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Biomedical subjects

X Ma

Publications and source records attributed to X Ma.

At least 343 records · Page 19Linked to original sources
PubMed1994

Indole alkaloids from Peschiera laeta that enhance vinblastine-mediated cytotoxicity with multidrug-resistant cells.

Coronaridine [1], conoduramine [2], and voacamine [3], three indole alkaloids isolated from Peschiera laeta, have been found to enhance the cytotoxic response mediated by vinblastine [4] with multidrug-resistant KB cells. Inhibition of vinblastine binding with membrane vesicles isolated from this cell line was also assessed, and the bisindole alkaloids conoduramine [2] and voacamine [3] were found to be more potent inhibitory agents than the monomeric alkaloid, coronaridine [1]. Thus, these compounds appear to function by binding with P-glycoprotein.

Alkaloids↗
PubMed1994

Stability study of drug-loaded proteinoid microsphere formulations during freeze-drying.

Drug-loaded proteinoid microspheres were freeze-dried to facilitate shipping and handling and to enable long term storage. Heparin was chosen as the model drug in developing the optimum lyophilization process. The factors influencing the integrity of either heparin-loaded or unloaded ('empty') proteinoid microspheres during freeze-drying were determined, with emphasis on: selecting an optimum freezing and resuspending temperature; choosing an appropriate cryoprotectant and its optimum concentration in the formulation; and, designing a suitable method for formulating the microspheres. Freezing at/below -70 degrees C was found to minimize damage to the microspheres. Addition of sugars, such as trehalose and lactose, as cryoprotectants, further increased the stability of the heparin-loaded microspheres during freeze-drying. The optimum trehalose or lactose concentrations were determined to be 5% (w/v). Using the optimumized lyophilization process described in this manuscript, microspheres remained intact during freeze-drying. The freeze-dried microspheres were stable for at least three months post-lyophilization.

Cryoprotective Agents↗
PubMed1994

Generation of alginate-poly-l-lysine-alginate (APA) biomicrocapsules: the relationship between the membrane strength and the reaction conditions.

Alginate-poly-l-lysine-alginate (APA) microcapsules have proven effective in protecting enclosed live cells from immune rejection following transplantation into experimental animals, thereby eliminating the need for immunosuppressive therapy. However, in order for the capsules to remain intact for extended periods in vivo, the thickness of the membrane material must be optimized. In this study, the membrane thickness was examined as an indicator of membrane strength and measured under different reaction conditions. The thickness was found to increase 1) from 4.6 microns to 6.6 microns with an increase in the concentration of sodium alginate from 1.25 (w/v) to 2.0% (w.v); 2) from 4.2 microns to 6.2 microns with an increase in the concentration of the calcium solution from 20 mM to 100mM; 3) from 3.9 microns to 10.3 microns with an increase in the concentration of poly-l-lysine (PLL) from 0.02% (w/v) to 0.08% (w/v); and 4) from 2.3 microns to 7.4 microns with an increase in the reaction time with the PLL from two to seven minutes. On the other hand, membrane thickness decreased 1) from 9.8 microns to 8.6 microns with an increase of the pH in the PLL solution from 5.8 to 9.2; 2) from 13.2m to 5.8 microns with an increase in the molecular weight of PLL from 14,000 to 57,000; 3) from 8.4 microns to 6.0 microns with an increase in the treatment time with 0.9 (w/v) NaCl solution from zero to fifteen minutes and; 4) from 7.5 microns to 6.1 microns with an increase in the treatment time of the second sodium alginate coating from zero to ten minutes. Membrane thickness was inversely proportional to capsule volume expansion during membrane synthesis. By replacing calcium chloride by calcium lactate and eliminating the use of CHES in the construction of capsule membranes, we improved the strength and biocompatibility of our capsules, as evidenced by marked improvements in the survival rates of diabetic mice treated with islet transplants enclosed in the new capsules. These results indicate that it is possible to obtain optimal membrane thickness for a given purpose by creating specific reaction conditions under which membranes are synthesized.

Alginates↗
PubMed1994

Electrophysiologic effects of total flavones of Hippophae rhamnoides L on guinea pig papillary muscles and cultured rat myocardial cells.

The effects of total flavones of Hippophae rhamnoides L (TFH) were evaluated using conventional microelectrode technic. After administration of TFH 100-200 mg.L-1, the action potential duration of 50% repolarization (APD50) was shortened both in cultured rat myocardial cells and in guinea pig papillary muscles. The slope of phase 4 of depolarization (SP4) in cultured rat myocardial cells was decreased and the contractile force (CF) in guinea pig papillary muscles was weakened. Arrhythmias evoked by strophantin G in guinea pig papillary muscles were suppressed by TFH 100 mg.L-1. These findings suggested that the influence of TFH on myocardial cells may be resulted mainly from its inhibition of Ca2+ influx and its interference with intracellular Ca2+ reservoir.

Action Potentials↗
PubMed1993

Acute 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure results in enhanced tyrosylphosphorylation and expression of murine hepatic cyclin dependent kinases.

An increase in tyrosine phosphorylation of two hepatic S9 proteins migrating at 34 and 33 kDa that cross-reacted with anti-PSTAIR antibody on immunoblots was seen 24 h after administration of a single dose of 0.25, 0.5, 1 or 2 micrograms 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)/kg to C57BL/6J female mice. Two hepatic S9 proteins migrating at 34 and 33 kDa that cross-reacted with anti-cdc2 C-terminus antibody on immunoblots were observed in corn oil control mice; increased expression of these proteins was seen with increasing doses of TCDD. A maximal increase in expression of 3-times the control was observed at 1 and 2 micrograms TCDD/kg for both p34 and p33. The stimulation of enhanced tyrosylphosphorylation and expression of cyclin dependent kinases p34cdc2 and p3cdk2 by TCDD is consistent with a mechanism of action of TCDD toxicity associated with stimulation of cellular proliferation.

Animals↗
PubMed1993

Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells.

Natural killer cell stimulatory factor or interleukin 12 (NKSF/IL-12) is a heterodimeric cytokine produced by monocytes/macrophages, B cells, and possibly other accessory cell types primarily in response to bacteria or bacterial products. NKSF/IL-12 mediates pleiomorphic biological activity on T and NK cells and, alone or in synergy with other inducers, is a powerful stimulator of interferon gamma (IFN-gamma) production. IL-10 is a potent inhibitor of monocyte-macrophage activation, that inhibits production of tumor necrosis factor alpha (TNF-alpha), IL-1 and also IFN-gamma from lymphocytes acting at the level of accessory cells. Because TNF-alpha and IL-1 are not efficient inducers of IFN-gamma, the mechanism by which IL-10 inhibits IFN-gamma production is not clear. In this paper, we show that IL-10 is a potent inhibitor of NKSF/IL-12 production from human peripheral blood mononuclear cells activated with Staphylococcus aureus or lipopolysaccharide (LPS). Both the production of the free NKSF/IL-12 p40 chain and the biologically active p70 heterodimer are blocked by IL-10. NKSF/IL-12 p40 chain mRNA accumulation is strongly induced by S. aureus or LPS and downregulated by IL-10, whereas the p35 mRNA is constitutively expressed and only minimally regulated by S. aureus, LPS, or IL-10. Although IL-10 is able to block the production of NKSF/IL-12, a powerful inducer of IFN-gamma both in vitro and in vivo, the mechanism of inhibition of IFN-gamma by IL-10 cannot be explained only on the basis of inhibition of NKSF/IL-12 because IL-10 can partially inhibit IFN-gamma production induced by NKSF/IL-12, and also, the IFN-gamma production in response to various stimuli in the presence of neutralizing antibodies to NKSF/IL-12. Our findings that antibodies against NKSF/IL-12, TNF-alpha, or IL-1 beta can significantly inhibit IFN-gamma production in response to various stimuli and that NKSF/IL-12 and IL-1 beta can overcome the IL-10-mediated inhibition of IFN-gamma, suggest that IL-10 inhibition of IFN-gamma production is primarily due to its blocking production from accessory cells of the IFN-gamma-inducer NKSF/IL-12, as well as the costimulating molecule IL-1 beta.

Antigen-Presenting Cells↗
PubMed1993

Mutagenic analysis of AMP nucleosidase from Escherichia coli. Deletion of a region similar to AMP deaminase and peptide characterization by mass spectrometry.

AMP nucleosidase (EC 3.2.2.4) from Escherichia coli and AMP deaminase (EC 3.5.4.6) from bakers' yeast are proposed to regulate cellular AMP levels under allosteric control of the activator ATP and the inhibitor, PO4. Both enzymes contain catalytic sites which bind AMP and regulatory sites which bind ATP. The deduced amino acid sequences of the proteins revealed only one region of homology in which six of eight amino acids are identical. A similar sequence is found in glyceraldehyde-3-phosphate dehydrogenase, phoE, ras proteins, RNA polymerase, K(+)-ATPase, nucleolin, and other proteins expected to have nucleotide or phosphate binding properties. In the crystal structure of glyceraldehyde-3-phosphate dehydrogenase, this sequence is part of the NAD(+)-binding site. The function of these amino acids was explored with a deletion mutant of AMP nucleosidase. The protein was over-produced in a pTZ construct using the AMP nucleosidase promoter which resulted in approximately 30% of the total protein as the desired enzyme. The mutation was characterized by DNA sequence analysis and by direct analysis of the peptides using high performance liquid chromatography-mass spectrometry. Deletion of amino acids 128-135, corresponding to DGSELTLD, produced an enzyme with a 20-fold decrease in Vmax but with smaller changes in substrate saturation kinetics, activation by MgATP, inhibition by inorganic phosphate, and inhibition by the tight-binding inhibitor, formycin 5-phosphate. The deletion mutant of AMP nucleosidase exhibits hysteresis in establishing a steady-state rate of product formation which is most pronounced in the absence of MgATP. These results establish that the sequence DGSELTLD in E. coli AMP nucleosidase is not required for binding of AMP, MgATP, or inorganic phosphate. However, the mutant enzyme has a structural defect related to the polymerization state which delays the onset of catalysis and decreases the catalytic efficiency.

AMP Deaminase↗
PubMed1993

Purification and characterization of a nuclear DNA-binding factor complex containing topoisomerase II and chromosome scaffold protein 2.

In a search for factors that influence the process of erythroid differentiation at the molecular level, we have identified UB2, a nuclear protein factor that was originally observed for its ability to bind to a very specific and highly conserved sequence motif present in human, mouse, rabbit, and chicken beta-globin genes, as well as carbonic anhydrase I, c-myb, and the immunoglobulin heavy chain enhancer region. It was also observed for its appearance in undifferentiated but not differentiated mouse erythroleukemia cells. Purification of UB2 by DEAE-cellulose chromatography and repeated passages through a DNA affinity column, revealed a complex pattern with three major components of 170, 116, and 48 kDa, respectively. The 170-kDa protein was identified as topoisomerase (topo) II by Western blot analysis, catalytic assays, and antibody interference with UB2 binding. The complex topo II in UB2, however, has a more stringent sequence requirement for DNA binding than does topo II. The 116-kDa protein has been determined to be a proteolytic product of topo II. The chromosome scaffold protein 2 (135 kDa) copurified with UB2, and anti-scaffold protein 2 serum inhibited UB2 binding to DNA.

Animals↗
PubMed1993

Interaction of heparin with polyallylamine-immobilized surfaces.

A new method to bind ionically and remove heparin from solution and dilute serum is described. Utilizing cellulose diacetate (CA) as the polymer substrate, a cationic polymer chain--poly(allylamine)-PALA--was immobilized directly onto the CA surface and onto the surface using poly(ethylene glycol) (PEG) spacer groups. The ionic interaction between the anionic heparin molecule and the cationic PALA polymer is specific and effective to remove heparin from the bulk solution. The binding properties of heparin onto the PALA and PEG-PALA surfaces were examined. The effects of PEG spacers on heparin binding onto the PALA-immobilized surface were investigated by varying the Mw of PEG spacers. PALA (Mw 8500)-immobilized surfaces exhibited enhanced heparin binding. The maximum heparin binding was observed in the region of PEG Mw 2000-4000. For the high-molecular-weight PALA (Mw 50,000)-immobilized surfaces, heparin binding was independent of the molecular weight of PEG. PEG spacers were also evaluated for their ability to prevent or decrease protein (albumin) adsorption. It was observed that at high albumin concentrations, the adsorption of proteins decreased with increasing chain length of PEG, up to Mw 3400. These observations suggest that low-molecular-weight PALA (Mw 8500)-immobilized CA surfaces with PEG spacers (Mw 3400) may provide increased heparin binding capacity and decreased protein adsorption.

Adsorption↗
PubMed1993

Sequence and gene structure of the hepatitis E virus isolated from Myanmar.

Hepatitis E virus (HEV) is a causative agent of enterically transmitted non-A, non-B hepatitis. Hepatitis E occurs not only in sporadic forms but also in epidemic outbreaks in the developing world. We have revealed the nucleotide and predicted amino acid sequences of full cDNA of HEV isolated from sporadic hepatitis E of Myanmar. The genome is 7194 nucleotides long, followed by a poly(A) tail, and has three open reading frames. The nonstructural gene is located in the 5' terminus, while the structural gene is situated in the 3' terminus. Our HEV strain has 98.5% nucleic acid identity with the HEV strain cloned by workers at Genelabs Incorporated from Myanmar. The difference is point nucleotide substitutions. There is a high degree of nucleotide relatedness among HEVs isolated from the same geographical location.

Amino Acid Sequence↗
PubMed1993

Nursing diagnoses for psychiatric patients in China.

This report describes the development of a preliminary Chinese psychiatric classification system with nine categories and 211 separate diagnoses, and the corresponding diagnostic interview that enables psychiatric nurses in China to systematically assess the presence or absence of the various diagnoses. Using this interview with 30 representative inpatients, the authors identified 152 of the 211 (72%) diagnoses. On average, each patient was given 26 different nursing diagnoses. The interrater reliability of the four nurses who independently coded the diagnostic interview for these 30 patients was excellent: in over 77% of the identified diagnoses the generalized kappa was greater than 0.75. These findings show that further work on this classification of nursing diagnoses is warranted.

Adolescent↗
PubMed1993

An epidemic outbreak of hepatitis E in Yangon of Myanmar: antibody assay and animal transmission of the virus.

An epidemic outbreak of hepatitis E occurred in an army recruit camp of Yangon, Myanmar, in October 1989. One hundred and eleven patients among 600 residents were hospitalized. As high as 83.7% of these patients were positive for the acute phase antibody against hepatitis E virus by an enzyme-linked immunosorbent assay developed in our laboratory. Also, 30.6% of 49 symptom-free residents examined were positive for the antibody. We prepared a stool extract from six patients and inoculated it into 10 rhesus monkeys for a series of three sub-passages. All of them developed acute biochemical hepatitis along with an elevation of antibody levels. A rechallenge with viruses of the present outbreak failed to provoke hepatitis in two monkeys that had previously recovered from acute hepatitis caused by an isolate of sporadic hepatitis E of the same area. Similarly, the rechallenge of the sporadic strain did not induce hepatitis in two monkeys that had been previously infected with the epidemic virus. These data suggested that the subjects would obtain neutralizing antibodies against the hepatitis E virus once infected, and many adult inhabitants of the endemic area had no protective antibodies and were still susceptible to hepatitis E infection.

Adult↗
PubMed1993

Porcine pancreatic islets: isolation, microencapsulation, and xenotransplantation.

To provide a plentiful source of pancreatic islets for future clinical transplants into diabetic patients, we have developed a simple and reliable method to isolate porcine islets of a high degree of purity. Porcine pancreata were perfused and digested with collagenase, and the islets were then purified on dextran density gradients. In order to avoid any damage to the islets, no mechanical devices nor any strenuous treatment was employed. As many as 5 x 10(5) islets were isolated from a single porcine pancreas. Islets were encapsulated in alginate-polylysine-alginate membranes with the aid of an electrostatic droplet generator. In vitro studies demonstrated that the isolated islets secreted insulin in response to glucose and 3-isobutyl-L-methylxanthine (IBMX) challenge for at least 4 weeks. Perifusion studies showed that the kinetics of insulin release from the encapsulated islets was similar to that exhibited by free islets. In in vivo studies, 18 diabetic BALB-c mice were transplanted with 1,500-2,500 encapsulated islets each. In 13 recipients, the diabetic condition was reversed for at least 85 days. When capsules were removed from 2 transplant recipients, their diabetic condition quickly recurred.

Animals↗
PubMed1993

[Recording intracellular potentials of viscero-nociceptive neurons in posterior group of thalamic nuclei (PO) and their labeling with HRP intracellularly in cats].

Intracellular potentials (ICP) of the neurons in PO evoked by stimulating greater splanchnic nerve (GSN) were recorded in 42 cats. The resting potentials and action potentials induced by stimulating GSN were 53.74 +/- 11.68 mV and 29.44 +/- 9.22 mV respectively. Owing to intensity of the stimulation we can record EPSP and single spike or polyspikes on it. The action potentials derived from stimulating GSN (related action potentials) were inhibited by injection of morphine intravenously. On the basis of recording ICP, we also labeled 2 related neurons with stimulation of GSN and 2 unrelated ones. Differences between two kinds of the neurons were compared preliminarily. The ICP of the related neuron with stimulating GSN were inhibited by stimulating Amygdala too. Factors influencing amplitude of action potential in intracellular recording, criteria of determining nociceptive neuron and inhibition derived from stimulating Amygdala were also discussed.

Action Potentials↗