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Biomedical subjects

M He

Publications and source records attributed to M He.

At least 145 records · Page 8Linked to original sources
PubMed1995

Inhibition of (S)-warfarin metabolism by sulfinpyrazone and its metabolites.

Sulfinpyrazone markedly potentiates the anticoagulant effect of warfarin. The increased clotting time is accompanied by a marked decrease in the clearance of (S)-warfarin by virtue of a decrease in the P4502C9-catalyzed formation clearance to its major and inactive metabolite (S)-7-hydroxywarfarin. These data suggested that the mechanism of the drug interaction might be mediated through the inhibition of the catalytic activity of P4502C9 by sulfinpyrazone. However, initial human liver microsomal studies indicated that the in vitro Ki, for inhibition of (S)-7-hydroxywarfarin formation by sulfinpyrazone is at least 25-fold higher than the therapeutic concentration of sulfinpyrazone in vivo. This result implied that other inhibitors probably contribute to the interaction. Kinetic studies conducted on sulfinpyrazone and two major metabolites, sulfinpyrazone sulfide and sulfinpyrazone sulfone, in microsomes prepared from three human livers give mean Ki's of 230 microM, 17 microM, and 73 microM respectively. Because sulfinpyrazone and its sulfide metabolite attain comparable plasma concentrations during the course of therapy, our inhibition results suggest that the sulfide metabolite is likely to be the primary species responsible for the inhibition of P4502C9-catalyzed formation of (S)-7-hydroxywarfarin and the decrease in (S)-warfarin clearance in vivo.

Drug Synergism↗
PubMed1995

[PCNA expression and AgNORs image analysis as factors in judging the prognosis of lung cancer].

The expression of PCNA and AgNORs count or AgNORs' area in 86 cases of primary lung cancer and 10 cases of inflammatory pseudotumor were studied by S-P immunostaining of monoclonal antibody (PC-10) and image analysis system. The results showed that the proliferating index (PI) of PCNA positive cells was closely related to AgNORs count and AgNORs' area measured. Both were correlated with the histological type, and the TNM status, decreased with the increasing degree of histological differentiation. Survival analysis showed that the PI and the results of AgNORs image analysis in surgical patients who survived over 5 years were significantly lower than those who survived less than 3 years. The results suggest that immunostaining of PCNA and AgNORs image analysis are useful in judging the malignant degree and the prognosis of primary lung cancer.

Adenocarcinoma, Bronchiolo-Alveolar↗
PubMed1995

Characterization of a progesterone-binding, three-domain antibody fragment (VH/K) expressed in Escherichia coli.

The heavy chain variable region (VH) and the kappa light chain of the anti-progesterone monoclonal antibody (mAb) DB3, have been expressed as a single-chain three-domain polypeptide, designated VH/K, and secreted into the periplasmic space of Escherichia coli (E. coli). The linker sequence was derived from the VH-CH1 elbow region. The C kappa domain provides a sensitive detection tail for Western blotting and enzyme-linked immunosorbent assay (ELISA). Periplasmic extracts of transformed E. coli contained material that bound progesterone and related steroids with similar specificity and affinity to DB3, and displayed the DB3 idiotype and kappa chain epitopes. Reference to the crystal structure of DB3 suggests that all the characteristics of the combining site interaction with steroids are retained in the bacterially expressed material. Western blotting demonstrated material with a molecular weight equivalent to three domains after reduction, but six domains in the unreduced state, suggesting that the VH/K polypeptide is assembled in the periplasm as a disulphide-bridged dimer. The VH/K construct provides a novel route to expression of antibody combining sites in E. coli for antibody engineering.

Animals↗
PubMed1995

Clinical application of monoclonal antibody-drug conjugate to immunotargeting chemotherapy of bladder cancer.

Monoclonal antibody-drug conjugate was applied in a clinical trial for patients with bladder cancer. Monoclonal antibody HB7A from a mouse splenocyte immunized against human bladder cancer was used as a drug carrier. The anti-cancer drug adriamycin (ADR) was bound to HB7A through a dextran (DEX) bridge to form the conjugate HB7A-DEX-ADR. The in vitro cytotoxic effect of the conjugate on BIU-87 bladder cancer cells was similar to that of free ADR and the mixture of HB7A and ADR. Seven patients with bladder cancer were given HB7A-DEX-ADR intravenously. The immunoperoxidase studies of the resected specimens showed that HB7A was localized specifically in cancer, and histological studies revealed degenerative and necrotic changes of the tumor cells. Patients receiving the conjugate did not experience serious side effects. This study suggests that immunotargeting chemotherapy with HB7A-DEX-ADR is well tolerated by patients and its cytotoxicity on tumor is substantial.

Aged↗
PubMed1995

[A new technique of esophagogastrostomy above the aortic arch--covering and suspending of the anastomotic area with mediastinal pleura flap].

The postoperative anastomotic leakage is the most severe complication and the principal cause of death after resection of esophageal carcinoma. Especially, anastomosis above the aortic arch is accompanied by high incidence of anastomotic leakage usually with fatal prognosis. The authors improved the conventional method of anastomosis by using a mediastinal pleura flap to cover and suspend the anastomotic area with excellent result. From January of 1990 to April of 1995, 210 patients with esophageal carcinoma were so treated. No anastomotic leakage or stricture developed. Only one patient died of extensive myocardial infarction with an overall mortality of 0.47%. The major merit of this procedure was that the mediastinal pleura flap could cover the area of anastomosis and sustain the dragging force acting on this area, providing a better condition for the anastomosis to heal and thus reducing the possibility of anastomotic leakage.

Adult↗
PubMed1994

The actin gene family in the oriental fruit fly Bactrocera dorsalis. Muscle specific actins.

The actin protein is a critical protein in eukaryotic cells. Four actin genes, constituting what appear to be a set of muscle specific actin genes, have been isolated from the genome of the oriental fruit fly Bactrocera dorsalis. DNA sequences have been determined for the coding as well as 3' and 5' flanking regions for each of these genes. These genes have also been characterized in terms of RNA expression patterns, and comparisons have been made to actin genes from other species. Consistent with other actins, there is a high degree of amino acid sequence conservation in the coding regions of these genes. However, even within the coding regions codon usage patterns in the oriental fruit fly are quite different from some other well characterized species. In addition, the DNA sequences in the intermediate 3' and 5' flanking regions exhibit virtually no detectable sequence homology both within and between species. In terms of introns, three of the four actin genes from the oriental fruit fly described here have a single intervening sequence. Two of these genes share the same intron position with the two muscle specific actin genes act79B and act88F from Drosophila melanogaster and with one muscle specific actin gene CcA1 from the Mediterranean fruit fly, Ceratitis capitata. Another gene from the oriental fruit fly shares the same intron position as the muscle specific actin gene act57B from D. melanogaster. Such conservation of intron positioning between species is highly unusual among previously characterized actin genes. Using unique sequences found in the 3' untranslated regions, gene specific probes have also been constructed. These have been used to detect the expression patterns of individual genes in a temporal and spatial manner. Each of the four genes examined here show differential patterns of expression. The patterns indicate that all four genes are most likely to encode muscle specific actins.

Actins↗
PubMed1994

Oligodendroglial signal transduction systems are developmentally regulated.

Studies from several laboratories indicate that oligodendroglia exhibit signal transduction systems that can be activated by classical neurotransmitters. Previous studies from this laboratory indicate that oligodendroglia express neuroligand receptors linked to the regulation of Ca(2+)i. Experiments presented in this article were designed to determine if developmental processes that influence the ability of oligodendroglia to respond to neuroligands with an increase in Ca2+i proceed either in vitro or in vivo. Findings support the view that developmental processes markedly affected the sensitivity of these cells to both purinergic and cholinergic receptor agonists, whereas their responsiveness to either histamine or bradykinin appeared relatively stable over time. Approximately 90 and 75% of oligodendroglia responded to ATP or carbachol, respectively, after 4 days in vitro, whereas < 10% of these cells responded to either of these neuroligands after 8 days in vitro. The decrease in the percentage of oligodendroglia responding to ATP, but not carbachol, could be prevented by including dibutyryl cyclic AMP in the culture medium during the final 4 days in vitro. However, once the loss in responsiveness to ATP had occurred, it could not be reversed by exposure to dibutyryl cyclic AMP. Developmental changes in the ATP sensitivity of oligodendroglia occurred in cells expressing galactocerebroside and myelin basic protein. The neuroligand sensitivity of oligodendroglia isolated from either neonatal, 2-, 3-, or 5-week-old spinal cord was examined to determine if developmental changes in oligodendroglial Ca2+ regulation occurred in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1994

Expression and export in Escherichia coli of fusion proteins containing carboxy-terminally located honeybee prepromelittin.

The aim of this work was to express a eukaryotic pre-protein in Escherichia coli so that it could be obtained intact, without cleavage, by bacterial leader peptidase. To this end, cDNA coding for honeybee prepromelittin was ligated to the 3' end of genes coding for truncated forms of either Protein A or beta-galactosidase (beta-Gal) under the control of inducible promoters, with an oligonucleotide coding for the Factor Xa cleavage site at the junction between the two proteins. The Protein A fusion was expressed in good yield, and about 80% of it formed inclusion bodies. The prepromelittin section of the Protein A fusion caused some export of the intact fusion protein to the growth medium. The prepromelittin beta-Gal fusion was expressed in low yield and became associated with the E. coli cytoplasmic membrane. Its expression was toxic to E. coli. Thus, the synthesis of a full-length eukaryotic pre-protein in E. coli is best achieved when the fusion protein forms inclusion bodies.

Animals↗
PubMed1994

Expression and secretion of alpha-amylase and glucoamylase in Saccharomyces cerevisiae.

alpha-Amylase genes of Bacillus licheniformis and glucoamylase cDNA of Aspergillus niger were ligated to a E. coli-yeast shuttle vector. The resultant plasmid was used to transform Saccharomyces cerevisiae to construct starch-degrading yeast strain. The results of enzyme activity assay and enzyme property analysis show that alpha-amylase and glucoamylase genes have been expressed simultaneously in yeast under the control of promoters and terminators of yeast MF-alpha 1 factor and PGK genes and over 99% of enzyme activities were secreted to the medium. The engineered yeast strain hydrolyses 97% of the starch (10%) in the medium after 6 days. The recombinant plasmid exists stably in yeast.

Aspergillus niger↗
PubMed1994

Two novel human serine/threonine kinases with homologies to the cell cycle regulating Xenopus MO15, and NIMA kinases: cloning and characterization of their expression pattern.

Using polymerase chain reaction (PCR)-based methods, we have isolated cDNA clones of two new members of serine/threonine kinases, STK1 and STK2, from a cDNA library constructed from the BT-20 human breast cancer cell line. STK1 is transcribed as a 1.4 kilobase (kb) mRNA encoding for a protein of 346 amino acids. Based on amino acid sequence analysis, STK1 is 86% identical to the Xenopus p40mo15, a cdc2-related serine/threonine kinase recently found to be the activating kinase for p34cdc2 and p33cdk2. Thus, STK1 is most likely the human homologue of MO15. An alternatively spliced STK1 message expressed variably in cell lines and in primary carcinomas generates a predicted 58 amino acid protein that lacks the kinase domain. STK2 is transcribed into a 4.0 kb mRNA encoding for an 841 residue protein which exhibits 50% identity in the kinase domain with the mouse nek1 gene product, the relative of the fungal G2-M regulator, nimA. STK1 and STK2 display a variable pattern of expression among a series of primary carcinomas as well as cancer cell lines. Both STK1 and STK2 were expressed at the highest levels in the heart but were also detected in all other organs tested. In embryonal tissues, lower levels of expression were noted. Using cell cycle inhibitors, we have shown that both STK1 and STK2 mRNA levels remain relatively invariant through the cell cycle. Chromosomal assignment has localized STK1 on chromosome 2pcen-2p15, a region implicated in hereditary non-polyposis colorectal carcinoma, and STK2 on chromosome 3p21.1, a region frequently showing chromosomal alterations in renal cells carcinomas.

Amino Acid Sequence↗
PubMed1993

Changes in levels of pancreatic endoplasmic reticulum proteins that function in translocation and maturation of secretory proteins in response to cholecystokinin.

Two pathways operate to target newly-synthesised proteins to the endoplasmic reticulum. In one, the signal recognition particle attaches to the signal sequences of nascent chains on ribosomes and slows or stops translation until contact is made with the docking protein at the membrane. The second operates via molecular chaperons. The pathways converge at the level of a 43 kDa signal binding protein integrated into the membrane, where translocation through a proteinaceous pore is initiated. In the lumen, proteins fold and disulphide formation is catalysed by the enzyme protein disulphide isomerase. The heavy chain binding protein may attach to unassembled or unfolded proteins and prevent their exit from the ER to the Golgi. Cholecystokinin (CCK) treatment increases the biosynthesis and secretion of pancreatic proteins, increases the levels of PDI and the 43 kDa binding protein, and reduces levels of BiP. These proteins may be possible targets for genetic manipulation to improve processing of heterologous proteins from cultured mammalian cells.

Acids↗
PubMed1993

Specificity of factor Xa in the cleavage of fusion proteins.

The precursor protein honey bee prepromelittin has been expressed as a fusion protein in Escherichia coli joined to the C-terminus of a truncated form of the bacteriophage gene 10 protein via an engineered recognition sequence for Factor Xa. Factor Xa was found to cleave poorly at the engineered site, giving a low yield of the required prepromelittin. In contrast, cleavage on the C-terminal side of the sequence VLGR at residue 67 in the gene 10 sequence proceeded in high yield. Factor Xa may be inhibited by adjacent hydrophobic sequences on the C-terminal side of a potential cleavage site.

Amino Acid Sequence↗