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Y Yu

Publications and source records attributed to Y Yu.

At least 19 recordsLinked to original sources
PubMed1992

The iron-responsive element binding protein. Purification, cloning, and regulation in rat liver.

The iron-responsive element binding protein (IRE-BP) is a cytosolic protein that binds a highly conserved sequence in the untranslated regions of mRNAs involved in iron metabolism including ferritin, transferrin receptor, and erythroid 5-aminolevulinate acid synthase. This conserved sequence is termed the iron-responsive element and is necessary for the post-transcriptional regulation of these mRNAs by iron. The rat liver IRE-BP was purified to homogeneity by chromatographic methods and partial amino acid sequence was obtained. A cDNA was isolated from a rat liver cDNA library and sequenced. The amino acid sequence deduced from the cDNA sequence corresponds to a protein of 889 amino acids with a predicted molecular weight of 97.946. The NH2-terminal sequence obtained by Edman degradation matched the deduced amino acid sequence obtained from the cDNA, confirming the translational start site. Rat liver IRE-BP shares 95% identity with human IRE-BP and 98% identity with mouse IRE-BP indicating that the IRE-BPs have remained highly conserved during evolution. The 5'-untranslated region is at least 236 nucleotides and contains interesting structural features including two direct repeats, an inverted repeat, and three small open reading frames. The rat IRE-BP mRNA is approximately 3600 nucleotides and is expressed in a variety of rat tissues including liver, spleen, and gut. Over the course of 16 h following an intraperitoneal injection of iron in rats. IRE-BP RNA binding activity decreases to 50% of control levels. The decrease in IRE-BP RNA binding activity in extracts from iron-treated rats is reversible by pretreatment of the extracts with reducing agents. The steady-state levels of IRE-BP mRNA remain constant during iron treatment. These data suggest that the decrease in IRE-BP RNA binding activity by iron in rat liver is due to post-translational changes in the RNA binding affinity of the IRE-BP and not due a decrease in the transcription of the IRE-BP gene or to the destabilization of the IRE-BP mRNA.

Amino Acid Sequence↗
PubMed1992

Mechanistic studies of the biosynthesis of 3,6-dideoxyhexoses in Yersinia pseudotuberculosis. Purification and stereochemical analysis of CDP-D-glucose oxidoreductase.

An NAD(+)-dependent CDP-D-glucose oxidoreductase which catalyzes the first step of the biosynthesis of CDP-ascarylose (CDP-3,6-dideoxy-L-arabino-hexose), converting CDP-D-glucose to CDP-4-keto-6-deoxy-D-glucose, was isolated from Yersinia pseudotuberculosis. A protocol consisting of DEAE-cellulose, Matrex Blue-A, hydroxylapatite, DEAE-Sephadex, Sephadex G-100, and NAD(+)-agarose column chromatography was used to purify this enzyme 6000-fold to homogeneity. This enzyme consists of two identical subunits, each with a molecular weight of 42,500. Using CDP-D-glucose as the substrate, the Km and Vmax of this catalysis were determined to be 222 microM and 8.3 mumols mg-1 min-1, respectively. Unlike most other oxidoreductases of its class which have a tightly bound NAD+, this highly purified CDP-D-glucose oxidoreductase showed an absolute requirement of NAD+ for its activity. Using chemically synthesized (6S)- and (6R)-CDP-D-[4-2H,6-3H]glucose as substrates, a stereochemical analysis showed this enzymatic reaction involves an intramolecular hydrogen migration from C-4 to C-6, and the displacement of C-6 hydroxyl group by the C-4 hydrogen occurs with inversion. Thus, despite the low cofactor affinity, this enzyme undergoes a mechanism consistent with that followed by other members of its type. Such a mechanistic and stereochemical convergency found for all sugar oxidoreductases so far characterized suggests the presence of a common progenitor of this class of enzyme.

Amino Acids↗
PubMed1992

Epidermal growth factor stimulates cAMP accumulation in cultured rat cardiac myocytes.

We have previously shown that epidermal growth factor (EGF) augments cAMP accumulation in the heart and stimulates cardiac adenylyl cyclase via a G protein mediated mechanism (Nair et al., 1989). More recently, employing an antibody against the carboxy-terminus decapeptide of Gs alpha, we have demonstrated that Gs alpha mediates the effects of EGF on cardiac adenylyl cyclase (Nair et al., 1990). Since the heart comprises of a variety of cell types, the purpose of the studies presented here was to determine whether or not the effects of EGF on adenylyl cyclase were mediated in cardiac myocytes or noncardiomyocytes. Therefore, cultures of ventricular cardiomyocytes and noncardiomyocytes from neonatal rat hearts were established and characterized. Apart from the differences in cellular morphology, cardiomyocytes but not the noncardiomyocytes employed in our studies expressed the alpha- and beta-myosin heavy chain (MHC) mRNA and the beta-MHC protein. Additionally, as described previously, treatment of cardiomyocytes with thyroid hormone increased alpha-MHC mRNA and decreased the expression of beta-MHC mRNA, indicating that the cardiomyocytes employed in our studies were responding in a physiologically relevant manner. EGF in a time-dependent manner increased cAMP accumulation in the cardiomyocytes but not in noncardiomyocytes. Maximum and half-maximum effects were observed at 100 nM and 2 nM concentrations of EGF, respectively. As determined by the presence of immunoreactive EGF receptors and tyrosine phosphorylation of the 170 kDa protein in membranes of cardiomyocytes and noncardiomyocytes, both the cell populations contained functional EGF receptors. Therefore, the differential effects of EGF on cAMP accumulation in the two cell populations appear to be due to differential coupling of the EGF receptors to the adenylyl cyclase system rather than the absence of EGF receptors in noncardiomyocytes. Consistent with our previous findings in isolated membranes and perfused rat hearts, EGF-elicited increase in cAMP accumulation in cardiomyocytes did not involve activation of beta-adrenoreceptors and was abolished by prior treatment of cells with cholera toxin. Overall, our findings demonstrate that EGF-elicited increase in cAMP accumulation in the heart is the reflection of changes in cAMP content of cardiomyocytes and not noncardiomyocytes.

Animals↗
PubMed1992

Serum levels of macrophage colony-stimulating factor in patients with ovarian cancer undergoing second-look laparotomy.

OBJECTIVE: The purpose of this study was to evaluate the prognostic significance of macrophage colony-stimulating factor serum levels in patients with ovarian cancer undergoing second-look laparotomy. STUDY DESIGN: The presurgical serum levels of macrophage colony-stimulating factor from 33 consecutive patients with ovarian cancer undergoing second-look laparotomy were determined and compared with those of 50 healthy control subjects. Mean differences in groups were evaluated with the Student t test. RESULTS: Sixteen of 33 patients had a positive result at second look and a mean serum macrophage colony-stimulating factor level of 2.31 +/- 1.45 ng/ml. Seventeen of 33 patients had a negative result at second look and a mean macrophage colony-stimulating factor level of 1.90 +/- 0.86 ng/ml (p greater than 0.05). The mean macrophage colony-stimulating factor level in the control group was 1.20 +/- 0.51 ng/ml. This was statistically lower than the mean level found in patients with a negative second-look result (p less than 0.05). CONCLUSION: Regardless of tumor status, serum macrophage colony-stimulating factor levels tend to be elevated at the time of second-look laparotomy.

Adult↗
PubMed1992

Studies on compartmentation and turnover of murine retrovirus envelope proteins.

Several aspects of turnover and degradation of cell membrane proteins were studied in an NIH 3T3 cell clone expressing the env gene of Moloney murine leukemia virus ts1. Both internalization and shedding of the extracellular domain of the envelope protein gp70 occurred at the cell surface, albeit, in the case of shedding, only a very small fraction of gp70 was shed. The turnover rate of gp70 at the cell surface was similar to that of the same protein in the postendoplasmic reticulum intracellular compartment. In the presence of L-methionine methyl ester, the transmembrane domain of the envelope protein Prp15E was degraded faster than gp70.

3T3 Cells↗
PubMed1992

How the degenerate signal is recognized and delivered in the course of protein translocation across biological membranes.

Protein translocation across a membrane is generally directed by a degenerate signal peptide encoded in the nascent polypeptide chains. Effective signal recognition and the subsequent traversal of the polypeptide chain through the membrane are ensured by the mechanism of cotranslational translocation and/or by chaperone molecules which universally bind to nascent polypeptides and keep them under-structured and translocation-competent. In order to decipher the degenerate signal with a high degree of fidelity, the cells seem to have evolved a multistep signal recognition and relay mechanism, by which a weak, less-specific interaction between individual signal peptides and a defined signal recognition factor can take place, while the highly specific signal delivery can be achieved by only allowing the correct, effective signals to pass through all check-points along the relay cascade. In addition, a "double recognition" and energy-dependent "proof-reading" mechanism may be involved in single steps of such weak-bond interactions.

Biological Transport, Active↗
PubMed1991

Construction and characterization of expression systems for the env gene of ts1, a mutant of Moloney murine leukemia virus-TB.

A temperature-sensitive mutant of the Moloney murine leukemia virus-TB, ts1, causes hindlimb paralysis and immunodeficiency in mice. At the restrictive temperature, the envelope precursor polyprotein, gPr80env, is inefficiently processed intracellularly, and this is associated with the neurovirulence of ts1. To test the hypothesis that expression of the envelope proteins of ts1 alone without infectious virus production can induce paralysis, it is necessary to use either transmissible retroviral expression vectors or microinjection of eukaryotic gene expression plasmid to introduce the env gene of ts1 into germlines of mice. In this study, we have constructed three retrovirus vectors and three gene expression plasmids, all of which contain the env gene of ts1. By comparing the different expression systems, we found that one construct, pts1-env(F) can express the envelope proteins at a level comparable to the level expressed in ts1-infected cells. Furthermore, the expressed envelope proteins of pts1-env(F)-transfected cells possess the phenotypes of the proteins expressed by the env gene of ts1.

Animals↗
PubMed1991

Functional analysis of the CPS I upstream sequences with a cat assay.

Expression plasmids (pKCPS-CAT) containing carbamyl phosphate synthetase (CPS I) upstream sequences of different lengths were constructed, and the function and characteristics of the sequences were studied with the CAT assay. Results showed that the CPS I upstream sequences exerted highly tissue-specific control on CPS I gene expression, and the -113 approximately -38 bp region relative to the cap site was found to be indispensable for CPS I gene transcription. The -1700 approximately -161 bp region contains sequences which confer an enhancing effect on CPS I gene transcription. Dexamethasone and thioproline (a differentiation inducer) showed enhancing effects on CPS I gene transcription in hepatoma cells. These results would have significance in studies on the gene regulation of CPS I associated with the mechanism of hepatocyte differentiation and carcinogenesis.

Antineoplastic Agents↗
PubMed1991

Studies on antipeptic ulcer agents: the quantitative structure-activity relationship analysis of heterocyclic aldehyde N4-substituted phenyl (thio) semicarbazones.

Forty-five condensation products of furan-, pyrrole- and N-methyl pyrrole-alpha-carboaldehyde with N4-3- or N4-4-substituted phenyl semicarbazones and thiosemicarbazones were designed to optimize the antiulcer activity of a previously derived lead structure, formula II. Quantitative structure-activity relationships revealed that among the series of semicarbazones, increasing hydrophobicity and the introduction of electron-donating groups into the phenyl ring raise the antiulcer activity. Generally, semicarbazones are more active than the corresponding thiosemicarbazones. The wide gulf between the activity and toxicity of two derivatives (Compounds III and IV) necessitates further investigation of their pharmacological effects.

Anti-Ulcer Agents↗
PubMed1991

Induction of cytochrome P450 isozymes in human amnion FL cells and its application to the biological detection of mutagens.

Using AHH, EROD, ECOD and APND as marker enzymes and 3-MC, beta-NF, NE and PB as inducers, inducible cytP450 IA and IIB gene expression was demonstrated in the human amnion FL cell line; these cells possess a broad spectrum of drug-metabolizing enzymes. Maximum induction was observed following co-treatment with 3-MC and NE. Both constitutive and induced AHH were proved to have the characteristics of cytP448-dependent mixed-function oxygenases. Induced cytP450 isozyme activity remained at a high level for 24-36 h after removal of the inducer. The induced FL cells were demonstrated to activate common promutagens/procarcinogens in UDS and ADPRT-mediated decrease of NAD content assay systems. This new design can be used as a simplified mutagen screening system: no supplemental liver microsomal activation system is needed.

Amnion↗
PubMed1991

[Ultrastructural study of adenolymphoma].

Ultrastructure of 10 adenolymphomas of parotid gland were studied. The neoplastic epithelium consists of "oncocytes" which are arranged in pseudostratified pattern. Their cytoplasm is almost totally packed with a large numbers of hypertrophic mitochondria showing a variety of peculiar forms. The amount and alteration of the mitochondria gradually increase in the order of pyramidal cell, common columnar cell and degenerated columnar cell. This might be a process of transposition representing degeneration due to metabolic disorder in the tumor. The obvious alterations of mitochondria are found in the cases with not only "hot" nodule but also "cold" nodule shown in the studies of 99mTc scintigraphy. There seems no relationship between "hot" nodule and alteration of mitochondria.

Adenolymphoma↗
PubMed1991

[C fiber is not necessary in electroacupuncture analgesia, but necessary in diffuse noxious inhibitory controls (DNIC)].

Experiments were carried on rats. We applied capsaicin topically on sciatic nerve and used the techniques of extracellular recording and nerve trunk recording, Our results showed that the size of C compound action potentials in nerve trunk and C fiber response of spinal cord WDR neurons were decreased by at least 70% (mean) after topical application of capsaicin (250 micrograms) on the nerve, but A compound action potentials and A fiber response did not change significantly. It indicated that capsaicin blocked C fiber conduction selectively. Electroacupuncture (EA: 100 Hz, 0.1 ms, 3V) applied on left Zusanli (S36) and Sanyinjiao (Sp6) points inhibited C fiber response of spinal WDR neurons in the right side. The effect was similar to animal behavior analgesia elicited by EA. After applying capsaicin (250 micrograms) topically on left sciatic nerve, the inhibitory effect of EA on WDR neurons remained essentially intact (from 61.3 +/- 12.0% to 59.0 +/- 11.6%, n = 6, P greater than 0.05). It indicated that C fiber was not important in EA analgesia. Noxious heat (NH) applied on left hind paw by immersing the hind paw into 52 degrees C water inhibited C fiber response of spinal WDR neurons in the right side. It was called diffuse noxious inhibitory controls (DNIC). After applying capsaicin (250 micrograms) topically on left sciatic nerve, the inhibitory effect of NH on WDR neurons was dramatically decreased (from 77.7 +/- 8.5% to 8.1 +/- 8.9%, n = 6, P less than 0.001). It indicated that C fiber was important in DNIC. Both inhibitory effects of NH and EA were not changed by vehicle treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Action Potentials↗
PubMed1991

[Histopathology of experimental hepatic amebiasis].

The method of intrahepatic inoculation was successfully used to establish an experimental model of hepatic amebiasis in golden hamsters for studying its pathological morphology. Three types of macroscopic liver lesions, i.e., isolated abscess, multinodular abscess and ruptured lesion were described. On light microscopy the alteration and the proliferation type lesions were found as distinct pathological characteristics of the early and advanced hepatic amebiasis respectively. The development pattern of hepatic amebiasis and the pathogenicity of Entamoeba histolytica were discussed.

Animals↗
PubMed1990

Structural requirements of iron-responsive elements for binding of the protein involved in both transferrin receptor and ferritin mRNA post-transcriptional regulation.

The synthesis of both transferrin receptor (TfR) and ferritin is regulated post-transcriptionally by iron. This is mediated by iron responsive elements (IREs) in the 5'- and 3'-untranslated regions, respectively, of TfR and ferritin mRNAs. Although these IREs have different sequences, they both form a characteristic stem-loop. We used competition assays and partial peptide mapping of UV-crosslinked ferritin and TfR IRE-protein complexes to show that the cytosolic protein binding to the ferritin 5'-IRE, the iron-responsive element binding protein (IRE-BP), also binds to TfR 3'-IREs. To identify the structural requirements necessary for RNA-protein binding, ferritin IRE RNAs were synthesized which contained altered secondary structures and base substitutions. Affinities of these RNAs for IRE-BP were assayed in RNA-protein binding gels. Substitutions disrupting base-pairing of the stem prevented IRE-BP binding. Substitutions which restored base-pairing also restored IRE-BP binding. We conclude that the IRE-BP binds to both ferritin and TfR IREs and recognizes a particular IRE conformation.

Animals↗
PubMed1990

Brain structures important for solving a sawdust-digging problem in the rat.

Young rats subjected to bilateral lesions to one of 37 different brain sites were initially trained to traverse a narrow runway to reach a goal box containing water. The test involved blocking a portion of the runway with sawdust and determining whether the animals would burrow through the sawdust to gain access to the goal box. Lesions to 30 brain sites produced deficient performance on the sawdust-digging problem. This study, combined with earlier investigation of 11 additional brain sites, shows that 41 of 48 total brain sites are involved in this task. These findings suggest that underlying brain mechanisms include a nonspecific mechanism and several specific mechanisms, such as visuospatial-attentional, visual discrimination, vestibular-proprioceptive-kinesthetic discrimination, response flexibility, but much less place learning mechanisms.

Animals↗
PubMed1990

Heparinless extracorporeal bypass for treatment of hypothermia.

In an attempt to assess the changes occurring to the coagulation profile during internal active core rewarming with partial cardiopulmonary bypass (CPB) without heparin anticoagulation, five pigs were anesthetized, and a model for severe to moderate hypothermia was created. Femoral-femoral bypass with Bio-Pump, heat exchanger, and a membrane oxygenator were used during the rewarming for 64.8 +/- 8.5 minutes. There were no statistically significant changes in platelet count, platelet index, activated clotting time (ACT), partial thromboplastin time (PTT), prothrombin time (PT), fibrinogen, fibrinogen index and fibrin split products (p greater than 0.05). There were no thromboembolic sequelae seen at autopsy. The components of the CPB circuit showed no signs of formation of aggregates or thrombi. The results of this study are attributed to the nonthrombogenic, atraumatic design of the Bio-Pump and the enhanced physiological fibrinolysis seen in the first hour of CPB. We concluded that heparinless CPB may serve as a safe alternative for active core rewarming for severe to moderate hypothermia.

Animals↗