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Biomedical subjects

T Song

Publications and source records attributed to T Song.

At least 19 recordsLinked to original sources

Cerebellar peduncular myelinolysis in a patient receiving hemodialysis.

Here, we report the observation of extrapontine lesions, in addition to the pontine lesions previously documented in a diagnosed case of hemodialysis-associated osmotic demyelination syndrome due to end-stage renal disease. The patient exhibited lesions on bilateral middle cerebellar peduncles, and had been receiving regular hemodialysis as treatment for end-stage renal disease. He presented with progressive gait disturbance and postural instability. Accompanying symptoms included peduncular hallucinations and mild cognitive dysfunction. Brain MRI revealed high signal intensity in the area of bilateral cerebellar peduncles on the diffusion and T2-weighted images, with a decreased signal intensity noted on the ADC map. The ataxic form of osmotic myelinolysis syndrome is quite rare. The involvement of the cerebellar peduncles in extrapontine myelinolysis in a patient with end stage renal disease has not, to our knowledge, been previously reported. Here, we describe the MRI findings and clinical features associated with this unique case, and include a review of the relevant literature.

Aged↗

Morphological study of the interaction between M21 melanoma and lymphatic endothelium.

In order to study the interaction between melanoma and lymphatic endothelial cells (LECs) and to investigate the mechanism of lymphatic metastasis, M21 melanoma cells were seeded on the confluent LECs monolayer and the alterations of both cells were observed. The results showed that tumor cells could both adhere by pseudopodia to LECs at the site near the intercellular junction and on the apical surface. The adhesion of the melanoma cells induced the endothelial junction dissolution and endothelial retraction, which allowed the passage of the tumor cells through the opened gap and attached to the subendothelial matrix, then the tumor cells invaded and migrated under the LECs monolayer. These findings suggest that tumor cells could metastasize through the lymphatic vessel by destroying intercellular junctions or the LECs directly.

Animals↗

Abnormal spectra alteration observed in Triton calibration method for measuring [Ca2+]i with fluorescence indicator, fura-2.

We compared two commonly used calibration methods for measuring the concentration of intracellular free calcium ([Ca2+]i) by ratiometric fluorescence dye, fura-2 in mouse neuroblastoma-rat glioma hybrid cells (NG108-15). One calibration method, the Triton method, employs detergent Triton X-100, while the other, the Ionomycin method, uses a calcium-specific ionophore, Ionomycin. In the Triton method, we observed that at excitation 380 nm, the fura-2 fluorescence intensity of steady-state cells abnormally situated beyond the limiting intensity for calibration. By excitation scan, we demonstrated that this abnormality was caused by the change of fura-2 isosbestic points, which in turn was due to cell lysis after the addition of Triton X-100. This problem was resolved in the Ionomycin method by avoidance of cell lysis. Our results showed the correlation between inconsistent isosbestic points and cell lysis. As the basis for [Ca2+]i calibration, the proportionality between the fluorescence intensity and the concentration of dye species was impaired because of inconsistent isosbestic points. This inconsistency can be eliminated by a preliminary experiment of excitation scan to test the feasibility of different calibration methods.

Animals↗

Neurodegeneration in the Niemann-Pick C mouse: glial involvement.

A mouse model of Niemann-Pick type C disease has been found that exhibits neuropathology similar to the human condition. There is an age-related neurodegeneration in several brain regions and a lack of myelin in the corpus callosum in these mice. The purpose of the present study was to examine the Niemann-Pick mouse and determine whether: (1) microglia and astrocytes exhibit ultrastructural pathology similar to that found in neurons; (2) nerve fiber number is reduced when the myelin sheath is absent; and (3) the lysosomal hydrolase, cathepsin-D, is involved in the neurodegenerative process. Using light and electron microscopic methods, and immunocytochemistry, Niemann-Pick and control animals were examined at several ages. Cathepsin-D content was semi-quantitatively measured in neurons and glial cells in brain regions known to exhibit neurodegeneration, as was the density of glial fibrillary acidic protein-labeled astrocytes. The Niemann-Pick mouse exhibited: (1) an age-related increase in inclusion bodies in microglia and astrocytes, similar to that observed within neurons; (2) an almost complete absence of myelin in the corpus callosum by 7-8 weeks of age, along with a 30% reduction in the number of corpus callosum axons; (3) a mild age-related increase in cathepsin-D content within nerve cells in many brain regions. However, the cathepsin-D elevation was greatest in microglial cells; (4) an age-related increase in the number of microglial cells containing intense cathepsin-D immunoreactivity in both the thalamus and cerebellum. Both of these brain regions have been shown previously to exhibit an age-related loss of neurons; and (5) an increase in the number of reactive astrocytes immunostained for glial fibrillary acidic protein, especially in the thalamus and cerebellum. These data indicate that glial cells are a major target for pathology in the Niemann-Pick mouse. The lack of myelin within the corpus callosum may be related to the loss of nerve fibers in this structure. The increase in cathepsin-D-laden microglial cells, in brain regions previously shown to undergo neurodegeneration, is consistent with a role for microglia in the phagocytosis of dead neurons and in actively contributing to the neurodegenerative process. The activation of astrocytes in regions that undergo neurodegeneration is also consistent with a role for these glial cells in the neurodegenerative process.

Animals↗

Fine-scale mapping of a locus for severe bipolar mood disorder on chromosome 18p11.3 in the Costa Rican population.

We have searched for genes predisposing to bipolar disorder (BP) by studying individuals with the most extreme form of the affected phenotype, BP-I, ascertained from the genetically isolated population of the Central Valley of Costa Rica (CVCR). The results of a previous linkage analysis on two extended CVCR BP-I pedigrees, CR001 and CR004, and of linkage disequilibrium (LD) analyses of a CVCR population sample of BP-I patients implicated a candidate region on 18p11.3. We further investigated this region by creating a physical map and developing 4 new microsatellite and 26 single-nucleotide polymorphism markers for typing in the pedigree and population samples. We report the results of fine-scale association analyses in the population sample, as well as evaluation of haplotypes in pedigree CR001. Our results suggest a candidate region containing six genes but also highlight the complexities of LD mapping of common disorders.

Alleles↗

The alternative sigma factor SigH regulates major components of oxidative and heat stress responses in Mycobacterium tuberculosis.

Mycobacterium tuberculosis is a specialized intracellular pathogen that must regulate gene expression to overcome stresses produced by host defenses during infection. SigH is an alternative sigma factor that we have previously shown plays a role in the response to stress of the saprophyte Mycobacterium smegmatis. In this work we investigated the role of sigH in the M. tuberculosis response to heat and oxidative stress. We determined that a M. tuberculosis sigH mutant is more susceptible to oxidative stresses and that the inducible expression of the thioredoxin reductase/thioredoxin genes trxB2/trxC and a gene of unknown function, Rv2466c, is regulated by sigH via expression from promoters directly recognized by SigH. We also determined that the sigH mutant is more susceptible to heat stress and that inducible expression of the heat shock genes dnaK and clpB is positively regulated by sigH. The induction of these heat shock gene promoters but not of other SigH-dependent promoters was markedly greater in response to heat versus oxidative stress, consistent with their additional regulation by a heat-labile repressor. To further understand the role of sigH in the M. tuberculosis stress response, we investigated the regulation of the stress-responsive sigma factor genes sigE and sigB. We determined that inducible expression of sigE is regulated by sigH and that basal and inducible expression of sigB is dependent on sigE and sigH. These data indicate that sigH plays a central role in a network that regulates heat and oxidative-stress responses that are likely to be important in M. tuberculosis pathogenesis.

Bacterial Proteins↗

[Clinical and experimental study on the ovulation promoting effect of yangjing decoction].

OBJECTIVE: To observe the effect of Yangjing Decoction (YJD) to promote ovulation and to explore its mechanism. METHODS: The clinical effect of YJD in treating patients of functional non-ovulation and changes of gonadotropic hormone, sex hormone and diameter of main follicle before and after treatment were observed. In experimental study, the influence of YJD on monoamine neurotransmitter of hypothalamus, endorphin (beta-EOP), serum sex hormone, trace element, endometrium and histomorphology of ovary were observed. RESULTS: In the 36 patients observed clinically, the ovulation rate was 61.1%, the serum contents of follicular stimulation hormone (FSH), luteinizing hormone (LH) and estradiol (E2) increased and the diameter of main follicle enlarged significantly after treatment (P < 0.01, P < 0.001). The experimental study showed that the totals of follicle at different levels, the corpus luteum and the proportion of superior follicle in ovary were improved obviously in the YJD group as compared with those in the control group. After YJD treatment the hypothalamus contents of monoamine neurotransmitter, including norepinephrine (NE) and dopamine (DA) increased, the 5-hydroxytryptamine (5-HT) 5-hydroxyindoleaceticacid (5-HIAA) and beta-EOP decreased significantly, and the NE/DA ratio and NE/5-HT ratio elevated obviously (P < 0.05, P < 0.001). The serum levels of E2, P and trace elements, including Zn, Cu and Fe increased significantly (P < 0.05, P < 0.01). CONCLUSION: YJD is an effective recipe in promoting ovulation, the effect may be achieved through the multi-linked, multi-systemetic synthetic effect of hypothalamus, hypophysis and ovary, etc.

Adult↗

[The clinical and pathologic study of embeded supernumerary teeth].

OBJECTIVE: To analyse the clinic and pathologic features of 100 embedded supernumerary teeth, to find out the rule of cystic change of supernumerary teeth and its relationship to malocclusion, and to present the methods of therapy. METHODS: Analysis of clinical data, X-ray manifestation,comparison of the findings on operation and pathological changes demonstrated the correct diagnosis of supernumerary teeth. RESULTS: On statistics and analysis,66% of the crowns of the supernumerary teeth were showed different sizes of circular photic shades,but only 35% were proved to be cystic change by biopsies. CONCLUSION: This study showed that 35% had cystic change among 100 cases,so if the diagnosis can be made in these cases with indication of operation the extraction of the supernumerary teeth must be done as soon as possible.

English Abstract↗

Protein kinase C mediates insulin-inhibited Ca2+ transport and contraction of vascular smooth muscle.

Insulin acutely inhibits contraction of primary cultured vascular smooth muscle (VSM) cells from canine femoral artery by inhibiting contractile agonist-induced Ca2+ influx. Insulin also inhibits contraction at step(s) distal to intracellular Ca2+ concentration (Ca2+i) by stimulating cyclic guanosine monophosphate (GMP) production. We wished to see whether these effects of insulin are mediated by protein kinase C (PKC). Ca2+ influx was assessed by measuring the rate of fluorescence quenching of intracellular fura 2 by extracellular Mn2+. We found that 10 micromol/L serotonin (5-HT) stimulated Mn2+ influx 3-fold, and 1 nmol/L insulin inhibited the 5-HT-stimulated component of Mn2+ influx by 63% (P < .05), but insulin had no effect in the presence of 1 micromol/L staurosporine, an inhibitor of PKC. In the absence of insulin, preincubating cells with 0.1 micromol/L phorbol 12-myristate 13-acetate (PMA) for 5 min inhibited the 5-HT-stimulated component of Mn2+ influx by 69% (P < .05). Insulin inhibited cell contraction induced by raising Ca2+i to supraphysiologic levels with ionomycin by 75% (P < .05). We also noted that 10(-6) mol/L calphostin C, another PKC inhibitor, or 16-h preincubation with PMA completely blocked this effect of insulin. Finally, 10-min exposure to insulin or PMA increased cyclic GMP production in ionomycin-treated cells by 50% and 64%, respectively (both P < .05). We conclude that insulin inhibits VSM cell contraction by inhibiting 5-HT-stimulated Ca2+ influx and also at step(s) distal to Ca2+i by a PKC-dependent mechanism.

Animals↗

Insulin increases NO-stimulated guanylate cyclase activity in cultured VSMC while raising redox potential.

Insulin acutely stimulates cyclic guanosine monophosphate (cGMP) production in primary confluent cultured vascular smooth muscle cells (VSMC) from canine femoral artery, but the mechanism is not known. These cells contain the inducible isoform of nitric oxide (NO) synthase (iNOS), and insulin-stimulated cGMP production in confluent cultured cells is blocked by the NOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA). In the present study, it is shown that iNOS is also present in freshly dispersed VSMC from this artery, indicating that iNOS expression in cultured VSMC is not an artifact of the culture process. Insulin did not stimulate NOS activity in primary confluent cultured cells because it did not affect citrulline or combined NO(-)(3)/NO(-)(2) production. To see whether insulin required the permissive presence of NO to stimulate cGMP production, iNOS and basal cGMP production were inhibited with L-NMMA, and the cells were incubated with or without 1 nM insulin and/or the NO donor, S-nitroso-N-acetyl-D,L-penicillamine (SNAP) at a concentration (0.1 microM) that restored cGMP production to the basal value. In the presence of L-NMMA, insulin no longer affected cGMP production but when insulin was added to L-NMMA plus SNAP, cGMP production was increased by 69% (P < 0.05 vs. L-NMMA plus SNAP). Insulin, which increases glucose uptake by these cells, increased the cell lactate content and the lactate-to-pyruvate ratio (LPR) by 81 and 97%, respectively (both P < 0.05), indicating that the hormone increased aerobic glycolysis and the redox potential. The effects of insulin on LPR and cGMP production were blocked by removing glucose or by adding 2-deoxyglucose to the incubation media and were duplicated by the reducing substrate, beta-hydroxybutyrate. We conclude that insulin does not acutely affect iNOS activity in these VSMC but it does augment cGMP production induced by the NO already present in the cell while increasing aerobic glycolysis and the cell redox potential.

Aerobiosis↗

Isoflavones in retail and institutional soy foods.

A national sampling plan was developed to select the most widely used isoflavone-containing foods in the United States. Foods were selected based on their retail volume and sampled in five geographical areas representing seven metropolitan areas. Isoflavones were analyzed from composite samples, raw and cooked, and reported by brand. Quality control measures were evaluated throughout the study. Isoflavone levels ranged from 1 microg/g in soy sauces to 540 microg/g in tempeh. Soymilk and tofu represented the major portion of soy foods evaluated. These data will appear in the electronic version of USDA Handbook No. 8 of Food Composition Data in 1999.

Isoflavones↗

Spontaneous CSF rhinorrhoea secondary to a middle cranial fossa defect.

Spontaneous cerebrospinal fluid rhinorrhoea is a clinical entity that is difficult to both diagnose and treat. A case in a 56-year-old woman with a defect in the lateral wall of the sphenoid sinus and middle cranial fossa floor is presented. Pathogenesis and management of this rare condition are discussed. Copyright 1999 Harcourt Publishers Ltd.

Journal Article↗

XspI, a new type II restriction endonuclease from a Xanthomonas species.

A new type II restriction endonuclease, XspI, was purified 11-fold in seven steps to homogeneity from Xanthomonas sp. strain YK1 grown aerobically in Luria broth. The final specific activity of the purified enzyme was 3890 micrograms of lambda DNA digested per h per mg of protein. The molecular weight of the native enzyme was determined to be 54,000. Sodium dodecyl sulfate-gel electrophoresis revealed two identical subunits of molecular weight 29,000. The purified enzyme was most active at 37 degrees C in the presence of 100 mM KCl and 5 mM MgCl2 under pH range from 7.5 to 8.0. The enzyme was stable at least for 1 h at 37 degrees C, but was inactivated after 20 min at 65 degrees C. XspI recognized the tetranucleotide sequence, 5'-CTAG-3', and cleaved it between C and T, like its isoschizomers MaeI and BfaI. The ability of the source organism to grown aerobically, together with the heat inactivation of the enzyme, confers practical advantages upon XspI over its known isoschizomers.

Base Sequence↗

Soy isoflavone analysis: quality control and a new internal standard.

Development of a database of the soy isoflavone content of foods requires accurate and precise evaluation of different food matrixes. To evaluate accuracy, we estimated recoveries of both internal and external standards in 5 different soyfoods weekly. Standards were evaluated daily for system quality assurance. To evaluate sample precision, we analyzed soybeans and soymilk bimonthly for within-day precision and over 4 d for day-to-day precision. CVs should be < or = 8%. We validated our methods for single and multiple recovery concentrations by using our new internal standard, 2,4,4'-trihydroxydeoxybenzoin, and the external standards daidzein, genistein, and genistin. Concentrations of 12 isoflavone isomers, 3 aglycones (daidzein, genistein, and glycitein), and 9 glucosides (daidzin, genistin, glycitin, acetyldaidzin, acetylgenistin, acetylglycitin, malonyldaidzin, malonylgenistin, and malonylglycitin) were measured in a variety of soybeans and soyfoods. The extraction methods used depended on soyfood type. The HPLC conditions for soy isoflavone analysis were improved, leading to good separation with a short analysis time (60 min/sample). A data bank of concentration and distribution of isoflavones in different soybean products was assembled. A wide range of isoflavone concentrations, from < 50 microg/g to > 20,000 microg/g, was found in different soy products. The glucoside forms are almost twice the molecular weight of the aglycones; reported isoflavone concentrations should be normalized to the aglycone mass (or an isoflavonoid equivalent) rather than a simple sum of all isomers.

Chromatography, High Pressure Liquid↗

Insulin inhibits vascular smooth muscle contraction at a site distal to intracellular Ca2+ concentration.

Several hypertensive states are associated with resistance to insulin-induced glucose disposal and insulin-induced vasodilation. Insulin can inhibit vascular smooth muscle (VSM) contraction at the level of the VSM cell, and resistance to insulin's inhibition of VSM cell contraction may be of pathophysiological importance. To understand the VSM cellular mechanisms by which insulin resistance leads to increased VSM contraction, we sought to determine how insulin inhibits contraction of normal VSM. It has been shown that insulin lowers the contractile agonist-stimulated intracellular Ca2+ (Ca2+i) transient in VSM cells. In this study, our goal was to see whether insulin inhibits VSM cell contraction at steps distal to Ca2+i and, if so, to determine whether the mechanism is dependent on nitric oxide synthase (NOS) and cGMP. Primary cultured VSM cells from canine femoral artery were bathed in a physiological concentration of extracellular Ca2+ and permeabilized to Ca2+ with a Ca2+ ionophore, either ionomycin or A-23187. The resultant increase in Ca2+i contracted individual cells, as measured by photomicroscopy. Preincubating cells with 1 nM insulin for 30 min did not affect basal Ca2+i or the ionomycin-induced increase in Ca2+i, as determined by fura 2 fluorescence measurements, but it did inhibit ionomycin- and A-23187-induced contractions by 47 and 51%, respectively (both P < 0.05). In the presence of 1.0 microM ionized Ca2+, ionomycin-induced contractions were inhibited by insulin in a dose-dependent manner. In the presence of ionomycin, insulin increased cGMP production by 43% (P < 0.05). 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (10 microM), a selective inhibitor of guanylate cyclase that blocked cGMP production in these cells, completely blocked the inhibition by insulin of ionomycin-induced contraction. It was found that the cells expressed the inducible isoform of NOS. NG-monomethyl-L-arginine or NG-nitro-L-arginine methyl ester (0.1 mM), inhibitors of NOS, did not affect ionomycin-induced contraction but prevented insulin from inhibiting contraction. We conclude that insulin stimulates cGMP production and inhibits VSM contraction in the presence of elevated Ca2+i. This inhibition by insulin of VSM contraction at sites where Ca2+i could not be rate limiting is dependent on NOS and cGMP.

Animals↗

Sensory reinnervation of autologous tissue TRAM flaps after breast reconstruction.

The use of the transverse rectus abdominis musculocutaneous (TRAM) flap has come to the forefront for breast reconstruction following mastectomy. Despite our ability to create surgically a supple breast mound, simulate the nipple with local skin flaps, and pigment the skin to create an areola, one of the last drawbacks has been the reestablishment of normal sensation. Some patients have anecdotally reported some sensory return in the reconstructed breast mound. We sought to quantitate the pattern of sensory return in TRAM flaps in 24 patients to identify factors that favor sensory reinnervation of the flap. Patients were recalled for sensory testing after unilateral or bilateral breast reconstruction following mastectomy for cancer or premalignant mastopathy. The interval from surgery varied from 3 to 41 months. Sensation was evaluated using the Semmes-Weinstein monofilament test, hot/cold recognition, and vibratory sensation measured in 16 segments of the reconstructed breast mound and compared to the opposite, unoperated breast or to volunteer controls. Thirty-four flaps were evaluated. The Semmes-Weinstein measurements demonstrated measurable sensation in 32 of 34 flaps with 2 flaps developing sensation equal to the control unoperated breast. The return of hot recognition occurred in 21 of 34 flaps, cold recognition in 22 of 34 flaps, and vibratory sensation in 26 of 34 flaps. Our findings suggest that excellent sensory return occurs in the majority of patients via nerve ingrowth into the flap from the mastectomy bed. It would appear that a natural breast reconstruction with some sensation can be a reality for the majority of patients in the absence of additional complex surgical maneuvers such as nerve preservation or nerve-nerve coaptation.

Breast↗

Dihydroxyacetone synthase from a methanol-utilizing carboxydobacterium, Acinetobacter sp. strain JC1 DSM 3803.

Acinetobacter sp. strain JC1 DSM 3803, a carboxydobacterium, grown on methanol was found to show dihydroxyacetone synthase, dihydroxyacetone kinase, and ribulose 1,5-bisphosphate carboxylase, but no hydroxypyruvate reductase and very low hexulose 6-phosphate synthase, activities. The dihydroxyacetone synthase was found to be expressed earlier than the ribulose 1,5-bisphosphate carboxylase. The dihydroxyacetone synthase was purified 19-fold in eight steps to homogeneity, with a yield of 9%. The final specific activity of the purified enzyme was 1.12 micromol of NADH oxidized per min per mg of protein. The molecular weight of the native enzyme was determined to be 140,000. Sodium dodecyl sulfate-gel electrophoresis revealed a subunit of molecular weight 73,000. The optimum temperature and pH were 30 degrees C and 7.0, respectively. The enzyme was inactivated very rapidly at 70 degrees C. The enzyme required Mg2+ and thiamine pyrophosphate for maximal activity. Xylulose 5-phosphate was found to be the best substrate when formaldehyde was used as a glycoaldehyde acceptor. Erythrose 4-phosphate, glycolaldehyde, and formaldehyde were found to act as excellent substrates when xylulose 5-phosphate was used as a glycoaldehyde donor. The Kms for formaldehyde and xylulose 5-phosphate were 1.86 mM and 33.3 microM, respectively. The enzyme produced dihydroxyacetone from formaldehyde and xylulose 5-phosphate. The enzyme was found to be expressed only in cells grown on methanol and shared no immunological properties with the yeast dihydroxyacetone synthase.

Acinetobacter↗