Biomedical subjects

L Ma

Publications and source records attributed to L Ma.

At least 253 records · Page 14Linked to original sources

PubMed1999

Changes in corneal endothelial apical junctional protein organization after corneal cold storage.

PURPOSE: Understanding the mechanisms regulating corneal endothelial permeability during storage and recovery is of critical importance both to improving Eye Banking practices and preventing corneal transplant failure. The goal of this study was to determine the effects of cold storage on the organization of apical junctional complex (AJC) proteins and their relationship to F-actin in corneal endothelium. METHODS: Immunostaining using antibodies to the AJC proteins, ZO-1, cadherin, and alpha- and beta-catenin was performed on 16 eye bank corneas and four cat corneas after 2-8 days of storage at 4 degrees C in Optisol-GS, and compared with fresh corneas. The 3-D in situ localization of the AJC proteins was then determined by using laser confocal microscopy. AJC organization also was assessed after stored human corneas were further incubated at 37 degrees C in Optisol-GS or in serum-free media. RESULTS: In normal human and cat corneas, F-actin was organized into dense peripheral bands (DPBs) along the apical cell border. The tight-junction protein, ZO-1, and the adherens junction proteins, cadherin and alpha- and beta-catenin, each formed a uniquely discontinuous hexagonal apical band with the largest gaps occurring at the Y-junctions between adjacent endothelial cells. In stored eye bank and cat corneas, cells lost their normal hexagonal F-actin staining pattern and appeared rounded and distorted, with increased cytoplasmic staining and incomplete and condensed DPBs. Similar distortions were observed in the apical bands of cadherin, catenin, and ZO-1 staining between endothelial cells. Gaps in staining at the endothelial Y-junctions were significantly enlarged; corresponding gaps also were observed with phalloidin staining. These changes were reversed after overnight incubation at 37 degrees C in either serum-free media or Optisol-GS. Quantitative analysis demonstrated a significant increase in the size of the Y-junctional gaps (p < 0.0001) after cold storage of cat corneas as compared with fresh corneas. CONCLUSION: These results suggest that disruption of the F-actin cytoskeleton and AJC may explain, in part, the loss of function (corneal swelling) after prolonged cold storage.

Animals↗

PubMed1999

The presence of a heterotrimeric G protein and its role in signal transduction of extracellular calmodulin in pollen germination and tube growth

The role of heterotrimeric G proteins in pollen germination, tube growth, and signal transduction of extracellular calmodulin (CaM) was examined in lily pollen. Two kinds of antibodies raised against animal Gzalpha, one against an internal sequence and the other against its N terminus, cross-reacted with the same 41-kD protein from lily pollen plasma membrane. This 41-kD protein was also specifically ADP ribosylated by pertussis toxin. Microinjection of the membrane-impermeable G protein agonist GTP-gamma-S into a pollen tube increased its growth rate, whereas microinjection of the membrane-impermeable G protein antagonist GDP-beta-S and the anti-Galpha antibody decreased pollen tube growth. The membrane-permeable G protein agonist cholera toxin stimulated pollen germination and tube growth. Anti-CaM antiserum inhibited pollen germination and tube growth, and this inhibitory effect was completely reversed by cholera toxin. The membrane-permeable heterotrimeric G protein antagonist pertussis toxin completely stopped pollen germination and tube growth. Purified CaM, when added directly to the medium of plasma membrane vesicles, significantly activated GTPase activity in plasma membrane vesicles, and this increase in GTPase activity was completely inhibited by pertussis toxin and the nonhydrolyzable GTP analogs GTP-gamma-S and guanylyl-5'-imidodiphosphate. The GTPase activity in plasma membrane vesicles was also stimulated by cholera toxin. These data suggest that heterotrimeric G proteins may be present in the pollen system where they may be involved in the signal transduction of extracellular CaM and in pollen germination and tube growth.

Journal Article↗

PubMed1999

Inhibition of calcium/calmodulin-dependent protein kinase II in rat hippocampus attenuates morphine tolerance and dependence.

Learning and memory have been suggested to be important in the development of opiate addiction. Based on the recent findings that calcium/calmodulin-dependent protein kinase II (CaMKII) is essential in learning and memory processes, and morphine treatment increases CaMKII activity in hippocampus, the present study was undertaken to examine whether inhibition of hippocampal CaMKII prevents morphine tolerance and dependence. Here, we report that inhibition of CaMKII by intrahippocampal dentate gyrus administration of the specific inhibitors KN-62 and KN-93 to rats significantly attenuated the tolerance to the analgesic effect of morphine and the abstinence syndrome precipitated by opiate antagonist naloxone. In contrast, both KN-04 and KN-92, the inactive structural analogs of KN-62 and KN-93, failed to attenuate morphine tolerance and dependence, indicating that the observed effects of KN-62 and KN-93 are mediated through inhibition of CaMKII. Furthermore, administration of CaMKII antisense oligonucleotide into rat hippocampal dentate gyrus, which decreased the expression of CaMKII specifically, also attenuated morphine tolerance and dependence, while the corresponding sense oligonucleotide of CaMKII did not exhibit such inhibitory effect. Moreover, the KN-62 treatment abolished the rewarding properties of morphine as measured by the conditioned place preference. These results suggest that hippocampal CaMKII is critically involved in the development of morphine tolerance and dependence, and inhibition of this kinase may have some therapeutic benefit in the treatment of opiate tolerance and dependence.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗

PubMed1999

Pharmacokinetics of the antiviral agent beta-D-2',3'-didehydro-2',3'-dideoxy-5-fluorocytidine in rhesus monkeys.

The values of the pharmacokinetic parameters of the nucleoside antiretroviral agent beta-D-2',3'-didehydro-2',3'-dideoxy-5-fluorocytidine (D-D4FC) in rhesus monkeys were determined with a two-compartment model after the administration of a single dose. The average values for the terminal half-life, renal clearance, and total systemic clearance for the intravenous administration route were 3.6 h and 0.31 and 0.43 liter.kg-1.h-1, respectively. The oral bioavailability of D-D4FC averaged 41%. For the intravenous administration route, 76% of the compound was recovered intact in the urine within 8 h, indicating that D-D4FC was eliminated mainly by renal excretion. D-D4FC was detected in the cerebrospinal fluid (CSF) at similar concentrations after administration by both the intravenous and oral routes. D-D4FC levels in plasma and CSF were higher than the median effective concentration for human immunodeficiency virus type 1 in vitro.

Animals↗

PubMed1999

The Gal3p-Gal80p-Gal4p transcription switch of yeast: Gal3p destabilizes the Gal80p-Gal4p complex in response to galactose and ATP.

The Gal3, Gal80, and Gal4 proteins of Saccharomyces cerevisiae comprise a signal transducer that governs the galactose-inducible Gal4p-mediated transcription activation of GAL regulon genes. In the absence of galactose, Gal80p binds to Gal4p and prohibits Gal4p from activating transcription, whereas in the presence of galactose, Gal3p binds to Gal80p and relieves its inhibition of Gal4p. We have found that immunoprecipitation of full-length Gal4p from yeast extracts coprecipitates less Gal80p in the presence than in the absence of Gal3p, galactose, and ATP. We have also found that retention of Gal80p by GSTG4AD (amino acids [aa] 768 to 881) is markedly reduced in the presence compared to the absence of Gal3p, galactose, and ATP. Consistent with these in vitro results, an in vivo two-hybrid genetic interaction between Gal80p and Gal4p (aa 768 to 881) was shown to be weaker in the presence than in the absence of Gal3p and galactose. These compiled results indicate that the binding of Gal3p to Gal80p results in destabilization of a Gal80p-Gal4p complex. The destabilization was markedly higher for complexes consisting of G4AD (aa 768 to 881) than for full-length Gal4p, suggesting that Gal80p relocated to a second site on full-length Gal4p. Congruent with the idea of a second site, we discovered a two-hybrid genetic interaction involving Gal80p and the region of Gal4p encompassing aa 225 to 797, a region of Gal4p linearly remote from the previously recognized Gal80p binding peptide within Gal4p aa 768 to 881.

Adenosine Triphosphate↗

PubMed1999

Cigarette smoking delays ulcer healing: role of constitutive nitric oxide synthase in rat stomach.

Epidemiological studies have shown that cigarette smoking is associated with peptic ulceration. This study aims to investigate the mechanisms by which cigarette smoking delays ulcer healing in rats. Gastric ulcers were induced by applying acetic acid to the luminal surfaces in rats. Twenty-four hours later, rats were exposed to different concentrations of cigarette smoke (0, 2, or 4%) for a 1-h period once daily for 3 or 6 days. Cigarette smoke exposure delayed ulcer healing and decreased gastric blood flow and angiogenesis at the ulcer margin. These changes were accompanied by a significant reduction of constitutive nitric oxide synthase (cNOS) activity but not PGE2 production and vascular endothelial growth factor levels. Administration of L-arginine (10 mg/kg iv) completely reversed the adverse actions on ulcer healing, gastric blood flow, and angiogenesis in the mucosa at the ulcer margin but partially restored angiogenesis in granulation tissues. In conclusion, cigarette smoke exposure delays ulcer healing through depression of gastric blood flow and angiogenesis at the ulcer margin. Reduction of cNOS expression and activity is suggested to be involved in these ulcerogenic processes.

Animals↗

PubMed1999

Hoxa-10 regulates uterine stromal cell responsiveness to progesterone during implantation and decidualization in the mouse.

Hoxa-10 is an AbdominalB-like homeobox gene that is expressed in the developing genitourinary tract during embryogenesis and in the adult uterus during early pregnancy. Null mutation of Hoxa-10 in the mouse causes both male and female infertility. Defective implantation and decidualization resulting from the loss of maternal Hoxa-10 function in uterine stromal cells is the cause of female infertility. However, the mechanisms by which Hoxa-10 regulates these uterine events are unknown. We have identified two potential mechanisms for these uterine defects in Hoxa-10(-/-) mice. First, two PGE2 receptor subtypes, EP3 and EP4, are aberrantly expressed in the uterine stroma in Hoxa-10(-/-) mice, while expression of several other genes in the stroma (TIMP-2, MMP-2, ER, and PR) and epithelium (LIF, HB-EGF, Ar, and COX-1) are unaffected before implantation. Further, EP3 and EP4 are inappropriately regulated by progesterone (P4) in the absence of Hoxa-10, while PR, Hoxa-11 and c-myc, three other P4-responsive genes respond normally. These results suggest that Hoxa-10 specifically mediates P4 regulation of EP3 and EP4 in the uterine stroma. Second, since Hox genes are implicated in local cell proliferation, we also examined steroid-responsive uterine cell proliferation in Hoxa-10(-/-) mice. Stromal cell proliferation in mutant mice in response to P4 and 17beta-estradiol (E2 was significantly reduced, while epithelial cell proliferation was normal in response to E2. These results suggest that stromal cell responsiveness to P4 with respect to cell proliferation is impaired in Hoxa-10(-/-) mice, and that Hoxa-10 is involved in mediating stromal cell proliferation. Collectively, these results suggest that Hoxa-10 mutation causes specific stromal cell defects that can lead to implantation and decidualization defects apparently without perturbing epithelial cell functions.

Animals↗

PubMed1999

Microtubule bending and breaking in living fibroblast cells.

Microtubules in living cells frequently bend and occasionally break, suggesting that relatively strong forces act on them. Bending implies an increase in microtubule lattice energy, which could in turn affect the kinetics and thermodynamics of microtubule-associated processes such as breaking. Here we show that the rate of microtubule breaking in fibroblast cells increases approximately 40-fold as the elastic energy stored in curved microtubules increases to > approximately 1 kT/tubulin dimer. In addition, the length-normalized breaking rate is sufficiently large (2.3 breaks x mm(-1) x minute(-1)) to infer that breaking is likely a major mechanism by which noncentrosomal microtubules are generated. Together the results suggest a physiologically important, microtubule-based mechanism for mechanochemical information processing in the cell.

3T3 Cells↗

PubMed1999

Differential serodiagnosis for cystic and alveolar echinococcosis using fractions of Echinococcus granulosus cyst fluid (antigen B) and E. multilocularis protoscolex (EM18).

Echinococcus granulosus cyst fluid and E. multilocularis protoscolex extract were fractionated by a single step of preparative isoelectric focusing, resulting in an antigen B-rich fraction (8-kD) and an Em18-rich fraction, respectively. The usefulness of both fractions for differential serodiagnosis of cystic (CE) and alveolar (AE) echinococcosis was evaluated by a large-scale immunoblot analysis on a battery of 354 serum samples. These included 66 from AE patients originating from four different endemic areas, 173 from CE patients originating from seven different endemic areas, 71 from patients with other parasitic diseases, 15 from patients with hepatomas, and 29 from healthy individuals. In an immunoblot with the antigen B-rich fraction, 92% (158 of 173) of the CE sera as well as 79% (52 of 66) of the AE sera reacted with the 8-kD subunit. No cross-reactivity occurred with any sera from patients with cysticercosis, other parasitic diseases, or with hepatomas, or from healthy controls. In an immunoblot with the Em18-rich fraction, all but two sera from AE patients (64 of 66, 97%) recognized Em18, and only nine of 34 CE sera from China reacted with it. All other (139) CE sera from six other countries were negative as were all (115) other non-echinococcosis sera. These findings indicate that antigen B (8-kD) is not species-specific for E. granulosus but is genus-specific for Echinococcus, and that the Em18 antigen is a reliable serologic marker for species-specific differentiation of AE from CE.

Animals↗

PubMed1999

Prostaglandin E-prostanoid-3 receptor activation of cyclic AMP response element-mediated gene transcription.

The prostaglandin E-prostanoid (EP)3 receptor signals primarily through the inhibitory G protein Gi, thereby decreasing intracellular cAMP levels. To study the signal transduction properties of the rabbit EP3 receptor, five splice variants were expressed in HEK293tsA201 cells: 72A, 74A, 77A, 80A and the novel splice variant NT, which lacks the C-terminal sequence. The ability of the EP3 receptor splice variants to modulate expression of a beta-galactosidase reporter gene under the control of a promoter containing cAMP response elements (CRE) was assessed. Each splice variant induced sulprostone-mediated increase in beta-galactosidase enzymatic activity with EC50 ranging from 0.8 nM for the NT splice variant to 3.1 nM for the 77A splice variant. Substitution of either Asp338 with Ala, or Arg329 with Ala or Glu in the 77A splice variant resulted in a loss of receptor-evoked increases in beta-galactosidase activity, whereas substitution of Lys300 with alanine had no effect on signal transduction. These phenotypes correlate with the inhibition of cAMP generation by direct cAMP measurement. Signal transduction was insensitive to pretreatment of cells with pertussis toxin, suggesting that a nonGi/Go pathway is activated by the EP3 receptor. Direct measurement of second messenger levels confirmed that there was no increase in cAMP levels mediated by the 77A splice variant, however, there was a modest increase in intracellular Ca2+. Partial blockade of the reporter activity with kinase inhibitors demonstrates that CRE activation is mediated in part by a Ca2+-dependent kinase pathway. These data suggest that the EP3 receptor signals through a novel cAMP response element binding protein/CRE pathway.

Alternative Splicing↗

PubMed1999

[Molecular characterization of glucose-6-phosphate dehydrogenase variants in four ethnic groups in Yunnan province of China].

OBJECTIVE: In order to understand the molecular evolution, race origin and the relationship between the G6PD gene structure and clinical symptoms, the authors identified the molecular characterization of glucose-6-phosphate dehydrogenase and determined the G6PD gene frequency in four ethnic groups in Yunnan province of China. METHODS: The point mutations of G6PD were detected by Mismatch-PCR/RE, SSCP,ARMS,DNA sequence and so on. G6PD gene frequency was determined by Hardy-Weinberg Law. RESULTS: G6PD G1388A, G1376T,A95G mutations were determined in Bai and Dai people for the first time and G1388A also in Harni people by DNA sequence. G6PD C1024T were detected in Dai population by Mismatch-PCR/RE. The gene frequency of G6PD in Bai population in Dali city is 0.0113, and the incidence is 1.19% which are different from those in Dai population. CONCLUSION: G6PD G1388A,G1376T, A95G and C1024T are the mutations in national minorities as well as in the Han people. The results suggest that different national minorities of China may have the same ancestor. The incidence of G6PD deficiency and the G6PD gene frequency in Bai population are different from those in Dai population. The distribution of G6PD deficiency in Yunnan is associated with the distribution of malaria epidemic in that province.

China↗

PubMed1999

Unalloyed pyrolytic carbon for implanted mechanical heart valves.

Materials for implanted heart valves face challenges unmatched for most mechanical applications. The valves must function without interruption for lifetimes in the order of 10(9) cycles in a corrosive, hostile environment. A particular carbon has been widely used for the past three decades. The mechanical behavior of this isotropic pyrolytic carbon (PyC), with and without silicon alloying, was studied in this research program. Several questions were addressed: the fatigue strength at lifetimes of 10(9) cycles, the threshold for crack growth, the validity of fracture mechanics for PyC, and the statistics of ultra-high survival under cyclic stress. It was found that cyclic stressing within the scatter band of the static strength did not initiate cracks. Furthermore, artificially induced cracks did not grow under cyclic stressing below a threshold stress intensity factor. This threshold stress intensity factor was valid over a wide range of crack lengths. No failures occurred when batches of 29 specimens were tested above the service stress at 6 x 10(8) cycles. Cyclic stressing did not reduce the static strength. The service stress is a small fraction of the fracture strength; thus, a very high probability of survival can be ensured by a proof test of the assembly.

Biocompatible Materials↗

PubMed1999

The temperature dependence of protoporphyrin IX production in cells and tissues.

The formation of protoporphyrin IX (PpIX) in human skin during topical application of 5-aminolevulinic acid (ALA) was found to be strongly temperature dependent, with an activation energy of about 17 kcal/mol. This temperature dependence is mainly related to porphyrin production and not to ALA penetration into the skin. The penetration of ALA into mouse and human skin was almost temperature independent. The activation energy of PpIX production in mouse skin was practically identical with that in human skin. The activation energy of ALA uptake by cells in vitro was about 10 kcal/mol and that for PpIX production was about 13 kcal/mol. The latter activation energy was within the error limits similar to that for the activity of the enzyme porphobilinogen deaminase, suggesting that this enzyme might represent a rate-limiting step for PpIX production in living tissue.

Administration, Topical↗

PubMed1999

Site-directed mutagenesis analysis of draTG genes and their downstream region from Azospirillum brasilense Yu62.

draT and draG genes are involved in posttranslational regulation of nitrogenase activity of Azospirillum brasilense Yu62. Both genes and their downstream region were mutagenized by Kmr cassette insertions. Analysis of mutations introduced into the draTG region on the A. brasilense Yu62 chromosome showed that mutants affected in draT were incapable of regulating nitrogenase activity in response to ammonium. In contrast, a mutant with an insertion in draG was still capable of ADP-ribosylating dinitrogenase reductase in response to ammonium but was no longer able to recover activity after ammonium depletion. Analysis of mutations introduced into the draTG downstream region (the mutagenized site is about 2 kb downstream from draG) showed that the mutant had higher nitrogenase activity than the wild strain while growing in nitrogen-free medium and medium with 2 mmol/L ammonium. These results reveal that there is no gene required for nitrogen fixation in this mutagenized region, but it is possible that there are genes which play a role in regulating nitrogen fixation. The results of monitoring the expression of transcriptional nifH-lacZ gene fusion in the mutant YZ4 showed that the transcriptional regulation of nif gene in the mutant YZ4 was the same as that in the wild type.

Azospirillum brasilense↗

PubMed1999

[A rational selection of retroperitoneal lymphadenectomy for advanced epithelial ovarian cancer].

OBJECTIVE: To evaluate the rational application of retroperitoneal lymphadenectomy to advanced epithelial ovarian cancer. METHODS: 42 patients of advanced epithelial ovarian cancer were treated by retroperitoneal lymphadenectomy. Two groups were divided according to the residual disease post-operation. A: 26 patients with the residual disease < 2 cm; B: 16 patients > or = 2 cm. The regime of combined chemotherapy was the same in the two groups after operation. Clinical stage and pathologic grade showed no difference. RESULTS: The 5-year survival rate was 53.8% (14/26) in A and 12.5% (2/16) in B. There was a significant difference between the two groups (P < 0.001). CONCLUSIONS: The survival rate could be greatly improved for advanced epithelial ovarian cancer through retroperitoneal lymphadenectomy when the residual disease was smaller than 2 cm. This procedure would not be performed if the residual disease was larger than or equal to 2 cm.

Adolescent↗

PubMed1999

[Ailing No. I in treating 62 cases of acute promyelocytic leukemia].

OBJECTIVE: To obtain further knowledge on the effect, mechanism and toxic side-effect of Ailing No. I (AL), a preparation of As2O3, in treating acute promyelocytic leukemia (APL). METHODS: AL was used in induction and consolidation treatment of 62 cases of APL, the effect of treatment and the changes of clinical symptoms, peripheral blood and bone marrow pictures, T-lymphocyte subset, and immunofunctions of patients after treatment were observed. RESULTS: Twenty-seven cases out of the 31 patients who received induction treatment got complete remission (CR), the CR rate being 87.1%. Of the 31 cases who received consolidation treatment, the longest remission period was 37 months, and 7 cases relapsed within one month to 2 years. Peripheral white blood cell count raised after one week but lowered after 4 weeks of induction treatment, but in receiving consolidation treatment, it started to lower after one week and raised after 4 weeks of treatment. No obvious change on hemoglobin level was observed. Platelet count significantly increased after treatment. The treatment showed no obvious inhibition on bone marrow. The T-lymphocyte subsets and immunoglobulin level were changed insignificantly after treatment. Various side-effects were shown clinically, particularly those digestive tract involved symptoms were prominent, and liver damage was shown in a certain degree. CONCLUSION: AL has good and protracted effect in treating APL with high CR rate, the mechanism may be related with its cytotoxicity and action of differentiation induction.

Adolescent↗

PubMed1999

[Clinical observation on treatment of postcardiotomic complications with Chinese herbal medicine based on syndrome differentiation with angiocardiopathy].

OBJECTIVE: To study the effect of Chinese herbal medicine (CHM) based on Syndrome Differentiation on postcardiotomic complications in patients with angiocardiopathy. METHODS: Aimed at the frequently encountered postcardiotomic complications including fever, cough and expectoration, belching, abdominal distension, palpitation, short breath, etc. CHM treatment was applied in combination with routine western drugs treatment (cardiac tonic, diuretics, vascular dilatator and anticoagulant). RESULTS: Twenty out of 22 patients with protracted fever and irresponsive to multi-antibiotics therapy were cured, the other one with hydrothorax received other therapy and the another one with drug fever was natural cured after stopping medication. Among 23 patients complicated mainly with respiratory symptoms, 17 were cured and 6 improved, among 15 with digestive symptoms, 12 cured and 3 improved, and among 7 with cardiovascular symptoms, 3 cured, 2 improved and 2 ineffective. CONCLUSION: CHM has good effect on postcardiotomic complications, it could improve the functional recovery of heart and lung.

Adolescent↗

PubMed1999

[Nested polymerase chain reaction in detecting Mycobacterium tuberculosis DNA in bioptic tissue from patients with endobronchial tuberculosis].

OBJECTIVE: To evaluate the nested polymerase chain reaction (NPCR) technique in diagnosing endobronchial tuberculosis. METHODS: NPCR method was used to detect Mycobacterium tuberculosis DNA in bioptic tissue from 67 patients with endobronchial tuberculosis, and the results were compared with pathologic examination, brushing smear, sputum smear and culture for Mycobacterium tuberculosis after bronchoscopy. The controls were 43 patients with lung cancer. RESULTS: The positive rates of detecting Mycobacterium tuberculosis were 13%, 19%, 22%, 15% and 76% respectively by pathologic examination, brushing smear, sputum smear, culture after bronchoscopy and NPCR methods. Significant differences were found between NPCR and the other 4 methods (all Ps < 0.01). No positive result for Mycobacterium tuberculosis was found in 43 controls by NPCR. CONCLUSIONS: NPCR is a useful method for diagnosing patients with endobronchial tuberculosis, especially for those with normal demonstration in X-ray chest film, negative sputum and nonspecific pathologic changes in endobronchial biopsy.

Biopsy↗