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3.
Toxicol Lett ; 165(3): 221-9, 2006 Sep 10.
Article in English | MEDLINE | ID: mdl-16797889

ABSTRACT

It had been observed that the chromaffin cells of bovine adrenal medulla contain high levels of neuropathy target esterase (NTE), the esterase whose inhibition and aging is associated with induction of the organophosphorous induced delayed neuropathy. In this study, total esterase and NTE activities, and their inhibition kinetics by OPs are characterized in adrenal medulla of several species in order to find the best source for chromaffin cells. Total esterase activity in membrane fraction of bovine, equine, porcine, ovine and caprine were 6100+/-840, 4200+/-270, 5000+/-120, 28800+/-3000, and 10800+/-2400mU/gtissue, respectively (mean+/-S.D., n=3-4). NTE represented around 70%, 24%, 58%, 10% and 24% of the total esterases in the same tissues, respectively. It was deduced that NTE represents between 69% and 89% of the "B-activity" (activity resistant to 40microM paraoxon) in the membrane fraction of all species. The mipafox I(50) calculated for 30-min inhibition of NTE at 37 degrees Celsius ranged between 7.4 and 12microM. These values are in the range of that for brain NTE in hen (the usual model for testing OP delayed neurotoxicity). Considering that bovine adrenal medulla contains high NTE activity, that it represents a high proportion of total activity, it is easier to dissect than adrenal medulla from equine, caprine or ovine, and is more readily available than species cited previously, and that its inhibitory properties are similar to the classical hen brain model, it is deduced that bovine adrenal medulla is the most appropriate source of chromaffin cells to study OP toxicity, with porcine as the second alternative. The kinetic properties of chromaffin cell cultures from bovine and porcine were in accordance with their properties in homogenate and subcellular fractions, and they displayed an appropriate stability and viability of the primary culture to be used in in vitro toxicological studies for both mechanistic and testing purposes.


Subject(s)
Chromaffin Cells/drug effects , Organophosphorus Compounds/pharmacology , Organophosphorus Compounds/toxicity , Animals , Carboxylic Ester Hydrolases/metabolism , Cattle , Cells, Cultured , Goats , Horses , Isoflurophate/analogs & derivatives , Isoflurophate/pharmacology , Isoflurophate/toxicity , Paraoxon/pharmacology , Paraoxon/toxicity , Sheep , Species Specificity , Swine
4.
Rev. toxicol ; 22(1): 44-48, ene.-abr. 2005. tab
Article in Es | IBECS | ID: ibc-66485

ABSTRACT

Durante los tres últimos cursos académicos la asignatura de Biotoxinas de la Universidad Miguel Hernández de Elche ha sido impartida utilizando un sistema que combinó metodologías docentes expositivas clásicas con otras centradas principal-mente en el trabajo del estudiante. Así, inicialmente los profesores presentaron en forma de lección magistral participativa los temas esenciales para el desarrollo de la asignatura. Posteriormente el profesor tutorizó la elaboración de trabajos por parte de los alumnos que voluntariamente accedieron a ello. Dichos trabajos fueron presentados posteriormente al resto de la clase en forma de seminario. El trabajo elaborado y su presentación constituyeron los principales elementos empleados en la evaluación del aprendizaje del alumno. Los alumnos que no elaboraron trabajo fueron evaluados mediante un examen tradicional. Los resultados obtenidos demuestran una gran aceptación por parte de los alumnos, ya que se registró un progresivo aumento del número de matrículas así como una excelente media de asistencia a clase y a los seminarios. Los resultados académicos también fueron satisfactorios, ya que la tasa de éxito fue siempre del 100% y la de rendimiento siempre fue superior al 80%. Además de los buenos resultados obtenidos la metodología empleada tiene la ventaja de que se adapta a los requerimientos del futuro Espacio Europeo de Educación Superior


During the last three academic years the subject called Biotoxins has been taught at the Universidad Miguel Hernández de Elche using a system that combines classical expositive methodologies with others that are mainly focused on the work of the student. Initially, the teachers performed magisterial lessons the most important themes necessary for the understanding of the subject. Afterwards, the teachers supervised the development of individual academic work voluntarily carried out by the students. These works were then presented to the class in seminary format and together with the presentation of the seminary, they were the main elements employed in the learning evaluation of each student. Students that did not perform the voluntary work were evaluated through a classical exam. The obtained results splay a good acceptance of the subject among the students as the number of students who signed up for the course increased every academic year. The number of students that regularly attended the teaching sessions was also very high. The academic results were also very satisfactory as the success ratio was always 100%, while the yield ratio was always higher than 80%. In addition to the good obtained results, this methodology also has the additional advantage that it meets the requirements of the European Higher Education System


Subject(s)
Toxicology/education , Toxins, Biological/analysis , Toxicology/trends , European Union , Universities , Schools, Medical/trends
5.
Toxicol Lett ; 151(1): 163-70, 2004 Jun 15.
Article in English | MEDLINE | ID: mdl-15177651

ABSTRACT

Based on the high level of phenyl valerate esterase activities, and in particular of neuropathy target esterase (NTE) found in bovine adrenal medulla, chromaffin cells culture have been proposed as an alternative model for the study of organophosphorus neurotoxicity. Organophosphorus-induced polyneuropathy is a syndrome related to the inhibition and further modification by organophosphorus compounds of NTE (a protein that displays phenyl valerate esterase activity resistant to mipafox and sensitive to paraoxon). Total phenyl valerate esterase activities found in homogenate, particulate and soluble fractions of bovine adrenal medulla were 5200+/-35, 5000+/-280 and 1700+/-260 mU/g tissue, respectively. Cultured chromaffin cells displayed a total hydrolysing activity of 41+/-5 mU/10(6) cells. Homogenates of bovine adrenal medulla displayed only about 6% of activity sensitive to paraoxon. Most of the phenyl valerate esterase activity inhibited by mipafox (a neuropathy inducing compound) was found in particulate fraction. Cultured chromaffin cells displayed kinetics of inhibition by mipafox similar to the kinetics displayed by homogenates of bovine adrenal medulla. We conclude that NTE could be assayed in this system by only using one inhibitor (mipafox) instead of two (paraoxon and mipafox). Also, the proposal is supported of using chromaffin cells as in vitro model for the study of the role of NTE and related esterases in organophosphorus-induced polyneuropathy.


Subject(s)
Adrenal Medulla/drug effects , Chromaffin Cells/drug effects , Isoflurophate/analogs & derivatives , Isoflurophate/toxicity , Organophosphorus Compounds/toxicity , Paraoxon/toxicity , Adrenal Medulla/enzymology , Adrenal Medulla/metabolism , Animals , Carboxylic Ester Hydrolases/antagonists & inhibitors , Carboxylic Ester Hydrolases/metabolism , Cattle , Chromaffin Cells/enzymology , Chromaffin Cells/metabolism , Enzyme Inhibitors/pharmacokinetics , Isoflurophate/pharmacokinetics , Neurotoxicity Syndromes/etiology , Organophosphorus Compounds/pharmacokinetics , Paraoxon/pharmacokinetics , Toxicity Tests/methods
6.
J Anal Toxicol ; 27(5): 290-3, 2003.
Article in English | MEDLINE | ID: mdl-12908942

ABSTRACT

This work was performed to adapt the manual laboratory method of measuring serum paraoxonase activity using a routine automatized method in the clinical laboratory and to study the distribution of paraoxonase activity in a large population from Alcoy, a region of Spain. The serum samples for the study were obtained from extractions of blood from 2891 individuals, distributed by sex and age groups, in a routine check in a primary care facility of Alcoy. Paraoxonase activity was assayed by measuring the release of p-nitrophenol according to a previously published method adapted to an automatized analyzer. The mean paraoxonase activity recorder was 70.2 +/- 16.5 IU/L. Paraoxonase activity in children (both males and females) was significantly lower (p < 0.0005) than in older individuals. Paraoxonase activity detected in males and females older than 56 was slightly lower than that detected in younger individuals, although in this case the difference was not statistically significant. The paraoxonase activity shows higher mean values in females than in males (p < 0.0005). Human paraoxonase activity shows a unimodal distribution pattern in the studied population, which is in contrast with other studies showing bimodal distribution.


Subject(s)
Aryldialkylphosphatase/blood , Insecticides/blood , Paraoxon/blood , Adolescent , Adult , Autoanalysis , Female , Humans , Hydrolysis , Male , Middle Aged , Nitrophenols/metabolism , Spain
7.
J Biol Chem ; 274(51): 36656-62, 1999 Dec 17.
Article in English | MEDLINE | ID: mdl-10593969

ABSTRACT

Ras proteins are small GTPases playing a pivotal role in cell proliferation and differentiation. Their activation depends on the competing action of GTPase activating proteins and guanine nucleotide exchange factors (GEF). The properties of two dominant-negative mutants within the catalytic domains of the ras-specific GEF, CDC25(Mm), are described. In vitro, the mutant GEF(W1056E) and GEF(T1184E) proteins are catalytically inactive, are able to efficiently displace wild-type GEF from p21(ras), and strongly reduce affinity of the nucleotide-free ras x GEF complex for the incoming nucleotide, thus resulting in the formation of a stable ras.GEF binary complex. Consistent with their in vitro properties, the two mutant GEFs bring about a dramatic reduction in ras-dependent fos-luciferase activity in mouse fibroblasts. The stable ectopic expression of the GEF(W1056E) mutant in smooth muscle cells effectively reduced growth rate and DNA synthesis with no detectable morphological changes.


Subject(s)
Muscle, Smooth, Vascular/physiology , Mutation , Signal Transduction , ras-GRF1/genetics , Animals , Cell Line , Mice , Rats , Signal Transduction/genetics , ras-GRF1/metabolism
8.
Appl Environ Microbiol ; 65(9): 4211-5, 1999 Sep.
Article in English | MEDLINE | ID: mdl-10473436

ABSTRACT

Interest in the production of L-(+)-lactic acid is presently growing in relation to its applications in the synthesis of biodegradable polymer materials. With the aim of obtaining efficient production and high productivity, we introduced the bovine L-lactate dehydrogenase gene (LDH) into a wild-type Kluyveromyces lactis yeast strain. The observed lactic acid production was not satisfactory due to the continued coproduction of ethanol. A further restructuring of the cellular metabolism was obtained by introducing the LDH gene into a K. lactis strain in which the unique pyruvate decarboxylase gene had been deleted. With this modified strain, in which lactic fermentation substituted completely for the pathway leading to the production of ethanol, we obtained concentrations, productivities, and yields of lactic acid as high as 109 g liter(-1), 0.91 g liter(-1) h(-1), and 1.19 mol per mole of glucose consumed, respectively. The organic acid was also produced at pH levels lower than those usual for bacterial processes.


Subject(s)
Genetic Engineering , Kluyveromyces/enzymology , Kluyveromyces/genetics , L-Lactate Dehydrogenase/genetics , Lactic Acid/metabolism , Animals , Bioreactors , Cattle , Hydrogen-Ion Concentration , L-Lactate Dehydrogenase/metabolism , Plasmids , Pyruvate Decarboxylase/genetics , Pyruvate Decarboxylase/metabolism , Transformation, Bacterial
9.
FEMS Microbiol Lett ; 171(2): 133-40, 1999 Feb 15.
Article in English | MEDLINE | ID: mdl-10077837

ABSTRACT

Introduction of the Lactobacillus casei lactate dehydrogenase (LDH) gene into Saccharomyces cerevisiae under the control of the TPI1 promoter yielded high LDH levels in batch and chemostat cultures. LDH expression did not affect the dilution rate above which respiro-fermentative metabolism occurred (Dc) in aerobic, glucose-limited chemostats. Above Dc, the LDH-expressing strain produced both ethanol and lactate, but its overall fermentation rate was the same as in wild-type cultures. Exposure of respiring, LDH-expressing cultures to glucose excess triggered simultaneous ethanol and lactate production. However, the specific glucose consumption rate was not affected, indicating that NADH reoxidation does not control glycolytic flux under these conditions.


Subject(s)
Glucose/pharmacology , L-Lactate Dehydrogenase/genetics , NAD/metabolism , Saccharomyces cerevisiae/enzymology , Saccharomyces cerevisiae/genetics , Aerobiosis , Cloning, Molecular , Fermentation/drug effects , Gene Expression Regulation, Bacterial/drug effects , Gene Expression Regulation, Fungal/drug effects , Genes, Bacterial/physiology , Glycogen/metabolism , L-Lactate Dehydrogenase/metabolism , Lacticaseibacillus casei/genetics , Oxidation-Reduction , Saccharomyces cerevisiae/growth & development
10.
FEBS Lett ; 440(3): 291-6, 1998 Dec 04.
Article in English | MEDLINE | ID: mdl-9872389

ABSTRACT

The role of two residues within the catalytic domain of CDC25Mm, a mouse ras-specific guanine nucleotide exchange factor (GEF), was investigated by site-directed mutagenesis. The function of the mutant proteins was tested in vivo in both a Saccharomyces cerevisiae cdc25 complementation assay and in a mammalian fos-luciferase assay, and in in vitro assays on human and yeast Ras proteins. Mutants CDC25Mm(E1048K) and CDC25Mm(S1122V) were shown to be (partly) inactive proteins, similar to their yeast homologs. Mutant CDC25Mm(S1122A) showed higher nucleotide exchange activity than the wild type protein on the basis of both in vitro and in vivo assays. Thus, alanine and valine substitutions at position 1122 within the GEF catalytic domain originate mutations with opposite biological properties, indicating an important role for position 1122 in GEF function.


Subject(s)
Cell Cycle Proteins/metabolism , Phosphoprotein Phosphatases/metabolism , 3T3 Cells , Animals , CHO Cells , Catalytic Domain , Cell Cycle Proteins/genetics , Cricetinae , Genes, fos , Guanine Nucleotides/metabolism , Mice , Mutagenesis, Site-Directed , Phosphoprotein Phosphatases/genetics , Saccharomyces cerevisiae , Transfection , ras Proteins/metabolism , ras-GRF1
11.
Toxicol Lett ; 71(1): 47-51, 1994 Mar.
Article in English | MEDLINE | ID: mdl-8140588

ABSTRACT

Neuropathy target esterase (NTE) is a protein suggested to be involved in the initiation mechanism of organophosphorus-induced delayed neuropathy (OPIDP). We previously described two different forms of NTE activity in hen sciatic nerve: a particulate form (P-NTE) representing 40-50% of total NTE activity in sciatic nerve, and a remaining soluble component (S-NTE). In brain tissue on the other hand, more than 90% of NTE activity was recovered as P-NTE. In this work we studied the in vivo inhibition of both NTE forms with different doses of mipafox and the results were compared with sensitivity to mipafox in vitro. The highest dose with no observable neuropathic effects (1.5 mg/kg mipafox p.o.) inhibited 33% P-NTE and 55% S-NTE activity. The difference between P-NTE and S-NTE activity was statistically significant (P < 0.001, n = 9). Higher doses (3 mg/kg) induced neuropathy and inhibited NTE more than 75%, but differences between P- and S-NTE were not significant (P > 0.5). The greater inhibition of S-NTE than P-NTE in vivo contrasts with the observation that S-NTE is less sensitive in vitro.


Subject(s)
Carboxylic Ester Hydrolases/antagonists & inhibitors , Isoflurophate/analogs & derivatives , Sciatic Nerve/enzymology , Animals , Ataxia/chemically induced , Chickens , Female , Isoflurophate/administration & dosage , Isoflurophate/toxicity , Reflex/drug effects , Sciatic Nerve/drug effects , Solubility
12.
Exp Cell Res ; 210(2): 353-7, 1994 Feb.
Article in English | MEDLINE | ID: mdl-8299730

ABSTRACT

The CDC25Mm gene codes for Ras-guanine nucleotide exchange factors. Four different full-length cDNA clones derived from the same gene and coding for proteins of different sizes that have in common the last 661 amino acids have been isolated from mouse brain. In order to investigate the expression of the products of this gene in different tissues we have prepared two polyclonal antibodies directed toward two different regions of the protein comprised in the last C-terminal 472 amino acids. While in most of the tested tissues we have been unable to definitely identify CDC25Mm products, in NIH3T3 fibroblasts we have found a poorly expressed 120-kDa protein. In the mouse brain we have identified two proteins of 140 and 58 kDa. While the former is expressed in the adult mouse, the latter is present in the embryo and persists for few days after birth. This finding suggests that differential expression of various forms of CDC25Mm may be involved in brain development.


Subject(s)
Aging/metabolism , Brain/metabolism , Protein Biosynthesis , 3T3 Cells , Animals , Blotting, Western , Brain/growth & development , Cell Line, Transformed , Embryo, Mammalian , Embryonic and Fetal Development/physiology , Gene Expression Regulation , Guanine Nucleotide Exchange Factors , Mice , Molecular Weight , Proteins/analysis , Proteins/genetics , ras Guanine Nucleotide Exchange Factors , ras-GRF1
13.
Chem Biol Interact ; 87(1-3): 369-81, 1993 Jun.
Article in English | MEDLINE | ID: mdl-8343994

ABSTRACT

NTE (neuropathy target esterase) is considered to be the target for organophosphorus-induced delayed polyneuropathy and is operationally measured by radiolabelling or by determining its esteratic activity as the paraoxon-resistant mipafox-sensitive phosphorylable site(s). From electrophoresis and density gradient centrifugation using radiolabelling techniques, several phosphorylable sites have been described in hen brain that are paraoxon-resistant mipafox-sensitive; however, only the majority electrophoresis band (155 kDa) shows properties related with the aging reaction. Kinetic criteria have also suggested two components of brain NTE (NTEA and NTEB). Most brain NTE is recovered in the particulate microsomal fraction and only about 1% in soluble fraction. In sciatic nerve about 50%/50% activity is recovered as soluble (S-NTE) or particulate (P-NTE) forms. A similar distribution were observed in hen, cat, rat and young chick. The fixed time inhibition curves show that P-NTE is more sensitive to mipafox, DFP and hexyl-DCP than S-NTE, while the reverse is true for methamidophos. P-NTE fits properly to one sensitive component while S-NTE fits better to two sensitive component models, except in the case of methamidophos. In vivo, significant differences in the inhibition of P- and S-NTE by mipafox were found only when using low non-neuropathic dosing. The possible significance of different NTE forms are discussed.


Subject(s)
Carboxylic Ester Hydrolases/metabolism , Isoenzymes/metabolism , Aging/metabolism , Animals , Brain/enzymology , Carboxylic Ester Hydrolases/antagonists & inhibitors , Carboxylic Ester Hydrolases/physiology , Chickens , Female , Humans , Isoenzymes/antagonists & inhibitors , Isoenzymes/physiology , Peripheral Nerves/enzymology
14.
Chem Biol Interact ; 87(1-3): 417-23, 1993 Jun.
Article in English | MEDLINE | ID: mdl-8343997

ABSTRACT

NTE inhibitors cause different toxicological consequences (protection, induction or potentiation/promotion of neuropathy) depending on the order of dosing. These effects might be explained in terms of several phosphorylable sites with 'allosteric irreversible' behaviour. Brain neuropathy target esterase (NTE) has been preinhibited with phenylmethylsulphonyl fluoride (PMSF) (0, 5, 10, 15, 30 and 60 microM) or with diisopropylphoshoro fluoridate (DFP) (0, 0.2, 0.5, and 1 microM) at 37 degrees C for 30 min. After washing by centrifugation, tissues were then reinhibited with a range of PMSF (0 to 80 microM) or DFP (0 to 1 microM) concentrations. The slopes of the inhibition curves (log % activity vs. concentration) of pretreated tissues were identical to those of the non-pretreated tissues, with non-distinguishable I50 values. It is concluded that allosteric effects are not likely to be involved in membrane-bound NTE of hen brain.


Subject(s)
Brain/enzymology , Carboxylic Ester Hydrolases/antagonists & inhibitors , Animals , Binding Sites , Carboxylic Ester Hydrolases/metabolism , Chickens , Female , Isoflurophate/metabolism , Isoflurophate/pharmacology , Kinetics , Phenylmethylsulfonyl Fluoride/metabolism , Phenylmethylsulfonyl Fluoride/pharmacology , Sensitivity and Specificity
15.
Arch Toxicol ; 67(6): 416-21, 1993.
Article in English | MEDLINE | ID: mdl-8215911

ABSTRACT

One of the main detoxification mechanisms of organophosphorus (OP) compounds is hydrolysis by OP hydrolysing enzymes (OP-hydrolases) or phosphoric triester hydrolases. We previously reported an OP-hydrolase from hen plasma which hydrolyses O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP). In this study, a total of 18 cations, as well as several thiol blocking reagents, ethylenediaminetetraacetic acid (EDTA) and mipafox (N,N'-diisopropyl phosphorodiamidofluoridate) were assayed as activators or inhibitors of the HDCP hydrolysing activity of hen plasma in vitro. Of the 18 inorganic cations only 1 M Na+ caused any inhibition. Most of the cations, including Ca2+, exerted no detectable effect; however, 1 mM Cu2+ was found to produce an activation of up to 263%, with a lesser activation of up to 168% for 1 mM Zn2+. The thiol blocking reagents methyl vinyl ketone (MVK) and N-ethylmaleimide (NEM) inhibited the enzyme in a time-dependent manner, the maximum effect depending upon concentration in the case of NEM, but not in the case of MVK; however, 5,5'-dithiobis (2-nitrobenzoic acid) caused inhibition that was concentration dependent but which was independent of time. Other thiol blocking reagents such as p-hydroxymercuribenzoic acid (sodium salt), phenylmercuric acetate, iodoacetic acid (sodium salt) and iodoacetamide produced only slight inhibition, as did EDTA. Finally, the OP compound mipafox exerted no detectable effect.


Subject(s)
Hydrolases/antagonists & inhibitors , Hydrolases/blood , Metals/pharmacology , Animals , Aryldialkylphosphatase , Butanones/pharmacology , Calorimetry , Cations/pharmacology , Chickens , Copper/pharmacology , Dithionitrobenzoic Acid/pharmacology , Edetic Acid/pharmacology , Enzyme Activation , Ethylmaleimide/pharmacology , Female , Hydrolysis , Kinetics , Organophosphorus Compounds/blood , Phosphoric Monoester Hydrolases/blood , Zinc/pharmacology
16.
Toxicol Appl Pharmacol ; 117(2): 218-25, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1471154

ABSTRACT

Diisopropyl phosphorofluoridate (DFP), mipafox, cresylsaligenyl phosphate, and phenylsaligenyl phosphate were applied to a 1.5-cm segment of the common trunk of the sciatic nerve in adult hens. At doses of 18-182 micrograms mipafox and 9-110 micrograms DFP, inhibition of neuropathy target esterase (NTE) for the treated segment was over 80%, whereas for the adjacent distal and proximal segments inhibition was under 40%, 15 min after application. NTE was not affected in the peripheral distal terminations arising from the common sciatic nerve (peroneal branches), contralateral sciatic nerve, brain, and spinal cord. A 24-hr study suggested a displacement of the activity-free region toward more distal segments of the nerve. All animals treated with 55 and 110 micrograms DFP or 110 micrograms mipafox lost a characteristic avian retraction reflex in the treated leg 9-15 days after dosing, suggesting peripheral neurological alterations. Only hens dosed at the maximum dose in both extremities presented alterations in motility (Grade 1 or 2 on a 0-8 scale), suggesting no significant central nervous system alterations. Electron microscopy of peroneal branches showed axon swelling and accumulation of smooth endoplasmic reticulum similar to animals dosed systemically (s.c.) with 1-2 mg/kg DFP. The branches also contained granular and electron-dense materials, as well as some intraaxonal and intramyelinic vacuolization. Clinical effects were not observed in animals protected with a 30 mg/kg (s.c.) dose of phenylmethanesulphonyl fluoride. It is concluded that the peripheral neurological effects of local dosing correlate with the specific modification of NTE in a segment of sciatic nerve and that the axon is a more likely target than the perikaryon or nerve terminal in the triggering mechanism of this axonopathy.


Subject(s)
Carboxylic Ester Hydrolases/antagonists & inhibitors , Isoflurophate/analogs & derivatives , Isoflurophate/pharmacology , Peripheral Nerves/drug effects , Sciatic Nerve/drug effects , Animals , Chickens , Female , Locomotion/drug effects , Peroneal Nerve/drug effects , Sciatic Nerve/physiology , Sciatic Nerve/ultrastructure
17.
Aten Primaria ; 9(9): 483-6, 1992 Jun 01.
Article in Spanish | MEDLINE | ID: mdl-1525309

ABSTRACT

Evaluate the assistencial drug-dependence program. DESIGN. Transversal retrospective study. Primary care (Unitat de Prevenció i Seguiment de Drogodependéncies). PATIENTS OR OTHER PARTICIPANTS. Sixty patients treated from 23.02.90 to 31.06.91. MEASUREMENTS AND MAIN RESULTS. It was made 165 interviews. Sixty patients were treated: 2 (3.3%) with methadone substitution and 58 (96.6%) started the "drug free program". In this program, 7 (12%) patients accomplished hospital desintoxication and 51 (88%) home desintoxication. At the end of the period control, 48 patients were in "drug free program". At the beginning of treatment 32 (53.3%) patients were 22-27 years old. The most usual illegal drugs at consultation were: cannabis 57 (95%), cocaine 53 (88.3%), heroine 51 (85%) and benzodiazepines 50 (83.3%). Twenty-five patients (41.6%) needed derivation to another health services. From 51 patients addicted to intravenous heroin, only ten (19.6%) were positive to HIV antibodies. CONCLUSIONS. The increase of patients number who started and remained the "drug free program" and the low percent of patients who stopped treatment show that the assistant program in primary care is working. This program has been appropriate to know the characteristics of this group in our area: low age at initiation, progressive use of more than one drug and low prevalence of HIV positivity.


Subject(s)
Primary Health Care , Substance-Related Disorders/prevention & control , Adolescent , Adult , Humans , Program Evaluation
18.
J Neurochem ; 55(4): 1258-65, 1990 Oct.
Article in English | MEDLINE | ID: mdl-2398358

ABSTRACT

Neuropathy target esterase (NTE) is the suggested "target" molecule involved in the initiation of organophosphorus-induced delayed polyneuropathy. Sciatic nerve NTE was separated into particulate (P-NTE) and soluble (S-NTE) fractions by ultracentrifugation at 100,000 g for 1 h in 0.32 M sucrose and compared with the corresponding brain extract. Total sciatic NTE activity was 80-100 nmol/min/g tissue from which 50-60% was recovered in the soluble supernatant fraction and the remaining 40-50% in the pellet fraction. About 90% of brain tissue activity (approximately 1,800 nmol/min/g tissue) was recovered as P-NTE. A similar distribution was obtained when more drastic centrifugation without sucrose was performed. P-NTE and S-NTE were distributed with the membrane and cytosolic markers assayed, respectively, glucose-6-phosphatase, Na+,K(+)-ATPase, 5'-nucleotidase, phospholipids, and lactate dehydrogenase. When the pH during the centrifugation was increased from 6.4 to 11, recovered P-NTE activity decreased from 1,750 to 118 nmol/min/g tissue for brain and from 31 to 12 nmol/min/g for sciatic nerve. However, S-NTE activity and total nonfractionated control activity were only slightly affected by the same pH treatment. The distribution pattern encountered may be better understood as representing two different proteins than an equilibrium between soluble and membrane-bound portions of a single protein, with P-NTE activity depending on a membrane factor from which it is separated through fractionation at high pH.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Brain/enzymology , Carboxylic Ester Hydrolases/metabolism , Sciatic Nerve/enzymology , Animals , Carboxylic Ester Hydrolases/isolation & purification , Cell Fractionation/methods , Chickens , Cytosol/enzymology , Female , Hydrogen-Ion Concentration , Kinetics , L-Lactate Dehydrogenase/metabolism , Phospholipids/analysis , Ultracentrifugation/methods
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