Search PubMed⌕ Search

Biomedical subjects

M M Wu

Publications and source records attributed to M M Wu.

At least 19 recordsLinked to original sources
PubMed2002

Biosynthesis and secretion of pituitary hormones: dynamics and regulation.

Production and secretion of hormones by the pituitary involve highly orchestrated intracellular transport and sorting steps. Hormone precursors are routed through a series of compartments before being packaged in secretory granules. These highly dynamic carriers play crucial roles in both prohormone processing and peptide exocytosis. We have employed the ACTH-secreting AtT-20 cell line to study the membrane sorting events that confer functionality (prohormone activation and regulated exocytosis) to these secretory carriers. The unique ability of granules to promote prohormone processing is attributed to their acidic interior. Using a novel avidin-targeted fluorescence ratio imaging technique, we have found that the trans-Golgi of live AtT-20 cells maintains a mildly acidic (approximately pH 6.2) interior. Budding of secretory granules causes the lumen to acidify to <pH 6.0, which is both necessary and sufficient to trigger SPC3-mediated proteolytic conversion of proopiomelanocortin to ACTH. Investigation of the pH regulatory mechanism indicates that the trans-Golgi and secretory granules maintain different pH values by distinct sorting of key membrane transporters. Mathematical modeling of our data suggests that the decreasing pH values of organelles of the regulated secretory pathway is established by gradually increasing the density of active H+ pumps from the ER to Golgi while concomitantly decreasing the H+ permeability from ER to Golgi to secretory granules. An in vitro assay was developed to study the formation of processing-competent secretory granules from their processing-incompetent precursor trans-Golgi compartment. Our data suggest that ARF1-mediated sorting of proton pumps and leaks during early stages of granule formation confers processing competency to the resulting organelle. Once formed, these young granules continue to undergo membrane remodeling which results in dynamic changes in their exocytotic behavior. Two SNAREs, VAMP4 and synaptotagmin IV, enter newly formed granules but are removed from the maturing granule membrane by vesicle budding. Sorting of these proteins is correlated with the acquisition of Ca2+-triggered exocytosis and a decrease in unregulated exocytotic rate. Thus, biosynthesis and secretion of pituitary hormones are dynamically regulated by intracellular sorting events that govern the functions of their secretory carriers.

Animals↗
PubMed2001

Mechanisms of pH regulation in the regulated secretory pathway.

A precise pH gradient between organelles of the regulated secretory pathway is required for sorting and processing of prohormones. We studied pH regulation in live endocrine cells by targeting biotin-based pH indicators to cellular organelles expressing avidin-chimera proteins. In AtT-20 cells, we found that steady-state pH decreased from the endoplasmic reticulum (ER) (pH(ER) = 7.4 +/- 0.2, mean +/- S.D.) to Golgi (pH(G) = 6.2 +/- 0.4) to mature secretory granules (MSGs) (pH(MSG) = 5.5 +/- 0.4). Golgi and MSGs required active H(+) v-ATPases for acidification. ER, Golgi, and MSG steady-state pH values were also dependent upon the different H(+) leak rates across each membrane. However, neither steady-state pH(MSG) nor rates of passive H(+) leak were affected by Cl(-)-free solutions or valinomycin, indicating that MSG membrane potential was small and not a determinant of pH(MSG). Therefore, our data do not support earlier suggestions that organelle acidification is primarily regulated by Cl(-) conductances. Measurements of H(+) leak rates, buffer capacities, and estimates of surface areas and volumes of these organelles were applied to a mathematical model to determine the H(+) permeability (P(H+)) of each organelle membrane. We found that P(H+) decreased progressively from ER to Golgi to MSGs, and proper acidification of Golgi and MSGs required gradual decreases in P(H+) and successive increases in the active H(+) pump density.

Animals↗
PubMed2001

Lower BMI cut-off value to define obesity in Hong Kong Chinese: an analysis based on body fat assessment by bioelectrical impedance.

There is increasing evidence suggesting that the cut-off values for defining obesity used in the Western countries cannot be readily applied to Asians, who often have smaller body frames than Caucasians. We examined the BMI and body fat (BF) as measured by bioelectrical impedance in 5153 Hong Kong Chinese subjects. We aimed to assess the optimal BMI reflecting obesity as defined by abnormal BF in Hong Kong Chinese. Receiver operating characteristic curve (ROC) analysis was used to assess the optimal BMI predicting BF at different levels. The mean age and SD of the 5153 subjects (3734 women and 1419 men) was 51.5 (SD 16.3) years (range: 18.0-89.5 years, median: 50.7 years). Age-adjusted partial correlation (r) between BMI and BF in women and men were 0.899 (P < 0.001) and 0.818 (P < 0.001) respectively. Using ROC analysis, the BMI corresponding to the conventional upper limit of normal BF was 22.5-23.1 kg/m(2), and the BMI corresponding to the 90 percentiles of BF was 25.4-26.1 kg/m(2). Despite similar body fat contents, the BMI cut-off value used to define obesity in Hong Kong Chinese should be lower as compared to Caucasians. We suggest a BMI of 23 kg/m(2) and 26 kg/m(2) as the cut-off values to define overweight and obesity respectively in Hong Kong Chinese.

Absorptiometry, Photon↗
PubMed2001

Association of blood arsenic levels with increased reactive oxidants and decreased antioxidant capacity in a human population of northeastern Taiwan.

Arsenic is a notorious environmental toxicant known as both a carcinogen and an atherogen in human beings, but the pathogenic mechanisms are not completely understood. In cell culture studies, trivalent arsenic enhanced oxidative stress in a variety of mammalian cells, and this association may be closely associated with the development of arsenic-related diseases. To investigate the effect of arsenic exposure on oxidative stress in humans, we conducted a population study to determine the relationships of blood arsenic to reactive oxidants and antioxidant capacity at the individual level. We recruited 64 study subjects ages 42-75 years from residents of the Lanyang Basin on the northeast coast of Taiwan, where arsenic content in well water varies from 0 to > or = 3,000 microg/L. We used a chemiluminescence method, with lucigenin as an amplifier for measuring superoxide, to measure the plasma level of reactive oxidants. We used the azino-diethyl-benzthiazoline sulphate method to determine the antioxidant capacity level in plasma of each study subject. We determined arsenic concentration in whole blood by hydride formation with an atomic absorption spectrophotometer. The average arsenic concentration in whole blood of study subjects was 9.60 +/- 9.96 microg/L (+/- SD) with a range from 0 to 46.50 microg/L. The level of arsenic concentration in whole blood of study subjects showed a positive association with the level of reactive oxidants in plasma (r = +0.41, p = 0.001) and an inverse relationship with the level of plasma antioxidant capacity (r = -0.30, p = 0.014). However, we found no significant association (p = 0.266) between levels of plasma reactive oxidants and antioxidant capacity. Our results also show that the lower the primary arsenic methylation capability, the lower the level of plasma antioxidant capacity (p = 0.029). These results suggest that ingestion of arsenic-contaminated well water may cause deleterious effects by increasing the level of reactive oxidants and decreasing the level of antioxidant capacity in plasma of individuals. Persistent oxidative stress in peripheral blood may be a mechanism underlying the carcinogenesis and atherosclerosis induced by long-term arsenic exposure.

Adult↗
PubMed2001

Body mass index profile in Hong Kong Chinese adults.

INTRODUCTION: Obesity has now become an epidemic with increasing prevalence in most parts of the world. However, information on the problem of obesity in Asians is limited. SUBJECTS AND METHODS: From April 1996 to August 1997, 17,121 Chinese adult subjects from the community of Hong Kong presented themselves voluntarily at the United Christian Nethersole Community Health Service Centers for primary health care check-up. Demographic data including height and weight were documented. Their body mass indexes (BMI) were measured. The prevalence rates of overweight and obese subjects were calculated according to the 1995 World Health Organization criteria. RESULTS: There were 4781 (27.9%) men and 12,340 (72.1%) women. The mean age (+/- SD) was 51.3 +/- 16.0 years (range 20 to 96 years, median 49.8 years). Of the 17,121 subjects, 4939 (28.85%) were overweight and 656 (3.83%) were obese. Based on the 1997 Hong Kong Census statistics of the overall Hong Kong population data, the age-standardised prevalence rates of overweight and obese subjects were 30.52% and 3.19% in men and 22.14% and 3.35% in women, respectively. Of the 10,937 subjects of working age (20 to 65 years), the prevalence rates of overweight and obese subjects were 28.98% and 3.55%, respectively. Of the 6648 subjects aged > or = 60 years, the prevalence rates of overweight and obese subjects were 36.31% and 4.81%, respectively. CONCLUSIONS: In Hong Kong, the age-standardised prevalence rate of overweight and obese subjects were 30.5% and 3.2% in men and 22.1% and 3.4% in women, respectively. This information provides some useful estimations of the extent of an important public health problem, obesity, in Hong Kong.

Adult↗
PubMed2000

4-aminopyridine, a specific blocker of K(+) channels, inhibited inward Na(+) current in rat cerebellar granule cells.

The effects of 4-aminopyridine (4-AP), a specific blocker of outward K(+) current, on voltage-activated transient outward K(+) current (I(K(A))) and inward Na(+) current (I(Na)) were investigated on cultured rat cerebellar granule cells using the whole cell voltage-clamp technique. At the concentration of 1-5 mM, 4-AP inhibited both I(K(A)) and I(Na). It reduced the amplitude of peak Na(+) current without significant alteration of the steady-state activation and inactivation properties. The inhibitory effect was not enhanced by repeated depolarizing pulses (0.5 or 0.1 Hz), suggesting that the binding affinity of 4-AP on Na(+) channels is state-independent. In contrast, the effect of 4-AP on Na(+) channels appeared to be voltage-dependent, the weaker inhibition occurred at more depolarization. Moreover, 4-AP slowed both the activation and inactivation kinetics of Na(+) current. These effects were similar to those induced by alpha-scorpion toxin and sea anemone toxins on Na(+) channels in other cell model. Our data demonstrate for the first time that 4-AP is able to block not only A-type K(+) channels, but also Na(+) channels in rat cerebellar granule cells. It is concluded that the inhibition exerted by 4-AP on Na(+) current likely differs from that provoked by local anesthetics. The possibility that the binding site of neurotoxin receptor 3 may be involved is discussed.

4-Aminopyridine↗
PubMed2000

Organelle pH studies using targeted avidin and fluorescein-biotin.

BACKGROUND: Mammalian organelles of the secretory pathway are of differing pH. The pH values form a decreasing gradient: the endoplasmic reticulum (ER) is nearly neutral, the Golgi is mildly acidic and the secretory granules are more acidic still ( approximately pH 5). The mechanisms that regulate pH in these organelles are still unknown. RESULTS: Using a novel method, we tested whether differences in H(+) 'leak' and/or counterion conductances contributed to the pH difference between two secretory pathway organelles. A pH-sensitive, membrane-permeable fluorescein-biotin was targeted to endoplasmic-reticulum- and Golgi-localized avidin-chimera proteins in HeLa cells. In live, intact cells, ER pH (pH(ER)) was 7.2 +/- 0.2 and Golgi pH (pH(G)) was 6.4 +/- 0.3 and was dissipated by bafilomycin. Buffer capacities of the cytosol, ER and Golgi were all similar (6-10 mM/pH). ER membranes had an apparent H(+) permeability three times greater than that of Golgi membranes. Removal of either K(+) or Cl(-) did not affect ER and Golgi H(+) leak rates, or steady-state pH(G) and pH(ER). CONCLUSIONS: The Golgi is more acidic than the ER because it has an active H(+) pump and fewer or smaller H(+) leaks. Neither buffer capacity nor counterion permeabilities were key determinants of pH(G), pH(ER) or ER/Golgi H(+) leak rates.

Avidin↗
PubMed2000

Risk of internal cancers from arsenic in drinking water.

The U.S. Environmental Protection Agency is under a congressional mandate to revise its current standard for arsenic in drinking water. We present a risk assessment for cancers of the bladder, liver, and lung from exposure to arsenic in water, based on data from 42 villages in an arseniasis-endemic region of Taiwan. We calculate excess lifetime risk estimates for several variations of the generalized linear model and for the multistage-Weibull model. Risk estimates are sensitive to the model choice, to whether or not a comparison population is used to define the unexposed disease mortality rates, and to whether the comparison population is all of Taiwan or just the southwestern region. Some factors that may affect risk could not be evaluated quantitatively: the ecologic nature of the data, the nutritional status of the study population, and the dietary intake of arsenic. Despite all of these sources of uncertainty, however, our analysis suggests that the current standard of 50 microg/L is associated with a substantial increased risk of cancer and is not sufficiently protective of public health.

Adult↗
PubMed2000

A rapid method for evaluating microbicidal activity of dentifrice formulations against salivary bacteria ex vivo.

A new rapid ex vivo method was developed for evaluating the short-term bactericidal activity of dentifrices against salivary microorganisms. Dentifrice aliquots of 0.25 or 1.0 g were rapidly dispersed into 3.5 mL of freshly pooled human saliva, and 1.0 mL aliquots of the dentifrice-saliva suspension were collected after 30 or 60 seconds of exposure, diluted in neutralizing broth and plated on non-selective agar media for enumeration of surviving total cultivable microflora. Eight experimental dentifrices containing increasing amounts (0 to 2.6%) of a fixed ratio of essential oils (thymol, menthol, methyl salicylate and eucalyptol) were dispersed in saliva at a 0.25:3.5 (w/v) dentifrice:saliva ratio. Recoverable CFUs/mL at 30 sec. were reduced in a dose-responsive manner from > 10(6) to < 10(4). Additional tests using both 0.25 and 1.0 g amounts of dentifrice prototypes containing 2.1% of the essential oil mixture showed that the experimental dentifrices exhibited highly significant (approximately 2.5 log) reductions in viable recoverable microorganisms relative to essential oil-free placebos after both 30- and 60-second exposures. When compared to in vitro models previously used to evaluate the antimicrobial activity of dentifrices, this method provides a rapid, reproducible, more biologically representative means of screening dentifrice formulations for microbicidal activity using dilutions and exposure times approximating those achieved during toothbrushing. However, since other factors may influence microbicidal activity of dentifrice formulations in vivo, conclusions drawn using this model require clinical confirmation.

Analysis of Variance↗
PubMed2000

Swelling-activated chloride currents in embryonic chick heart cells.

AIM: To characterize a swelling-activated chloride current, I(Cl, swell), in white Leghorn chick heart cells and the effects of chlorpromazine (CPZ) effects. METHODS: The patch-clamp technique in the whole-cell configuration was used. RESULTS: Hyposmotic swelling elicited I(Cl, swell) in white Leghorn chick heart cells. The current amplitude increased from (452 +/- 200) pA to (849 +/- 373) pA with a reduction of osmolarity from 300 mmol.L-1 to 270 mmol.L-1, 4',4-Diisothiocyanostilbene-2,2'-disulphonic acid (DIDS) 100 mumol.L-1 decreased I(Cl, swell) from (1196 +/- 505) pA to (830 +/- 328) pA in hyposmotic solution. In white Leghorn chick heart cells I(Cl, swell) was not induced by CPZ 30 mumol.L-1, which is different from the case of E coli spheroplast. CONCLUSION: Swelling-activated chloride current was activated by hyposmotic swelling in white Leghorn chick heart cells. The mechanism for activating the current is different from that of mechanosensitive ion channels of E coli.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed2000

Anticalculus effect of two zinc citrate/essential oil-containing dentifrices.

PURPOSE: To assess the feasibility of incorporating zinc citrate, a known anticalculus ingredient, into a dentifrice formulation containing a fixed combination of essential oils, a known antiplaque/antigingivitis agent. MATERIALS AND METHODS: This randomized, parallel, double-blind study evaluated the potential of two essential oil dentifrice formulations containing different levels of zinc citrate (1.0% and 2.0% ZCT) to reduce supragingival calculus formation compared to a marketed control dentifrice, Crest Regular. Following a 3-month pre-test phase, subjects received a dental prophylaxis, were stratified into three balanced groups on the basis of Volpe-Manhold calculus scores and brushed twice daily with their assigned dentifrice for 3 months. RESULTS: One hundred ninety-six evaluable subjects completed all phases of the study. ANCOVA revealed that the 1.0% ZCT and 2.0% ZCT essential oil dentifrice formulations provided significant reductions in calculus formation of 26.4% and 29.0% (P< 0.001), respectively, compared to the control dentifrice, Crest Regular. The magnitude of calculus reductions is similar to those levels obtained by other zinc salt formulations.

Adolescent↗
PubMed2000

Essential oils in an antiplaque and antigingivitis dentifrice: a 6-month study.

PURPOSE: To determine if a dentifrice formulated with the fixed combination of essential oils possessed antiplaque and antigingivitis properties. MATERIALS AND METHODS: This study was a blinded, randomized, controlled clinical trial. The 6-month study had clinical examinations at baseline, 3 and 6 months. The examinations consisted of three indices; a plaque index, the Modified Gingival Index and a bleeding index. RESULTS: At 6 months, the essential oil-containing dentifrice group had statistically significantly (P < 0.001) lower whole-mouth and interproximal mean plaque index scores exhibiting mean reductions of 18.3% and 18.1% relative to the vehicle control. At 6 months, the essential oil-containing dentifrice group had statistically significantly (P < 0.001) lower whole-mouth and interproximal mean gingival index scores compared to the vehicle control with reductions of 16.2% and 15.5%, respectively. Finally, for the gingival bleeding index, the essential oil-containing dentifrice group had statistically significantly (P < 0.001) lower whole-mouth and interproximal mean bleeding index scores and reductions of 40.5% and 46.9% at 6 months compared to the vehicle control.

Adolescent↗
PubMed1999

DNA repair methyltransferase (Mgmt) knockout mice are sensitive to the lethal effects of chemotherapeutic alkylating agents.

We have generated mice deficient in O6-methylguanine DNA methyltransferase activity encoded by the murine Mgmt gene using homologous recombination to delete the region encoding the Mgmt active site cysteine. Tissues from Mgmt null mice displayed very low O6-methylguanine DNA methyltransferase activity, suggesting that Mgmt constitutes the major, if not the only, O6-methylguanine DNA methyltransferase. Primary mouse embryo fibroblasts and bone marrow cells from Mgmt -/- mice were significantly more sensitive to the toxic effects of the chemotherapeutic alkylating agents 1,3-bis(2-chloroethyl)-1-nitrosourea, streptozotocin and temozolomide than those from Mgmt wild-type mice. As expected, Mgmt-deficient fibroblasts and bone marrow cells were not sensitive to UV light or to the crosslinking agent mitomycin C. In addition, the 50% lethal doses for Mgmt -/- mice were 2- to 10-fold lower than those for Mgmt +/+ mice for 1,3-bis(2chloroethyl)-1-nitrosourea, N-methyl-N-nitrosourea and streptozotocin; similar 50% lethal doses were observed for mitomycin C. Necropsies of both wild-type and Mgmt -/mice following drug treatment revealed histological evidence of significant ablation of hematopoietic tissues, but such ablation occurred at much lower doses for the Mgmt -/- mice. These results demonstrate the critical importance of O6-methylguanine DNA methyltransferase in protecting cells and animals against the toxic effects of alkylating agents used for cancer chemotherapy.

Alkylating Agents↗
PubMed1999

Fluconazole prophylaxis prevents intra-abdominal candidiasis in high-risk surgical patients.

OBJECTIVE: To evaluate the efficacy and safety of intravenous fluconazole for the prevention of intra-abdominal Candida infections in high-risk surgical patients. DESIGN: Randomized, prospective, double-blind, placebo-controlled study. SETTING: Two university-affiliated hospitals in Switzerland. PATIENTS: Forty-nine surgical patients with recurrent gastrointestinal perforations or anastomotic leakages. INTERVENTIONS: Prophylaxis with intravenous fluconazole (400 mg per day) or placebo continued until resolution of the underlying surgical condition. MEASUREMENTS AND MAIN RESULTS: Patients were evaluated daily, and specimens for culture were obtained three times per week during prophylaxis. The primary study end points were the frequency of and the time to intra-abdominal Candida infections. Secondary end points were the frequency of candidiasis (intra-abdominal and extra-abdominal) and the emergence or persistence of Candida colonization. Among patients who were not colonized at study entry, Candida was isolated from surveillance cultures during prophylaxis in 15% of the patients in the fluconazole group and in 62% of the patients in the placebo group (relative risk, 0.25; 95% confidence interval, 0.07 to 0.96; p = .04). Candida peritonitis occurred in one of 23 patients (4%) who received fluconazole and in seven of 20 patients (35%) who received placebo (relative risk, 0.12; 95% confidence interval, 0.02 to 0.93; p = .02). In addition, one catheter-related Candida albicans sepsis occurred in a fluconazole-treated patient. Thus, overall, candidiasis developed in two fluconazole patients and seven placebo patients (relative risk, 0.25; 95% confidence interval, 0.06 to 1.06; p = .06). C. albicans accounted for 87% of the Candida species isolated before or during prophylaxis, and all C. albicans strains were susceptible to fluconazole. Fluconazole was well tolerated, and adverse events occurred at similar frequencies in both treatment groups. CONCLUSIONS: Fluconazole prophylaxis prevents colonization and invasive intra-abdominal Candida infections in high-risk surgical patients.

Abdomen↗
PubMed1999

The effect of having myopic parents: an analysis of myopia in three generations.

PURPOSE: The etiology of myopia has been attributed to both genetic and environmental factors. The purpose of this paper is to attempt to distinguish between genetic and environmental factors in three generations of subjects from three locations. METHODS: Noncycloplegic autorefraction and distance visual acuity were measured in 3131 Chinese children 7 to 17 years old from Hong Kong, Tianjin, and Ban Chau. Data on the refractive status of the parents and grandparents were collected using questionnaires. Myopia was defined operationally as spherical equivalent refractive error (SER) of at least -1.00 D and uncorrected vision worse than logMAR 0.18 (6/9). RESULTS: The odds of having myopia for the grandparents', parents', and children's generations were 0.06, 0.26, and 0.35, respectively. Having myopic parents increased the odds ratio for having myopia (odds ratio 1.85 for children's generation and 6.71 for parents' generation), showing a genetic influence. However, the odds of having myopia also increased in offspring of nonmyopic parents between generation 2 and 3, suggesting an environmental influence. Results of prevalence and odds ratios from the three locations also demonstrated an environmental effect on development of myopia. CONCLUSION: We propose that myopic development in Chinese follows a multifactorial and polygenic model in which the genetic input has remained constant while the environmental influence has increased over the last three generations.

Adolescent↗
PubMed1998

A chemical and genetic approach together define the biological consequences of 3-methyladenine lesions in the mammalian genome.

DNA-damaging agents produce a plethora of cellular responses that include p53 induction, cell cycle arrest, and apoptosis. It is generally assumed that it is the DNA damage produced by these agents that triggers such responses, but there is limited direct evidence to support this assumption. Here, we used DNA alkylation repair proficient and deficient isogenic mouse cell lines to demonstrate that the signal to trigger p53 induction, cell cycle arrest, and apoptosis in response to alkylating agents does emanate from DNA damage. Moreover, we established that 3-methyladenine, a relatively minor DNA lesion produced by most methylating agents (which form mainly 7-methylguanine), can specifically induce sister chromatid exchange, chromatid and chromosome gaps and breaks, S phase arrest, the accumulation of p53, and apoptosis. This study was made possible by the generation of 3-methyladenine DNA glycosylase null mutant cells by targeted homologous recombination and by the chemical synthesis of a methylating agent that almost exclusively produces 3-methyladenine DNA lesions. The combined use of these two experimental tools has defined the biological consequences of 3-methyladenine, a DNA lesion produced by endogenous cellular metabolites, environmental carcinogens, and chemotherapeutic alkylating agents.

Adenine↗