Alterations of energy metabolism and glutathione levels of HL-60 cells induced by methacrylates present in composite resins.

OBJECTIVES: Methacrylic compounds such as 2-hydroxyethyl methacrylate (HEMA), triethylene glycol dimethacrylate (TEGDMA) and bisphenol A glycerolate (1 glycerol/phenol) dimethacrylate (Bis-GMA) are largely present in auto- or photopolymerizable composite resins. Since the polymerization reaction is never complete, these molecules are released into the oral cavity tissues and biological fluids where they could cause local adverse effects. The aim of this work was to verify the hypothesis that the biological effects of HEMA, TEGDMA and Bis-GMA - at a non-cytotoxic concentration - depend on the interaction with mitochondria and exert consequent alterations of energy metabolism, GSH levels and the related pathways in human promyelocytic cell line (HL-60). METHODS: The biological effects of methacrylic monomers were determined by analyzing the following parameters: GSH concentration, glucose-6-phosphate dehydrogenase (G6PDH) and glutathione reductase (GR) activity, oxygen and glucose consumption and lactate production along with cell differentiation and proliferation. RESULTS: All monomers induced both cellular differentiation and decrease in oxygen consumption. Cells treated with TEGDMA and Bis-GMA showed a significant enhancement of glucose consumption and lactate production. TEGDMA and HEMA induced GSH depletion stimulating G6PDH and GR activity. CONCLUSIONS: All the monomers under study affect the metabolism of HL-60 cells and show differentiating activity. Since alterations in cellular metabolism occurred at compound concentrations well below cytotoxic levels, the changes in energy metabolism and glutathione redox balance could be considered as potential mechanisms for inducing clinical and sub-clinical adverse effects and thus providing useful parameters when testing biocompatibility of dental materials.

Plasma levels of coenzyme Q(10), vitamin E and lipids in uremic patients on conservative therapy and hemodialysis treatment: some possible biochemical and clinical implications.

Coenzyme Q(10) (CoQ(10)), vitamin E, total cholesterol, HDL-cholesterol (HDLC) and triglycerides were measured in the plasma of 62 patients with kidney failure, 46 under hemodialysis treatment and 16 under conservative therapy, and 95 controls. The sum of LDL-cholesterol (LDL-C) and VLDL-cholesterol (VLDL-C) was also calculated for each patient. The ratio CoQ(10)/LDL-C+VLDL-C in both conservative therapy and hemodialysis populations was significantly lower (P<0.001) compared with normal controls and remained unchanged after the dialysis treatment. On the contrary the ratio vitamin E/LDL-C+VLDL-C was normal but decreased significantly (P<0.02) after each dialysis. Since coenzyme Q is the main inhibitor of the prooxidant action of vitamin E, it was hypothesized that its decrease in both the populations examined could make the lipoproteins of these patients more vulnerable to a peroxidative attack.

Determination of copper, zinc, and selenium in human plasma and urine samples by potentiometric stripping analysis and constant current stripping analysis.

Potentiometric stripping analysis and constant current stripping analysis are proposed as routine methods for analysis of copper, zinc and selenium in plasma and urine samples. The analytical performance of these methods is comparable with that reported for atomic absorption spectrometry. However the low cost, greater simplicity of the apparatus, and the facility of execution make this methodology a valid candidate for routine application in Clinical Chemistry laboratories.

Multimodal approach in invasive fetal therapy.

BACKGROUND: Integrated fetal therapy is a new approach to prenatal management consisting of a combination of invasive procedures which complement each other to provide as much information as possible on the fetal compartments. METHODS: We carried out a study on 50 fetuses of singleton pregnancies undergoing invasive procedures -- at least three per fetus -- for diagnostic and therapeutical purposes. A total of two hundred and fifty invasive procedures were adopted. The study population was divided into two groups, those studied between 1988 and 1992 and those studied between 1993 and 1995. RESULTS: The diagnostic and therapeutic utility of complementary invasive procedures in fetuses with nonimmune fetal hydrops and urinary tract malformations was assessed. In fetuses with nonimmune fetal hydrops integrated invasive procedures markedly affected the fetal-neonatal survival rate, whereas in those with urinary tract malformations scheduled for postnatal surgery these procedures made it possible to limit intrauterine renal damage. CONCLUSIONS: Complementary invasive procedures in NIFH fetuses particularly influence the fetal-neonatal survival rate. Since urinary tract malformations must be treated by postnatal surgery, complementary invasive procedures serve to limit intrauterine renal damage in the meantime and to reduce cesarean section rate.

Ontogeny of the fetal immune system: study on pregnancies with Rh-isoimmunization and nonimmune fetal hydrops.

This study aims at observing and comparing the antigen expression of some fetal T- and B-lymphocyte subpopulations in Rh-isoimmunization, which determines anemic hypoxia in the fetus, and nonimmune fetal hydrops (NIFH) which, even if there are some etiological factors involved, causes hipoxic hypoxia in the fetus. Twelve fetuses were studied by way of 30 fetal blood samples obtained by ultrasound-guided cordocentesis between the 20th and 36th gestational week. Twenty-four blood samples in all where taken from the eight fetuses with Rh-isoimmunization. Six blood samples were obtained from the four fetuses with NIFH. The lymphocyte phenotypes studied by monoclonal antibodies and flow cytometry were the following: CD3, CD4, CD8, expression of T-lymphocyte subpopulations; BsIg, CD19, expression of B-lymphocyte subpopulations. We observed a near-normal maturation process in fetuses with Rh isoimmunization, whereas in fetuses with NIFH we observed inhibition and/or delayed expression of T-lymphocytes. An early and increased B-lymphocyte activation marked a cooperation between the two systems in the early gestational periods.

Enzymatic determinations in acute rejection after liver transplantation: preliminary report on necrosis index.

The catalytic activities of some mitochondrial and cytoplasmic enzymes were measured in plasma from 19 patients after orthotopic liver transplantation, in order to detect and monitor the evolution of hepatocellular damage and to predict liver rejection. The enzymatic activities determined were: mitochondrial isoenzyme of aspartate aminotransferase, glutamate dehydrogenase, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltranspeptidase and alkaline phosphatase. The results of all enzymatic activities were normalized by expressing them as multiples of the upper limit of the relevant reference range and then the necrosis index (NI) has been calculated. The proposed NI consists of percent ratio of the normalized mitochondrial enzymatic activities over the sum of cytoplasmic and mitochondrial normalized activities. We observed that NI values higher than 30% correctly identified all but two acute rejection events which were documented by liver biopsies showing a diagnostic sensitivity of 90%, specificity of 78% and a predictive value of 90%.

Biochemical parameters to assess viability of blood stored for transfusional use.

Blood collected in citrate-phosphate-dextrose-adenine (CPDA-1) containing transfusional bags, was weekly tested throughout a 35 day period. Biochemical assays included plasmatic glucose, electrolytes, free Hb, acid-base balance and hemogasanalysis. Intraerythrocytic ATP and 2,3 DPG were also determined. Results show that an almost total 2,3 DPG depletion occurs during the first three weeks, whereas intracellular ATP are about 50% of the initial values, at the same time. Lowering of pH is also maximal at the third week. pCO2 variation pattern is biphasic: an early increase due to HCO3- titration by lactic acid arising from red cells glycolysis, followed by a decrease probably due to plastic bag permeability to CO2 itself. The percentage of O2Hb also rises during blood storage: this might be the combined result of increase in pO2 and decrease in 2,3 DPG content. A rise of free Hb was obtained; extracellular K+ levels underwent a sixfold increase in 35 days. The mechanism of relative variations of these parameters and the gas transport are discussed. Some of these parameters might be used as routine controls to asses viability and functional status of stored red cells for transfusional use.

Long-term retrospective control procedures for monitoring analytical performance of laboratory instruments.

We propose a statistical procedure for long-term quality-control of laboratory instruments, including daily, day-to-day, and monthly evaluations. The procedure is based on the unique and unequivocal interpretation of five results for control sera by calculation of a Reliability Index and further manipulations of this unitless parameter. This method, which we have tested during the past two years, allows for monitoring analytical performance and making comparisons with results of interlaboratory surveys. The monthly analytical variability, expressed as "total error," is an indicator of the clinical usefulness of analytical results.

Subcellular distribution of some specific enzymatic activities in human leucocytes.

The activities and subcellular distribution of the following enzymes: NADH oxidase, alkaline phosphatase, beta-glucuronidase and ATPase, were assayed in human mononuclear and polymorphonuclear leucocytes and in particular the contamination and integrity of the mitochondrial fractions were evaluated with this new separation procedure. Results show that maximal contamination was found to be that from lysosomal beta-glucuronidase especially in polymorphonuclear leucocyte mitochondria fractions. Furthermore oligomycin-sensitive ATPase data suggest that mitochondria do not decrease in number or lose their integrity to a great extent. Controversial p-nitrophenyl-phosphatase activity was also found to be present in polymorphonuclear and mononuclear leucocytes granular-soluble fractions.