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Biomedical subjects

Y Ma

Publications and source records attributed to Y Ma.

At least 451 records · Page 25Linked to original sources

Separation and purification of peptides by high-speed counter-current chromatography.

Preparative separations of peptides have been accomplished using high-speed counter-current chromatography. This has been made possible by the use of a particular solvent system that does not exhibit solvent carryover at high speed flow and centrifugation conditions. The solvent system composed of tert.-butyl methyl ether, n-butanol, acetonitrile and aqueous trifluoroacetic acid which can be adjusted in volume ratios and percent acid is employed in two instruments for counter-current chromatography. The cross-axis coil planet centrifuge was used for separation of various dipeptides. Superior resolution was obtained with the multi-layer coil planet centrifuge in the separation of six dipeptides and in the preparative purification of two synthetic peptides.

Amino Acid Sequence↗

Susceptibility to tumors induced by polyoma virus is conferred by an endogenous mouse mammary tumor virus superantigen.

A dominant gene carried in certain inbred mouse strains confers susceptibility to tumors induced by polyoma virus. This gene, designated Pyvs, was defined in crosses between the highly susceptible C3H/BiDa strain and the highly resistant but H-2k-identical C57BR/cdJ strain. The resistance of C57BR/cdJ mice is overcome by irradiation, indicating an immunological basis. In F1 x C57BR/cdJ backcross mice, tumor susceptibility cosegregates with Mtv-7, a mouse mammary tumor provirus carried by the C3H/BiDa strain. This suggests that Pyvs might encode the Mtv-7 superantigen (SAG) and abrogate polyoma tumor immunosurveillance through elimination of T cells bearing specific V beta domains. DNA typing of 110 backcross mice showed no evidence of recombination between Pyvs and Mtv-7. Strongly biased usage of V beta 6 by polyoma virus-specific CD8+ cytotoxic T lymphocytes in C57BR/cdJ mice implicates T cells bearing this Mtv-7 SAG-reactive V beta domain as critical anti-polyoma tumor effector cells in vivo. These results indicate identity between Pyvs and Mtv-7 sag, and demonstrate a novel mechanism of inherited susceptibility to virus-induced tumors based on effects of an endogenous superantigen on the host's T cell repertoire.

Animals↗

Determination of vitamin A in dried human blood spots by high-performance capillary electrophoresis with laser-excited fluorescence detection.

We have developed a high-performance capillary electrophoresis (HPCE) method to analyze the retinol (vitamin A) concentration as retinol-retinol binding protein (holo-RBP) from microvolumes of serum (5-10 microliters) or one to two drops (approximately 20 microliters) of blood collected and air-dried on blood collection filter paper. A 0.64-cm diameter disk was cut from the dried whole blood specimens and the samples were dissolved in a pretreatment buffer and filtered. Filtrate was injected onto the HPCE column for analysis. The separation was carried out in a 60 cm x 50 microns I.D. fused-silica capillary and the running voltage was 20 kV. A He-Cd laser with a wavelength of 325 nm was used for excitation, and the fluorescence of the holo-RBP complex was monitored at 465 nm by a photodiode. A virtual linear relationship was obtained for the retinol concentrations between HPCE and HPLC for 28 serum samples, 19 dried venous blood samples and 9 capillary dried blood spot samples, indicating that valid measures of serum retinol can be obtained from one to two drops of capillary blood collected on filter paper. The absolute detection limit for retinol by HPCE is below 3 micrograms/l. The method is very useful for vitamin A level screening, especially for children and premature new-born babies.

Chromatography, High Pressure Liquid↗

The human type 1 inositol 1,4,5-trisphosphate receptor from T lymphocytes. Structure, localization, and tyrosine phosphorylation.

Inositol 1,4,5-trisphosphate receptors (IP3R) are intracellular calcium release channels involved in diverse signaling pathways. An IP3R is thought to play a role in mobilizing calcium required for activation of T lymphocytes. The IP3R is a tetrameric structure comprised of four approximately 300-kDa subunits encoded by a approximately 10-kilobase mRNA. In the present study we determined the structure of the human type 1 IP3R expressed in T lymphocytes (Jurkats). The IP3R in human T cells had a predicted molecular mass of 308 kDa and was most similar to the non-neuronal form of the rodent type 1 IP3R. Two putative tyrosine phosphorylation sites were identified, one near the amino terminus and one near the putative channel pore at the carboxyl terminus. During T cell activation the IP3R was tyrosine phosphorylated. A site-specific anti-IP3R antibody was used to localize the carboxyl terminus of the IP3R to the cytoplasm in T cells.

Amino Acid Sequence↗

Cloning and expression of a novel truncated calcium channel from non-excitable cells.

Calcium entry, via a dihydropyridine-sensitive pathway, is required for differentiation in murine erythroleukemia cells (MELC). Calcium channel currents have been identified physiologically in some non-excitable cells, but little is known regarding the structure of these channels. We show that a truncated form of the alpha 1 subunit of the cardiac voltage-gated calcium channel (dihydropyridine receptor, DHPR) is expressed in MELC. This MELC calcium channel lacks the first four transmembrane segments of the DHPR (IS1 to IS4). A MELC calcium channel/cardiac DHPR chimera, co-expressed with the alpha 2 and beta subunits of the DHPR, forms a functional calcium channel in Xenopus oocytes.

Acetamides↗

Bone mineral metabolism in T lymphocyte-deficient and -replete strains of rat.

The immune and skeletal systems are known to interact. We have repeatedly shown that in contrast to in vitro data, the administration of T lymphocyte immunosuppressants, such as cyclosporin A, leads to an increase in bone resorption and a high turnover osteopenia. The purpose of this study was to characterize the bone metabolism of the T lymphocyte deficient Rowett athymic homozygous (rnu/rnu) nude rat. We wished to determine whether these rats share the bone abnormalities of cyclosporin A-treated rats. Eleven 10-week-old Sprague-Dawley rats and 12 similarly aged nude rats were studied over a 4-week period. Metaphyseal cancellous bone histomorphometry was similar in the two groups of rats and only differed with regard to percentage eroded perimeter (lower in nude rats, p = 0.0008) and longitudinal growth rate (49% lower in nude rats, p < 0.001). The nude rats had less body mass (p < 0.001) but nevertheless gained the same percentage of their body weight over the study period. The athymic rats had lower levels of serum, 1,25-dihydroxyvitamin D (p < 0.014) and serum osteocalcin(p < 0.009), and at the age of 14 weeks the nude rats had lower concentrations of serum creatinine (p = 0.001) and blood ionized calcium (p = 0.0002), yet serum PTH was similar throughout. RNA isolated from the contralateral tibias revealed that the nude group had lower steady-state levels of osteocalcin mRNA despite similar rates of bone formation. In its entirety, the data suggest that T cell deficiency per se is not necessarily associated with high turnover osteopenia.

Animals↗

Partial maintenance of extra cancellous bone mass by antiresorptive agents after discontinuation of human parathyroid hormone (1-38) in right hindlimb immobilized rats.

The current study employs the immobilization (IM) rat model to induce osteopenia, parathyroid hormone (PTH) as the anabolic agent to restore bone mass, and 17 beta-estradiol, calcitonin, or risedronate as the maintenance agents to answer the following questions: How much cancellous bone loss occurs when PTH is withdrawn? Which antiresorptive or antiactivation agent maintains bone best? Ideally, what tissue-level histomorphometric conditions maintain added bone? Six-month-old female rats were treated with 200 micrograms PTH/day subcutaneously at 30 days post-IM for 75 days. Then PTH treatment was stopped and switched to a vehicle (no treatment), 10 micrograms calcitonin/kg/day, 10 micrograms 17 beta-estradiol/kg/day, or 5 micrograms risedronate twice weekly for another 15 days (early response) or 60 days (late response). The rats had their right hindlimb throughout the study. The current report deals only with the maintenance phase involving 92 animals. Bone histomorphometry was performed on the secondary spongiosa of the right proximal tibia metaphysis (PTM). Cessation of PTH treatment followed by vehicle administration for 15 days resulted in partial loss of trabecular bone area and thickness from stimulated bone resorption and the fall of all formation indices. By contrast, all three antiresorptive agents maintained the cancellous bone mass during the same period. However, after prolonged withdrawal of PTH for 60 days, we found that 17 beta-estradiol and calcitonin maintained the cancellous bone slightly better than no treatment, while risedronate partially protected it from the mechanostat-induced bone loss. The risedronate treatment retained 71% of the PTH-added bone while calcitonin retained 48%, estrogen 42%, and no treatment 32%. The favorable histomorphometry profile for maintenance was the sustained reduction in bone resorption and turnover and normal age-related bone balance. We concluded that 1) cessation of PTH treatment will result in the loss of two-thirds of the added bone in 60 days; 2) currently, risedronate at the dose level employed as a maintenance agent is far superior to 17 beta-estradiol or calcitonin because of its long retention in bone; however, a longer observation period might result in less difference; and 3) the ideal tissue-level histomorphometry continues depressing bone resorption and turnover and maintains a normal age-related bone balance. Furthermore, we found the "lose, restore plus add, and maintain (LRAM)" concept was successful in maintaining most of the PTH-induced extra bone by risedronate for 60 days. It was far superior to 17 beta-estradiol or calcitonin. Possibly the last two agents would be effective in maintaining a normal amount of bone but not in preserving an excessive amount of bone. Nevertheless, the current study further emphasizes that clinicians should consider using the LRM treatment strategy when they plan to treat osteoporosis with bone anabolic agents.

Animals↗

Evidence for a cholesterol-lowering gene in a French-Canadian kindred with familial hypercholesterolemia.

We describe a four-generation kindred with familial hypercholesterolemia (FH) in which two of the eight heterozygotes for a 5-kb deletion (exons 2 and 3) in the low density lipoprotein (LDL) receptor gene were found to have normal LDL-cholesterol levels. In our search for a gene responsible for the cholesterol-lowering effect in this family, we have studied variation in the genes encoding the LDL receptor, apolipoprotein (apo) B, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, apoAI-CIII-AIV, and lipoprotein lipase. The analysis showed that it was unlikely that variation in any of these genes was responsible for the cholesterol-lowering effect. Expression of the LDL receptor, as assessed in vitro with measurements of activity and mRNA levels, was similar in normo and hyperlipidemic subjects carrying the deletion. Analysis of the apo E isoforms revealed that most of the e2 allele carriers in this family, including the two normolipidemic 5-kb deletion carriers, were found to have LDL-cholesterol levels substantially lower than subjects with the other apo E isoforms. Thus, this kindred provides evidence for the existence of a gene or genes, including the apo e2 allele, with profound effects on LDL-cholesterol levels.

Cholesterol↗

Clinical Utility of Electrocardiographic ST-Segment Area for Predicting Unsatisfactory Outcomes Following Thrombolytic Therapy.

The bedside surface 12-lead electrocardiogram is a mainstay in the early diagnostic evaluation of patients with suspected acute myocardial infarction. The presence of ST-segment elevation exceeding 1.0 mm in two or more anatomically associated leads is a reliable marker of myocardial injury and, when considered along with concomitant ST-segment depression, reflects the extent of myocardial injury. Mounting evidence also suggests that prolonged repolarization is a marker of injury and predicts the likelihood of malignant ventricular arrhythmias. We questioned whether a measure of both ST-segment duration and deviation (ST-deviation area) would offer additional prognostic information. Methods/Results: Admission electrocardiograms from 200 consecutive patients with ischemic chest pain accompanied by ST-segment elevation in whom thrombolytic therapy was given within 6 hours from symptom onset were analyzed. The sum of ST-segment elevation (Sigma ST elevation) and ST-segment deviation (Sigma ST deviation) were calculated, as was the sum of ST-segment deviation area (Sigma ST deviation area). All ST measurements were performed 60 msec after the J point. Computerized planimetry was used to calculate ST-segment area. Sigma ST deviation and Sigma ST deviation area remained constant over time. Patients with large deviations (Sigma ST elevation > 20 mm (odds ratio 2.14, p = 0.02) and Sigma ST deviation area > 150 (odds ratio 1.92, p = 0.02) had a higher incidence of in-hospital unsatisfactory clinical outcome (defined as death, congestive heart failure, cardiogenic shock, recurrent myocardial infarction, or the need for coronary revascularization). These relationships were present for both inferior and anterior infarctions. Sigma ST deviation area correlated closely with Sigma ST elevation (r = 0.92; p = 0.0001) and significantly but much less strongly with the sum of Q waves (r = 0.18; p = 0.01). By univariate analysis, only site of infarction (p = 0.01), Sigma ST deviation area (p = 0.04), and the sum of Q waves (p = 0.005) were identified as predictors of a poor clinical outcome. The sum of Q waves was identified by multivariate analysis as the best independent predictor of an unsatisfactory clinical outcome. Conclusions: A clinician's ability to provide optimal care is influenced strongly by the availability of diagnostic and prognostic information. In the evaluation of patients with acute myocardial infarction, ST-segment deviation area derived from the admission surface electrocardiogram can be used to risk-stratify patients. The full clinical potential of this measure is unknown and will require further evaluation.

Journal Article↗

[Selenium content of tea from different provinces of the People's Republic of China].

In the People's Republic of China there are wide regions where many people suffer from strong selenium deficiency. In order to elucidate the possibilities to compensate this state of need by the national drink, the selenium contents of 31 tea samples from nine provinces of the PR China were determined by hydride generation atomic absorption spectrometry (HG-AAS). The Se contents of the ten analyzed tea samples from one region with soils containing high selenium amounts range from 83 ng/g to 7530 ng/g. These results are in contrast to the Se contents of the remaining tea samples from other regions, which range from 28 to 573 ng/g. The extractable part of selenium in the tea drink ranges between 7% and 27% without significant difference among different tea samples. The accuracy of Se determination was tested by using one tea standard reference material. The daily consumption of one "Se tea" examined in this study assures that people who suffer from a Se deficiency resume enough of it. Drinking normal quantities of this tea an intoxication by Se is not to be expected.

China↗

Impact of the high tyrosine fraction in complementarity determining regions: measured and predicted effects of radioiodination on IgG immunoreactivity.

Although iodine-131 is the most widely used radionuclide for radioimmunotherapy, direct radiolabeling methods yield decreased immunoreactivity of the antibody as a function of increased iodine incorporation. We have studied the amino acid sequences of a therapeutic IgG (HuM195), and in particular its complementarity determining regions (CDR), in order to correlate the iodination of tyrosine residues in the antigen binding site with changes in immunoreactivity. The CDR contained an overabundance of tyrosines relative to an expected random distribution of amino acids. In contrast, lysine residues that can be used for ligand attachment were evenly distributed throughout the IgG. HuM195 was first trace labeled with 111In and then labeled with stable 127I at various specific activities. The immunoreactivity of each product was determined using the 111In tracer. The immunoreactivity measured after varying levels of iodination fit a theoretical curve that was generated based on the assumption that a single iodine incorporation anywhere on a tyrosine residue in a CDR destroys the immunoreactivity of the antibody. Similar theoretical curves for antibody fragments (Fab, Fv) suggest an even faster decrease in immunoreactivity with increasing iodination. A review of the sequences of other therapeutic IgG shows that a similar overabundance of tyrosine residues is found in the CDRs. Using enzyme digestion, the distribution of iodine on different parts of the antibody was also studied. The iodinated residues were distributed non uniformly throughout the IgG, with the heavy chain variable region tyrosines having a higher propensity for iodine incorporation than tyrosines in the other regions of the IgG. The common abundance of tyrosine in the CDR of IgG and its correlation with loss of function have important implications for therapeutic use of high specific activity radioiodinated monoclonal antibodies or fragments.

Amino Acid Sequence↗

Poly(binaphthyl-20-crown-6) as receptor based molecular selective potentiometric electrodes for catecholamines and other 1,2-dihydroxybenzene derivatives.

A novel type of poly(crown ether) electrode that is capable of selectively determining some 1,2-dihydroxybenzenes has been developed. A lipophilic macrocyclic crown ether, binaphthyl-20-crown-6, is electrochemically deposited on a platinum disc electrode. The film obtained is used as a sensory element for a potentiometric electrode for the determination of some neurotransmitters, namely, catecholamines. The new electrode is also capable of discriminating the steric shapes of 1,2-dihydroxybenzene moieties. The response of the new electrode is based on the principle of 'host-guest' chemistry. The potentiometric response is dependent on the pH of the solution and the nature of the buffer medium. The new sensor electrode has a useful analytical range of 1.5 x 10(-8) M-2 x 10(-5) M with a linear dynamic range between about 1 x 10(-7) M-5 x 10(-4) M with a 'super-Nernstian' slope of 110-130 mV/decade. The detection limit in phosphate buffer (0.1 M, pH 9.4) is ca. 3 x 10(-8) M for catecholamine. The sensor electrode is virtually insensitive towards interference from most inorganic ions and circumvents the interference from ascorbic acid, which is often found using amperometric methods in biological samples. A partial response mechanism of the present electrode is discussed, supported by results of electron dispersive analysis by x-rays (EDAX).

Animals↗

Co-treatment of PGE2 and risedronate is better than PGE2 alone in the long-term treatment of ovariectomized-induced osteopenic rats.

We studied the effects of prostaglandin (PGE2) and risedronate (Ris) alone or in combination in 3.5-month-old intact and ovx-induced osteopenic rat skeletons to determine whether PGE2 plus Ris was more anabolic than PGE2 alone. Six mg PGE2/kg/d and 5 micrograms Ris/kg/2x/wk alone or in combination were given to sham-ovx and ovx rats for 30 or 90 days beginning 60 days post operation. Secondary spongiosa of proximal tibial metaphyses (PTM) was studied. Ovariectomy (ovx) induced dramatic bone loss. Ris increased bone mass in sham-ovx rats and prevented further bone loss in ovx rats. PGE2 treatment for 30 days added extra bone in sham-ovx rats and no further increase after 90 days treatment. Thirty days of PGE2 alone treatment restored the bone mass in ovx rats to the level of sham-ovx rats, but the restored bone was partially lost by 90 days of treatment. Co-treatment for 30 days produced the same amount of bone mass in both sham-ovx and ovx rats as PGE2 alone did. However, unlike the PGE2 alone treated, co-treatment animals continued to form more bone for 90 days. The difference in tissue-level histomorphometry between co-treatment and PGE2 alone was that the former depressed the bone resorption and turnover. These findings indicated that the long-term administration of PGE2 alone cannot maintain or continue to add bone mass in ovx rats but that co-treatment of a PGE2 with an anti-resorptive or activation agent can resist the influence of the mechanostat induced bone loss as well as continue to add bone.

Aging↗

Equal vibrotactile sense thresholds of the fingers and its diagnostic significance for hand-arm vibration syndrome.

The vibrotactile sense thresholds (VSTs) of the middle fingers of 60 healthy persons and 97 patients with Hand-Arm Vibration Syndrome (HAVS) or subclinical HAVS were measured quantitatively. Intermittent vibratory irritations were adopted, with vibration stimulus frequencies at 8, 16, 31.5, 63, 125, 250, and 500 Hz. The equal VST contours of the fingers were mapped. Results showed that the VSTs of the normal group were not correlated with sex or handedness. From 8 Hz to 250 Hz the equal VST contours of the normal group were relatively flat; at more than 250 Hz the contours began an abrupt ascent. The VST values had a logarithmic rising tendency with the increasing age of subjects. In the equal VST contours the frequency of the most sensitive threshold value was 125 Hz in the normal group and 8 Hz in the HAVS group. The patients' VST values were higher than that of the healthy persons. The vibrotactilegram showed that the VST values of the patient groups first shifted at high frequencies and VST loss displayed a "V"-type hollow at 125 Hz and 250 Hz. The quantitative test method of VST was a valuable auxiliary detection method for HAVS. The "V"-type hollow of VST was an early clinical manifestation of HAVS.

Adult↗