Characteristics And Outcome Influence Of Increased Plasma Triglyceride Level In Severely Burned Adult Patients.

The aims of this study were to investigate the profile of serum triglyceride level and its influence on outcomes in adult patients with severe burns. An observational study was conducted on 62 patients with burn extent from and over 20% TBSA. Results indicated that serum triglyceride level steadily increased from 1.9mmo/l on the 3rd day to 2.5 mmol/l on the 14th day before reducing on the 21st day after burn. Remarkably higher triglyceride level was seen in patients with full thickness burn area >20% TBSA and in inhalation injury (p < .05). Liver size significantly increased over time and was greater in increased triglyceride patients, but the difference was not significant (p > .05). In addition, patients with elevated serum triglyceride level had significantly higher rates of multiple organ failure and death compared with the remaining group. Further studies need to be conducted to understand and determine intervention for increased plasma triglyceride levels in severely burned patients.

Le but de cette étude était d'évaluer les triglycéridémies et leur influence sur le devenir d'adultes sévèrement brûlés. Il s'agit d'une étude observationnelle réalisée auprès de 62 patients brûlés sur > 20% SCT. La triglycéridémie augmente régulièrement, de 1,9 mmol/L à J3 jusqu'à 2,5 mmol/L à J14 pour diminuer à partir de J21. Des taux particulièrement élevés étaient observés en cas d'atteinte profonde et d'inhalation de fumées (p < 0,05). La taille du foie augmentait au cours du temps et semblait plus élevés chez les patients hypertriglycéridémiques, sans être significative. En outre, les patients hypertriglycéridémiques développaient plus fréquemment une défaillance multiviscérale et leur mortalité était plus élevée. D'autres études sont nécessaires pour comprendre le mécanisme de cette hypertriglycéridémie et proposer une conduite à tenir pour ces patients.

Influence of inhalation injury on resting energy expenditure and plasma metabolic hormones in adult burn patients.

The aim of this study was to investigate the influence of inhalation injury on resting energy expenditure (REE) and some plasma metabolic hormones in adult burn patients. A prospective study was conducted on 16 adult burn patients admitted to the burn intensive care unit, National Burn Hospital, Vietnam. Eight patients with inhalation injury were matched with 8 non-inhalation injury patients by burn extent and age. REE measurements were obtained within 48h of admission and every week after burn. Plasma levels of epinephrine and cortisol were determined on admission and on the 7th day after burns. The results showed that, apart from REE at admission, all values of REE were significantly higher than basal metabolic rate (BMR) at all time points (p < .005). Over time, REE of both groups significantly increased and reached peak values on the 7th day after burn (1964 ± 300Kcal/m2 and 1991.8 ± 467.8Kcal/m2; REE/BMR: 1.5 vs. 1.6 respectively). These values then steadily reduced, but no remarkable differences of REE and REE/BMR were seen between the two groups at any time point (p > .05). In addition, plasma concentrations of epinephrine and cortisol were not significantly different in each group and between the two groups of patients with and without inhalation injury. In conclusion, inhalation injury may not affect metabolic response state in adult burn patients as measured by REE and metabolic hormones.

Le but de ce travail était d'étudier l'influence de l'inhalation de fumées sur la dépense énergétique de repos (DER) et le niveau plasmatique de certaines hormones « métabolique ¼ chez des adultes brûlés. Elle a été réalisée sur 16 patients hospitalisés dans le service de réanimation de l'hôpital brûlologique national du Viêt- Nam, 8 ayant inhalé de la fumée, 8 n'en ayant pas inhalé, appariés sur la surface brûlée. La DER était mesurée dans les 48 h de l'admission puis toutes les semaines. Les taux plasmatiques de cortisol et d'adrénaline étaient mesurés à l'admission et à J7. Mise part celle mesurée à l'admission, DER était systématiquement supérieure à celle calculée selon la formule de Harris- Benedict (p < 0,005), avec un pic à J7 (1964 +/- 300 Cal/m2 chez les patients avec inhalation de fumées, 1991,8 +/- 467,8 chez les autres soit 1,5 et 1,6 fois la DER supposée. Les valeurs mesurées diminuaient ensuite sans jamais qu'il y ait de différence entre les groupes. Pas plus que ne différaient les taux de cortisol ni d'adrénaline. L'inhalation de fumée semble donc de pas avoir d'impact sur la réponse métabolique, mesurée par calorimétrie ou évaluée sur les taux hormonaux.

Function of NAD glycohydrolase in ADP-ribose uptake from NAD by human erythrocytes.

The function of the ectoenzyme NAD glycohydrolase (NADase) in ADP-ribose uptake from extracellular NAD was studied in human erythrocytes that express relatively high NADase activity (adult erythrocytes) and erythrocytes expressing very low activity (newborn erythrocytes). The rates of ADP-ribose uptake from NAD in human erythrocytes were correlated with their NADase activities. In contrast, there was no significant difference in the rates of ADP-ribose uptake among these cells when incubated with ADP-ribose. These results indicate that ecto-NADase may have a role as supplier of ADP-ribose for its uptake into the cells and that the cleavage of NAD by NADase is necessary for the ADP-ribose uptake by human erythrocytes. From ADP-ribose uptake studies at 37 degrees C a Km of 0.7 +/- 0.05 microM and a Vmax of 2.04 +/- 0.1 pmol/min per microliter cell water was found for the uptake of [3H]ADP-ribose. The thiol-reactive reagents p-chloromercuribenzene sulfonic acid and N-ethylmaleimide inhibited the uptake ADP-ribose with IC50 values of 50 +/- 4 and 750 +/- 25 mM, respectively. Since efflux of [3H]ADP-ribose was negligible, the ADP-ribose transport system appears to be unidirectional. The unidirectionality was supported by the evidence that transported ADP-ribose was rapidly degraded to AMP which is impermeable to the membrane.

Glycosylphosphatidylinositol-anchored NAD glycohydrolase is released from peritoneal macrophages activated by interferon-gamma and lipopolysaccharide.

We have previously shown that an ectoenzyme, NAD glycohydrolase (NADase) could be solubilized by treatment with bacterial phosphatidylinositol phospholipase C (PIPLC). However, it is unknown whether endogenous PIPLC can cleave this ectoenzyme. In this study, we used mouse peritoneal exudate macrophages which have been known to have relatively high activity of NADase. The results show that release of ecto-NADase was markedly increased when mouse peritoneal macrophages were costimulated with interferon-gamma (IFN-gamma) and bacterial lipopolysaccharide (LPS), compared to unstimulated cells. This increase was preceded by markedly enhanced activity of endogenous glycosylphosphatidylinositol phospholipase C (GPIPLC). The cross-reacting determinant (CRD) of the glycosylphosphatidylinositol anchor in released NADase from activated macrophages was detected by immunoblotting with anti-CRD antibody. Taken together, ecto-NADase is release from peritoneal exudate macrophages during IFN-gamma/LPS-induced activation and endogenous GPIPLC is involved in the NADase release from the activated macrophages.

Increase of NAD glycohydrolase activity in uterine cervix cancers is caused by infiltration of lymphocytes.

CD38 is a type II transmembrane glycoprotein which is expressed by hematopoietic and nonhematopoietic cells in human. It has two functions of ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities and the sum of these two enzyme activities is identical with NAD glycohydrolase (NADase) activity. The levels of NADase activity in human cervical carcinoma and normal cancer tissue were measured. With a total of 12 patients with cervical cancer and 11 women with normal cervix, cancer tissues were found to have significantly higher NADase and ADP-ribosyl cyclase activities than the control group. Moreover, immunoblot analysis showed an increase of immunoreactivity against CD38 in cervical cancer tissues compared with normal tissues. Immunohistochemical data indicated that the increase of CD38 expression was due to increased infiltration of lymphocytes.

The activation of PI 3-K and PKC zeta in PMA-induced differentiation of HL-60 cells.

The human myelocytic leukemia cell line HL-60 is a useful model for the study of cellular differentiation. Phorbol 12-myristate 13-acetate (PMA) induces the monocyte/macrophage-like differentiation of HL-60 cells and results in growth arrest, increasing adherence. In PMA-induced differentiation of HL-60 cells, phosphoinositide 3-kinase (PI 3-K) activity was measured as phosphatidylinositol3P recovery from phosphatidylinositol by in vitro kinase assay. PI 3-K activity was increased in HL-60 cells that were stimulated by 20 nM PMA and the activity was inhibited by pretreatment with 20 microM LY294002, a specific inhibitor of PI 3-K. Members of the protein kinase C (PKC) family have been suggested to be one of the downstream targets of PI 3-K. PKC zeta is one of the atypical PKCs, non-diacylglycerol-responsive PKCs, and the activity was measured by the ability of phosphorylation onto myelin basic protein. PMA also induced the activation of PKC zeta during monocytic differentiation of HL-60 cells, and LY294002-pretreated cells failed to induce PKC zeta activation. The activity of PI 3-K is essential for PKC zeta activation, and LY294002 blocks both monocytic differentiation of HL-60 cells and activation of PKC zeta during PMA-induced cell differentiation. This implies that activated PI 3-K subsequently stimulates the PKC zeta in the process of PMA-induced monocytic differentiation.

Polymorphism of the angiotensin-converting enzyme gene in patients with cerebral infarction in Koreans.

The relationship between cerebrovascular disease and an insertion/deletion (I/D) polymorphism in the angiotensin-converting enzyme (ACE) gene is still being debated. The frequency of the DD genotype of the ACE gene was significantly higher in subjects with than those without cerebral infarction in Japan. The aim of the present study was to assess the relationship between ACE gene polymorphism and the development of cerebral infarction in a population from Korea. We examined its possible role as a risk factor in patients with cerebral infarction. The association between ACE gene polymorphism and cerebral infarction was examined in 106 patients with cerebral infarction and 498 controls without cerebral infarction. Frequencies of the genotypes and alleles of the ACE gene were investigated. The ACE genotype was analyzed by the polymerase chain reaction (PCR). The frequency of D allele was 37.7% in patients and 39.1% in controls (chi2 = 0.128, p = 0.720). The frequencies of the genotypes of the ACE gene were II: 39.6%, ID: 45.3%, and DD: 15.1% in patients, and II: 37.1%, ID: 47.6%, and DD: 15.3% in controls (chi2 = 0.127, p = 0.721). There was no significant difference in the frequency of the DD genotype of the ACE gene, and we did not find any association between ACE polymorphism and cerebral infarction. These results indicate that ACE polymorphism is not a risk factor for the development of cerebral infarction in a Korean population.

Auto-ADP-ribosylation of NAD glycohydrolase from Neurospora crassa.

NAD glycohydrolase (NADase; EC 3.2.2.5) is an enzyme that catalyzes hydrolysis of NAD to produce ADP-ribose and nicotinamide. We recently demonstrated that self-inactivation of NADase from rabbit erythrocytes was due to an auto-ADP-ribosylation. In the present study, a mechanism of self-inactivation of NADase from Neurospora crassa by its substrate was investigated by using intact mycelia of N. crassa and purified NADase, which had molecular characteristics different from mammalian NADases. The results suggested that inactivation of NADase from N. crassa was also due to an auto-ADP-ribosylation. These findings indicate that the auto-modification of NADase is one of the universal phenomena to regulate enzyme functions.

The nitric oxide-producing properties of Solanum lyratum.

We examined the effect of Solanum lyratum Thunb. (Solanaceae) (SL) on the production of nitric oxide (NO). Stimulation of mouse peritoneal macrophages with SL after the treatment of recombinant interferon-gamma (rIFN-gamma) resulted in increased NO synthesis. SL had no effect on NO synthesis by itself. When SL was used in combination with rIFN-gamma, there was a marked cooperative induction of NO synthesis in a dose-dependent manner. The optimal effect of SL on NO synthesis was shown 6 h after treatment with rIFN-gamma. The increased production of NO from rIFN-gamma plus SL-stimulated cells was decreased by the treatment with staurosporin. In addition, synergy between rIFN-gamma and SL was mainly dependent on SL-induced tumor necrosis factor-alpha (TNF-alpha) secretion. All the preparations of SL were endotoxin free. The present results indicate that the capacity of SL to increase NO production from rIFN-gamma-primed mouse peritoneal macrophages is the result of SL-induced TNF-alpha secretion via the signal transduction pathway of PKC activation.

Effect of Vitex rotundifolia on immediate-type allergic reaction.

We investigated the effect of aqueous extract of Vitex rotundifolia (L.) (Verbenaceae) fruits (VRFE) on the immediate-type allergic reactions in vivo and in vitro. VRFE (10(-4)-1.0 g/kg) dose-dependently inhibited systemic allergic reaction induced by compound 48/80. When VRFE was employed in a systemic allergic reaction test, the plasma histamine levels were reduced in a dose-dependent manner. VRFE (5x10(-1) and 1.0 g/kg) inhibited passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) IgE. VRFE (10(-3)-1.0 mg/ml) also dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 or anti-DNP IgE. Moreover, VRFE (10(-3) mg/ml) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results suggest that VRFE may be beneficial in the regulation of immediate-type allergic reaction.

Inhibition of tumor necrosis factor-alpha-induced apoptosis by Asparagus cochinchinensis in Hep G2 cells.

A human hepatoma cell line, Hep G2 cells, is a reliable system for the study of alcohol-induced hepatotoxicity. In this study, we investigated the effect of an aqueous extract of Asparagus cochinchinensis(MERRIL) (Liliaceae) roots (ACAE) on ethanol (EtOH)-induced cytotoxicity in Hep G2 cells. ACAE (1-100 microg/ml) dose-dependently inhibited the EtOH-induced tumor necrosis factor-alpha (TNF-alpha) secretion. ACAE (1-100 microg/ml) also inhibited the EtOH and TNF-alpha-induced cytotoxicity. Furthermore, we found that ACAE inhibited the TNF-alpha-induced apoptosis of Hep G2 cells. These results suggest that ACAE may prevent the EtOH-induced cytotoxicity through inhibition of the apoptosis of Hep G2 cells.

Inhibition of stem cell factor- and nerve growth factor-induced morphological change by wortmannin in mast cells.

Recombinant murine stem cell factor (rmSCF) or recombinant murine nerve growth factor (rmNGF) induced the morphological change of large numbers of rat peritoneal mast cells (RPMC). We investigated the role of phosphatidylinositol 3'-kinase (PI3-kinase) in receptors-mediated morphological change in RPMC. Exposure of RPMC to PI3-kinase inhibitor, wortmannin, before the addition of rmSCF and rmNGF antagonized those factors-induced morphological change. These results suggest that the Pl3-kinase is involved in the signal transduction pathway responsible for morphological change following stimulation of rmSCF and rmNGF and that wortmannin blocks these responses.

Expression of glycosylphosphatidylinositol-anchored NAD glycohydrolase in differentiated HL60 cells by phorbol ester.

Human leukemic HL60 cells are known to express NAD glycohydrolase (NADase) activity following differentiation into macrophage-like cells by 12-O-tetradecanoylphorbol-13-acetate (TPA) or granulocyte-like cells by retinoic acid (RA) treatment. Recently, it was reported that 46 kDa human leukocyte antigen, CD38, expressed by RA-differentiated HL60 cells contained NADase, ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities. In the present study we questioned whether the NADase activity found in TPA-differentiated HL60 cells is similar to that found in RA-treated cells. Herein we demonstrate that, unlike what is observed following RA treatment, the NADase activity of TPA differentiated cells associates with a 65 kDa glycosylphosphatidylinositol-anchored NADase.

Interleukin-3 or immunoglobulin E promotes expression of protein kinase C delta gene in murine mast cells.

We reported previously that protein kinase C delta (PKCdelta) is the main isoenzyme in various types of murine mast cells. In the present study we investigated the regulation of expression of PKCdelta gene in murine mast cells in vitro and in vivo. The mRNA expressions of PKCdelta were promoted in response to interleukin-3 (IL-3) or immunoglobulin E (IgE) in mouse mastocytoma P-815 cells. In addition we have evaluated the mast cells which express PKCdelta mRNA in IgE-dependent passive cutaneous anaphylaxis reaction, using in situ hybridization with the antisense riboprobe in skin. These results indicate that mast cell activation can induce a marked promotion in steady state levels of PKCdelta mRNA.

Taxol and related compounds in Korean native yews (Taxus cuspidata).

The concentrations of taxol and related compounds in the bark and needles of Taxus cuspidata grown on Mt. Jiri, Mt. Sobaek, and Cheju Island, and T. cuspidata var. latifolia on Ullung Island in Korea were determined by high performance liquid chromatography (HPLC). The taxane content significantly varied with the location and plant part. The taxol content in the bark of native yews from Mt. Jiri and Mt. Sobaek was high when compared to that reported for Pacific yew (T. brevifolia), whereas bark from trees on Cheju and Ullung islands contained a much lower level. Surprisingly, the needles from Cheju and Ullung islands contained a much higher level of taxol (0.022% and 0.0173%, respectively) than those of intermountain locations (0.0058% to 0.0085%), on the basis of dry weight. The bark and needles of T. cuspidata var. latifolia on Ullung Island also contained relatively high concentrations of 10-deacetylbaccatin III, 0.0497% and 0.0545%, respectively, and indicated that environmental factors may affect the quantity. Taxol in the needles was confirmed by electrospray mass spectrometry. These results suggest that foliage from yew trees growing in their natural habitats on Cheju and Ullung islands may provide a renewable source for taxol.

Action of Sosiho-Tang on systemic and local anaphylaxis by anal administration.

The herbal formulation Soshiho-Tang (SS-Tang) has been used against allergic disease for generations, and still occupies an important place in traditional medicine in Korea. Previously, we reported that SS-Tang potently inhibited mast cell- mediated anaphylaxis when orally administered. In this study, we investigated the effect of SS-Tang by anal administration in anaphylaxis responses. SS-Tang dose-dependently inhibited compound 48/80-induced systemic anaphylaxis with doses of 10(-4) to 1 g/kg 1 h before anally administered. Of special note, SS-Tang inhibited systemic anaphylaxis completely with a dose of 1 g/kg. SS-Tang reduced plasma histamine levels induced by compound 48/80 significantly. However, the mortality was 100% when SS-Tang was administered after compound 48/80 treatment. SS-Tang (10(-1) g/kg) also inhibited passive cutaneous anaphylaxis activated by anti-dinitrophenyl IgE antibody by 30.9%. These results provide evidence that anal therapy of SS-Tang may be beneficial in the treatment of systemic and local anaphylaxis.

Inhibitory effect of anaphylactic reaction of Sosiho-Tang.

Mast cells synthesize and secrete chemical mediators which play an central role in anaphylactic reactions. Compound 48/80 is a condensation product of formaldehyde with paramethoxyphenylethylamine that reliably induces the release of chemical mediators in the mast cell granules. Aggregation of the high-affinity Fc receptor also stimulates the mast cells. The objective of the current study was to determine the effect of Sosiho-Tang (SS-Tang) on mast cell-mediated anaphylactic reaction. SS-Tang completely inhibited systemic anaphylaxis induced by compound 48/80 in mice. SS-Tang inhibited local anaphylaxis induced by anti-dinitrophenyl (DNP) IgE. In addition SS-Tang concentration-dependently inhibited histamine release in mast cells induced by compound 48/80 or anti-DNP IgE. These results indicate that SS-Tang may contain compounds with actions that inhibit mast cell degranulation.

Inhibitory effect of anaphylactic shock by caffeine in rats.

Caffeine is known to reduce evoked histamine secretion, but the effects of caffeine on anaphylactic shock have not been clarified. We have investigated the effects of caffeine on anaphylactic shock in rats. Systemic anaphylactic shock by compound 48/80 injection was monitored for 1 h. An IgE-dependent local anaphylactic shock was generated by sensitizing the skin with anti-dinitrophenyl (DNP) IgE followed 48 h later with an injection of antigen. Caffeine inhibited compound 48/80-induced anaphylatic shock to 40% with a dose of 1 mg/kg. Caffeine (0.1 mg/kg) inhibited to 56.4+/-0.4% passive cutaneous anaphylactic shock activated by anti-DNP IgE. Caffeine (5-20 mM) significantly inhibited histamine release from rat peritoneal mast cells (RPMCs) activated by compound 48/80 or anti-DNP IgE. Especially, caffeine (20 mM) inhibited by 96.7+/-0.5% histamine release activated by compound 48/80. Moreover, caffeine (1-20 mM) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMCs. The level of cAMP in RPMCs, when caffeine (20 mM) was added, increased significantly after 5-60 min compared with that of a normal control. These results indicate that caffeine inhibits immediate-type allergic reactions by inhibition of mast cell degranulation in vivo and in vitro.

Regulation of NAD+ glycohydrolase activity by NAD(+)-dependent auto-ADP-ribosylation.

NAD+ glycohydrolase (NADase; EC 3.2.2.5) is an enzyme that catalyses hydrolysis of NAD+ to produce ADP-ribose and nicotinamide. Its physiological role and the regulation of its enzymic activity have not been fully elucidated. In the present study, the mechanism of self-inactivation of NADase by its substrate, NAD+, was investigated by using intact rabbit erythrocytes and purified NADase. Our results suggest that inactivation of NADase was due an auto-ADP-ribosylation reaction. ADP-ribosylated NADase of rabbit erythrocytes was deADP-ribosylated when incubated without NAD+, and thus enzyme activity was simultaneously restored. These findings suggest that reversible auto-ADP-ribosylation of NADase might regulate the enzyme's activity in vivo.

Significance of ecto-cyclase activity of CD38 in insulin secretion of mouse pancreatic islet cells.

Cyclic ADP-ribose (cADPR), a product of CD38, has a second messenger role for in intracellular Ca(2+) mobilization from microsomes of pancreatic islets as well as from a variety of other cells. ADP-ribosylation of CD38 by ecto-mono ADP-ribosyltransferase in activated T cells results in apoptosis as well as inactivation of its activities. We, therefore, examined the effect of ADP-ribosylation of CD38 in mouse pancreatic islet cells. NAD-dependent inactivation and ADP-ribosylation of CD38, intracellular concentrations of cADPR and Ca(2+), and insulin secretion were measured following incubation of mouse pancreatic islet cells with NAD. ADP-ribosylation of CD38 inactivated its ecto-enzyme activities, and abolished glucose-induced increase of cADPR production, intracellular concentration of Ca(2+), and insulin secretion. Taken together, ecto-cyclase activity of CD38 to produce intracellular cADPR seems to be indispensable for insulin secretion.