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Biomedical subjects

A F Hernández

Publications and source records attributed to A F Hernández.

15 recordsLinked to original sources

Immunohistochemical evidence for the expression and induction of paraoxonase in rat liver, kidney, lung and brain tissue. Implications for its physiological role.

Studies on the localization of paraoxonases (PON's) are of interest because of its involvement in both the detoxication of activated organophosphorus pesticides and in the prevention of peroxidative damage to phospholipids and cholesteryl-esters in LDL and HDL particles and cell membranes during the atherogenic process. In the present study, we have investigated the cellular localization of PON1 by immunohistochemistry in different rat tissues. The protein was mainly detected in the endothelial lining of every tissue studied (liver, kidney, lung and brain). Besides, it was found in hepatocytes from the centrolobular region of the liver, in the glomeruli and basal pole of the proximal convoluted tubule of the kidney, in cells from bronchiolar epithelium and type I pneumocytes of the lung, and in leptomeningeal cells, ependymal cells and ventricular side of choroid plexus cells of the brain. However, neurons and glia lacked immunostaining. After 3-methylcholanthrene induction an increase in the intensity of immunostaining was observed in the same areas, as well as an additional staining in midzonal hepatocytes. On the basis of the tissue distribution observed for PON1, it is proposed that this enzyme might have a function related to the inactivation of oxidative stress by-products (either at a cellular level or blood-vessel wall) and other environmental chemicals. At present it has not yet been established whether the paraoxonase detected in the various tissues is truly a product of the PON1 gene or could represent products of the PON2 or PON3 genes.

Animals↗

Distribution profiles of paraoxonase and cholinesterase phenotypes in a Spanish population.

The paraoxonase/arylesterase phenotype was measured in a Spanish population as previous studies have reported that the polymorphic variation in serum paraoxonase activity may affect the metabolism of organophosphates in individuals at risk of chronic intoxication. The prevalence of congenital deficiency in serum cholinesterase was also established in order to ascertain whether individuals with a congenital defect would be at a higher risk against a potential organophosphate exposure. We consider it useful to incorporate these two biomarkers into the health programme of agricultural workers with the purpose of monitoring workers who spray organophosphate pesticides, as they provide reliable indications of early-stage effects related to biochemical alterations that might precede overt clinical pictures.

Adolescent↗

Identification of two rat liver proteins with paraoxonase activity: biochemical evidence for the identity of paraoxonase and arylesterase.

The existence of two or more enzyme forms with paraoxonase activity has been reported in sheep, rabbit, human and rat serum and recently in mouse and rat liver. In this study we describe the presence of two peaks with paraoxonase activity (M1 and M2) after non-specific affinity chromatography of rat liver microsomes on Cibacron Blue 3GA. The first peak (M1) was obtained during the washing of the column and coeluted with albumin. The second active peak (M2) was eluted with 1 M NaCl. The characterization of each peak was determined by SDS/PAGE electrophoresis and Western-blotting. A comparison of both active fractions on the basis of kinetic parameters, heat inactivation and pH stability, calcium requirement and inhibition by EDTA and several metals was performed. Our results support the fact that two proteins capable of hydrolyzing paraoxon are present in rat liver microsomes. Furthermore, during the purification to homogeneity of rat liver paraoxonase we have performed a study of its hydrolytic ability against three different substrates: paraoxon, phenylacetate and phenyl thioacetate (Paraoxonase (PON), Arylesterase (ArE), Phenyl thioacetate esterase (PTase)). The elution profile in different chromatographic steps, as well as the activity ratios from the crude extract throughout the purification process, heat inactivation and effect of inhibitors were used as identity criteria for the three hydrolytic activities. Our results show evidence for the hydrolysis of paraoxon and phenylacetate by the same protein from rat liver (paraoxonase).

Animals↗

Inhibition of paraoxonase activity in human liver microsomes by exposure to EDTA, metals and mercurials.

Inhibition of paraoxon hydrolase (paraoxonase) activity by 'in vitro' exposure to EDTA, Mg2+, Co2+, Ba2+, La3+, Zn2+, Cu2+, Hg2+, p-hydroxymercuribenzoate (p-OH-MB) and phenyl mercuric acetate (PMA) was investigated in human liver microsomes. Enzyme activity was totally inhibited by 1 mM EDTA in a time-dependent manner, in contrast to previous data obtained in rat liver where an EDTA-resistant fraction was detected. The possible influence of postmortem changes in these results was checked in a parallel experiment using rat livers with different postmortem intervals. From our results the existence in human liver of an EDTA-resistant fraction cannot be discarded. Ba, La and PMA showed immediate inhibition. By contrast the other compounds tested were time-dependent inhibitors. Ba and Zn showed the highest IC50 values. Cu and mercurials (Hg, p-OH-MB, PMA) were the most potent inhibitors of human liver paraoxonase. Kinetic analysis (Lineweaver-Burk and Dixon plots) indicated that different inhibitors exhibit different inhibition patterns: competitive (EDTA, Ba, La, Cu, p-OH-MB and PMA), non competitive (Zn) and mixed (Hg). The pretreatment of sample with dithiothreitol (DTT) protects against the inhibitory effect of mercurials. Furthermore after inhibition by mercurials the activity was restored by DTT. These results confirmed the essential role of the -SH groups to maintain the catalytic activity of paraoxonase and suggest the existence of two types of -SH groups that could differ in their localization.

Animals↗

Simultaneous death of twins. An environmental hazard or SIDS?

The sudden and simultaneous death of twin infants is described. The search for hazards in the home and the postmortem investigation provided an insight into the possible mechanism of death. Instead of a diagnosis of simultaneous sudden infant death syndrome (SIDS), the infants were determined to be victims of a combination of several environmental factors that led suddenly to death. Strong circumstancial evidence of nonnatural (accidental) death included sublethal levels of carbon monoxide (CO), overwrapping, and mechanical obstruction of upper airways. The literature concerning the phenomenon of simultaneous twin death is also reviewed.

Accidents↗

Primary hyperparathyroidism due to parathyroid carcinoma.

Most cases of primary hyperparathyroidism are due to either a parathyroid adenoma or to parathyroid hyperplasia. Parathyroid carcinoma is a very rare cause of hyperparathyroidism. Although the diagnosis of parathyroid carcinoma is usually established based on pathological criteria of vascular and capsular invasion, some clinical and biochemical features differentiate it from benign forms of hyperparathyroidism. We report the case of a middle-aged woman with a long standing history of nephrolithiasis, who presented with a palpable neck mass, weight loss, severe hypercalcemia and hypophosphatemia, as well as very high serum levels of intact parathyroid hormone. Surgical neck exploration revealed a large tumor that invaded trachea, esophagus, reccurrent laryngeal nerve, right apical pleura and right carotid artery. Pathological examination confirmed the invasive nature of the tumor. Along with the case report, we review the literature and discuss the diagnostic and therapeutic options of this rare condition.

Carcinoma↗

Increased risk of suicide with exposure to pesticides in an intensive agricultural area. A 12-year retrospective study.

Several reports have suggested that exposure to agricultural pesticides (mainly chronic exposure to organophosphates) produces depression, and depression is a major risk factor for suicide. A retrospective epidemiological study of 251 suicide cases was undertaken to explore the possible relationship between the high suicide rates in an intensive agricultural area, and a specific group of population at risk, namely farmers with chronic exposure to pesticides, who are at risk to develop mood disorders (mainly depression). Our data show that the suicide rate in that area is significantly higher than the suicide rates from other geographic areas with very similar socioeconomic and demographic features. In addition, the mortality from suicide in this population (farmers) does differ significantly from that of the rest of the population.

Agricultural Workers' Diseases↗

Clinical and biochemical changes in greenhouse sprayers chronically exposed to pesticides.

1 This study was conducted with the aim of evaluating the impact on health produced by the use of different types of pesticides in greenhouses. It is based on the need to practice and develop biological monitoring techniques to assess exposure and predict health risk in workers occupationally exposed to pesticides. 2 Two groups of greenhouse workers with either high or low exposure to a combination of pesticides was taken in Almería, a Spanish province where cultures under plastic are very extended. 3 One hundred and five sprayers were interviewed to collect information about symptoms and signs related to past exposures. Each pesticide sprayer was examined by a physician, and a blood sample was drawn for plasma and red blood cell cholinesterases, complete blood count, and liver and renal function tests. 4 Exposure of workers to a combination of pesticides resulted in 37% of the workers showing toxic signs and symptoms. The main toxic effect observed were a high incidence of spontaneous abortion, depression, and certain neurologic disorders like headache, tremor and paraesthesia. 5 The major analytical change was a decrease of the mean corpuscular haemoglobin concentration in 38% of the cases. However, no significant decrease in both serum and erythrocyte cholinesterase activities was observed. 6 The sprayers were not usually aware of the potential hazards of pesticides and did not try their best to maintain personal hygiene.

Adult↗

Partial purification of paraoxonase from rat liver.

A method for the partial purification of rat liver paraoxonase is presented. The method consists of the following steps: preparation of microsomes, solubilization with Triton X-100, adsorption on hydroxylapatite and chromatography on DEAE-52 cellulose. A partially purified preparation of rat liver paraoxonase has been obtained, showing a specific activity of 422 mU/mg with a yield of about 22% and a purification factor of 77-fold.

Animals↗

Rat liver paraoxonase: subcellular distribution and characterization.

The subcellular localization and some biochemical properties of rat liver paraoxonase have been studied in order to establish a correlation with plasma enzyme. The whole paraoxonase activity was found in the microsomal fraction. Rat plasma and liver paraoxonase showed similar optimum pH (8.5), Km (0.4 mM) and calcium requirement, but differed in the response to several inhibitors.

Animals↗

Characterization of paraoxonase activity in pericardial fluid: usefulness as a marker of coronary disease.

In this study, the presence of paraoxonase activity in pericardial fluid was demonstrated. A comparison of some properties, such as optimum pH, stability versus pH, heat inactivation, effect of inhibitors, isoelectric point and kinetic parameters (Km and Vmax), between plasma and pericardial fluid paraoxonase was made. The properties studied were practically identical. The enzyme activity in pericardial fluid was tested as a marker in the postmortem diagnosis of myocardial infarction. The paraoxonase activity in the myocardial infarction group (47 cases) was lower than in the control group (40 cases), but the difference was not significant.

Aryldialkylphosphatase↗

Decrease of phosphofructokinase activity in relation to the pathogenesis of triorthocresyl-phosphate-induced delayed neuropathy.

The in vivo effect of a single dose of the neuropathic compound triorthocresyl-phosphate (TOCP) on phosphofructokinase (PFC, E.C. 2.7.1.11) and its relation with the initiation step (inhibition and aging of neuropathy target esterase, NTE) in the TOCP-induced delayed neuropathy have been studied. Hens were treated with a neurotoxic dose of TOCP (500 mg/kg, p.o.) and with a protective compound (Phenylmethanesulfonyl fluoride, PMSF, 30 mg/kg s.c.) in different combinations: TOCP, TOCP + PMSF, PMSF + TOCP and PMSF. PFK activity was determined in brain and sciatic nerve 1, 3, 7 and 15 days after treatment. PFK activity decreased in sciatic nerve 15 days after dosing with TOCP or TOCP + PMSF. When animals were dosed with the protective agent (PMSF) alone or before administering the neurotoxic compound, PFK activity was unaltered and clinical signs of neuropathy were absent. The data presented here suggest that phosphofructokinase is involved in the pathogenesis of the neuropathy induced by TOCP.

Acetylcholinesterase↗