Biomedical subjects

L Ao

Publications and source records attributed to L Ao.

27 records · Page 2Linked to original sources

PubMed1997

Multiple mitogen-activated protein kinases are regulated by hyperosmolality in mouse IMCD cells.

Inner medullary collecting duct (IMCD) cells adapt to a hypertonic environment by synthesizing transporters that allow for accumulation of organic osmolytes. To examine for activation of additional mitogen-activated protein (MAP) kinases, extracts of IMCD-3 cells subjected to a hypertonic medium (600 mosmol/kgH2O) for 15 min were fractionated by Mono Q fast-performance liquid chromatography and assayed with the epidermal growth factor receptor [EGFR-(662-681)] peptide as substrate. Three peaks of activity were identified. Western blotting revealed that these peaks coincided with Jun NH2-terminal kinase (JNK), extracellular signal-regulated protein kinases, ERK1 and ERK2, and p38 MAP kinase. To assess the functional significance of ERK2 activation in IMCD-3 cells, the effect of PD-098059, an inhibitor of the upstream regulatory protein kinase MAP/ERK kinase (MEK) was assessed. PD-098059 inhibited ERK activation by hypertonicity. Yet, the stimulation of inositol uptake, a marker of adaptation, after 16 h was unaltered. Direct measurements of JNK activity [phosphorylation of GST-cJun-(1-79)] revealed a marked (20- to 40-fold) increase in activity as medium osmolality was increased from 300 to 900 mosmol/kgH2O with either NaCl or mannitol. Urea induced a more modest increase in activity. The response is prompt and detected as early as 2 min after exposure, reaching a maximum activation at 10-15 min. Downregulation of cellular protein kinase C (PKC) by chronic exposure to phorbol esters only minimally attenuated the JNK response to hyperosmolality, indicating a lack of involvement of PKC. We conclude that, in IMCD-3 cells, inhibition of ERK activation by hyperosmolality does not prevent osmoregulatory increase in inositol transport. This is not consistent with a role for ERKs in the response. The roles for JNK and p38 have not been ruled out, and these pathways may represent the initiating event in the subsequent transcription of organic osmolyte transporter genes and adaptation to extracellular hypertonicity.

Animals↗

PubMed1997

A new direction for cleft palate research.

Successful outcomes in the treatment of complete unilateral cleft-lip and palate (CUCL/P) and complete bilateral cleft lip and palate (CBCLP) are not obtained universally, despite significant improvements in surgical techniques over the past three decades. In particular, deficient palatal growth may occur even when expert surgical treatment is rendered. The factors which contribute most significantly to unfavorable growth outcomes remain obscure.

Journal Article↗

PubMed1997

[Lead exposure level in umbilical cord blood and its related factors].

Six hundred and five specimens of umbilical cord blood were collected from Yangpu District, Shanghai, and blood lead levels were determined with graphite stove atom absorption spectrophotometry in 348 of them to study fetal exposure to lead and its related factors in the people of Shanghai. Survey on household social and environmental health status was conducted in families with the babies whose umbilical cord blood lead were above the 70th percentile and below the 30th percentile with a face-to-face questionnaire. Data were analyzed with multiple regression for the factors which affected the lead level in umbilical cord blood significantly. Results showed that blood lead levels in 348 cases were normally distributed, with a range of 0.08 to 0.84 mumol/L, a mean of 0.44 mumol/L and a standard deviation of 0.15 mumol/L. Specimens with cord blood lead level exceeding the safety criteria of 0.48 mumol/L accounted for 40.8 percent of the total. The study also found that passive smoking during pregnancy, exposure to lead in family members, one's living room adjacent to the major traffic roads, pollution by coal-burning smoke in the environment of one's residence, use of coal as domestic fuel, and eating preserved eggs during pregnancy all were risk factors for lead exposure. Multiple logistic regression analysis revealed that contribution of passive smoking to cord blood lead level was still statistically significant after adjusting other confounding factors. It concluded that environmental lead pollution could cause ad-verse effects on fetal development.

China↗

PubMed1996

A single endotoxin challenge induces delayed myocardial protection against infarcation.

Sublethal endotoxemia attenuates cardiac functional injury from global ischemia but it is unknown whether endotoxemia can protect myocardium against infarction. Furthermore, increases in myocardial catalase and heat shock protein (HSP) following endotoxemia have been associated with cardiac ischemic protection. We therefore hypothesized that a 72-hr pretreatment with endotoxin (ETX) would reduce myocardial tissue necrosis in association with augmented catalase activity and stress protein expression. Rabbits were treated with normal saline or lipopolysaccharide (Salmonella typhimurium) at 10, 5, and 1 microgram/kg doses. Three days after saline or ETX injection they were subjected to 45 min of coronary artery occlusion followed by 3 hr of reperfusion. Area of necrosis (tetrazolium staining) was normalized to anatomic risk zone size (Evans blue staining). Catalase activity was measured by a standard assay and HSP 72 was assessed by immunohistochemistry. During regional ischemia and reperfusion there were no differences in heart rate or mean arterial blood pressure between groups. ETX treated rabbits had the same risk zone size as controls. Infarct size was reduced in the ETX treated rabbits at the 10 and 5 microgram/kg doses compared with control rabbits (17.5 +/- 1.5% and 22.2 +/- 3.1% vs 45.3 +/- 2.5%; P < 0.05) but no protective effect was observed at the 1.0 micrograms/kg dose (38.0 +/- 4.6%; P > 0.05 vs control). Catalase activity was not different between control and ETX (5 microgram/kg) treated groups (997.8 +/- 59.1 U/g vs 1099.6 +/- 69.3 U/g myocardium; P > 0.05) but endotoxin induced expression of myocardial HSP 72. We conclude that a single challenge with endotoxin can induce delayed myocardial protection against infarction in vivo. This delayed cardioprotective response involves enhanced stress protein expression without changes in myocellular catalase activity.

Animals↗

PubMed1996

Norepinephrine induces cardiac heat shock protein 70 and delayed cardioprotection in the rat through alpha 1 adrenoceptors.

OBJECTIVE: Previous studies have demonstrated that norepinephrine (NE) confers immediate cardioprotection against ischemia and reperfusion injuries. The present study tests the hypothesis that in vivo treatment with NE induces delayed cardioprotection against postischemic dysfunction in the rat heart which is associated with expression of c-fos and heat shock protein (HSP) 70 in the myocardium. METHODS: Rats were treated with NE (3.1 mumol/kg, ip) and hearts isolated and perfused with a modified Langendorff technique 2 or 24 h after injection. Left ventricular developed pressure (LVDP) and left ventricular end-diastolic pressure (LVEDP) were recorded during 25 min normothermic global ischemia and 40 min reperfusion. Total RNA was extracted from left ventricular tissue at various time points and Northern hybridization was applied to detect c-fos and HSP70 mRNAs. Expression and distribution of c-fos and HSP72 proteins in the myocardium were examined by immunohistochemical staining. RESULTS: In vivo NE treatment improved postischemic recovery of both LVDP and LVEDP in the hearts isolated at 24 h after treatment but not in those isolated at 2 h. LVDP was 68.9 +/- 4.8 mmHg in NE 24 h group at the end of reperfusion in comparison to 43.6 +/- 2.9 mmHg in saline control group (P < 0.01). Pretreatment with prazosin abolished NE-induced cardioprotection while propranolol pretreatment had no effect. Northern analysis demonstrated rapid and transient increases in c-fos and HSP70 mRNAs in the myocardium after NE treatment. Accumulation of c-fos protein was observed at 3 h and increased amount of HSP72 protein was demonstrated at 24 h in the myocardium. Pretreatment of rats with prazosin eliminated NE-induced increase in cardiac HSP70 mRNA. CONCLUSIONS: The results suggest that in vivo treatment with NE upregulates the expression of c-fos and HSP70 in the myocardium and induces delayed protection against postischemic myocardial dysfunction in the isolated rat heart. Induction of both the expression of cardiac HSP70 and the delayed cardioprotection by NE appears to be mediated by alpha 1 adrenoceptors.

Adrenergic alpha-Antagonists↗

PubMed1996

Endotoxin induces cardiac HSP70 and resistance to endotoxemic myocardial depression in rats.

Endotoxin (bacterial lipopolysaccharide, LPS) depresses myocardial function. However, heat shock and sublethal LPS can confer cardiac resistance to postischemic dysfunction. We hypothesized that a prior exposure to LPS stress induces the expression of cardiac heat shock protein 70 (HSP70) and resistance to endotoxemic myocardial depression. Moreover, induction of HSP70 by hyperthermia should also increase cardiac resistance to LPS toxicity. LPS (500 micrograms/kg ip) depressed rat left ventricular developed pressure (LVDP) maximally at 6 h (58.4 +/- 3.72 vs. 101 +/- 1.46 mmHg in saline control, P < 0.01), and myocardial contractile function recovered at 24 h. In rats pretreated with LPS 24 h earlier, subsequent LPS exposure did not depress LVDP (97.0 +/- 3.53 mmHg at 6 h, P < 0.01 vs. single exposure). Both LPS and hyperthermia (42 degrees C, 15 min) induced HSP72 mainly in the cardiac interstitial cells, including macrophages at 24 h after treatment. When hyperthermia-pretreated animals were similarly challenged with LPS, myocardial depression at 6 h was partially abrogated (LVDP 80.1 +/- 5.67 vs. 62.2 +/- 4.91 mmHg in sham+LPS group, P < 0.01). We conclude that LPS induces HSP70 in rat heart and that an exposure to LPS or heat stress confers cardiac resistance to endotoxemic myocardial depression.

Animals↗

PubMed1995

Mechanisms of immature myocardial tolerance to ischemia: phenotypic differences in antioxidants, stress proteins, and oxidases.

BACKGROUND: Previous work has suggested tolerance to ischemic injury in newborn myocardium. Although various mechanisms for this protection have been proposed, a link between oxidant-antioxidant factors, stress protein expression, and protection from cardiac ischemia/reperfusion (I/R) injury has not been made in newborn myocardium. We hypothesized total newborn myocardial resistance to I/R is related to decreased oxygen radical producing potential, increased free radical scavenging capacity and augmented stress protein expression. The purposes of the study were to examine in newborn and adult rat hearts (1) functional recovery from I/R, (2) catalase and xanthine oxidase (XO) activities, and (3) heat shock protein 72 (HSP 72) expression. METHODS: Isolated rat hearts (7 to 10 days versus 60 days) were perfused on a nonworking Langendorff apparatus at 60 mm Hg (Krebs-Henseleit buffer, pH 7.4, 37 degrees C) and subjected to 20 minutes of global ischemia and 40 minutes of reperfusion. Left ventricular developed pressure was recorded by using a left ventricular catheter. Catalase and XO were measured by means of standard assays, and HSP 72 was assessed with in situ immunohistochemistry. RESULTS: Newborn rat hearts had greater percentage functional recovery of left ventricular developed pressure after I/R (66.0% +/- 4.2% versus 44.3% +/- 3.5%; p < 0.05). The newborn myocardium also had increased catalase activity (1027.9 +/- 20.6 units/gm versus 707.3 +/- 38.7 units/gm; p < 0.05), whereas the activity of XO was decreased relative to the adult (0.23 +/- 0.01 mU/gm versus 7.6 +/- 1.4 mU/gm; p < 0.05). Furthermore, the expression of HSP 72 was greater in the newborn than the adult control. CONCLUSIONS: Relative to adult hearts, newborn rat hearts are more tolerant to a global ischemic insult followed by reperfusion. This improved functional recovery is associated with decreased oxidant production potential (XO), increased scavenging capacity (catalase), and augmented stress protein expression (HSP 72).

Aging↗

PubMed1995

Preconditioning of isolated rat heart is mediated by protein kinase C.

Catecholamines have been implicated in the phenomenon of ischemic preconditioning. We have previously demonstrated that ischemic preconditioning against postischemic mechanical dysfunction in the isolated rat heart is mediated by the alpha 1-adrenergic receptor. The purpose of this study was to delineate the signal transduction of preconditioning distal to the alpha 1-adrenergic receptor. Our results suggest that (1) transient ischemia and alpha 1-adrenergic receptor-induced preconditioning is inhibited by protein kinase C (PKC) antagonists, (2) functional protection against global ischemia/reperfusion injury can be induced by infusion of diacylglycerol, the second messenger of the alpha 1-adrenergic pathway, and (3) transient ischemia and alpha 1-adrenergic preconditioning are both characterized by similar translocation of PKC-delta to the sarcolemma of myocardium. These findings suggest that PKC is an effector of preconditioning in the isolated rat heart.

Animals↗

PubMed1995

Expression of GTPase-deficient Ras inhibits vasopressin signaling in cultured cortical collecting duct cells.

Cross-talk between signaling pathways is increasingly recognized as integral to cellular function. We investigated whether the mitogen-activated protein kinase (MAPK) pathway alters vasopressin (AVP) stimulation of protein kinase A (PKA) by specifically studying the role of Ras. Mouse cortical collecting duct cells (M-1) were transfected with a cDNA encoding oncogenic Ras. Transfection was confirmed by Western blot analysis and functionally by enhanced basal MAPK activity. When compared with basal MAPK activity of 26.4 +/- 6.6 pmol/mg/min in controls, basal MAPK activity varied widely in Ras-transfected clones from 29.0 +/- 6.6 to 96.6 +/- 13.4 pmol/mg/min. Clones that functionally expressed activated Ras displayed complete abolition of AVP-stimulated PKA activity, whereas those that failed to express elevated basal MAPK activity showed intact AVP-stimulated PKA. The correlation between expression of high basal MAPK activity and inhibition of AVP-induced PKA yielded a correlation coefficient of -0.92 (P = 0.009). Exposure to 10 microM forskolin or 1 microgram/ml cholera toxin resulted in comparable activation of PKA in all clones. We found no correlation between PKC activity of the clones and PKA inhibition. To assess whether the observed effect was due to one known Ras target, cells were transfected with constitutively activated Raf. M-1 cells expressing activated Raf exhibited elevated MAPK activity. The Raf clones showed no impairment of AVP-stimulated PKA activity. We conclude that expression of activated Ras is inhibitory of AVP-induced PKA activation in the M-1 cortical collecting duct cell line at a site proximal to G alpha s protein. The failure of Raf to influence AVP signaling indicates that the action of Ras is through a pathway independent of this Ras target.

Amino Acid Sequence↗