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Biomedical subjects

D M Wu

Publications and source records attributed to D M Wu.

At least 19 recordsLinked to original sources

Accuracy of direct digital radiography in the study of the root canal type.

OBJECTIVES: To determine the accuracy of direct digital radiography (DDR) in the study of the root canal type. A sub aim was to assess intraexaminer and interexaminer agreement for this feature using DDR system. METHODS: 100 recently extracted permanent maxillary first premolars from Chinese population were used. Standardized periapical DDR images were taken from a buccolingual and mesiodistal direction. Then the specimens were accessed, injected with dye, demineralized, dehydrated and finally were cleared. The DDR images were evaluated based on the root canal type by two independent trained post-graduate students (examiner A and B) twice at an interval of 2 weeks. The cleared teeth were examined under 5 x magnification using a dental operating microscope by another experienced endodontist, and the data of root canal type were collected. RESULTS: The Kappa values for the agreement between the examiner A and the clearing technique were 0.3793 and 0.329, between the examiner B and the clearing technique were 0.2481 and 0.2184. While the Kappa values between the two observations of each examiner were 0.7704 and 0.7725, between the two examiners they were 0.537 and 0.4793, respectively. CONCLUSIONS: The intraexaminer agreement was good, and the interexaminer agreement was moderate, but the agreement between either DDR examiner and the clearing technique was poor, indicating the limited value of DDR alone when studying root canal type.

Adolescent↗

Determinants and distributions of plasma total homocysteine concentrations among school children in Taiwan.

BACKGROUND: Plasma total homocysteine (tHcy) level is an independent risk factor for cardiovascular disease (CVD) even among children. The purpose of this study is to evaluate the determinants and distributions of plasma tHcy levels and the relationship between plasma tHcy, folate and vitamin B12 levels among school children in Taipei. METHODS: After multi-stage sampling, we randomly selected 1234 school children (609 boys and 625 girls) with the mean age of 13 years (from 12 to 15 years) in this study. Fasting plasma tHcy levels were measured using an ABBOTT IMx analyzer (Axis Biochemicals ASA, Oslo, Norway). Plasma folate and vitamin B12 levels were measured by ACS:180 automated chemiluminescence analyzer (Bayer, Tarrytown, NY, USA). RESULTS: The distribution of plasma tHcy levels were skewed to the right with the mean values of 10.50 and 8.95 micromol/l and medians of 9.67 and 8.474 micromol/l for boys and girls, respectively. Plasma tHcy concentrations were lower in younger children and progressively increased with increasing age. Boys had significantly higher plasma tHcy levels than girls (10.50 +/- 4.134 vs. 8.95 +/- 2.61 micromol/l, p < 0.01) and lower plasma folate levels (6.05 +/- 2.85 vs. 6.39 +/- 2.58 nmol/l, p < 0.01), and vitamin B12 levels (444.8 +/- 158.4 vs. 495.0 +/- 181.5 pmol/l, p < 0.001). Plasma tHcy levels were significantly positively associated with anthropometric measures in boys; but these characteristics attenuated and became insignificant after adjusting for other potential confounders in girls. Plasma tHcy levels were negatively associated with plasma folate and vitamin B12 levels even after adjusting for BMI and other potential confounders in both genders. CONCLUSIONS: From this study, the distributions of tHcy levels were skewed to the right and the boys had higher plasma tHcy levels than girls. Plasma tHcy levels were significantly positively associated with BMI among boys. Further studies are needed to evaluate the relationship between tHcy and CVD risk factors among children for the better prevention of heart disease in early life.

Adolescent↗

Familial resemblance of adiposity-related parameters: results from a health check-up population in Taiwan.

The mechanisms of obesity is still unclear, however, genetic and environmental factors are two major possible causes of obesity. The purpose of this study was to assess the degree of familial resemblance of different obesity-related parameters in a health check-up population in Taiwan. We measured body mass index (BMI), waist-to-hip ratio (WHR) and percentage of body fat (BFAT) anthropometrics in 1724 members of 431 families participating in the MJ Health Screening program. Each family contributed four members, i.e. father, mother, son, and daughter. All the participants were free from coronary heart disease, hypertension, diabetes, dyslipidemia, and generally in good health. The degree of familial aggregation was measured by heritability that was calculated based on age-adjusted familial (parent-offspring, sibling, spouse) correlations. The maximal heritability estimates were 39, 30 and 35% for BMI, WHR and BFAT, respectively. For WHR, the correlation between spouses was not significant and the heritability appears to be predominantly due to genetic causes. Furthermore, for BMI and BFAT, the spouse correlations were 0.08 and 0.11, respectively. The heritabilities for BMI and BFAT were mostly explained by genetic factors and familial environmental factors such as dietary habits or physical activity. The familial resemblance of various obesity-related parameters was moderate in a health check-up population in Taiwan. While the heritability for WHR appears to be mainly due to genetic factors, the familial resemblance for BMI and percentage of BFAT may involve both genetic and familial environmental factors.

Adiposity↗

Viral load of high-risk human papillomavirus in cervical squamous intraepithelial lesions.

OBJECTIVES: This case-control study was conducted to investigate the role of viral load of high-risk human papillomaviruses (HPVs) in the development of cervical squamous intraepithelial lesions (SILs) and invasive cancers. METHODS: A total of 30 female cases who had histological evidence of low-grade SIL (n=10) or high-grade SIL and above (n=20) were identified as the case group at the Tri-Service General Hospital, Taipei between September 1998 and March 1999. In addition, 80 female controls who had normal cervical cytology were enrolled and individually matched on age (+/-5 years) and date of recruitment to each case. Cervical swabs collected from study subjects were tested for the positivity and viral load of high-risk HPVs by Hybrid Capture II assay. Additionally, subjects completed a risk factor questionnaire. RESULTS: Among sex behavioral factors studied, younger age at first intercourse was associated with a significantly elevated risk of cervical SIL and invasive cancers. With respect to HPV infection, high-risk HPV DNA was present in 70% (21/30) of case and 21% (17/80) of control subjects, resulting in an odds ratio (OR) of 6.6 [95% confidence interval (C.I.)=2.6-17.0]. Moreover, women who had a high viral load were at significantly greater risk for cervical SIL and invasive cancers than those who were infected with a low viral load (OR=18.0, 95% C.I.=3.0-108.5). CONCLUSIONS: Among the variables tested, infection with a high viral load of high-risk HPVs is the strongest determinant for cervical SIL and cervical cancers in Taiwan.

Adult↗

Nitric oxide/cGMP-induced inhibition of calcium and chloride currents in retinal pericytes.

In the CNS, contractile pericytes positioned on endothelium-lined lumens appear to play a role in regulating capillary blood flow. This function may be particularly important in the retina where pericytes are more numerous than in other tissues. Despite the importance of pericytes, knowledge of the effects of vasoactive molecules, such as nitric oxide (NO), on the physiology of these cells is limited. Since it is likely that ion channels play a role in the response of pericytes to signaling molecules from other cells, we used the perforated-patch configuration of the patch-clamp technique to record the whole-cell currents of pericytes located on microvessels freshly isolated from the rat retina. We found that voltage-gated calcium currents and calcium-activated chloride currents were inhibited during exposure to the NO donor, sodium nitroprusside (SNP). 8-Bromo-cyclic guanosine monophosphate (cGMP) mimicked these effects. In contrast, neither SNP nor the cGMP analog significantly affected the potassium or nonspecific cation conductances, which establish the resting membrane potential of retinal pericytes. Consistent with endogenous NO suppressing pericyte channel activity, exposure of isolated microvessels to an inhibitor of NO synthase increased the calcium and chloride currents. Since our experiments indicate that chloride channel activity is dependent, in part, upon the function of voltage-gated calcium channels, we postulate that a NO/cGMP-mediated inhibition of calcium channels reduces calcium influx and, thereby, lessens the opening of the calcium-activated chloride channels. This may be one mechanism by which NO decreases the contractile tone of pericytes.

8-Bromo Cyclic Adenosine Monophosphate↗

Joint effects of alcohol consumption and cigarette smoking on atherogenic lipid and lipoprotein profiles: results from a study of Chinese male population in Taiwan.

BACKGROUND: The present study examines the effect of joint exposure to cigarette smoking and alcohol intake on serum levels of total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and triglyceride (TG) among Chinese male adults in Taiwan. METHODS: A sample of 3311 men aged 20-59 years who reported having stable smoking and drinking behaviors during the period between January 1995 and December 1996 was selected from a periodic health checkup population. Serum lipids and lipoprotein cholesterol fractions were measured on fasting blood samples taken from participants. Statistical methods of analysis of variance and analysis of covariance were conducted to examine the associations of different smoking-drinking behavioral patterns with lipid and lipoprotein levels. RESULTS: In the observed population, the percentages of men who had stable cigarette smoking and alcohol consumption behaviors were 39.5% (1,307/3,311) and 27.0% (895/3,311), respectively. Mean values of TC and TG increased significantly and monotonically with increasing levels of cigarette smoking and alcohol consumption. In addition, alcohol intake was significantly associated with increased HDL-C and reduced LDL-C levels in a dose-dependent manner. More interestingly, the effect of alcohol consumption on LDL-C (negative) and TG (positive) levels was substantially greater for heavy smoker (>20 cigarettes/day) than for light smokers (< or = 20 cigarettes/day) and non-smokers, while alcohol intake exerted a strong positive influence on HDL-C concentration regardless of levels of cigarette smoking. CONCLUSIONS: In this Chinese male population, cigarette smoking and alcohol consumption were confirmed to have similar effects on lipid and lipoprotein levels as in Caucasians. More interestingly, a significance of joint exposure to smoking and drinking in predicting lipid and lipoprotein levels was evident. These data indicate the importance of multifactorial interventions to obtain more favorable lipid and lipoprotein levels in the population.

Adult↗

Relationship between plasma leptin levels and lipid profiles among school children in Taiwan--the Taipei Children Heart Study.

Leptin, an adipose tissue-derived of gene product, is important in energy metabolism. However, the role of leptin in the metabolism of lipids is still not clear in humans. The purpose of this study was to evaluate the association of plasma leptin concentrations and lipid profiles among school children in Taiwan. After multistage sampling of 85 junior high schools in Taipei, we randomly selected 1264 children (617 boys and 647 girls) aged 12-16 years for this study. We measured the anthropometric variables, lifestyle factors and biochemical parameters among these children. Anthropometric measurements included body height (BH) and weight (BW) and we calculated body mass index (BMI) as the ratio of the BW to the square of the BH, expressed in kg/m2. Plasma leptin levels were measured by radioimmunoassay. We also measured lipid profiles including serum total cholesterol (CHOL), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C), apolipoprotein-A1 (Apo-A1), apolipoprotein-B (Apo-B), and lipoprotein(a) (Lp(a)) levels, and calculated low density lipoprotein-cholesterol (LDL-C) levels and CHOL to HDL-C ratio (TCHR). Girls had higher leptin, CHOL, TG, HDL-C, (LDL-C), Apo-A1, Apo-B, and Lp(a) levels and lower BMI than boys did. Plasma leptin concentrations were significantly positively correlated with TG, LDL-C, and Apo-B, but negatively with HDL-C and Apo-A1 in both the genders. Children with higher plasma leptin levels (>75th percentiles) have significantly higher TG, HDL-C, LDL-C, TCHR, and Apo-B than those with relatively lower leptin levels. In multivariate regression analyses, the association between plasma leptin level and lipid profiles (such as CHOL, TG, and Apo-B) were still significant (p < 0.05) even after adjusting for BMI among boys. However, this association became attenuated and insignificant among girls. Finally, in the model that included the standard covariates, plasma leptin was the most predictive of CHOL, TG and Apo-B levels among those school children in Taiwan. Our results suggest that plasma leptin and BMI were independently associated with the lipids and lipoprotein profiles among Taiwanese Children. In both genders, children in the top 25% of the leptin distribution have more adverse lipid and lipoprotein profiles.

Adolescent↗

Prevalence and clustering of cardiovascular risk factors among healthy adults in a Chinese population: the MJ Health Screening Center Study in Taiwan.

OBJECTIVE: To gain insight into the prevalence and clustering of multiple cardiovascular risk factors in a healthy Chinese adult population in Taiwan. DESIGN: A cross-sectional study was carried out in 1996. SUBJECTS: A total of 46,603 subjects (23,485 men and 23,118 women) who were aged 20--59 y and attended a private health screening center for health examination of their own volition. MEASUREMENTS: Multiple cardiovascular risk factors including cigarette smoking, overweight (23 kg/m(2)< or =body mass index (BMI)<25 kg/m(2)) and obesity (BMI> or =25 kg/m(2)), lipid disorder (a ratio of total cholesterol level to the level of high density lipoprotein cholesterol>5 or use of lipid-lowering drugs), hypertension (systolic blood pressure> or =140 mmHg or diastolic blood pressure> or =90 mmHg or use of anti-hypertensive medications), and diabetes mellitus (fasting serum plasma glucose level> or =126 mg/dl or use of anti-diabetic medications) were determined. RESULTS: In comparison to women, men had a higher prevalence of current smoking (42.1 vs 5.6%), overweight (25.1 vs 17.1%) and obesity (33.1 vs 21.5%), lipid disorder (45.1 vs 19.6%), hypertension (17.4 vs 13.2%), as well as diabetes mellitus (4.1 vs 3.4%). The prevalence of men or women having two or more of the cardiovascular risk factors of interest was 54.3 and 21.7%, respectively. With advancing age, the prevalence of risk factors became greater for both genders. More importantly, the clustering of risk factors increased monotonically with increasing BMI levels for men and women. CONCLUSIONS: The prevalence and clustering of cardiovascular risk factors are commonplace in this healthy Chinese adult population. Considering the significant association between clustering of risk factors under study and BMI levels, this study gives an indication that population-based multifactorial interventions may work out favorably for specific groups.

Adult↗

Genetic polymorphisms of glutathione S-transferases M1 and T1 associated with susceptibility to aflatoxin-related hepatocarcinogenesis among chronic hepatitis B carriers: a nested case-control study in Taiwan.

This study was conducted to investigate the modifying effect of glutathione S-transferase (GST) M1 and T1 polymorphisms on aflatoxin-induced hepatocarcinogenesis among chronic hepatitis B virus surface antigen (HBsAg) carriers. A total of 79 HBsAg-positive cases of hepatocellular carcinoma (HCC) diagnosed between 1991 and 1997 were identified and individually matched to one or two HBsAg-positive controls on age, gender, residence and date of recruitment from the same cancer screening cohort in Taiwan. Blood samples were tested for hepatitis B and C viral markers by enzyme immunoassay and for aflatoxin B(1) (AFB(1))-albumin adducts by competitive enzyme-linked immunosorbent assay. GSTM1 and GSTT1 genotypes were determined by PCR. There was a statistically significant relationship between detectable levels of AFB(1)-albumin adducts in serum and risk of HCC among chronic HBsAg carriers, with an adjusted odds ratio (OR) of 2.0 [95% confidence interval (CI) 1.1-3.7]. In addition, the effect of aflatoxin exposure on HCC risk was more pronounced among chronic HBsAg carriers with the GSTT1 null genotype (OR 3.7, 95% CI 1.5-9.3) than those who were non-null (OR 0.9, 95% CI 0.3-2.4). The interaction between serum AFB(1)-albumin adduct level and GSTT1 genotype was statistically significant (P = 0.03). For GSTM1 the effect of aflatoxin exposure on HCC risk in those with the null genotype was also greater (adjusted OR 2.8, 95% CI 1.0-7.8) than in those with the gene present (adjusted OR 1.8, 95% CI 0.8-4.5), but the difference was not significant (P = 0.91). Notably, when the interaction between aflatoxin exposure and GSTT1 genotype was considered, aflatoxin exposure by itself was not a significant determinant of HCC risk among chronic HBsAg carriers. These results demonstrate the importance of gene-environment interactions in the multifactorial development of HCC.

Adult↗

Cones regenerate from retinal stem cells sequestered in the inner nuclear layer of adult goldfish retina.

PURPOSE: To determine whether retinal progenitor cells in the inner nuclear layer give rise to regenerated cones after laser ablation of photoreceptors in adult goldfish retina. METHODS: Using a technique developed previously in this laboratory, photoreceptors in the retina of adult goldfish were ablated with an argon laser. The mitotic marker, bromodeoxyuridine, was used to label proliferating and regenerated cells, which were identified with cell-specific markers. RESULTS: Cells proliferating locally within lesion included microglia, Müller glia, and retinal progenitors in the inner nuclear layer (INL). The nuclei of both Müller glia and associated retinal progenitors migrated from the inner to the outer nuclear layer. The proliferating retinal progenitors, which express Notch-3 and N-cadherin, regenerated cone photoreceptors and then rod photoreceptors. CONCLUSIONS: Previous work has demonstrated that photoreceptors in the goldfish retina regenerate selectively after laser ablation, but the source of regenerated cones has not been identified. The results reported here provide support for the existence of retinal stem cells within the adult fish retina that are capable of regenerating cone photoreceptors. The data also support the involvement of Müller glia in the production of regenerated cones.

Animals↗

Effects of calcitonin gene-related peptide and BIBN4096BS on myocardial ischemia in anesthetized rats.

AIM: The cardioprotective effect of calcitonin gene-related peptide (CGRP) was investigated in an ischemia rat model. METHODS: Ischemia-reperfusion injury was provoked by 60 min left main coronary artery occlusion followed by 60 min of reperfusion in anesthetized rats. The transverse slices of ventricles were stained by 2,3,5-triphenyltetrazolium chloride to determine the infarct area. Plasma creatine phosphokinase levels were determined by means of a creatine phosphokinase (CPK) kit. A radioimmunoassay was used to determine plasma CGRP levels. RESULTS: Intravenous infusion of CGRP (1 nmol . kg-1 . h-1) 10 min before occlusion until the end of reperfusion reduced infarct size by 89 %+/- 5 %. The reduction in infarct size was accompanied by a decrease in circulating levels of creatine phosphokinase. Infusion of the same dose of CGRP commencing from the start of reperfusion until its end induced a 40 % +/- 3 % reduction of the infarct size. The cardioprotective effects of CGRP were blocked by the novel CG RP antagonist BIBN4096BS (20 nmol . kg-1 . h-1). Although cardiac ischemia resulted in an almost 50 % increase in plasma CGRP levels in blood sampled from right cardiac ventricle, intravenous infusion of the CGRP antagonist BIBN4096BS before occlusion until the end of reperfusion had no statistically significant effect on the infarct size. CONCLUSION: The present study demonstrates that CGRP is a potent myocardial protective substance.

Animals↗

[Inhibition of Na+/Ca2+ exchange by tetrapeptide FMRFa in intact rat ventricular myocytes].

AIM: To study the effects of Phe-Met-Arg-Phe-NH2(FMRFa) on Na+/Ca2+ exchange and its specificity for Na+/Ca2+ exchange in rat ventricular myocytes. METHODS: Na+/Ca2+ exchange current and other currents of ion channels were measured using whole cell voltage clamp techniques. RESULTS: A dose-related inhibition of tetrapeptide FMRFa on Na+/Ca2+ exchange was observed in rat ventricular myocytes. Inward and outward INa+/Ca2+ were inhibited by 60.1% and 56.5%, respectively, at highest concentration (100 mumol.L-1) and its IC50 were 20 mumol.L-1 and 34 mumol.L-1 in inward and outward INa+/Ca2+, respectively. Inward and outward INa+/Ca2+ were inhibited 38.7% and 34.9%, respectively, at FMRFa 5 mumol.L-1. FMRFa 5 mumol.L-1 and 20 mumol.L-1 did not affect L-type calcium current, sodium current, transient outward current and inward rectifier potassium current. CONCLUSION: These data indicate that FMRFa is a specific inhibitor of Na+/Ca2+ exchange in intact rat ventricular myocytes.

Animals↗

The notch ligand jagged-1 represents a novel growth factor of human hematopoietic stem cells.

The Notch ligand, Jagged-1, plays an essential role in tissue formation during embryonic development of primitive organisms. However, little is known regarding the role of Jagged-1 in the regulation of tissue-specific stem cells or its function in humans. Here, we show that uncommitted human hematopoietic cells and cells that comprise the putative blood stem cell microenvironment express Jagged-1 and the Notch receptors. Addition of a soluble form of human Jagged-1 to cultures of purified primitive human blood cells had modest effects in augmenting cytokine-induced proliferation of progenitors. However, intravenous transplantation of cultured cells into immunodeficient mice revealed that human (h)Jagged-1 induces the survival and expansion of human stem cells capable of pluripotent repopulating capacity. Our findings demonstrate that hJagged-1 represents a novel growth factor of human stem cells, thereby providing an opportunity for the clinical utility of Notch ligands in the expansion of primitive cells capable of hematopoietic reconstitution.

Animals↗

Isolation and characterization of human CD34(-)Lin(-) and CD34(+)Lin(-) hematopoietic stem cells using cell surface markers AC133 and CD7.

Recent evidence indicates that human hematopoietic stem cell properties can be found among cells lacking CD34 and lineage commitment markers (CD34(-)Lin(-)). A major barrier in the further characterization of human CD34(-) stem cells is the inability to detect this population using in vitro assays because these cells only demonstrate hematopoietic activity in vivo. Using cell surface markers AC133 and CD7, subfractions were isolated within CD34(-)CD38(-)Lin(-) and CD34(+)CD38(-)Lin(-) cells derived from human cord blood. Although the majority of CD34(-)CD38(-)Lin(-) cells lack AC133 and express CD7, an extremely rare population of AC133(+)CD7(-) cells was identified at a frequency of 0.2%. Surprisingly, these AC133(+)CD7(-) cells were highly enriched for progenitor activity at a frequency equivalent to purified fractions of CD34(+) stem cells, and they were the only subset among the CD34(-)CD38(-)Lin(-) population capable of giving rise to CD34(+) cells in defined liquid cultures. Human cells were detected in the bone marrow of non-obese/severe combined immunodeficiency (NOD/SCID) mice 8 weeks after transplantation of ex vivo-cultured AC133(+)CD7(-) cells isolated from the CD34(-)CD38(-)Lin(-) population, whereas 400-fold greater numbers of the AC133(-)CD7(-) subset had no engraftment ability. These studies provide novel insights into the hierarchical relationship of the human stem cell compartment by identifying a rare population of primitive human CD34(-) cells that are detectable after transplantation in vivo, enriched for in vitro clonogenic capacity, and capable of differentiation into CD34(+) cells. (Blood. 2000;95:2813-2820)

AC133 Antigen↗

Characterization of chemokine receptors expressed in primitive blood cells during human hematopoietic ontogeny.

Chemokines are capable of regulating a variety of fundamental processes of hematopoietic cells that include proliferation, differentiation, and migration. To evaluate potential chemokine signaling pathways important to the regulation of primitive human hematopoietic cells, we examined chemokine receptor expression of highly purified subpopulations of uncommitted human blood cells. CXCR1-, CXCR2-, CXCR4-, and CCR5-expressing cells were detected by flow cytometry among human blood subsets depleted of lineage-restricted cells (Lin(-)) derived from adult bone marrow, mobilized peripheral blood, cord blood (CB), and circulating fetal blood. Although these chemokine receptors could be detected on Lin(-) cells throughout human development, only CXCR4 could be detected in CD34(-)CD38(-)Lin(-) and CD34(+)CD38(-)Lin(-) subfractions enriched for stem cell function, suggesting that independent of ontogeny, CXCR4-mediated signals are critical to primitive hematopoiesis. Distinct to other stages of human hematopoietic development, primitive CB cells expressed higher levels of CXCR1, CXCR2, CCR5, and CXCR4 on both CD34(-)CD38(-)Lin(-) and CD34(+)CD38(-)Lin(-) subsets. Isolation of these fractions revealed expression of additional chemokine receptors CCR7, CCR8, and Bonzo (STRL133), whereas BOB (GPR15) could not be detected. Our study illustrates that rare uncommitted hematopoietic cells express chemokine receptors not previously associated with primitive human blood cells. Based on these results, we suggest that signaling pathways mediated by chemokine receptors identified here may play a fundamental role in hematopoietic stem cell regulation and provide alternative receptor targets for retroviral pseudotyping for genetic modification of repopulating cells.

Adult↗

Class III anti-arrhythmia drug E-4031 potentiates Na+/Ca2+ exchange current in rat ventricular myocytes.

AIM: To study the effects of E-4031 on the Na+/Ca2+ exchange currents (INa/Ca). METHODS: The quasi-steady state current-voltage relationship from the isolated rat ventricular myocytes was measured using whole-cell voltage-clamp techniques with a ramp pulse protocol. RESULTS: At potential of mV, E-4031 5, 10, and 20 mumol.L-1 increased Ni(2+)-sensitive current from (0.48 +/- 0.12), to (0.78 +/- 0.20), (0.96 +/- 0.16), and (1.15 +/- 0.13) pA/pF, respectively; tetradecanoylphorbol acetate (TPA) 50 nmol.L-1 increased Ni(2+)-sensitive current from (0.60 +/- 0.16) to (1.33 +/- 0.25) pA/pF. Tamoxifen 20 mumol.L-1 completely prevented the current changes induced by E-4031 and TPA. CONCLUSION: E-4031 stimulates the Na+/Ca2+ exchange via a protein kinase C-dependent pathway.

Animals↗

Physiology of rat retinal pericytes: modulation of ion channel activity by serum-derived molecules.

1. Pericytes, which are contractile cells located on the outer wall of microvessels, are thought to be particularly important in the retina where the ratio of these cells to vascular endothelial cells is the highest of any tissue. Retinal pericytes are of interest since they may regulate capillary blood flow and because their selective loss is an early event in diabetic retinopathy, which is a common sight-threatening disorder associated with dysfunction of the blood-retinal barrier. 2. Although a breakdown in the vascular endothelial barrier is a frequent pathophysiological event, knowledge of the effects of blood-derived molecules on pericyte function is limited. Based on the premise that ion channels play a vital role in cellular function, we examined the effect of serum on the ionic currents of retinal pericytes. To do this, we used the perforated-patch configuration of the patch-clamp technique to monitor the whole-cell currents of pericytes located on freshly isolated rat retinal microvessels. 3. Exposure to serum reversibly activated inward and outward currents in virtually all of the sampled retinal pericytes. Two types of sustained conductances were induced by serum. These were a calcium-permeable non-specific cation (NSC) current and a voltage-dependent potassium current. In addition, exposure to serum increased the activity of chloride channels which caused transient depolarizing currents. 4. Associated with the activation of these conductances, the membrane potential showed a sustained decrease of 10 +/- 2 mV from -56 mV to -46 mV and, also, transient depolarizations to near -30 mV. The serum-induced depolarizations can activate the voltage-gated calcium channels expressed by the retinal pericytes. 5. Calcium-permeable NSC channels appear to play a critical role in the response of pericytes to serum-derived molecules. Consistent with this, activation of the chloride and potassium channels was sensitive to SK&F 96365, which is a blocker of NSC channels. In addition, chloride and potassium channel activation was dependent on extracellular calcium. 6. The effects of serum on the activity of channels in retinal pericytes were qualitatively mimicked by insulin-like growth factor-1 (IGF-1), which is a normal constituent of the blood. 7. There are significant differences in the effects of serum on retinal pericytes compared with vascular smooth muscle cells. Serum activated sustained conductances in retinal pericytes but not in the vascular smooth muscle cells. This suggests a fundamental difference in the mechanisms by which serum-derived molecules affect these two types of cells. 8. We conclude that serum-derived molecules, such as IGF-1, can activate several types of ion channels in retinal pericytes. These changes in channel activity are likely to influence pericyte function at sites of a breakdown in the blood-retinal barrier.

Animals↗

Processing and release of IL-16 from CD4+ but not CD8+ T cells is activation dependent.

IL-16 is synthesized as a precursor molecule of 68 kDa (pro-IL-16) that is processed by caspase-3, a member of the IL-1 converting enzyme (ICE) family. This cleavage results in a 13-kDa carboxy terminal peptide, which constitutes the bioactive secreted form of IL-16. We have previously reported constitutive IL-16 mRNA expression and pro-IL-16 protein in CD4+ and CD8+ T cells. Although bioactive IL-16 protein is present in unstimulated CD8+ T cells, there is no bioactive IL-16 present in CD4+ T cells. Along these lines, unstimulated CD8+ T cells contain active caspase-3. In the current studies we investigated the regulation of IL-16 protein and mRNA expression in CD4+ T cells and determined the kinetics of secretion following stimulation of the TCR. CD4+ T cells release IL-16 protein following antigenic stimulation, and this release is accelerated in time by costimulation via CD28. However, CD3/CD28 costimulation did not alter IL-16 mRNA appearance or stability in either CD4+ or CD8+ T cells. The secretion of bioactive IL-16 from CD4+ T cells correlated with the appearance of cleavage of pro-caspase-3 into its 20-kDa active form. Thus, resting CD8+ T cells contain active caspase-3 that is capable of cleaving pro-IL-16, whereas CD4+ T cells require activation for the appearance of active caspase-3. The mechanism of release or secretion of bioactive IL-16 is currently unknown, but does not correlate with cellular apoptosis.

Animals↗