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Biomedical subjects

Y Wei

Publications and source records attributed to Y Wei.

At least 307 records · Page 17Linked to original sources
PubMed1995

[Toxicity of drugs on nasal mucocilia and the method of its evaluation].

Effect of solutions or suspensions of eight drugs including analgin, paracetamol, propafenone hydrochloride, propranolol hydrochloride, ephedrine hydrochloride, gentamycin sulfate, sodium deoxycholate and hydrocortisone on ciliary movement were evaluated with in vitro or in situ toad palate model and scanning electron microscope. In vitro toad palate model: 0.2 ml of test drug solution or suspension was applied to a piece of freshly dissected upper palate of toad. The mucocilia were examined with an optical microscope and the lasting time of ciliary movement was recorded after drug application. The upper palate was rinsed with physiological saline when the ciliary movement stopped. The lasting time of ciliary movement after rinsing was then recorded again. In situ palate model: 0.5 ml of test drug solution or suspension was applied to the upper palate of toad for 30 min, and rinsed with physiological saline. The palate was dissected out and the operation was carried out in a similar manner. The results showed that the in situ toad palate model is a satisfactory method for studying the ciliotoxicity of drugs. The in vitro toad palate model is unsuitable for suspension and gel. The results of the eight drugs revealed that ciliary movement is frequently affected by many drugs and, therefore, care must be taken in developing any nasal dosage form to ensure its least ciliotoxicity.

Acetaminophen↗
PubMed1995

[A quantitative study on alpha-1 adrenergic receptor in the liver of Wistar rats during the process of CCl4-induced hepatic cirrhosis].

In this study, radioligand binding analysis was used to quantitatively measure the concentration and affinity of alpha-1 adrenergic receptor in the liver of Wistar rats during the process of CCl4-induced liver cirrhosis. It was found that compared with normal controls, the maximal binding capacity (Bmax) of alpha-1 adrenergic receptor in the liver of 13 pre-cirrhotic rats decreased from 132.8 +/- 25.0fmol/mg to 96.9 +/-20.8fmol/mg of protein (P < 0.01) with increased dissociation constant (Kd, from 0.1633 +/- 0.0322nM to 0.3079 +/- 0.0786nM, P < 0.01). The Bmax in 20 cirrhotic rats decreased significantly from 132.8 +/- 25.0fmol/mg to 71.9 +/- 17.7fmol/mg of protein (P < 0.01), while Kd values increased from 0.1633 +/- 0.0322nM to 0.4012 +/- 0.0965nM (P < 0.01). These findings indicate that the decrease of binding capacity and affinity of alpha adrenergic receptor may play a role in the disturbances of metabolism of catecholamine often seen in some cirrhotic patients and have implications in the pathogenesis of portal hypertension.

Animals↗
PubMed1995

[Study on animal models of immune mediate motoneuron disease].

Swine motoneurons (SMN) were isolated from fresh spinal cords of pigs. Homogenates of these SMN or fresh anterior horns of pigs (SAH) as immunogens were inoculated to guinea pigs or Lewis rats and Wistar rats respectively. The impairments of motion were observed in the immunized guinea pig four months after the fifth inoculation of SMN, in the immunized Lewis rats after the first or second inoculation of SAH and in the Wistar rat after the third inoculation of SAH respectively. Degeneration and loss of motoneurons in the spinal cords of these symptomatic animals were found histologically. Antibodies against motoneurons of guinea pig and rat can be detected in sera of these symptomatic animals with immunocytochemical method respectively. In control guinea pigs, Lewis rats and Wistar rats there were no symptom, and did not found degeneration of motoneurons in the spinal cords of these control animals. Antibodies against motoneurons can not be detected in the sera of these control animals. The results indicated immune mediate guinea pig or rat models for MND can be established with pure SMN or impure SMN as immunogens. It was shown the conservative homology between antigenic structures of lower motoneurons in pigs and guinea pigs or rats. The pathogenesis in immunized animals with SAH is faster than that in immunized animals with SMN at least in terms of the appearance of symptoms. The Lewis rats produced symptoms first and the incidence ratio of symptomatic animals in Lewis rats was the highest. The immunized Wistar rats produced symptoms a little bit slower.

Animals↗
PubMed1994

Identification of the urokinase receptor as an adhesion receptor for vitronectin.

Urokinase receptors, expressed on surfaces of many cell types, focus to the pericellular space plasminogen-dependent proteolysis important in matrix remodeling and cell movement. We now report that the urokinase receptor (uPAR) is also a high affinity (Kd < 30 nM) receptor for vitronectin. Recombinant uPAR binds vitronectin in the absence of urokinase, but vitronectin binding is promoted by concurrent receptor binding of either urokinase or fragments thereof containing its uPAR binding domain. Stable epithelial cell transfectants expressing membrane-anchored uPAR, but not cells expressing soluble uPAR, become strongly adhesive with altered morphology in the absence of urokinase. These observations identify a new class of vitronectin receptor and imply a duality in function for the receptor that intrinsically links matrix adhesion to regulation of protease activity. Increases in urokinase receptor expression known to be associated with cellular activation and malignant transformation could modulate cellular trafficking and function by promoting attachment to vitronectin.

Adsorption↗
PubMed1994

Crystallization of the malonyl coenzyme A-acyl carrier protein transacylase from Escherichia coli.

The malonyl coenzyme A-acyl carrier protein transacylase, a single polypeptide chain of 358 amino acid residues and a molecular mass of 32 kDa, is a key component of the fatty acid synthase multienzyme complex. The elucidation of its three-dimensional structure will help in the understanding of the molecular basis of the biosynthesis of fatty acids, as well as of polyketides and related biologically active molecules. Three X-ray-quality crystal forms of the Escherichia coli fabD gene product encoding for malonyl coenzyme A-acyl carrier protein transacylase have been obtained using the hanging-drop method and ammonium sulfate as precipitant. Two are tetragonal and each contains two molecules in the asymmetric unit (form I: space group P4(3(1))2(1)2 with a = b = 83.9 A, c = 166.5 A and form II: space group P4 with a = b = 132.64 A, c = 38.85 A), whereas the third form belongs to the hexagonal system and contains one molecule in the asymmetric unit (space group P6(1(5)) with a = b = 68.52 A, c = 117.71 A). In each case, the diffraction pattern extends to approximately 2.0 A resolution using CuK alpha radiation from a rotating anode source.

Acyl-Carrier Protein S-Malonyltransferase↗
PubMed1994

Crystal structure, at 2.6-A resolution, of the Streptomyces lividans xylanase A, a member of the F family of beta-1,4-D-glycanases.

The crystal structure of the 32-kDa catalytic domain of the Streptomyces lividans xylanase A was solved by molecular isomorphous replacement methods and subsequently refined at 2.6-A resolution to a conventional crystallographic R factor of 0.21. This is the first successful structure determination of a member of the F family of endo-beta-1,4-D-glycanases. Unlike the recently determined xylanases of the G family (Wakarchuk, W. W., Campbell, R. L., Sung, W. L., Davoodi, J., and Yaguchi, M. (1994) Protein Sci. 3, 467-475), where the catalytic domains have a unique beta-sheet structure, the 32-kDa domain of the S. lividans xylanase A is folded into a complete (alpha/beta)8 barrel, the first such fold observed among beta-1,4-D-glycanases. The active site is located at the carbonyl end of the beta barrel. The crystal structure supports the earlier assignment of Glu-128 and Glu-236 as the catalytic amino acids (Moreau, A., Roberge, M., Manin, C., Shareck, F., Kluepfel, D., and Morosoli, R. (1994) Biochem. J., in press).

Binding Sites↗
PubMed1994

Structure of a myristoyl-ACP-specific thioesterase from Vibrio harveyi.

The crystal structure of a myristoyl acyl carrier protein specific thioesterase (C14ACP-TE) from a bioluminescent bacterium, Vibrio harveyi, was solved by multiple isomorphous replacement methods and refined to an R factor of 22% at 2.1-A resolution. This is the first elucidation of a three-dimensional structure of a thioesterase. The overall tertiary architecture of the enzyme resembles closely the consensus fold of the rapidly expanding superfamily of alpha/beta hydrolases, although there is no detectable homology with any of its members at the amino acid sequence level. Particularly striking similarity exists between the C14ACP-TE structure and that of haloalkane dehalogenase from Xanthobacter autotrophicus. Contrary to the conclusions of earlier studies [Ferri, S. R., & Meighen, E. A. (1991) J. Biol. Chem. 266, 12852-12857] which implicated Ser77 in catalysis, the crystal structure of C14ACP-TE reveals a lipase-like catalytic triad made up of Ser114, His241, and Asp211. Surprisingly, the gamma-turn with Ser114 in a strained secondary conformation (phi = 53 degrees, psi = -127 degrees), characteristic of the so-called nucleophilic elbow, does not conform to the frequently invoked lipase/esterase consensus sequence (Gly-X-Ser-X-Gly), as the positions of both glycines are occupied by larger amino acids. Site-directed mutagenesis and radioactive labeling support the catalytic function of Ser114. Crystallographic analysis of the Ser77-->Gly mutant at 2.5-A resolution revealed no structural changes; in both cases the loop containing the residue in position 77 is disordered.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1994

Staphylococcal protein A containing phospholipid monolayers on aqueous and solid surfaces.

The staphylococcal protein A containing monolayer is characterized on both aqueous and solid surfaces. The density of SpA in dipalmitoylphosphatidic acid (DPPA) monolayers is determined from ultraviolet spectroscopy. The influence of calcium on the SpA concentration in the monolayer is also investigated. The spectroscopic results indicate that the incorporation of SpA in the monolayer is influenced by the calcium concentration in the subphase, and more than 90% of adsorbed protein is excluded from the monolayer when calcium concentration is in the order of millimoles. The IgG binding activity of the SpA in the reconstituted membrane is influenced by the surface pressure at which the membrane is formed.

Calcium↗
PubMed1994

Evidence for long-range oncogene activation by hepadnavirus insertion.

Insertional mutagenesis of host genes, a common oncogenic strategy of slow transforming retroviruses, has recently been described for a DNA virus of the hepadnavirus group: the woodchuck hepatitis virus. This virus causes insertional activation of myc genes, mainly the intronless N-myc2 oncogene, in > 50% of woodchuck liver tumours. In most remaining tumours, N-myc2 is overexpressed without any apparent genetic alteration. To elucidate the role of the virus in such cases, we have cloned and analysed single integration sites in four woodchuck tumours carrying wild-type myc alleles. All sites were clustered within < 20 kb in a single locus, in which scarce unique sequences showed no detectable transcriptional activity. By fluorescent in situ hybridization, N-myc2 and the new locus (win) were localized to the same region of the long arm of the woodchuck X chromosome, and a 150-180 kb intervening distance was deduced from pulse-field gel analysis. The detection of viral integrations in win in additional tumours that produced abundant N-myc2 transcripts further substantiates the link between these two loci in woodchuck tumorigenesis. We propose that efficient activation of the N-myc2 promoter by the hepadnavirus enhancer acting over a long distance might operate in liver cell transformation.

Animals↗
PubMed1994

Characterization of modified staphylococcal protein A containing phospholipid monolayer on both solution and slide surfaces.

A method is described for incorporation of water-soluble protein Staphylococcal protein A (SpA) into phospholipid monolayer using covalent protein-lipid conjugates in detergent solution. The amphiphilic conjugates have solubility properties very similar to intergral membrane proteins. When the conjugates are applied into dipalmitoyl-phosphatidic acid monolayer, a protein containing monolayer is formed on subphase surface. The monolayer is transferred to pre-coated substrate surface to form an artificial membrane. Results show that unmodified SpA is readily ejected from the monolayer when compressing the monolayer but modified SpA incorporates into the monolayer stably. The incorporation of the protein is proportional to the lipid coupling degree. When the protein is excessively modified, the IgG binding activity of the SpA in the membrane is lost significantly.

Immunoglobulin G↗
PubMed1994

Crystallization of canine cardiac calsequestrin.

Calsequestrin is the major Ca2+ binding protein in the lumen of the sarcoplasmic reticulum membranes. Two X-ray quality crystal forms of canine cardiac calsequestrin were obtained by the hanging drop method using KCl as a precipitant. One form is monoclinic (space group P2(1), a = 73.4 A, b = 104.4 A, c = 60.2 A, beta = 120.4 degrees) with two molecules in the asymmetric unit and a solvent content of approximately 40%. The second form is trigonal (P3(1)21 or P3(2)21, a = b = 99.3 A, c = 89.8 A) with a single molecule in the asymmetric unit and 55% solvent content. Cross rotation function calculations show that despite the different space groups the packing of the molecules in both crystals is likely to be similar suggesting the existence of a stable dimer. The monoclinic crystals diffract beyond 3 A using a laboratory rotating anode source, while under the same conditions the trigonal crystals diffract only to approximately 4.5 A. This is the first report of successful preparation of X-ray quality crystals of a high capacity Ca2+ binding protein.

Animals↗
PubMed1994

Cloning and characterization of the t(X;11) breakpoint from a leukemic cell line identify a new member of the forkhead gene family.

Chromosome translocations involving 11q23 are associated with a number of different types of leukemia. These translocations fuse a gene encoding a putative transcription factor, HTRXI, to genes on other chromosomes. We report cloning and sequencing the t(X;11) breakpoint region from a cell line established from an infant with acute lymphocytic leukemia. The gene AFXI, on the X chromosome, is expressed in a variety of cell types. Sequence analysis indicates a high degree of homology between AFXI and the forkhead family of transcription factors. The high degree of identity within the forkhead region and the lack of homology outside that region suggest that AFXI represents a novel forkhead family member. It is predicted that a chimeric fusion protein with altered DNA binding activity will be the result of the translocation.

Amino Acid Sequence↗
PubMed1994

Crystallization and preliminary crystallographic studies of the precursor and mature forms of a neutral lipase from the fungus Rhizopus delemar.

A neutral lipase from the filamentous fungus Rhizopus delemar has been crystallized in both its proenzyme and mature forms. Although the latter crystallizes readily and produces a variety of crystal forms, only one was found to be suitable for X-ray studies. It is monoclinic (C2, a = 92.8 A, b = 128.9 A, c = 78.3 A, beta = 135.8) with two molecules in the asymmetric unit related by a noncrystallographic diad. The prolipase crystals are orthorhombic (P2(1)2(1)2(1), with a = 79.8 A, b = 115.2 A, c = 73.0 A) and also contain a pair of molecules in the asymmetric unit. Initial results of molecular replacement calculations using the refined coordinates of the related lipase from Rhizomucor miehei identified the correct orientations and positions of the protein molecules in the unit cells of crystals of both proenzyme and the mature form.

Crystallization↗
PubMed1994

Hepatic sinusoidal endothelial cell G1/S arrest correlates with severity of alcoholic liver injury in the rat.

BACKGROUND/AIMS: Capillarization of the hepatic sinusoid occurs in alcoholic liver disease. Because endothelial cell proliferation is relevant to capillarization, we used the intragastric feeding rat model to evaluate the relationship between pathological liver injury and endothelial cell proliferation. METHODS: Male Wistar rats (225-250 g) were fed liquid diets containing corn oil and ethanol for periods ranging between 1 week and 2 months. At the time the rats were killed, the severity of pathological injury and endothelial cell proliferation using anti-rat proliferating cell nuclear antigen/antibody was evaluated. RESULTS: Two distinctly different populations of proliferating sinusoidal lining cells were identified; one population showed relatively weak granular staining consistent with cells arrested at the G1/S boundary. The other population of cells showed strong granular staining of the nucleoplasm and nucleoli (cells in mid to late S phase). A strong correlation (r = 0.85; P < 0.01) was obtained between pathological severity and G1/S-arrested endothelial cells. There was no correlation between cells in S phase. CONCLUSIONS: The presence of an increased number of G1/S-arrested endothelial cells in animals with severe pathological change suggests that stimuli are present for both endothelial cell proliferation and G1/arrest. The identification of these stimuli could lead to a better understanding of the pathogenesis of alcoholic liver disease.

Animals↗
PubMed1994

An unusual buried polar cluster in a family of fungal lipases.

The stability of globular proteins arises largely from the burial of non-polar amino acids in their interior. These residues are efficiently packed to eliminate energetically unfavorable cavities. Contrary to these observations, high resolution X-ray crystallographic analyses of four homologous lipases from filamentous fungi reveal an alpha/beta fold which contains a buried conserved constellation of charged and polar side chains with associated cavities containing ordered water molecules. It is possible that this structural arrangement plays an important role in interfacial catalysis.

Amino Acid Sequence↗