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H M Huang

Publications and source records attributed to H M Huang.

At least 73 records · Page 4Linked to original sources
PubMed1997

Altered phospholipid metabolism in sodium butyrate-induced differentiation of C6 glioma cells.

We examined the changes in phospholipid metabolisms in sodium butyrate-treated C6 glioma cells. Treatment of 2.5 mM sodium butyrate for 24 h induced an increase in the activity of glutamine synthetase, suggesting that these cells were under differentiation. Similar treatment was associated with (i) increased arachidonic acid incorporation into phosphatidylcholine, and (ii) decreased arachidonic acid incorporation into phosphatidylinositol and (iii) phosphatidylethanolamine. These effects were subsequently investigated by examining the acylation process, de novo biosynthesis, and the agonist-stimulated phosphoinositides hydrolysis in these cells. Our results indicated that sodium butyrate stimulated the acylation of arachidonic acid into lysophosphatidylcholine, lysophosphatidylethanolamine, and lysophosphatidylinositol. The glycerol incorporation into these lipids was not affected, but the inositol incorporation into total chloroform extracts and Pl and phosphatidylinositol 4-phosphate was decreased in the sodium butyrate-treated cells. Moreover, the accumulation of the rapid histamine-stimulated phosphoinositide metabolites, i.e., inositol monophosphate, inositol diphosphate, and inositol triphosphate (IP3) was decreased in these cells. To elucidate whether the decreased inositol phosphates were due to a decrease in the phosphoinositides hydrolysis, we measured the transient IP3 production directly by a receptor-binding assay. Our results indicated that histamine-stimulated transient IP3 formations were decreased. Taken together, these results indicated that multiple changes by multiple mechanisms of phospholipid metabolisms were found in sodium butyrate-treated C6 glioma cells. The decreased IP3 formation and its subsequent action, i.e., Ca2+ mobilization, may play an early but pivotal role by which sodium butyrate induces C6 glioma cell differentiation.

Acylation↗
PubMed1997

The design and application of a pediatric centrifugal pump.

This centrifugal pump (CP) includes two parts: the blood pump and the driving apparatus. They are connected by six twin magnetic disc plates and driven by a magnetic DC motor (120W). The blood pump had six leaves deadlocked between two plastic discs. Six leaves were set at 30 degrees angles, separately. In the lower chamber of the CP, there was an inlay magnetic disc, which is connected with the disc leaves by an axis. This axis was sealed by silicon rubber and a ceramic ring. The priming volume of the blood chamber was 34 ml. In vitro testing showed that the free hemoglobin caused by the CP was much less than that caused by a roller pump after 180 min. The effect of this CP on blood cell damage was also studied in an animal model. Six goats were placed on cardiopulmonary bypass for 180 min. Perfusion flow rates were maintained between 1.5 and 2.5 L/min. The plasma free hemoglobin was lower in the CP group (6.04 mg/dL) than in the roller pump group (32.25 mg/dL), p < 0.01. The CP has been used in ten pediatric patients undergoing cardiopulmonary bypass surgery. The patients' ages were from three to five years, and body weights were from 15 to 20 kg. Perfusion flow rates were maintained between 1.8 and 2.5 L/min, and bypass times were from 30 to 50 min. The rotation speeds were from 2000 to 2500 rpm. All the patients recovered smoothly, and no hemoglobinuria occurred.

Animals↗
PubMed1996

Regulation of bradykinin-induced Ins(1,4,5)P3 formation by protein kinase C in human fibroblasts.

To better understand the molecular mechanisms that underlie the exaggerated bradykinin (BK)-stimulated release of Ins(1,4,5)P3 in fibroblasts from Alzheimer patients, the role of G-proteins, protein kinase C (PKC) and cyclic AMP in BK-induced Ins(1,4,5)P3 formation was determined. A role for G-proteins in the coupling of the BK receptor to intracellular signals was indicated by guanosine 5'-(3-O-thio)triphosphate (GTP gamma S) enhanced BK-stimulated Ins(1,4,5)P3 release. The coupling of G-proteins to Ins(1,4,5)P3 formation was sensitive to cholera toxin (CTX), but not pertussis toxin (PTX), and was not altered by PKC activation. The inhibition by CTX appeared to be secondary to its ability to increase cyclic AMP, because forskolin also inhibited the BK-mediated Ins (1,4,5)P3 release. Activation of PKC with TPA diminished the number of BK receptors by 33% and proportionally decreased BK-mediated Ins(1,4,5)P3 formation by 28%. The latter response was abolished by PKC inhibitors. Depletion of PKC by prolonged TPA treatment did not further alter the number of BK receptors but further decreased the Ins(1,4,5)P3 response by 65%. Thus, changes in PKC probably do not underlie the enhanced BK-induced Ins(1,4,5)P3 formation in AD fibroblasts, because both activation and depletion of the PKC diminished the Ins(1,4,5)P3 response.

Bradykinin↗
PubMed1996

Apoptosis of gamma/delta T cells in human ehrlichiosis.

Expansion of activated T cells expressing the T-cell receptor (TCR) gamma/delta, CD45RO, and HLA-DR antigens is a prominent feature of acute infection with Ehrlichia chaffeensis in humans. The fate of these activated cells and the resolution of the gamma/delta T-cell response with return to the usual alpha/beta T-cell populations in this disease are not clearly understood. At a morphologic level, apoptotic cells are present in the peripheral blood during the acute and resolution phases of the infection. Simple culture of density gradient-separated lymphocytes from the blood of patients with acute ehrlichiosis produced cell death rapidly in the media compared to alpha/beta T cells. This loss of viability after incubation was apparently mediated by apoptosis, based on flow cytometric and morphologic analyses. The results suggest that most primed (CD45RO+) and activated (HLA-DR+) gamma/delta T cells in acute ehrlichiosis might be subject to removal from the body by programmed or apoptotic cell death.

Apoptosis↗
PubMed1995

The role of signal transduction systems in mediating cell density dependent changes in tyrosine hydroxylase gene expression.

Cell density has been implicated in the regulation of neuronal gene phenotype. This study tested the interaction of signal transduction pathways and the expression of tyrosine hydroxylase (TH) mRNA with varying cell density. Increasing cell density in a parental, wild type PC12 cell line elevated steady state levels of TH mRNA. Three observations suggested that this induction is not related to the cyclic AMP dependent signalling pathway: (1) Forskolin stimulated the level of TH mRNA similarly at multiple densities. (2) PKA deficient mutant PC12 cell lines that have either one third (A123.7, AB11) or 3% (A126-1B2) of normal basal expression of TH mRNA still exhibit the same density induced elevation of TH mRNA levels as the wild type. (3) Different cell densities did not change cyclic AMP concentrations in the basal or in the receptor stimulated state. Increasing cell density did not change basal levels of inositol triphosphate (IP3) levels, which suggests that the phosphatidylinositol cascade (PI) is not responsible for density dependent changes in TH expression. Increasing confluence was highly correlated to [Ca2+]i in control (r = 0.70; P < 0.0001), A123.7 (r = 0.92; P < 0.001), AB11 (r = 0.72; P < 0.0001) and A126 (r = 0.42; P < 0.07). Taken together, the results show that neither cyclic AMP nor the PI cascade is involved in cell density induced changes in TH mRNA and suggest that altered [Ca2+]i may have a role.

Animals↗
PubMed1995

Increased inositol 1,4,5-trisphosphate accumulation correlates with an up-regulation of bradykinin receptors in Alzheimer's disease.

An alteration in signal transduction systems in Alzheimer's disease (AD) would likely be of pathophysiological significance, because these processes control normal brain functions. Previously, a diminished beta-adrenergic-mediated cyclic AMP response was found in cultured fibroblasts from AD patients. Because cross-talk between the phosphoinositide and cyclic AMP pathways exists, the phosphoinositide cascade was studied under conditions that were similar to those for studying the cyclic AMP response. Cells from AD patients and age-matched controls responded to bradykinin (BK) and released inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] in a time- and dose-dependent manner. The level of Ins(1,4,5)P3 increased rapidly and transiently in response to BK, peaked at 5 s, but still remained 116-132% above the basal level by 30 s. Although the temporal patterns were similar in both groups, the Ins(1,4,5)P3 concentrations in AD fibroblasts were 73 and 89% above levels in the age-matched controls at 5 and 10 s, respectively. Prostaglandin E1 also increased Ins(1,4,5)P3 formation, but this response was not different between the two groups. Although KD (affinity) values for the BK receptor were similar in both control and AD cells, the number of BK receptors (Bmax) was significantly elevated in AD fibroblasts (186.8 +/- 0.8 fmol/mg of protein) as compared with control fibroblasts (57.2 +/- 15.3 fmol/mg of protein). These results indicate that the elevated Ins(1,4,5)P3 production in response to BK in AD fibroblasts is positively correlated with an increase in the receptor numbers.

Aged↗
PubMed1994

The role of cytosolic free calcium in the regulation of pyruvate dehydrogenase in synaptosomes.

Calcium homeostasis and mitochondrial oxidative metabolism interact closely in brain and both processes are impaired during hypoxia. Since the regulation of the pyruvate dehydrogenase complex (PDHC) may link these two processes, the relation of cytosolic free calcium ([Ca2+]i) to the activation state of PDHC (PDHa) was assessed in isolated nerve terminals (i.e. synaptosomes) under conditions that alter [Ca2+]i. K+ depolarization elevated [Ca2+]i and PDHa and both responses required external calcium. Treatment with KCN, an in vitro model of hypoxia decreased ATP and elevated [Ca2+]i and PDHa. Furthermore, in the presence of KCN, PDHa became more sensitive to K+ depolarization as indicated by larger changes in PDHa than in [Ca2+]i. The calcium ionophore Br-A23187 elevated [Ca2+]i, but did not affect PDHa. K+ depolarization elevated [Ca2+]i and PDHa even if [Ca2+]i was elevated by prior addition of ionophore or KCN. Previous in vivo studies by others show that PDHa is altered during and after ischemia. The current in vitro results suggest that hypoxia, only one component of ischemia, is sufficient to increase PDHa. These data also further support the notion that PDHa is regulated by [Ca2+]i as well as by other factors such as ATP. Our results are consistent with the concept that PDHa in nerve endings may be affected by [Ca2+]i and that these two processes are clearly linked.

Animals↗
PubMed1994

Finite element analysis of cerebral contusion.

Finite element analysis was carried out to study the mechanism of cerebral contusion. Clinical findings indicate that most cerebral contusions in the absence of skull fracture occur at the frontal and temporal lobes. To explain these observations, cavitation and shear strain theories have long been advocated. Plane strain finite element models of a parasagittal section of the human head were developed in the present study. The model was first validated against a set of experimental results from the literature. Frontal and occipital impacts were then simulated, and pressure and shear stress distributions in the brain were compared. While comparable negative pressures always developed in the contrecoup regions, shear stress distributions remained nearly identical regardless of the impact direction, consistent with the clinically observed pattern for contusion. Therefore, shear strain theory appears to account better for the clinical findings in cerebral contusion.

Acceleration↗
PubMed1993

Altered beta-adrenergic receptor-stimulated cAMP formation in cultured skin fibroblasts from Alzheimer donors.

An alteration in signal transduction systems in Alzheimer's disease would likely be of pathophysiological significance, because these steps are critical to normal brain function. Since dynamic processes are difficult to study in autopsied brain, the current studies utilized cultured skin fibroblasts. The beta-adrenergic-stimulated increase in cAMP was reduced approximately 80% in fibroblasts from Alzheimer's disease compared with age-matched controls. The deficit in Alzheimer fibroblasts in response to various adrenergic agonists paralleled their beta-adrenergic potency, and enhancement of cAMP accumulation by a non-adrenergic agonist, such as prostaglandin E1, was similar in Alzheimer and control fibroblasts. Diminished adenylate cyclase activity did not underlie these abnormalities, since direct stimulation of adenylate cyclase by forskolin elevated cAMP production equally in Alzheimer and control fibroblasts. Cholera toxin equally stimulated cAMP formation in Alzheimer and control fibroblasts. Moreover, cholera toxin partially reduced isoproterenol-induced cAMP deficit in Alzheimer fibroblasts. Pertussis toxin, on the other hand, did not alter the Alzheimer deficits. The results suggest either that the coupling of the GTP-binding protein(s) to the beta-adrenergic receptor is abnormal or that the sensitivity of receptor is altered with Alzheimer's disease. Further, any hypothesis about Alzheimer's disease must explain why a reduced beta-adrenergic-stimulated cAMP formation persists in tissue culture.

Adenylate Cyclase Toxin↗
PubMed1993

Characterization of seven newly established nasopharyngeal carcinoma cell lines.

BACKGROUND: Nasopharyngeal carcinoma (NPC) has a relatively high incidence in Chinese living in Taiwan, Hong Kong, Singapore, and South China. To better understand this cancer, we have established several new NPC cell lines. EXPERIMENTAL DESIGN: We collected biopsy specimens from suspected NPC patients and divided each specimen into two parts: the first part was fixed for routine histopathologic examination, and the other part was put into culture medium for primary culture. Once the cell lines were established, they were extensively characterized. RESULTS: Seven NPC cell lines were established, and all have been passaged more than 100 times. Two lines were derived from keratinizing carcinomas and five from undifferentiated carcinomas. Electron microscopic examination revealed that both dark and light tumor cells contained intermediate filaments with clear desmosome formation. The average doubling time ranged from 10.7 to 16.3 hours. Karyotypic analysis showed multiple chromosome abnormalities with the average chromosome number between 84 and 95. Colony forming efficiency in soft agar was 18-42%. All cell lines could induce solid tumor mass formation when transplanted into nude mice, and the histopathological findings showed two keratinizing and five nonkeratinizing carcinomas. All cell lines contained less acidic keratin polypeptides than basic keratin polypeptides. Strong expression of vimentin in each single cell of all cell lines was also observed. The oncosuppressor retinoblastoma gene in each cell line showed no remarkable abnormality, but retinoblastoma protein was abnormally expressed in some interphase cells. The oncogenes, erbB and c-fgr, were both normally expressed. While the c-myc oncogene in all cell lines was overexpressed when compared with the Burkitt's lymphoma Raji Cell line, the c-myc DNA sequence in each cell line showed neither amplification nor rearrangement. CONCLUSIONS: The newly established seven NPC cell lines have been well characterized, and all showed overexpression of c-myc oncogene. The oncosuppressor retinoblastoma gene revealed no remarkable rearrangement, but its protein product was abnormal in certain interphase cells of each cell line.

Adult↗
PubMed1991

Interactions between inositol phosphates and cytosolic free calcium following bradykinin stimulation in cultured human skin fibroblasts.

The inositol triphosphate (IP3) that results from hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) is generally accepted to be responsible for the mobilization of intracellular calcium. However, some studies suggest that low concentrations of agonists elevate cytosolic free calcium concentration ([Ca2+]i) without IP3 formation. Thus, in the present studies, a comparison of the temporal response of inositol phosphates (IP3, IP2 and IP) and [Ca2+]i to a wide range of bradykinin concentrations was used to examine the relation of these two signal transduction events in cultured human skin fibroblasts (GM3652). In addition, the effects of alterations in internal or external calcium on the response of these second messengers to bradykinin were determined. Bradykinin stimulated accumulation of inositol phosphates and a rise of [Ca2+]i in a time- and dose-dependent manner. Decreasing the bradykinin concentration from 1 microM to 0.1 microM increased the time until the IP3 peak, and when the bradykinin concentration was reduced to 0.01 microM IP3 was not detected. [Ca2+]i was examined under parallel conditions. As the bradykinin concentration was reduced from 1 microM to 0.01 microM, the time to reach the peak of [Ca2+]i increased progressively, but the magnitude of the peak was unaltered. These two second messengers were variably dependent on external calcium. Although the bradykinin-stimulated initial spike of [Ca2+]i did not depend on extracellular calcium, the subsequent sustained levels of [Ca2+]i were abolished in calcium free medium. The bradykinin-stimulated inositol phosphate formation was not dependent on the extracellular calcium nor on the elevation of [Ca2+]i that was produced with Br-A23187. These results demonstrate that bradykinin-induced IP3 formation can be independent of [Ca2+]i and of external calcium, whereas changes in [Ca2+]i are partially dependent on external calcium.

Bradykinin↗
PubMed1991

Cytosolic free calcium concentrations in synaptosomes during histotoxic hypoxia.

Altered cytosolic free calcium concentrations ([Ca2+]i) accompany impaired brain metabolism and may mediate subsequent effects on brain function and cell death. The current experiments examined whether hypoxia-induced elevations in [Ca2+]i are from external or internal sources. In the absence of external calcium, neither KCl depolarization, histotoxic hypoxia (KCN), nor the combination changed [Ca2+]i. However, with external CaCl2 concentrations as small as 13 microM, KCl depolarization increased [Ca2+]i instantaneously while hypoxia gradually raised [Ca2+]i. The combination of KCN and KCl was additive. Increasing external calcium concentrations up to 2.6 mM exaggerated the effects of K+ and KCN on [Ca2+]i, but raising medium calcium to 5.2 mM did not further augment the rise. Diminishing the sodium in the media, which alters the activity and perhaps the direction of the Na/Ca exchanger, reduced the increase in [Ca2+]i due to hypoxia, but enhanced the KCl response. The changes in ATP following K+ depolarization, KCN or their combination in the presence of physiological calcium concentrations did not parallel alterations in [Ca2+]i, which suggests that diminished activity of the calcium dependent ATPase does not underlie the elevation in [Ca2+]i. Valinomycin, an ionophore which reduces the mitochondrial membrane potential, elevated [Ca2+]i and the effects were additive with K+ depolarization in a calcium dependent manner that paralleled the effects of hypoxia. Together these results suggest that hypoxia-induced elevations of synaptosomal [Ca2+]i are due to an inability of the synaptosome to buffer entering calcium.

Animals↗
PubMed1991

Selenium in Kashin-Beck disease areas.

In this article, the duplication portion technique was used to determine the daily intakes of selenium and ten other elements in the 24-h total diets collected in the typical Kashin-Beck endemic areas, i.e., Shanxi Province and Inner Mongolia Autonomous of China. The contents of Ca, Mg, Cu, Fe, Zn, Mn, Al, Sr, Ba, and P in freeze-dried samples were determined by ICP-AES. Se was determined by differential pulse catalytic polarography. The average Se contents in total diets of Shanxi Kashin-Beck endemic and nonendemic areas were 0.009 and 0.021 micrograms/g (dry weight), respectively (P less than 0.001), corresponding to the daily intakes for Se of 4.6 and 10.5 micrograms. After the Se-supplemented fertilizer was applied (225 g of Na2SeO3/ha), the average Se content in total diets of Kashin-Beck disease area was increased to 0.0336 micrograms/g, which corresponded to the average daily intake for Se of 16.8 micrograms. In Inner Mongolia Kashin-Beck endemic and nonendemic areas, the average Se contents in total diets were 0.006 and 0.017 micrograms/g, respectively (p less than 0.001), corresponding to the average daily intakes for Se of 3 and 8.5 micrograms. The contents of other ten elements in total diets in endemic and nonendemic areas were reported and compared.

China↗
PubMed1991

Cytosolic free calcium and ATP in synaptosomes after ischemia.

Elevations in cytosolic free calcium ([Ca2+]i) precede electrophysiological alterations due to ischemia in vivo. An in vitro model of these changes would help to elucidate their molecular basis. A model of postdecapitative ischemia was used to study these interactions. Nerve endings (i.e. synaptosomes) were isolated either immediately after decapitation or at various time periods after decapitation. Synaptosomal [Ca2+]i and ATP concentrations were determined during a basal period and following depolarization. K(+)-depolarization produced an initial spike of [Ca2+]i that was followed by a new equilibrium value. Ischemia elevated the basal [Ca2+]i and the new equilibrium [Ca2+]i after KCl but suppressed the [Ca2+]i spike. However, the difference between the basal [Ca2+]i and the new equilibrium [Ca2+]i after K(+)-depolarization did not vary with ischemia. Although ischemia reduced ATP, K(+)-depolarization did not alter ATP concentrations in either the controls or the ischemia group, which suggests that synaptosomal mitochondria can meet an energy demand after ischemia. ATP was inversely related to the basal or the new equilibrium [Ca2+]i following depolarization. These changes in [Ca2+]i may underlie the alterations in neurotransmitter release and cell death following ischemia. This appears to be a useful model in which to study the molecular basis of ischemia induced changes in [Ca2+]i.

Adenosine Triphosphate↗
PubMed1991

Sodium arsenite induces ATP depletion and mitochondrial damage in HeLa cells.

Our present results show that treatment with sodium arsenite apparently decreases cellular ATP levels in a dose- and time-dependent manner in HeLa S-3 cells. The reduction in ATP induced by sodium arsenite was possibly through mitochondrial damage, since treatment with sodium arsenite resulted in reduction of rhodamine 123 accumulation and disruption of the structure of the cristae in mitochondria. However, all of these changes could be reversed by removing sodium arsenite from the culture medium. The levels of ATP depletion were correlated with the killing effects of sodium arsenite in HeLa S-3 cells.

Adenosine Triphosphate↗
PubMed1991

The cellular basis of delirium and its relevance to age-related disorders including Alzheimer's disease.

A wide variety of conditions lead to delirium (i.e., metabolic encephalopathies) in human beings and animals. Despite the varied etiology the clinical consequences are relatively stereotyped which suggests that the diverse insults that cause delirium may act by common metabolic and cellular "final pathways." Related molecular and cellular mechanisms may be involved in aging and Alzheimer's disease, conditions that predispose to the development of delirium. Animal models of delirium better reflect age-related disorders such as Alzheimer's disease than those that impair a single neurotransmitter system such as the cholinergic system; the metabolic encephalopathies produce global cognitive disturbance, which is more typical of these disorders. Thus, research related to delirium has far-reaching implications for normal and abnormal brain function.

Acetylcholine↗