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Biomedical subjects

Y Tang

Publications and source records attributed to Y Tang.

At least 37 records · Page 2Linked to original sources

Binding of murine leukemia virus Gag polyproteins to KIF4, a microtubule-based motor protein.

A cDNA clone encoding a cellular protein that interacts with murine leukemia virus (MuLV) Gag proteins was isolated from a T-cell lymphoma library. The sequence of the clone is identical to the C terminus of a cellular protein, KIF4, a microtubule-associated motor protein that belongs to the kinesin superfamily. KIF4-MuLV Gag associations have been detected in vitro and in vivo in mammalian cells. We suggest that KIF4 could be involved in Gag polyprotein translocation from the cytoplasm to the cell membrane.

Animals

An exposed KID-like domain in human T-cell lymphotropic virus type 1 Tax is responsible for the recruitment of coactivators CBP/p300.

Human T-cell lymphotropic virus type 1 (HTLV-1) transcriptional activation is mediated by the viral transactivator, Tax, and three 21-bp repeats (Tax response element [TxRE]) located in the U3 region of the viral long terminal repeat (LTR). Each TxRE contains a core cyclic AMP response element (CRE) flanked by 5' G-rich and 3' C-rich sequences. The TxRE binds CREB (CRE-binding protein) and Tax to form a ternary complex and confers Tax-dependent transactivation. Recent data indicate that Tax functions as a specific link to connect CREB-binding protein (CBP)/p300 in a phosphorylation-independent manner to CREB/ATF-1 assembled on the viral 21-bp repeats. Glutathione S-transferase pull-down performed with Tax deletion mutants and peptide competition have localized the site in Tax critical for binding CBP/p300 to a highly protease-sensitive region around amino acid residues 81 to 95 (81QRTSKTLKVLTPPIT95) which lies between the domains previously proposed to be important for CREB binding and Tax subunit dimerization. Amino acid residues around the trypsin- and chymotrypsin-sensitive sites (88KVL90) of Tax bear resemblance to those in the kinase-inducible domain of CREB (129SRRPSYRKILNE140) surrounding Ser-133, which undergoes signal-induced phosphorylation to recruit CBP/p300. Site-directed mutagenesis of residues in this domain (R82A, K85A, K88A, and V89A) resulted in proteins which failed to transactivate from the HTLV-1 LTR in vivo. These mutants (K85A, K88A, and V89A) bind CREB with similar affinities as wild-type Tax, yet interaction with CBP/p300 is abrogated in various biochemical assays, indicating that the recruitment of CBP/p300 is crucial for Tax transactivation. A Tax mutant, M47, defective in the COOH-terminal transactivation domain, continued to interact with CBP/p300, suggesting that interactions with additional cellular factors are required for proper Tax function.

Amino Acid Sequence

MR imaging of the liver: comparison between single-shot echo-planar and half-Fourier rapid acquisition with relaxation enhancement sequences.

PURPOSE: To evaluate single-shot T2-weighted magnetic resonance sequences and their role in clinical practice in patients with hepatic lesions. MATERIALS AND METHODS: Prospective comparison of echo-planar and half-Fourier rapid acquisition with relaxation enhancement (RARE) imaging was performed in 80 patients with focal hepatic lesions. Spin-echo (SE) single-shot echo-planar (echo times, 47 and 80 msec) and half-Fourier RARE (echo time, 59 msec) images were compared with turbo SE (repetition time msec/echo time msec = 3,200-7,600/90) images. Quantitative, qualitative, and receiver operating characteristic (ROC) analyses were performed. RESULTS: For liver, signal-to-noise ratios on half-Fourier RARE images were significantly higher than those on echo-planar images (P < .01). For cystic lesions, contrast on half-Fourier RARE and echo-planar images was slightly higher than that on turbo SE images. For solid lesions, contrast on echo-planar images was better than that on half-Fourier RARE or turbo SE images. Artifacts including ghosting, bowel motion, susceptibility difference, and chemical shift were negligible on half-Fourier RARE images in all patients, whereas susceptibility difference and chemical shift of various degrees were seen on all echo-planar images. On the basis of ROC analyses, tumor detection rates were significantly higher with half-Fourier RARE and turbo SE images than with echo-planar images (P < .01). CONCLUSION: Echo-planar images provide sufficient contrast to allow detection of both solid and cystic masses, but severe artifacts preclude routine use. Half-Fourier RARE images are free from artifacts (chemical shift and susceptibility) and diagnostic performance with them is similar to that with turbo SE images.

Adult

Determination of concentration of cytosolic NADH-cytochrome b5 reductase in erythrocytes from normal Chinese adults, neonates and patients with hereditary methemoglobinemia by double-antibody sandwich ELISA.

NADH-cytochrome b5 reductase (b5R), present in various tissues of the body, is a redox enzyme of multiple functions. The deficiency of the enzyme leads to hereditary methemoglobinemia. With rabbit anti-b5R antibody for plate coating and enzyme-labeled anti-b5R monoclonal antibody as reporter, we have developed a sandwich ELISA procedure for the determination of b5R concentration. This procedure is sensitive to a wide range of linearity, and convenient in coping with large numbers of samples. Using this novel method, cytosolic b5R concentration in the erythrocytes of 30 normal Chinese adults was estimated to be 25.63+/-8.54 ng/mg Hb. It was found that the concentration of red cell soluble b5R of five newborns was significantly lower than that of normal adults and soluble b5R was undetectable in the erythrocytes of 4 patients with type I hereditary methemoglobinemia. Our results demonstrated that the reduced b5R activity in red cell cytosol of both neonates and type I hereditary methemoglobinemic patients results largely or mainly from the lowered b5R concentration. Our novel method might be further exploited for in-depth investigation of the relationship between the qualitative and quantitative changes of b5R with regard to physiological and pathological conditions.

Adult

Single-shot MR cholangiopancreatography of neonates, infants, and young children.

OBJECTIVE: This study was undertaken to assess the usefulness of MR cholangiopancreatography (MRCP) with a half-Fourier acquisition single-shot turbo spin-echo (HASTE) sequence as a noninvasive imaging technique to evaluate biliary or pancreatic disease in young children. SUBJECTS AND METHODS: We prospectively examined 45 children (mean age, 3 years) with MRCP using HASTE on a 1.5-T MR imaging unit. MRCP images were obtained in 1.4 sec without breath-hold. A body phased array coil and fat saturation technique were used for data collection. On the basis of the surgical (n = 19) and ERCP (n = 7) findings, clinical data, and follow-up observation, 21 children were deemed to have no significant abnormality. The other 24 children were found to have pancreaticobiliary disease, including congenital biliary atresia in five neonates, choledochal cyst in six infants, and anomalous connections between the bile and pancreatic ducts in four infants and nine young children. In children without abnormality, the MRCP images were graded for the depiction of normal structures. The findings obtained by MRCP were correlated with the surgical or ERCP results. RESULTS: HASTE MRCP showed the first branch of the intrahepatic duct, the common hepatic duct, the gallbladder, and the common bile duct in most children. In the 21 children with no abnormalities, the main pancreatic duct was visible in two of six neonates, in four of five infants, and in all 10 young children. The diagnostic accuracy of MRCP was 100% in patients with choledochal cysts, 100% in those with congenital biliary atresia, and 69% in those with anomalous connections between the bile and pancreatic ducts. CONCLUSION: MRCP using HASTE with a phased array coil is a noninvasive technique for revealing the pancreaticobiliary tract in young children.

Biliary Atresia

[Localization of the gene for 4 hereditary multiple exostoses families].

We investigated 11 families with hereditary multiple exostoses (EXT) by linkage analysis using 8 short-tandem-repeat (CA)n polymorphic markers on chromosomes 8, 11 and 19. The Lod score in four families indicated that the gene responsible for EXT is located in the pericentromeric region of chromosome 11.

Chromosome Mapping

Effect of the cannabinoid receptor SPECT agent, AM 281, on hippocampal acetylcholine release from rat brain slices.

The SPECT ligand AM 281, a less lipophilic analog of the cannabinoid receptor antagonist SR 141716A, robustly potentiated electrically-evoked release of acetylcholine from superfused hippocampal slices and prevented the inhibition of acetylcholine release by the cannabimimetic drug WIN 55212-2. These results, similar to earlier observations with SR 141716A, indicate that AM 281 is either a cannabinoid receptor antagonist or inverse agonist. Despite showing lower affinity than SR 141716A in hippocampal membrane binding experiments, AM 281 had slightly greater potency than SR 141716A in the hippocampal slice experiments, perhaps because of reduced drug absorption to slice membranes and to the apparatus.

Acetylcholine

Effects of muscarinic blockade in perirhinal cortex during visual recognition.

Stimulus recognition in monkeys is severely impaired by destruction or dysfunction of the perirhinal cortex and also by systemic administration of the cholinergic-muscarinic receptor blocker, scopolamine. These two effects are shown here to be linked: Stimulus recognition was found to be significantly impaired after bilateral microinjection of scopolamine directly into the perirhinal cortex, but not after equivalent injections into the laterally adjacent visual area TE or into the dentate gyrus of the overlying hippocampal formation. The results suggest that the formation of stimulus memories depends critically on cholinergic-muscarinic activation of the perirhinal area, providing a new clue to how stimulus representations are stored.

Animals

Sol-Gel Preparation and Electrorheological Activity of SiO2-TiO2 Composite Powders

The SiO2-TiO2 composite powders with various Si/Ti ratios were prepared by the sol-gel processing, using tetraisopropylorthotitane (Ti(OPri)4) and tetraethoxysilane (Si(OEt)4) as starting materials. The effect of the powder's heat history on the electrorheological (ER) activity of the resulting suspensions, which were prepared by dispersing the powders into silicon oil, was investigated. The infrared spectra, scanning electron microscopy, X-ray diffraction, and zeta potential were used to characterize the structural evolution of the as-prepared gel powders upon heating. The results show a remarkable reduction in gelation time, relative to the single-component systems, due to the combined effects of a higher reactivity of Ti(OPri)4 over Si(OEt)4 and a nucleophilic attack of the Ti atom by the as-hydrolyzed species of Si precursor to form an environment similar to alkali catalysis. It was found that gel powders would gradually eliminate the -OH groups and become denser in structure upon heating, leading to a decrease of ER activity of the resulting suspensions. However, the initial Si/Ti composite compositions have little effect on ER activity behavior. Copyright 1997 Academic Press. Copyright 1997Academic Press

Journal Article

Molecular modeling and 3D-QSAR studies on the interaction mechanism of tripeptidyl thrombin inhibitors with human alpha-thrombin.

The mechanism of inhibition of peptidyl inhibitors with thrombin was studied using molecular modeling, molecular mechanics, and CoMFA statistical analysis. A new procedure for the elucidation of binding conformations, BCSPL, is described and was employed to obtain the binding conformers of a series of 18 tripeptidyl thrombin inhibitors. Energetic studies and QSAR analysis of the BCSPL-derived conformers indicated a modest correlation between the calculated binding energies of the title compounds and their inhibitory activities to human alpha-thrombin. CoMFA analysis of the BCSPL alignment resulted in a satisfactory model of the thrombin active site.

Antithrombins

A stereological method for estimating the total length and size of myelin fibers in human brain white matter.

A practically unbiased stereological method to obtain estimates of the volume and total length of nerve fibers in brain white matter is described. The sampling scheme is designed so that the majority of brain white matter is left intact, thus providing the possibility for resampling and further analysis. Uniform sampling of one complete hemispherical white matter is performed. The volume fraction of nerve fibers in white matter is estimated by point counting. The total length of nerve fibers was estimated from the product of the volume of white matter, obtained with the Cavalieri principle, and the fiber length density, obtained from the isotropic, uniform random sections which were ensured by the isector. The size of nerve fibers was derived by measuring the profile diameter perpendicular to its longest axis. The influence of the postmortem fixation delay on nerve fiber parameters was investigated in one dog and one pig. The criteria for identification of nerve fiber profiles at light microscopy were evaluated using electron microscopy.

Adolescent

Inhibition of LPS-induced TNF-alpha production by calcitonin gene-related peptide (CGRP) in cultured mouse peritoneal macrophages.

The purpose of this study was to examine whether rCGRP has effects on TNF-alpha produced by mouse resident peritoneal macrophages. Macrophages were obtained from the peritoneal exudate of male Balb/c mouse. The cells were plated on culture dishes at a density of 2.5x10(5) cells per well and allowed to adhere for 2 hr. Pretreatment with rCGRP (10 nM-1 microM) for 24 hr, the macrophages were cultured with LPS 1 microg/ml for another 24 h. The medium was harvested for measuring TNF-alpha by ELISA kits. The results showed that rCGRP had no direct effects on TNF-alpha production, but it inhibited LPS-induced TNF-alpha production in a concentration-dependent manner. When rCGRP was at a concentration of 1 microM, the LPS-induced TNF-alpha production was inhibited by 39%. The effect of rCGRP was reversed by hCGRP(8-37) (10 microM), an antagonist of CGRP1 receptor. The LPS-induced TNF-alpha production from macrophages was also inhibited by forskolin 3 microM, an activator of adenylate cyclase. Furthermore, pretreatment with H-89 1 microM or Rp-cAMPS 100 microM, the inhibitors of cAMP-dependent protein kinase, the effect of rCGRP was abolished. These data suggest that the LPS-induced TNF-alpha production is inhibited by rCGRP via activation of cAMP responses in mouse resident peritoneal macrophages.

1-Methyl-3-isobutylxanthine

Mitochondrial involvement in post-tetanic potentiation of synaptic transmission.

Posttetanic potentiation (PTP) is an essential aspect of synaptic transmission that arises from a persistent presynaptic [Ca2+]i following tetanic stimulation. At crayfish neuromuscular junctions, several inhibitors of mitochondrial Ca2+ uptake and release (tetraphenylphosphonium or TPP+, carbonyl cyanide m-chlorophenylhydrazone or CCCP, and ruthenium red) blocked PTP and the persistence of presynaptic residual [Ca2+]i, while endoplasmic reticulum (ER) Ca2+ pump inhibitors and release channel activators (thapsigargin, 2,5-di-(tert-butyl)-1,4-benzohydroquinone or BHQ, and caffeine) had no effects. PTP apparently results from the slow efflux of tetanically accumulated mitochondrial Ca2+.

Animals

Neuron volume in the ventral horn in Wobbler mouse motoneuron disease: a light microscope stereological study.

Previous pathological reports have indicated that swollen and vacuolated motoneuron cell bodies are the most predominant feature characterising Wobbler mouse motoneuron disease, but there has been little supportive evidence using area measurements. The present study focuses on the possible role of changes in neuronal nuclear and perikaryal volumes in the cervical spinal cord ventral horn, using new and traditional stereological probes which provide unbiased estimates of volume. Semithin sections from the ventral horn of Wobbler mice and age and sex-matched phenotypically normal littermates were examined at 2 ages (young and old). The young Wobbler group had significantly larger volume weighted mean perikaryal volumes compared with age-matched controls, reflecting the presence of large swollen cells characteristic of this group; this situation was reversed in the control group. Number-weighted perikaryal volume estimates in the old Wobbler group were smaller than in age-matched controls. The variation in perikaryal volume was greatest in the young Wobbler group in which the coefficient of variation was 127%. The mean number weighted and volume weighted mean nuclear volumes were significantly smaller in the old Wobbler group compared with age-matched controls and young Wobbler groups. The application of new stereological probes has enabled us to document more precisely these changes in neuronal structure in the Wobbler mutant mouse.

Aging

Phlebotomine sandflies of Kenya (Diptera: Psychodidae). V. Phlebotomus (Paraphlebotomus) mireillae n.sp.

A new species of the sandfly subgenus Paraphlebotomus Theodor from Kenya is described and named Phlebotomus mireillae. The type locality is Utut Reserve, near Gilgil, Nakuru District. The description is based on 10 males and 10 females reared from individual egg batches of wild-caught females. Males of the new species differ from nine other species of the subgenus by the shape of the aedeagus, which is straight, not curved or hooked. They are separated from the remaining three species by other differences in the aedeagus, and in the style and coxite process. Female P. mireillae have backwardly pointing spines inside the distal quarter of the spermathecal ducts, a character not previously described for phlebotomine sandflies. Additional characters which distinguish females of the new species from others of the subgenus are differences in the pharyngeal armature and spermathecae and the lengths of the third antennal segment and labium.

Animals

Hydrated water molecules of pyrimidine/purine/pyrimidine DNA triple helices as revealed by FT-IR spectroscopy: a role of cytosine methylation.

Hydrated water molecules of pyrimidine/purine/pyrimidine DNA hairpin triplex was studied by a comparison of triplex (CC.AG6) formed by a host oligodeoxypyrimidine of 5'-d(TC)3T4(CT)3(CC) with a target hexadeoxypurine 5'-d(AG)3(AG6) strand and by triplexes (MM.AG6, MC.AG6, and CM.AG6) formed by oligonucleotides with the exact sequences as above except 5-methylcytosine replaced all (MM), 5' end half (MC), and 3' end half (CM) cytosine bases in CC via FT-IR spectroscopy in hydrated film. Results revealed that: (i) all these triplexes have a similar hydration pattern, in which water molecules probably bound in the N7 sites of adenines and guanines in the Crick-Hoogsteen groove, and to the methyl group of thymidines in the Watson-Hoogsteen groove. There are also some bound water molecules found at the O2 sites of thymines in both Watson-Crick and Crick-Hoogsteen grooves. (ii) In the CC.AG6 triplex the S-type sugars are always dominant in all hydrated states, whereas in MM.AG6 triplex the relative population of the N-type sugars is very close to that of the S-type between 86% and 66% of humidity. Furthermore, the sugar conformation in two partially modified triplexes (CM.AG6, and MC.AG6) are dominant by the N-type at lower humidity. This phenomenon might reflect that the degree of bound water varies among the binding sites of bases. (iii) The effect of introducing a methyl group on cytosine is to generate a spine of hydrophobic region in MM (MC and MC). The enlarging hydrophobic area not only increase the stability in solution, and also the stability in sodium hydrated films of the pyrimidine/purine/pyrimidine hairpin triplexes.

Cytosine

A role for HDJ-2/HSDJ in correcting subnuclear trafficking, transactivation, and transrepression defects of a glucocorticoid receptor zinc finger mutant.

All steroid receptors possess a bipartite nuclear localization signal sequence (NLS) that localizes within the second zinc finger of their DNA-binding domain. Fine-structure mapping of the rat glucocorticoid receptor (rGS) NLS identified a composite signal composed of three distinct proto-NLSs that function effectively when present in unique pairs. At least one of the rGR proto-NLSs appears to influence receptor trafficking within the nucleus, as revealed by a unique nuclear staining pattern of receptors possessing a point mutation (i.e., arginine at position 496; R496), at proto-NLS, pNLS-2. Specifically, carboxyl-terminal-truncated rGRs possessing various point mutations at R496 localized within a limited number of large foci in nuclei of transiently transfected COS-1 cells. R496 mutations did not affect subnuclear targeting when present in full-length rGR, reflecting a protective effect of the receptor's ligand-binding domain that can be exerted in cis and in trans. The effects of rGR R496 mutations on subnuclear targeting were not autonomous because we also observed a coincident localization of hsp70, the 70-kDa heat shock protein, within nuclear foci that include r496 mutant receptors. The elimination of R496 mistargeting by overexpression of an hsp70 partner (i.e., the DnaJ homologue, HDJ-2/HSDJ) suggests that the hsp70/DnaJ chaperone system is mobilized to specific sites within the nucleus in response to inappropriate targeting or folding of specific mutant receptors. HDJ-2/HSDJ overexpression also corrects defective transactivation and transrepression activity of R496 mutant GRs. Thus, molecular chaperones, such as members of the hsp70 and DnaJ families, may survey the nucleus for misfolded proteins and actively participate in their refolding into biologically active conformational states.

Amino Acid Sequence