synthesis rate, which might reduce the presence of those "killer factors" triggering apoptosis. EF-1 alpha involvement in cytoskeletal organization is another example, since cytoskeletal organization undergoes dramatic changes during apoptosis. Thus, this study has been planned to ascertain whether hypo- and hyperexpression of EF-1 alpha protein, achieved by constructing expression vectors with the EF-1 alpha cDNA in its antisense or sense orientation under the control of a cytomegalovirus promoter, can produce stable transfectants with either heightened or reduced responsiveness to apoptosis stimuli. Our results show the following: (1) induction of apoptosis by serum deprivation shows that antisense EF-1 alpha provides cells significant protection from apoptotic cell death and (2) EF-1 alpha overexpression causes a faster rate of cell death. These findings suggest that when EF-1 alpha protein is abundant the cells are proapoptosis, and vice versa in low abundance the cells are in the mode of antiapoptosis. Therefore, changes in levels of EF-1 alpha may be one of the global pivotal regulators modulating the rate of apoptosis.

3T3 Cells↗
PubMed1998

Overproduction and purification of glutaryl 7-amino cephalosporanic acid acylase.

The gene encoding glutaryl 7-amino cephalosporanic acid acylase (GL-7ACA acylase) from Pseudomonas sp. 130 has been cloned and expressed in Escherichia coli using a high-level expression system. The specific activity of the acylase in the crude extract of cells in this system is approximately 10 times that in the previous one. The overproduced enzyme can be easily isolated within 3 days to a purity of over 90% by a simple and inexpensive two-step preparative chromatographic method with an overall yield of nearly 50%. The deletion of the signal peptide and mutation in the alpha-subunit of the acylase have little influence on its posttranslational processing and its kinetic parameters.

Amidohydrolases↗
PubMed1998

Mechanism and specificity of lanthanide series cation transport by ionophores A23187, 4-BrA23187, and ionomycin.

A23187, 4-BrA23187, and ionomycin transport several lanthanide series trivalent cations at efficiencies similar to Ca2+, when compared at cation concentrations of approximately 10(-5) M, ionophore concentrations of approximately 10(-6) M, and a pH of 7.00. Selectivity sequences and the range of relative rates are as follows: A23187, Nd3+ > La3+ > Eu3+ > Gd3+ > Er3+ > Yb3+ > Lu3+ (approximately 34-fold); 4-BrA23187, Nd3+ > Eu3+ > Gd3+ > La3+ > Er3+ > Yb3+ > Lu3+ (approximately 34-fold); ionomycin, La3+ > Yb3+ > Nd3+ > Lu3+ > Er3+ > Eu3+ > Gd3+ (approximately 4-fold). At concentrations between 9 and 250 microM, La3+ is transported by an electroneutral mechanism, predominately through mixed complexes of the type (ionophore)2La-OH (A23187 and 4-BrA23187) or (ionophore)La-OH (ionomycin), when no membrane potential is present. For all three ionophores, an induced potential of approximately 160 mV accelerates transport by approximately 50-100%. However, measured values of H+/La3+ exchange indicate that only 4-BrA23187 displays a significant electrogenic activity under these conditions. At a La3+ concentration of 17 mM, transport by all three ionophores is electroneutral and apparently occurs through complexes of type (ionophore)3La (A23187 and 4-BrA23187) or (ionophore)La-OH (ionomycin). Analysis of these patterns in a context of comproportionation equilibria involving the transporting species and free La3+ indicates that the species containing three ionophore molecules are formed on the membrane when aqueous phase solution conditions would strongly favor a 1:1 complex, based upon previous studies in solution. The implications of this and other findings are discussed.

Aminoquinolines↗
PubMed1998

Determination of sulfadiazine and sulfamethoxazole by capillary electrophoresis with end-column electrochemical detection.

Capillary electrophoresis (CE) with end-column electrochemical detection (EC) of sulfadiazine (SDZ) and sulfamethoxazole (SMZ) is described. Under the optimum conditions, SDZ and SMZ were separated satisfactorily, and a highly sensitive and stable response was obtained at a potential of 1.1 V versus Ag/AgCl. Optimized end-column detection provides detection limits as low as 0.1 microM for both compounds, which corresponds to 0.024 and 0.021 fmol with peak efficiencies of 394,000 and 335,000 theoretical plates for SDZ and SMZ, respectively. The calibration graph was linear over three order of magnitude. The relative standard deviations (n = 12) of peak currents and migration times were 2.3 and 2.7%, and 0.8 and 1.3%, respectively, for the two compounds. The proposed method was applied to the analysis of tablets and human urine samples with satisfactory results.

Anti-Infective Agents↗
PubMed1998

Determinants of increased energy expenditure in HIV-infected women.

BACKGROUND: Little is known about sex-specific effects of HIV infection on energy expenditure. OBJECTIVE: We investigated the determinants of energy expenditure in HIV-infected women. DESIGN: Resting energy expenditure (REE), body composition, and hormonal and nutritional indexes were compared in 33 ambulatory, premenopausal HIV-infected women and 26 weight-matched, healthy premenopausal control subjects. REE was determined by indirect calorimetry and body composition by dual-energy X-ray absorptiometry (DXA), bioelectrical impedance analysis, and skinfold-thickness analysis. Hormonal indexes included leptin, testosterone, estradiol, and insulin-like growth factor I. RESULTS: HIV-infected subjects had a higher REE than control subjects [6794 +/- 1374 compared with 6011 +/- 607 kJ/d (1624 +/- 329 compared with 1437 +/- 145 kcal/d), P = 0.0096]. On average, REE was 119 +/- 23% of Harris-Benedict predictions in HIV-infected subjects compared with 102 +/- 9% for control subjects (P = 0.0007). In HIV-infected subjects, REE was highly correlated with fat-free mass (FFM) by DXA (R = 0.641, P < 0.001), but not with weight or disease status. The slope of the regression equation for REE and FFM was significantly greater (P = 0.027, analysis of covariance) for HIV-infected subjects [REE (kJ/d) = 203.5 (kg FFM) - 1237] than for control subjects [REE (kJ/d) = 77.4 (kg FFM) + 2923]. In a stepwise regression analysis, FFM was the most significant variable (P = 0.005), followed by free testosterone (P = 0.029), which together explained 49% of the variation in REE. The final equation was REE (kJ/d) = 230.8 (kg FFM) + 395.9 (free testosterone, pmol/L) - 3304. CONCLUSIONS: Energy expenditure was higher in HIV-infected women than in control women. FFM is the primary determinant of REE in HIV-infected women, but energy expenditure is greater per kg FFM in HIV-infected subjects than in control subjects, which may contribute to the wasting syndrome.

Absorptiometry, Photon↗
PubMed1998

Genotypic analysis of tumor suppressor genes PTEN/MMAC1 and p53 in head and neck squamous cell carcinomas.

OBJECTIVES: Tumor suppressor gene mutations in both p53 and PTEN/MMAC1 genomic DNA have been detected in many types of cancer. The purpose of this study was to investigate the presence and importance of PTEN/MMAC1 mutations in squamous cell carcinomas. METHODS: Exons of each gene were amplified after polymerase chain reaction (PCR) using genomic DNA derived from cell lines of squamous cell carcinoma of the head and neck (SCCHN) and snap-frozen biopsy specimens from primary established head and neck tumors. The amplified and purified DNA was then sequenced directly. RESULT: As anticipated, point mutations of the p53 gene were found in 80% of cell lines examined. A single base mutation in codon 151 was found in six of 10 cell lines studied. PTEN/MMAC1 gene mutations were found in neither the cell lines tested nor the tumor biopsy samples. CONCLUSION: This study, as well as a large volume of data, confirms that mutations of the p53 gene are frequent events in head and neck cancer cell lines. Although PTEN/MMAC1 gene mutations have been found in a variety of carcinomas, this gene was not found to be mutated in SCCHN cell lines or in primary squamous cell carcinomas of the head and neck. This information is useful for further studies of mutations in these cell lines.

Carcinoma, Squamous Cell↗
PubMed1998

Simultaneous genotyping and species identification using hybridization pattern recognition analysis of generic Mycobacterium DNA arrays.

High-density oligonucleotide arrays can be used to rapidly examine large amounts of DNA sequence in a high throughput manner. An array designed to determine the specific nucleotide sequence of 705 bp of the rpoB gene of Mycobacterium tuberculosis accurately detected rifampin resistance associated with mutations of 44 clinical isolates of M. tuberculosis. The nucleotide sequence diversity in 121 Mycobacterial isolates (comprised of 10 species) was examined by both conventional dideoxynucleotide sequencing of the rpoB and 16S genes and by analysis of the rpoB oligonucleotide array hybridization patterns. Species identification for each of the isolates was similar irrespective of whether 16S sequence, rpoB sequence, or the pattern of rpoB hybridization was used. However, for several species, the number of alleles in the 16S and rpoB gene sequences provided discordant estimates of the genetic diversity within a species. In addition to confirming the array's intended utility for sequencing the region of M. tuberculosis that confers rifampin resistance, this work demonstrates that this array can identify the species of nontuberculous Mycobacteria. This demonstrates the general point that DNA microarrays that sequence important genomic regions (such as drug resistance or pathogenicity islands) can simultaneously identify species and provide some insight into the organism's population structure.

Alleles↗
PubMed1998

Susceptibilities of neonatal respiratory isolates of Ureaplasma urealyticum to antimicrobial agents.

Twenty-one neonatal respiratory isolates of Ureaplasma urealyticum were serotyped, and their susceptibilities to ciprofloxacin, gentamicin, chloramphenicol, erythromycin, azithromycin, and doxycycline were tested. Most patient strains were Ureaplasma urealyticum bv. parvum. Chloramphenicol, doxycycline, and azithromycin had the lowest MICs. This data may be useful when designing prophylactic or therapeutic trials of antibiotics for chronic lung disease of the newborn.

Anti-Bacterial Agents↗
PubMed1998

Dynamics of segmental extracellular volumes during changes in body position by bioimpedance analysis.

Extracellular volume (ECV) of arms, trunk, and legs determined from segmental bioimpedance data in 11 healthy men (31.6 +/- 7 yr) obtained at the end of a 30-min equilibration phase in the supine body position was compared with ECV determined from whole body measurements (ECVWB). ECV was calculated from extracellular resistance (RECV) identified from the bioimpedance spectrum for a range of 10 frequencies. Whole body RECV (527.6 +/- 55.6 Omega) was equal to the sum of RECV in the arms, trunk, and legs (241.6 +/- 36. 3, 49.2 +/- 5.1, and 236.3 +/- 25.5 Omega, respectively). The sum of equilibrated ECV in arms (1.31 +/- 0.25 liters), trunk (10.08 +/- 1.65 liters), and legs (2.80 +/- 0.82 liters) was smaller than ECVWB (20.90 +/- 2.59 liters). In six subjects who changed from a standing to a supine body position, ECV decreased in arms (-2.59 +/- 2.51%, P = NS) and legs (-10.96 +/- 3.02%, P < 0.05) but increased in the trunk (+4.2 +/- 3.2%, P < 0.05). ECVWB also decreased (-4.98 +/- 1. 41%, P < 0.05). However, the sum of segmental extracellular volumes remained unchanged (-0.06 +/- 0.07%, P = NS). The sum of segmental ECVs is not sensitive to changes in body position, which otherwise interferes with the estimation of ECV in bioimpedance analysis when ECVWB is used.

Adult↗
PubMed1998

Pituitary adenomas: the effect of gamma knife radiosurgery on tumor growth and endocrinopathies.

Seventy-three patients have been treated with Gamma Knife radiosurgery (GKR) for pituitary adenomas. 12 had undergone surgery prior to GKR. Three had had previous radiation therapy. The prescription dose administered to the tumor margin ranged from 9 to 35 Gy. 65 patients were followed up for an average of 29 months. A slight expansion of tumor volume occurred in 3 cases. Otherwise, the tumor volume remained unchanged or decreased in the remainder. Endocrine changes were present in all except 3 cases. GKR was followed by a speedy decrease in raised serum hormone levels in the case of both growth hormone (GH)and adrenocortico-tropic hormone (ACTH). In 3 patients there was some visual deterioration associated with a slight increase in tumor volume. In 2 cases, the tumors were removed surgically. Some preliminary conclusions may be drawn. The dose required to correct an endocrinopathy may be higher than that required for control of tumor growth. The recommended prescription dose for endocrine-active adenomas may be more than 30 Gy. While clinical improvement may be noted in patients with raised serum prolactin levels (PRL), normalization of the endocrinopathy may be less readily achieved than in the case of raised GH and ACTH levels. Gamma Knife radiosurgery as a primary treatment of pituitary adenomas can be safe and effective.

Adenoma↗
PubMed1998

Purification and activity study of the A- and B-chains of cinnamomin, a type II ribosome-inactivating protein.

The strong hydrophobic interaction between the A- and B-chains of cinnamomin, a type II ribosome-inactivating protein, makes it difficult to separate A- and B-chains after the disulfide bond is broken. We failed to separate the A-chain from B-chain of cinnamomin using methods under usual conditions. A convenient method for purification of the A- and B-chains of cinnamomin on a large scale has been developed. We chose urea to weaken the non-covalent interaction between the A- and B-chains. In the presence of 4M urea, the A- and B-chains of the reduced cinnamomin are separated effectively by DEAE-cellulose chromatography. The purified A-chain still displays the RNA N-glycosidase activity and the B-chain loses the lectin activity. After refolding in vitro in the presence of lactose, the B-chain is renatured and the active B-chain with lectin activity can be further purified by Sepharose 4B affinity chromatography. From 80 mg of cinnamomin, 10 mg of A-chain (25%) and 38 mg of the B-chain (95%) were obtained. In addition, the intrinsic fluorescence spectra of the A- and B-chains were employed to study the structural changes in the active and the non-active forms of cinnamomin A- and B-chains.

Algal Proteins↗
PubMed1998

[Iodine deficiency and perceptive nerve deafness].

Iodine contents (ICS) in hairs of 381 children with perceptive nerve deafness (PND) and in table salt were determined in Yancheng prefecture and Shuyang county of Jiangsu province, east China from 1995 to 1996. The results showed that the ICS in hairs of deaf children were much lower than that of healthy children (P < 0.01). And the ICS in table salt in these regions were punder the standard content officially published. Presumably, iodine deficiency (ID) in these regions may be one of the major factors causing PND.

Adolescent↗