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Biomedical subjects

Y Yin

Publications and source records attributed to Y Yin.

At least 127 records · Page 7Linked to original sources
PubMed1995

Secretion and circular dichroism analysis of the C-terminal signal peptides of HlyA and LktA.

The secretion of the 107 kDa hemolysin A (HlyA) from Escherichia coli is mediated by membrane proteins hemolysin B (HlyB) and hemolysin D (HlyD). The signal for transport has been mapped to the C-terminal 60 amino acids of the HylA molecule. We have shown previously that the C-terminal 70 amino acids of leukotoxin (LktA) from Pasteurella hemolytica can substitute functionally for the HlyA signal sequence. This 70 amino acid peptide contains little primary sequence similarity to the HlyA signal sequence, and we have hypothesized that these signal sequences assume a similar higher-order structure which is recognized by the HlyB/D transporter. In the present study, we have expressed and purified small peptides containing the C-terminal 61 amino acids of HlyA and the C-terminal 70 amino acids of LktA. We show that these signal peptides are sufficient for secretion from E. coli in a HlyB/D dependent manner. Circular dichroism analyses show that both molecules exhibit common biophysical properties. In aqueous solution, they appear to be mainly unstructured, but in a membrane mimetic environment they assume a helical secondary structure. The conformational change observed for both peptides going from an aqueous to a membrane mimetic environment may be an important feature of these signal sequences necessary for their recognition and transport.

Bacterial Proteins↗
PubMed1995

The regulatory regions of the rice tungro bacilliform virus promoter and interacting nuclear factors in rice (Oryza sativa L.).

The Rice Tungro Bacilliform Virus (RTBV) promoter confers phloem-specific gene expression in transgenic rice plants. A series of promoter deletion mutants were fused with the Escherichia coli beta-glucuronidase A (uidA) reporter gene and introduced into transgenic rice plants. The RTBV promoter confers substantially stronger expression in shoots than in roots. A fragment of the promoter comprising nucleotides -164 to +45 relative to the transcriptional start site contains sufficient information for phloem-specific gene expression. Within this region, nucleotides -164 to -43 were essential for promoter function since deletion of this fragment dramatically reduced promoter activity. Gel-retardation assays identified two groups of rice nuclear factors (RNFG1 and RNFG2) that bind to the -164 to +45 promoter fragment. Competition and DNasel footprinting experiments indicated that RNFG1 bound to nucleotides -3 to +8 (Box I) while RNFG2 bound to nucleotides -53 to -39 (Box II). Interactions between the two groups of factors were observed. In addition, we found differences in the binding of nuclear factors from shoots versus from roots, in agreement with the different activities of the promoter in these two organs. It is proposed that binding of RNFG1 and RNFG2 between nucleotides -164 to +45 is essential for the tissue-specific expression of this promoter.

Base Sequence↗
PubMed1995

Colonic protein fermentation and promotion of colon carcinogenesis by thermolyzed casein.

Thermolyzed casein is known to promote the growth of aberrant crypt foci (ACF) and colon cancer when it is fed to rats that have been initiated with azoxymethane. We speculated that the promotion was a consequence of increased colonic protein fermentation (i.e., that the thermolysis of the casein decreases its digestibility, increases the amount of protein reaching the colon, and increases colonic protein fermentation and that the potentially toxic products of this fermentation promote colon carcinogenesis). We found that the thermolysis of casein reduces its digestibility and increases colonic protein fermentation, as assessed by fecal ammonium and urinary phenol, cresol, and indol-3-ol. Thermolysis of two other proteins, soy and egg white protein, also increases colonic protein fermentation with increased fecal ammonia and urinary phenols, and thermolysis of all three proteins increases the levels of ammonia and butyric, valeric, and i-valeric acids in the cecal contents. We found, however, that the increased protein fermentation observed with thermolysis is not associated with promotion of colon carcinogenesis. With casein, the kinetics of protein fermentation with increasing thermolysis time are clearly different from the kinetics of promotion of ACF growth. The formation of the fermentation products was highest when the protein was thermolyzed for one hour, whereas promotion was highest for protein that had been thermolyzed for two or more hours. With soy and egg white, thermolysis increased colonic protein fermentation but did not promote colon carcinogenesis. Thus, although thermolysis of dietary casein increases colonic protein fermentation, products of this fermentation do not appear to be responsible for the promotion of colon carcinogenesis. Indeed, the results suggest that protein fermentation products do not play an important role in colon cancer promotion.

Animals↗
PubMed1995

Mutations in rpoB gene of rifampicin resistant clinical isolates of Mycobacterium tuberculosis in Japan.

In Mycobacterium tuberculosis, involvement of alterations of the RNA polymerase beta subunit in resistance to rifampicin has been described by Telenti et al. To determine if the same correlation could be observed between the mutation of the rpoB gene and clinically isolated M. tuberculosis of the rifampicin-resistant phenotype in Japan, 47 strains of M. tuberculosis of the rifampicin-resistant phenotype, 17 of the rifampicin-susceptible phenotype, and 4 type strains were examined. A 411-base pair (bp) rpoB fragment was amplified by the polymerase chain reaction and subjected to solid phase direct sequencing. By comparing the nucleotides, mutation involving 8 conserved amino acids were identified in 44 of the 47 (93.6%) rifampicin-resistant isolates, but in none of the 17 sensitive isolates and 4 type strains. All mutations found were clustered within a region of 23 amino acids. Thus, similar to the results reported by Telenti et al., substitution of a limited number of highly conserved amino acids encoded by the rpoB gene appears to be the molecular mechanism responsible for resistance to rifampicin in Japanese clinical isolates of M. tuberculosis. Our results suggest that direct DNA sequencing of the rpoB gene may be a reliable method for identifying rifampicin-resistant M. tuberculosis strains among Japanese clinical isolates.

Amino Acid Sequence↗
PubMed1995

Ionic mechanism of acupuncture on improvement of learning and memory in aged mammals.

Memory impairment is one of the most frustrating problems for older people. Several acupoints were used for the treatment of memory loss in old rats, and the elemental mechanism of acupuncture therapy was studied with Inductively Coupled Plasma Spectroscopy. The result indicates that acupuncture improves learning and memory ability significantly in aged animals when compared with controls. This study demonstrated that elevations of eight essential elements (B, Ca, Cu, Fe, K, Mg, Na, and P) in the brain are the ionic basis for the therapeutic effect of acupuncture. A hypothetical model of the mechanism of acupuncture therapy is described.

Acupuncture Therapy↗
PubMed1995

Three-compartment wrist arthrography: direct comparison of digital subtraction with nonsubtraction images.

PURPOSE: To determine the diagnostic value of digital subtraction and fluoroscopic spot images in wrist arthrography, alone and in combination. MATERIALS AND METHODS: Randomized selective bilateral three-compartment wrist arthrography was performed in 64 patients. The fluoroscopic spot and digital subtraction images were read separately by two radiologists without knowledge of the clinical data. RESULTS: One hundred twenty communicating defects were found in 69 wrists in 42 patients. Fluoroscopic spot radiographs alone depicted defects in all 69 wrists, but precise localization was not possible in 10 defects. Subtraction images alone demonstrated 59 defects. The defects not seen with digital subtraction imaging were identified after wrist motion. Digital subtraction images demonstrated defects earlier during the joint injection and demonstrated the exact site of the 10 defects not localized on the nonsubtracted images. CONCLUSION: Digital subtraction and fluoroscopic spot images are complementary in wrist arthrography and, when used in combination, increase the ability to localize the sites of defects, although the value added by the subtraction technique is small.

Adolescent↗
PubMed1995

Giant-cell tumor of the distal phalanx of the hand in a child.

Giant-cell tumor of bone in the distal phalanx is rare, as is its clinical occurrence in children. A case of a 10-year-old girl with giant-cell tumor in the distal phalanx of the left thumb is reported, and a literature review of phalangeal giant-cell tumor of the hand is presented. The tumor was curetted, and after 2.5 years there was no recurrence.

Bone Neoplasms↗
PubMed1995

A 9-year-old girl with a bowed right forearm 6 months after a fracture.

The following case is presented to illustrate the roentgenographic and clinical findings of a condition of interest to the orthopedic surgeon. Initial history, physical findings, and roentgenographic examinations are found on the first page. The final clinical and roentgenographic differential diagnoses are presented on the following pages.

Cafe-au-Lait Spots↗
PubMed1995

[Peripheral anterior synechiae overlying the haptics of posterior chamber lenses].

To study the changes of the anterior chamber angle after intraocular lens implantation, 111 patients (130 eyes) with posterior chamber intraocular lens implantation at different post-operative periods were examined by gonioscopy. Peripheral anterior synechia overlying the position of the lens haptic (lens haptic PAS) was observed in up to 50.8% of eyes implanted with haptics vaulted anteriorly by 10%. The lens haptic PAS is easy to be identified, it is a broad based forward displacement of the peripheral iris adhesive to the anterior wall of the anterior chamber angle. The size of the PAS was of half to one o'clock position and occasionally it was of one and a half to two o'clock position. The differences between the rates of occurrence of lens haptic PAS were not significant at different post-operative periods (P > 0.05). 55.9% of the sulcus fixated and 42.3% of the capsular fixated eyes had lens haptic PAS (P > 0.05). There were more lens haptic PAS in eyes with horizontally oriented lens haptics (62.7%) than with lens haptics at vertical position (41.4%, P < 0.05). Regular gonioscopy for posterior chamber lens implantation was recommended.

Adult↗
PubMed1994

An autonomously propagating luciferase-encoding vector for Dictyostelium discoideum.

We have constructed a luc reporter vector for Dictyostelium discoideum using a 626-bp fragment from the nuclear-associated plasmid Ddp2. The ori from Ddp2 is localized within this fragment and was used to provide an autonomous replication sequence for the reporter vector. This reporter vector was stably retained in D. discoideum AX3K cells without alteration. The vector molecule was also found to exist in relatively low copy number compared to other Dictyostelium vectors in the transformed cells. We demonstrated the utility of this vector as a reporter vector with glycogen synthase promoter/luc fusions of varying sizes.

Animals↗
PubMed1994

Antagonism between T3 and amiodarone on the contractility and the density of beta-adrenoceptors of chicken cardiac myocytes.

3,3',5-Triiodothyronine (T3), at 10(-8) M, potentiated by 26.4-30.9% the isoproterenol-mediated inotropic effect in chick embryo cardiac myocytes in culture. Amiodarone (10(-6) M) decreased this response by 44.6% only in cells cultured with serum, where the T3 concentration was 10(-13) M. Amiodarone inhibited the potentiating effect of T3. Amiodarone alone had no influence on the beta-adrenoceptor density in cells cultured in serum-free medium. This confirms that the effects of amiodarone on cardiac beta-adrenoceptors are T3 dependent. T3 increased the density of beta-adrenoceptors through two concentration ranges, with an initial 30% increase between 10(-14) and 10(-11) M, followed by a second increase until 10(-7) M. Amiodarone not only inhibited the first positive effect of T3 but also decreased beta-adrenoceptor density far below the control value. The second positive T3 effect was also inhibited by 50% by amiodarone. This study suggests that T3 might increase the number of cell-surface beta-adrenoceptors and modify their cellular traffic through at least two mechanisms, one assumed to be non-genomic, the other being genomic, and that amiodarone could affect the two mechanisms differently.

Adrenergic beta-Antagonists↗
PubMed1994

Quantitative analysis in the characterization and optimization of protein crystal growth.

Protein crystal growth often depends on the combination of many different factors. Some affect protein solubility directly; others may act indirectly by causing conformational changes. Systematic characterization of these factors can be important for generating good crystals. It can also provide useful insight into the biochemical behavior of the protein being crystallized. Here we focus on statistical methods to achieve these two objectives. (1) Characterization of a protein system by analyzing patterns of crystal polymorphism under different levels of biochemical parameters, such as ligands and pH. Tests of the reproducibility of crystal growth experiments indicate that quantitative scales of crystal quality can be statistically significant. Analysis of variance for a replicated, full-factorial design in which four factors were tested at two levels has been used to demonstrate highly significant, biochemically relevant, two-factor interactions strongly implicating pH and ligand-dependent conformational changes. (2) Optimization of crystal growth via response-surface methods. 'Minimum predicted variance' designs provide for efficient response-surface experiments aimed at constructing quadratic models in several dimensions. We have used such models to improve crystal size and quality significantly for three forms of Bacillus stearothermophilus tryptophanyl-tRNA synthetase. In one case we can now avoid having to increase the size by repeated seeding, a difficult procedure that also produces unwanted growth of satellite crystals. Graphs of two-dimensional level surfaces reveal a number of ridges, where the same result is obtained for many combinations of the factors usually varied when trying to improve crystals. An important inference is that it may be better to sample simultaneously for the effects of protein concentration and supersaturation. For a system involving only one crystallizing agent, supersaturation can be approximated as the product of protein and precipitant concentrations. Use of this search direction significantly improves the performance of response-surface experiments. Advantages of growing crystals at stationary points of their response surfaces include better crystals and higher reproducibility, since crystal growth at stationary points is insulated from the deleterious effects of experimental fluctuations. This arises because the derivatives of the response are by definition zero with respect to the experimental variables. Quantitative analysis of appropriately designed crystal growth experiments can thus be a powerful way to characterize complex and interacting biochemical dependencies in macromolecular systems and optimize parameters important to the crystallography.

Journal Article↗
PubMed1994

Solution structure of the tetrameric minimum transforming domain of p53.

We report the solution structure of the minimum transforming domain (residues 303-366) of human p53 (p53tet) determined by multidimensional NMR spectroscopy. This domain contains a number of important functions associated with p53 activity including transformation, oligomerization, nuclear localization and a phosphorylation site for p34/cdc2 kinase. p53tet forms a symmetric dimer of dimers that is significantly different from a recent structure reported for a shorter construct of this domain. Phosphorylation of Ser 315 has only minor structural consequences, as this region of the protein is unstructured. Modelling based on the p53tet structure suggests possible modes of interaction between adjacent domains in full-length p53 as well as modes of interaction with DNA.

Amino Acid Sequence↗
PubMed1994

Disruption of glycogen phosphorylase gene expression in Dictyostelium: evidence for altered glycogen metabolism and developmental coregulation of the gene products.

Glycogen phosphorylase 1 and 2, the isozymes responsible for glycogen degradation, are encoded by separate genes in Dictyostelium. The two gene products display different transcriptional and translational expression and distinct post-translational regulation. Using DNA-mediated transformation, Dictyostelium clones which lacked either glycogen phosphorylase 1 or 2 (gp1 or gp2) expression were obtained. The loss of either enzyme did not change axenic growth patterns, developmental progression, or gross organismic morphology. In gp1- strains, glycogen accumulated to a 17- to 28-fold higher level during late stationary phase without any obvious detrimental effects. This implies that no alternative pathway for glycogen degradation is present in amoebae, and that glycogen metabolism is not critical for vegetative cell growth. Developmental glycogen concentrations were not altered significantly in any of the transformants, but in gp2- cells the posttranslational regulation of the intact gp1 enzyme was apparently modulated to compensate for the loss of gp2. Western blots of microdissected, lyophilized Dictyostelium slugs and early culminates showed that gp2 was found in both prestalk and prespore cells, with a slight enrichment in prespore cells. The gp1 protein was highly enriched in prestalk cells in the parental strain. In gp2- transformants, however, gp1 was detected in equal amounts in both cell types. The loss of gp2 led to a shift in the cell-type-specific expression pattern of gp1, presumably due to developmental coordinate regulation of gp1 and gp2 at the translational and/or transcriptional level.

Animals↗
PubMed1994

[Molecular cloning of alpha antigen like protein gene of Mycobacterium leprae and its over production in Escherichia coli].

I have constructed the genomic library of M. leprae Thai 53 strain, and cloned the alpha antigen like protein gene by plaque hybridization method by using M. leprae alpha antigen DNA fragment as probe which was characterized in the previous study, I have termed it as alpha 2 antigen gene. The alpha 2 antigen gene has been characterized by sequencing. By comparing the deduced amino acid sequence of alpha and alpha 2 antigen with 85 complex antigen of other mycobacteria. I have found the higher homology between alpha 2 antigen and 85A antigen and between alpha antigen and 85B antigen. We have constructed the over expression system of M. leprae alpha and alpha 2 antigen gene in E. coli using vector pMALc-RI. Recombinant alpha and alpha 2 antigen has been purified by amylose column chromatography at the purity of more than 95%. More than 6 mg and more than 10 mg of recombinant alpha and alpha 2 antigen has been obtained from 200 ml of liquid culture, respectively. ELISA tests have been performed with the sera of leprosy patient and healthy control against the recombinant alpha and alpha 2 antigens. The antibody titers in sera of leprosy patient against the two kinds of antigens were all much higher than healthy controls. The antibody titer against the alpha 2 antigen was higher than that against alpha antigen. Recombinant alpha and alpha 2 antigens in this study could be used as a new specific antigen for serodiagnosis of leprosy.

Amino Acid Sequence↗
PubMed1994

Dual regulation of the glycogen phosphorylase 2 gene Dictyostelium discoideum: the effects of DIF-1, cAMP, NH3 and adenosine.

Cell differentiation in Dictyostelium results in the formation of two cell types, stalk and spore cells. The stalk cells undergo programmed cell death, whereas spore cells retain viability. The current evidence suggests that stalk cell differentiation is induced by Differentiation Inducing Factor (DIF), while spore cell differentiation occurs in response to cAMP. We have discovered the first developmentally regulated Dictyostelium gene, the glycogen phosphorylase gene 2 (gp2) gene, that can be induced by both DIF-1 and cAMP, suggesting the possibility of a new group of developmentally regulated genes that have DIF-1 and cAMP dual responsiveness. The gp2 gene was found to be expressed in both prestalk/stalk cells and prespore/spore cells. The DIF-1 competence of the gp2 gene required uninterrupted development, whereas the cAMP-competence for the gene required only starvation. Both DIF-1 and cAMP induction of the gene could be inhibited by NH3, a factor that is thought to act as a developmental signal in Dictyostelium. Another developmental signal, adenosine, was found to repress the DIF-1 induction of the gp2 gene. Two introns in the gp2 gene were examined for their involvement in the regulation of the gene, but no regulatory function was detected. A model for the regulation of the gp2 gene during the development is proposed.

Adenosine↗