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Biomedical subjects

J Ma

Publications and source records attributed to J Ma.

At least 577 records · Page 32Linked to original sources

Transzygomatic approach to the tentorial incisura: surgical anatomy.

A simple transzygomatic approach to the middle fossa centered on the inferior retraction of the temporal muscle after transection of the zygomatic arch is described. This approach allows a very low basal exposure of the middle and temporal fossa, minimizing the amount of temporal lobe retraction needed to approach intradural and extradural lesion in this location. This approach is simple, easily performed, does not require extensive skull base removal, and still offers excellent visualization of the middle fossa and of the region of the tentorial incisura.

Journal Article↗

[Method of producing fusarin C in perlite-liquid culture medium].

For researching the biosynthesis labelled Fusarin C(Fc) by Fuscarium moniliforme, a more quick and convenient method of Fusarin C production and purification were established, and a good liquid culture medium consisted of different kinds organic matters (hydroxy proline, sucrose and glycerin), inorganic salts and perlite replaced corn grit medium. The perlite-liquid culture medium inoculated with the strain of F. moniliforme yields 936mg Fc/kg organic matter with in 14 days of incubation at 28 degrees C. As compared with the corn grit medium, the amount of Fc from perlite-liquid medium was more than that from corn grit medium (831mg Fc/kg corn grit). In all experiments both thin-layer chromatography and high-pressure liquid chromatography were used to confirm the presence of Fc. parameters which were important for the optimal biosynthesis of Fc included hydroxy proline and sucrose concentrations, incubated time/temperature and amount of perlite. The 40g of sucrose/L liquid culture was optimal concentration for Fusarin C production. Of three contained N-matter tested, hydroxy proline was the best sources of N-atom for Fusarin C. Under the absence of hydroxy proline, the Fc wasn't synthesized in perlite-liquid culture medium by F. moniliforme. A culture time/temperature study of Fc production was done, and the optimal Fc amounts was synthesized after incubation for 14 days at 28 degrees C on perlite-liquid culture medium.

Aluminum Oxide↗

[Production of CA125 in cell lines derived from human ovarian carcinoma: in relation to the cell cycle].

The association of the production of CA125 with the cell cycle was investigated in two cell lines derived from human ovarian cancer, one from a serous cystadenocarcinoma (HTOA) and the other from a mucinous cystadenocarcinoma (RMUG-s). HTOA and RMUG-s cells secreted CA125 at about 50 and 30U/ml/10(5) cell/24hr, respectively, in the logarithmic growth phase and at about 75 and 100U/ml/10(5) cell/24hr in the steady phase. Analysis by FCM revealed that cultures of both cell lines cultured for 7 days contained more cells in the G0/G1 phase and less cells in the S phase than those cultured for 3 days. The positive rate of immunologically stained DNA polymerase alpha was 31% in HTOA cells and 39% in RMUG-s cells after cultivation of the cells for 3 days. The addition of EGF at 0.01, 0.1 or 1.0nM did not affect the production of CA125 in HTOA or RMUG-s cells while the addition of NaBT at 1, 3 and 5mM raised production in both cell lines as the dose rose. With RMUG-s cells, the addition of EGF at 0.01nM to the culture media accelerated both logarithmic and steady phase growth without a significant change in the production of CA125. In contrast, the addition of NaBT at 1mM suppressed growth, but tended to increase the production of CA125 per cell. With the effect of EGF on the cell cycle of both cell lines, cells in the S phase increased by about 20% as compared with the control, 48 hours after its addition at 0.01nM. In contrast, after cultivation for 48 hours in the presence of 1mM NaBT, cells in the S phase were decreased while those in the G0/G1 phase increased. The results presented above suggested the possibility that some factors other than the cell cycle were involved in the production of CA125. There also is close correlation between cells in the G0/G1 phase and the production of CA125 in the culture of human ovarian cancer cells.

Antigens, Tumor-Associated, Carbohydrate↗

Rapid purification of a 110-kilodalton hemolysin of Actinobacillus pleuropneumoniae by monoclonal antibody-affinity chromatography.

An efficient, single-step method for purification of the 110-kilodalton (kDa) hemolysin of Actinobacillus pleuropneumoniae was developed. An immunoaffinity column was made by cross-linking murine monoclonal antibody 8C2 to the 110-kDa hemolysin of A pleuropneumoniae strain J45 serotype 5 to protein A-agarose beads. Purified hemolysin with high hemolytic activity was obtained after washing the column with phosphate-buffered saline solution, and eluting the hemolysin with 50 mM diethylamine, pH 11.0. The same column was also used to purify the hemolysin from A pleuropneumoniae strain 4074 serotype 1. The purification procedure could be completed within 5 hours, and almost 50% of the total hemolytic activity and hemolysin protein was recovered in pure form.

Actinobacillus pleuropneumoniae↗

[Expression of EGF receptors in cell lines derived from female genital cancers and enhancement of the therapeutic effect of anticancer drugs by EGF].

Manifestation of EGF receptors and enhancement of an anticancer agent by EGF were studied in cultured cells derived from female genital cancers. 1) The numbers of EGF-receptors of SKG-3a, RMUG-s, HUOA, A-431 and HEC-1 were 1.22 x 10(4), 6.94 x 10(4), 2.75 x 10(4), 5.25 x 10(5) and 0.92 x 10(4) sites per cell respectively. The values for the dissociation constant (Kd) of RMUG-s, SKG-3a, HUOA and HEC-1 were 340pM, 477pM, 989pM and 2,187pM, respectively. 2) All cell lines were stimulated by EGF at low concentrations and inhibited at higher concentrations. The growth stimulation rates for SKG-3a, HEC-1, HUOA and RMUG-s in the presence of 0.01 nM EGF at 48 hours were 8, 18, 21.1 and 3.7%, respectively. The growth inhibition rates for SKG-3a, A-431, HEC-1, HUOA and RMUG-s in the presence of 1.0 nM EGF at 48 hours were 32, 28, 25.8, 11.2 and 6.8%, respectively. 3) The antitumor effect of CDDP was enhanced by the presence of EGF at both a concentration of 0.01 nM (except for A-431) and a concentration of 1.0 nM, in all cell lines. 4) EGF receptors of RMUG-s and HUOA were decreased by CDDP. The values for RMUG-s and HUOA were 1.71 x 10(4), 0.52 x 10(4) (CDDP 0.4 microgram/ml) and 0.95 x 10(4), 0.29 x 10(4) sites per cell (CDDP 2.0 micrograms/ml), respectively. The effect of CDDP on EGF receptors was not recognized in SKG-3a and HEC-1. In brief, EGF receptors were significantly expressed in cell lines derived from female genital cancers.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Cycle↗

Detecting interactions between eukaryotic proteins in bacteria.

Few convenient genetic assays are available to study protein-protein interactions. This report describes a genetic scheme in E. coli to detect protein-protein interactions based on the concept of cooperative DNA binding of two interacting proteins. The yeast regulatory proteins GAL4 and GAL80, which are known to interact with each other, were used to test the scheme. A fusion protein, LexA-GAL80, was found to exert a cooperative effect on the DNA-binding activity of GAL4 as monitored by a bacterial repression assay.

Base Sequence↗

Oncofetal expression of the human intestinal mucin glycoprotein antigens in gastrointestinal epithelium defined by monoclonal antibodies.

A mucin preparation from a colonic adenocarcinoma was used to prepare monoclonal antibodies (MAbs) that reacted specifically either with normal adult small-intestine mucin antigen(s) (SIMA), or normal adult large-intestine mucin antigen(s) (LIMA). Both SIMA and LIMA show a unique oncofetal pattern of expression. Thus SIMA was expressed in early fetal stomach, large and small intestines but thereafter only in the normal small intestine. SIMA expression was detected immunohistochemically in cancers of the colorectum (82/112) and stomach (48/86). LIMA was detected in the stomach of the early fetus but thereafter only in the normal large intestine. LIMA expression was detected in 61/86 cancers of the stomach. Moreover, both SIMA and LIMA were expressed inappropriately in mucosa adjacent to tumors, indicative of the detection of possible pre-malignant epithelium. We used a sandwich ELISA and biochemical procedures to show that the SIMA and LIMA molecules were large extensively glycosylated multi-unit mucin glycoproteins that differed markedly from each other. SIMA, whether extracted from normal small-intestine or colonic cancers, had a molecular weight above 1.000 kDa, a mean buoyant density 1.33 g/ml and s value of 4.8. LIMA had a molecular weight above 10.000 kDa, a mean buoyant density 1.45 g/ml and an s value 9.5. The SIMA and LIMA epitopes were judged to be carbohydrate in nature by reason of their resistance to harsh physical chemical treatments or protease digestion, and sensitivity to periodate oxidation, neuraminidase or beta elimination. Only the SIMA epitope was sensitive to neuraminidase. In conclusion, MAbs to carbohydrate-dependent epitopes on SIMA and LIMA identify the oncofetal pattern of expression of these distinct intestinal mucin glycoproteins in colonic and gastric carcinoma. These MAbs will be useful in further studies of the significance of oncofetal mucin expression during carcinogenesis.

Adenocarcinoma, Mucinous↗

Generating yeast transcriptional activators containing no yeast protein sequences.

We previously reported that roughly 1% of the short peptides encoded by Escherichia coli genomic DNA fragments act as transcriptional activating regions in yeast when fused to GAL4(1-147), a DNA-binding portion of the yeast transcriptional activator GAL4 (ref. 1). Struhl questioned the conclusion that we had identified new transcriptional activating sequences that function in the absence of yeast transcriptional activating sequences. His criticism was based on two considerations: first, GAL4(1-147) contains an acidic segment (and subsequent experiments have shown that this region contains a weak activating region in vitro); second, attempts to isolate new activating regions failed when the DNA-binding domain of a bacterial repressor, LexA(1-87), was used as the DNA-binding unit. We report here a repeat of our original experiment using the complete LexA molecule LexA(1-202) as the DNA-binding region, instead of GAL4(1-147) or LexA(1-87). We find that, as in the original experiment, about 1% of the short peptides encoded by E. coli genomic fragments act as transcriptional activating regions when fused to intact LexA. All of the new activating regions whose sequences we determined bore an excess of acidic amino acids (see Table 1).

Amino Acid Sequence↗

Strategy for developing a genetically-engineered whole-virus vaccine against HIV.

The production of genetically-engineered, noninfectious virions of human immunodeficiency virus (HIV) represents a novel approach to the development of a safe and effective vaccine for the acquired immune deficiency syndromes (AIDS). Insofar as preparations of inactivated simian immunodeficiency virus (SIV) are now demonstrating protection in immunization-challenge studies in rhesus monkeys, a safe preparation of noninfectious HIV virions produced in a genetically-engineered cell line becomes a logical candidate vaccine for studies in humans. These particles, or pseudovirions, offer distinct advantages over the use of inactivated HIV for human AIDS vaccines. Guarantees of safety without the requirement for inactivation and their potential for structural modification for the modulation of immunogenicity are compelling reasons for the acceptance of HIV pseudovirions as a candidate vaccine in humans.

Acquired Immunodeficiency Syndrome↗

Dihydropyridine-sensitive skeletal muscle Ca channels in polarized planar bilayers. 1. Kinetics and voltage dependence of gating.

Rabbit skeletal muscle transverse tubule (T) membranes were fused with planar bilayers. Ca channel activity was studied with a "cellular" approach, using solutions that were closer to physiological than in previous studies, including asymmetric extracellular divalent ions as current carriers. The bilayer was kept polarized at -80 mV and depolarizing pulses were applied under voltage clamp. Upon depolarization the channels opened in a steeply voltage-dependent manner, and closed rapidly at the end of the pulses. The activity was characterized at the single-channel level and on macroscopic ensemble averages of test-minus-control records, using as controls the null sweeps. The open channel events had one predominant current corresponding to a conductance of 9 pS (100 mM Ba2+). The open time histogram was fitted with two exponentials, with time constants of 5.8 and 30 ms (23 degrees C). Both types of events were virtually absent at -80 mV. The average open probability (fractional open time) increased sigmoidally from 0 to a saturation level of 0.08, following a Boltzmann function centered at -25 mV and with a steepness factor of 7 mV. Ensemble averages of test-minus-control currents showed a sigmoidal activation followed by inactivation during the pulse and deactivation (closing) after the pulse. The ON time course was well fitted with "m3h" kinetics, with tau m = 120 ms and tau h = 1.2 s. Deactivation was exponential with tau = 8 ms. This study demonstrates a technique for obtaining Ca channel events in lipid bilayers that are strictly voltage dependent and exhibit most of the features of the macroscopic ICa. The technique provides a useful approach for further characterization of channel properties, as exemplified in the accompanying paper, that describes the consequences on channel properties of phosphorylation by cAMP dependent protein kinase.

Animals↗

Dihydropyridine-sensitive skeletal muscle Ca channels in polarized planar bilayers. 2. Effects of phosphorylation by cAMP-dependent protein kinase.

The effects of phosphorylation on the voltage-dependent properties of dihydropyridine-sensitive Ca channels of skeletal muscle were studied. Single channel currents were recorded upon incorporation of transverse tubule membranes into planar bilayers that were kept polarized at near physiological resting potential and subjected to depolarizing pulses under voltage clamp. Studies were conducted to analyze the properties of the channels at both the single channel and macroscopic level, using methods introduced in the preceding paper (Ma et al., 1991. Biophys. J. 60: 890-901.). Addition of the catalytic subunit of cAMP-dependent protein kinase to the cis (intracellular) side of the bilayers containing channels resulted in: (a) an increase in open channel probability at all voltages above -50 mV; (b) a leftward shift (by 7 mV) in the curve describing the voltage-dependence of activation; (c) an approximate twofold decrease in the rate of inactivation; and (d) an increase in the availability of the channel. These findings provide new insights at the single channel level into the mechanism of modulation of the dihydropyridine-sensitive Ca channels of skeletal muscle by signal transduction events that involve elevation in cAMP and activation of the cAMP-dependent protein kinase.

Animals↗

Antibody penetration of viable human cells. I. Increased penetration of human lymphocytes by anti-RNP IgG.

Antibody penetration of viable cells and interaction with intracellular antigens may have major consequences for immunopathological processes in connective tissue diseases. We have reported previously that antibody can penetrate viable human lymphocytes. To assess further the role of antinuclear antibodies in this process, peripheral blood lymphocytes (PBMC) were incubated with FITC-conjugated IgG fractions from sera containing anti-RNP (anti-RNP IgG), Ro(SS-A), La(SS-B) and dsDNA antibodies and control sera for 24 h. Using crystal violet to quench cell surface staining, intracellular fluorescence of viable lymphocytes was quantified on the flow cytometer. It was noted that anti-RNP IgG entered 46.4 +/- 7.2% of lymphocytes which was significantly higher than anti-Ro(SS-A) (29.9 +/- 4.1%, P less than 0.05), La(SS-B) (22.0 +/- 7.5%, P less than 0.01) IgG and control IgG (28.8 +/- 2.1%, P less than 0.05) and not statistically different from anti-dsDNA IgG (32.6 +/- 14.3%). Inhibition experiments showed that the increased number of cells penetrated by anti-RNP IgG was a specific process. Time-course studies showed that anti-RNP IgG entry into cells was different from pooled control IgG. With anti-RNP IgG, positive-staining lymphocytes gradually increased in number from 12 to 24 h incubation, whilst with pooled control IgG, the peak was reached within 5 min. Dual staining experiments suggested that whereas both anti-RNP IgG and pooled control IgG entered B and NK cells, anti-RNP IgG also entered T cells. Using IgG F(ab')2 and Fc fragments from either anti-RNP IgG or pooled control IgG to compete with their FITC-conjugated counterparts indicated that the entry of anti-RNP IgG into-viable cells appeared to involve both F(ab')2 and Fc fragments, and pooled control IgG depended exclusively on the Fc portion of IgG. Further investigation by incubating anti-RNP IgG with 35S-methionine-labelled monocyte-depleted PBMC (MD-PBMC) suggested that anti-RNP IgG might react with the corresponding antigens either on the cell surface or within the cytoplasm.

Animals↗

Intravenous recombinant tissue-type plasminogen activator in acute myocardial infarction.

The efficacy and safety of intravenously administered recombinant tissue-type plasminogen activator (rt-PA, Boehringer Ingelheim Corp.) was investigated in 10 patients with acute myocardial infarction (AMI). rt-PA was given as a 10 mg bolus dose followed by infusions of 50 mg, 20 mg and 20 mg in three successive hours. All patients underwent baseline coronary angiography before thrombolytic therapy. Ninety minutes after the initiation of rt-PA infusion, recanalization of infarct-related coronary arteries as confirmed by angiography was achieved in 7 patients. The largest reduction in circulating fibrinogen was observed 4 to 6 h after the start of rt-PA infusion--14.3%. Moderate hemorrhage at the sites of arterial puncture occurred in 2 cases, probably as a result of heparin anticoagulation. No other side effects occurred. So rt-PA is an effective and safe thrombolytic agent.

Aged↗

Synthesis of (+/-)-N-2-methylpropyl-N-1'-methylacetonyl-nitrosamine.

N-2-methylpropyl-N-1-methylacetonyl-nitrosamine (NMAMPA), first isolated from moldy and normal foods, was synthesized by chemical means. The structure and molecular weight of the synthetic NMAMPA were elucidated by spectrometry. A combination of spectral and chromatographic data indicated that the synthetic NMAMPA is identical to that obtained naturally. Synthetic NMAMPA is a racemic modification, and a carcinogen which can induce esophageal and forestomach cancer in rats.

Carcinogens↗

[Chemical constituents of Osbeckia chinensis L].

Five constituents, 2-furoic acid, succinic acid, ursolic acid, quercetin and daucosterol were isolated and identified from Osbeckia chinensis. All of them are reported to have been found from this plant for the first time.

Anti-Infective Agents↗