[Effects of folic acid on cerebrovascular and cancer risks]. / Effets de l'acide folique sur les risques cérébrovasculaires et oncologiques liés à l'homocystéine.

Homocysteine increase is associated with an elevated risk of cerebro-vascular (CV) disease as well as osteoporosis, dementia and depression. However, most secondary prevention trials did not show any CV benefit to decrease homocysteine levels through folate administration, with the possible exception of stroke. Reasons for these failures are analysed. Moreover, folate acid could decrease the risk of colon, breast and prostate cancers mainly in wine drinkers, whereas it increases the growth of preneoplasic cells of the latter cancers. In conclusion, folate acid does not benefit patients for secondary prevention of CV or malignant diseases but it still has to be proven that it could benefit patients for primary prevention.

Predictive value of microalbuminuria in medical ICU patients: results of a pilot study.

STUDY OBJECTIVES: To evaluate the predictive value of microalbuminuria in the development of acute respiratory failure (ARF) and multiple organ failure (MOF) in ICU patients. DESIGN: Prospective, observational study. SETTING: A 31-bed, mixed medicosurgical ICU in a university hospital. PATIENTS: All adult medical patients admitted to the ICU over a 2-month period, except those receiving nephrotoxic drugs, or those with urologic trauma resulting in frank hematuria or urinary infection, or with existing chronic renal disease (serum creatinine level > or 2.0 mg/dL). INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Urinary samples for microalbumin measurement were collected at hospital admission and at 8, 24, 48, 72, 96, and 120 h after hospital admission. The severity of illness was assessed by the APACHE (acute physiology and chronic health evaluation) II score calculated on the first ICU day, and the degree of organ dysfunction was assessed using the sequential organ failure assessment (SOFA) score. Acute respiratory failure (ARF) was defined as a SOFA respiratory score > or = 3. Patients were separated into two groups according to the trend in microalbuminuria levels over the first 48 h: patients in group 1 had increasing microalbuminuria levels, and patients in group 2 had decreasing microalbuminuria levels. Group 1 included 14 patients in whom microalbuminuria levels increased from 5.2 +/- 2.0 to 19.0 +/- 3.0 mg/dL. Group 2 included 26 patients in whom microalbuminuria levels decreased from 16.4 +/- 4.0 to 7.8 +/- 3.0 mg/dL. The hospital mortality rate was 43% in group 1 and 15% in group 2 (p < 0.05). The APACHE II score and the SOFA score were higher in group 1 than in group 2. The negative predictive value of increasing microalbuminuria was 100% for the development of ARF and 96% for MOF; the positive predictive value of increasing microalbuminuria was 57% for the development of ARF and 50% for MOF. CONCLUSIONS: Accurate identification of patients destined for ARF and MOF development may enable therapeutic strategies to be applied to limit the disease process. Trend analysis of urinary albumin excretion over the first 48 h of an ICU admission may provide a useful means of identifying such patients. Additional studies need to be performed in larger, mixed patient populations to confirm these findings.

Enzymic activities in two populations of purified rat islet beta-cells.

In terms of glucose sensing by pancreatic islet beta-cells, emphasis is currently placed on both the role of glucokinase, with negligible activity of low-Km hexokinase(s), and the prevalence of the oxidative over non-oxidative modality of glycolysis, a situation tentatively attributed, in part at least, to a low activity of lactate dehydrogenase. Conflicting information is available, however, on the activity of both low-Km hexokinase(s) and lactate dehydrogenase in purified beta-cell homogenates. This issue was reinvestigated, therefore, in two populations of purified rat islet beta-cells selected on the basis of their low (betaL) or high (betaH) content in reduced pyridine nucleotides. The size and protein content of betaH cells represented about twice that of betaL cells. Such was also the case for low-Km hexokinase(s), lactate dehydrogenase, mitochondrial FAD-linked glycerophosphate dehydrogenase, glutamate dehydrogenase and glutamate-alanine and glutamate-aspartate transaminases. Whether in betaH or betaL cells, the activity of low-Km hexokinase(s) was at least as high as or higher than that of glucokinase. In both betaH and betaL, the activity of lactate dehydrogenase exceeded that required to catalyze the full reduction of glucose-derived pyruvate to L-lactate, as estimated from the rate of D-glucose phosphorylation under physiological conditions. These findings thus argue against a low expression of either low-Km hexokinase(s) or lactate dehydrogenase as major determinants of the glucose-sensing device in beta-cells.

A method for the large-scale cloning of nuclear proteins and nuclear targeting sequences on a functional basis.

We describe here a selection strategy allowing the cloning of sequences that contain a functional nuclear targeting signal. Our method relies on the use of green fluorescent protein fusion proteins to identify nuclear targeting sequences. Transfected cells expressing nuclear protein fusions were isolated on the basis of their nuclear fluorescence using flow cytometry and the transfected DNAs were recovered after bacterial transformation with total DNA from pools of sorted cells. Starting from a cDNA expression library, in which only 1% of the expressed proteins were nuclear, we obtained a 70-fold enrichment in nuclear protein-encoding clones after a single round of selection. Among the 63 clones that have been partially sequenced to date, 25 (40%) corresponded to known nuclear proteins and 13 (20%) to previously uncharacterized sequences. Despite their ability to target the green fluorescent protein marker to the cell nucleus, about half of the cloned sequences did not encode canonical basic or bipartite nuclear localization signals. The method can thus be applied to the large-scale cloning of functional nuclear targeting sequences, which opens the way to a wide investigation of nuclear import mechanisms and to the identification of previously unknown nuclear proteins.

Differences in the time course of the metabolic response of B and non-B pancreatic islet cells to D-glucose and metabolized or non-metabolized hexose esters.

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Procalcitonin used as a marker of infection in the intensive care unit.

OBJECTIVE: To determine the value of procalcitonin (ProCT) as a marker of infection in critically ill patients. DESIGN: Prospective, observational study. SETTING: Medicosurgical department of intensive care (31 beds). PATIENTS: One hundred eleven infected and 79 noninfected patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: ProCT and C-reactive protein (CRP) concentrations were monitored daily. The best cutoff values for ProCT and CRP were 0.6 ng/mL and 7.9 mg/dL, respectively. Compared with CRP, ProCT had a lower sensitivity (67.6 vs. 71.8), specificity (61.3 vs. 66.6), and area under the receiver operating characteristic curve (0.66 vs. 0.78, p < .05). The combination of ProCT and CRP increased the specificity for infection to 82.3%. In the infected patients, plasma ProCT, but not CRP, values were higher in nonsurvivors than in survivors. Infected patients with bacteremia had higher ProCT concentrations than those without bacteremia, but similar CRP concentrations. ProCT levels were particularly high in septic shock patients. CONCLUSIONS: ProCT is not a better marker of infection than CRP in critically ill patients, but it can represent a useful adjunctive parameter to identify infection and is a useful marker of the severity of infection.

Galphaolf identification by RT-PCR in purified normal pancreatic B cells and in islets from rat models of non-insulin-dependent diabetes.

Recent reports using immunohistochemistry have shown that Galphaolf which shares 88% homology with Galphas was expressed in pancreatic islets. To test the specificity of the expression of this G protein isotype in rat islet cells, B and non-B cells were separated by flow cytometry. The expression of Galphaolf and adenylyl cyclases (AC) of types II, III, V, and VI was evaluated by reverse-transcriptase polymerase chain reaction (RT-PCR). Since alterations in the expression of AC III were recently reported in the GK rat (a model of non-insulin-dependent diabetes mellitus, NIDDM), we also have analyzed the mRNA expression of Galphaolf and AC isoforms in pancreatic islets from GK rats and from adult rats neonatally treated by streptozotocin (nSTZ rats), another model of NIDDM. Southern blots of amplicons generated with specific primers of Galphaolf revealed the presence of a 540-bp band only in B cells. AC of types II, III, V, and VI were expressed both in B and non-B cells. However, AC III mRNA was clearly more abundant in non-B than in B cells. Moreover, in B cells the expression of AC VI was higher than that of AC V, whereas equal expressions of AC V and AC VI were found in non-B cells. In GK rat islets, the mRNA expressions of Galphaolf, AC II, and AC III were clearly increased and no change in AC V and AC VI was found. In nSTZ rat islets, Galphaolf expression was barely detectable, but AC II and AC III mRNA levels were higher than those observed in controls. In conclusion, Galphaolf mRNA appeared specifically expressed in islet B cells and was increased in GK islets. The steady-state mRNA levels of AC II and AC III were clearly increased in the islets of the two rat models of NIDDM. Thus, alterations in the expression of G protein isotypes and AC isoforms could contribute to the diabetic phenotype.

Insulinotropic action of beta-L-glucose pentaacetate.

The metabolism of beta-L-glucose pentaacetate and its interference with the catabolism of L-[U-14C]glutamine, [U-14C]palmitate, D-[U-14C]glucose, and D-[5-3H]glucose were examined in rat pancreatic islets. Likewise, attention was paid to the effects of this ester on the biosynthesis of islet peptides, the release of insulin from incubated or perifused islets, the functional behavior of individual B cells examined in a reverse hemolytic plaque assay of insulin secretion, adenylate cyclase activity in a membrane-enriched islet subcellular fraction, cAMP production by intact islets, tritiated inositol phosphate production by islets preincubated with myo-[2-3H]inositol, islet cell intracellular pH, 86Rb and 45Ca efflux from prelabeled perifused islets, and electrical activity in single isolated B cells. The results of these experiments were interpreted to indicate that the insulinotropic action of beta-L-glucose pentaacetate is not attributable to any nutritional value of the ester but, instead, appears to result from a direct effect of the ester itself on a yet unidentified receptor system, resulting in a decrease in K+ conductance, plasma membrane depolarization, and induction of electrical activity.

Improved viability and metabolic behavior of hepatocytes after liver storage in the presence of a succinic acid ester.

BACKGROUND: Selected esters of succinic acid were recently proposed as novel nutrients to support ATP generation in cells endangered by an imbalance between the formation and breakdown of this adenine nucleotide. In the present study, a new ester, glycerol-1,2,3-trimethylsuccinate, was examined for its potential beneficial effect in the procedures preceding liver transplantation. METHODS: The viability and metabolic behavior of hepatocytes were examined after perfusion and storage of rat livers for 20 hr at 4 degrees C with a Belzer UW-CSS solution in the absence or presence or 2 mM glycerol-1,2,3-trimethylsuccinate. RESULTS: Although it failed to affect significantly the release of cellular enzymes during storage and the protein or glycogen content of the liver, and was unable to prevent the storage-induced decrease in both biosynthetic activity and D-[U-14C]glucose incorporation into glycogen in isolated hepatocytes, the ester restored to a close-to-normal value the viability of the hepatocytes and opposed the starvation-like effects of liver storage upon both the conversion of D-[U-14C]glucose to 14CO2 and radioactive amino acids and the de novo generation of 14C-labeled D-glucose from [2-14C]pyruvate. CONCLUSIONS: Because succinic acid esters are efficiently metabolized in several cell types, the present results suggest that such esters may have a wide field of application in transplantation procedures.

Importance of ionized magnesium measurement for monitoring of citrate-anticoagulated plateletpheresis.

BACKGROUND: The infusion of citrate during apheresis may affect the levels of ionized magnesium in the blood. Hypomagnesemia and concomitant hypocalcemia could influence the parathormone response and could be responsible for some of the symptoms observed during apheresis. STUDY DESIGN AND METHODS: The study reports measurement of ionized magnesium by the new ion-selective electrode technique in response to citrate infusion in 15 donors undergoing continuous flow high-yield plateletpheresis. The monitoring included measurement of ionized calcium and parathormone every 30 minutes during the 120-minute apheresis (plus the next 30 minutes to assess recovery). RESULTS: Ionized magnesium fell by 30 +/- 4 percent (mean +/- SD, p<0.01), which contrasts with minor changes in total concentrations. Comparison of variations in the levels of ionized and total magnesium found major formation of complexes during citrate infusion. Ionized calcium fell by 15 +/- 3 percent (p<0.01), while parathormone peaked at 356 +/- 114 percent (p<0.01) of initial value after 30 minutes. Ionized cations and parathormone recovered by more than 50 percent within 30 minutes of the end of apheresis. CONCLUSION: An acute and steep drop in ionized magnesium occurs during citrate administration. The measurement of ionized magnesium should be included in future prospective studies of donor safety and parathormone regulation during apheresis.

Pancreatic islet B-cell individual variability rather than subpopulation heterogeneity.

Isolated rat pancreatic islet B-cells loaded with the Ca2(+)-sensitive fluorochrome Fluo-3 were examined by single-step flow cytometry at increasing concentrations of D-glucose (1.0 to 20.0 mM). The near forward scatter of light was unaffected by changes in hexose concentration. The Fluo-3 fluorescent signal slightly decreased when the glucose concentration was raised from 1.0 to 5.0 mM, and progressively increased at higher hexose concentrations. The fluorescence attributable to endogenous NAD(P)H increased dramatically throughout the full range of D-glucose concentration, with a typical sigmoidal concentration-response relationship. No evidence for a bimodal frequency distribution of these variables was found, whether at low or high D-glucose concentrations. The dispersion of individual NAD(P)H measurements, as judged by either their coefficient of variation or the height of their modal peak, was less pronounced at high than at low D-glucose concentrations. These findings document vastly different concentration-response relationships for metabolic and ionic variables in glucose-stimulated B-cells. They confirm that all B-cells do not display an identical behavior, but argue against the existence of subpopulation heterogeneity in their responsiveness to D-glucose.

Pancreatic islet response to dicarboxylic acid esters in rats with type 2 diabetes: enzymatic, metabolic and secretory aspects.

This study aimed to compare the metabolic and secretory responses of pancreatic islets from animals with non-insulin-dependent diabetes to D-glucose with the effects of the methyl esters of succinic acid (SME) and glutamic acid (GME). The insulin secretory response to D-glucose was impaired in islets from rats with diabetes which was either inherited (Goto-Kakizaki (GK) rats) or acquired (streptozotocin-treated (STZ) rats). This coincided with a preferential alteration of oxidative relative to total glycolysis in intact islets and a selective defect of FAD-linked mitochondrial glycerophosphate dehydrogenase (m-GDH) in islet homogenates. This enzymatic defect was also found in purified B cells from STZ rats. It contrasted both with unaltered activities of glutamate dehydrogenase and succinate dehydrogenase in the islets of diabetic animals and with a normal or even increased activity of m-GDH in the livers of GK and STZ rats. The oxidation of [1,4-14C]SME and [U-14C]GME appeared decreased in islets of GK or STZ animals when compared with control rats, but no significant difference between control and diabetic rats was observed when the oxidative data were expressed relative to the rate of [U-14C]GME hydrolysis. Nevertheless, the absolute values for insulin release evoked by a non-metabolized analogue of L-leucine (BCH), by SME and by the association of BCH with either SME or GME were invariably lower in islets of GK and STZ rats than in those of control animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Preferential stimulation by D-glucose of oxidative glycolysis in pancreatic islets: comparison between B and non-B cells.

In purified pancreatic islet B cells, a rise in D-glucose concentration from 2.8 to 16.7 mM increased the production of both 14CO2 from D-[3,4-14C]glucose and 3HOH from D-[5-3H]glucose to a much greater relative extent than in purified non-B islet cells. Moreover, the paired ratio between D-[3,4-14C]glucose oxidation and D-[5-3H]glucose utilization was significantly increased, as a result of the rise in hexose concentration, in purified B cells, but not so in purified non-B cells. It is proposed, therefore, that a preferential stimulation by D-glucose of oxidative relative to total glycolysis represents an intrinsic attribute of insulin-producing cells, as distinct from other endocrine islet cells.

[Primary hyperoxaluria type 1, peroxisomal disease: therapeutic consequences]. / L'hyperoxalurie primitive de type 1, maladie des peroxysomes: conséquences thérapeutiques.

On the occasion of a combined liver-kidney graft doing well after 3 years, the molecular anomalies responsible for primary hyperoxaluria type 1 are discussed. This rare condition may be listed in the expanding group of hereditary diseases involving peroxisomes, cellular organelles with increasingly recognised functions. Recent progress in the molecular biology of this disease have led to the proposal of of new transplant strategies for its cure.

Computer simulation of a cytosolic calcium oscillator.

A new interpretation of existing data permits us to define a model capable of accounting for agonist-induced Ca2+ oscillations in the cytosol of electrically non-excitable cells. The model only requires one Ca2+ store, which contains Ca2+ channels controlled by inositol 1,4,5-trisphosphate and Ca2+. Computer simulations may generate different experimentally observed patterns of Ca2+ oscillations.