Gelatinasas como marcadores de consumo crónico de alcohol: un estudio piloto en Uruguay / Gelatinases as markers of chronic alcohol consumption: a pilot study in Uruguay / Gelatinasas como marcadores de consumo crônico de álcool: um estúdio piloto em Uruguai

El consumo crónico de alcohol en Uruguay es un problema creciente, sin embargo, las determinaciones de biomarcadores consensuados no se realizan sistemáticamente ni se investigan otros marcadores potenciales. Para validar la hipótesis de que las metaloproteinasas de matriz con actividad gelatinasa son biomarcadores de consumo crónico de alcohol, se evaluaron muestras de sangre de 100 alcohólicos que comenzaron a atenderse en la Unidad de Trastornos Relacionados con el Alcohol y de 50 donantes sanos no alcohólicos. Las muestras de alcohólicos presentaron actividad de gelatinasas que triplicaron la de los controles y aumentos pequeños pero significativos en los niveles de γ-glutamil transferasa, aspartato-aminotransferasa y volumen corpuscular medio. Los valores de transferrina deficiente en carbohidratos fueron menores en alcohólicos que en controles. Estos resultados permiten proponer a las gelatinasas como los indicadores más sensibles del consumo sostenido de alcohol en la población analizada, ya que las enzimas hepáticas y el volumen corpuscular medio muestran una tendencia acorde con la literatura pero no alcanzaron valores asociados a la patología. Dado que la transferrina deficiente en carbohidratos es considerada el biomarcador indirecto más sensible y específico de consumo crónico de alcohol, los valores menores obtenidos en alcohólicos respecto de controles sugieren problemas metodológicos que podrían subsanarse aplicando otras técnicas de medida o por la presencia de interferencias que deben ser identificadas. Finalmente, estos hallazgos justifican una extensión de este trabajo piloto, así como estudios adicionales centrados en la participación de las metaloproteinasas de matriz con actividad gelatinasa en las cascadas de daño asociadas al consumo crónico de alcohol.

Chronic alcohol consumption in Uruguay is a growing problem, however, determinations of consensual biomarker are not performed systematically neither potential markers are explored. To validate the hypothesis that matrix metalloproteinases with gelatinase activity are biomarkers of chronic alcohol consumption, blood samples of 100 alcoholics that began medical treatment at the Unidad de Trastornos Relacionados con el Alcohol and 50 healthy non-alcoholic donors were evaluated. Alcoholic samples showed gelatinase activity that tripled that of controls and small but significant increases in levels of γ-glutamyl transferase, aspartate-aminotransferase and mean cellular volume. Carbohydrate deficient transferrin values were lower in alcoholics than in controls. These results allow proposing gelatinases as the most sensitive indicators of sustained alcohol consumption in the population analyzed since hepatic enzymes and mean cellular volume showed a tendency consistent with the literature but did not reach values associated with the pathology. Since carbohydrate-deficient transferrin is considered the most sensitive and specific indirect biomarker of chronic alcohol consumption, lower values in alcoholics related to controls suggest methodological problems that could be solved by applying other measurement techniques or by the presence of yet unknown interferences. Finally, these findings justify an extension of this pilot work, as well as additional studies focused on the participation of matrix metalloproteinases with gelatinase activity in the cascades of damage associated with chronic alcohol consumption.

O consumo crônico de álcool no Uruguai é um problema crescente, no entanto, as determinações consensuais de biomarcadores não são realizadas sistematicamente ou os potenciais marcadores são explorados. Para validar a hipótese de que as metaloproteinases de matriz com atividade gelatinase são biomarcadores do consumo crônico de álcool, foram avaliadas amostras de sangue cd 100 alcoólatras que começaram a ser tratadas na Unidad de Trastornos Relacionados con el Alcohol e 50 doadores não-alcoólatras saudáveis. As amostras alcoólicas apresentaram atividade de gelatinase que triplicou a dos controles e pequenos más significativos aumentos nos níveis de γ-glutamil transferase, aspartato-aminotransferase e volume médio celular. Os valores de transferrina deficientes em carboidratos foram menores nos alcoolistas que nos controles. Esses resultados permitem que as gelatinases sejam propostas como os indicadores mais sensíveis do consumo sustentado de álcool na população analisada, uma vez que as enzimas hepáticas e o volume celular médio apresentam uma tendência consistente com a literatura, mas não alcançaram valores associados à patologia. Como a transferrina deficiente em carboidratos é considerada o biomarcador indireto mais sensível e específico do consumo crônico de álcool, os valores mais baixos em alcoólatras do que em controles sugerem problemas metodológicos que poderiam ser sanados pela aplicação de outras técnicas de mensuração pela presença de interferências que deben ser identificadas. Finalmente, esses achados justificam uma extensão deste trabalho piloto, bem como estudos adicionais voltados para a participação de metaloproteinases de matriz com atividade de gelatinase nas cascatas de danos associados ao consumo crônico de álcool.

Análisis de la interacción entre el polimorfismo Rs671 del gen ALDH2 y consumo de alcohol en individuos chilenos / Interaction between Rs671 polymorphism of the ALDH2 gene and alcohol consumption in Chilean individuals

El alcoholismo es un importante problema de salud pública. En los últimos años ha causado interés el metabolismo del alcohol, puesto que ha sido considerado un posible determinante biológico en la conducta de consumo. Variados estudios se han orientado a la búsqueda y comprensión de la influencia de polimorfismos, en genes que codifican para los principales sistemas enzimáticos que intervienen en el metabolismo hepático. El polimorfismo rs671 del gen que codifica la enzima ALDH2 ha sido asociado a un menor consumo de alcohol debido a la acumulación de acetaldehído en sangre. Diversos estudios indican que este polimorfismo es frecuente en países asiáticos y se considera un factor protector en los individuos que lo portan. Se incluyeron 207 individuos adultos no relacionados, a los cuales se les aplicó un cuestionario sobre consumo de alcohol. El polimorfismo rs671 fue analizado por la reacción de la polimerasa en cadena (PCR) seguida de restricción enzimática. Además, se determinaron los biomarcadores clásicos indirectos de consumo de alcohol, mediante técnicas enzimáticas y hematológicas. La frecuencia del genotipo homocigoto mutado AA para el polimorfismo rs671 fue 3,0% en sujetos consumidores de alcohol y 2,8% en el grupo no consumidor. La distribución de genotipos y las frecuencias alélicas para esta variante fueron semejantes entre los sujetos estudiados (p>0,05). Estos hallazgos sugieren que la variante rs671 del gen ALDH2 no está asociada al oconsumo de alcohol en los individuos estudiados.

Alcoholism is an important public health problem. In recent years, alcohol metabolism caused interest, since it has been considered a possible biological determinant of alcohol consumption behavior. Several studies have focused on finding and understanding the influence of polymorphisms affecting genes that encode for enzymatic systems involved in the hepatic metabolism. The rs671 polymorphism of the gene encoding ALDH2 has been associated with lower alcohol consumption by leading to acetaldehyde accumulation in blood. This genetic variant is frequently found in Asian population and has been considered as protector factor of alcoholism in these individuals. In the present study, 207 unrelated-adult individuals were included. Alcohol consumption was recorded using a structured questionnaire. The rs671 polymorphism was analyzed using polymerase chain reaction followed by enzymatic digestion. Furthermore, classical biomarkers for alcohol consumption were assessed using enzymatic and hematological techniques. The frequency of homozygote genotype for the A allele (AA) was 3 and 2.8% in those subjects defined as alcohol drinkers and non-alcohol drinkers respectively. The genotypes distribution and allelic frequencies were similar among the studied subject (p>0.05). These data suggest that rs671 ALDH2 gene polymorphism is not associated to alcohol consumption in the studied population.

Genetic polymorphism of alcohol-metabolizing enzyme and alcohol dependence in Polish men

Alcohol dependence poses a serious medical and sociological problem. It is influenced by multiple environmental and genetic factors, which may determine differences in alcohol metabolism. Genetic polymorphism of the enzymes involved in alcohol metabolism is highly ethnically and race dependent. The purpose of this study was to investigate the differences, if present, in the allele and genotype frequency of alcohol dehydrogenase 1B (ADH1B), ADH1C and the microsomal ethanol-oxidizing system (MEOS/CYP2E1) between alcohol-dependent individuals and controls and also to determine if these genotypes cause a difference in the age at which the patients become alcohol dependent. The allele and genotype frequencies of ADH1B, ADH1C, and CYP2E1 were determined in 204 alcohol dependent men and 172 healthy volunteers who do not drink alcohol (control group). Genotyping was performed by PCR-RFLP methods on white cell DNA. ADH1B*1 (99.3 percent) and ADH1C*1 (62.5 percent) alleles and ADH1B*1/*1 (N = 201) and ADH1C*1/*1 (N = 85) genotypes were statistically more frequent among alcohol-dependent subjects than among controls (99.3 and 62.5 percent, N = 201 and 85 vs 94.5 and 40.7 percent, N = 153 and 32, respectively). Differences in the CYP2E1 allele and genotype distribution between groups were not significant. The persons with ADH1C*1/*1 and CYP2E1*c1/*c2 genotypes became alcohol dependent at a considerably younger age than the subjects with ADH1C*1/*2, ADH1C*2/*2 and CYP2E1*c1/*c1 genotypes (28.08, 25.67 years vs 36.0, 45.05, 34.45 years, respectively). In the Polish men examined, ADH1C*1 and ADH1B*1 alleles and ADH1C*1/*1 and ADH1B*1/*1 genotypes favor alcohol dependence. The ADH1B*2 allele may protect from alcohol dependence. However, subjects with ADH1C*1/*1 and CYP2E1*c1/*c2 genotypes become alcohol dependent at a considerably younger age than the subjects with ADH1C*1/*2, ADH1C*2/*2 and CYP2E1*c1/*c1 genotypes.

Effects of Genetic Polymorphisms of Ethanol-Metabolizing Enzymes on Alcohol Drinking Behaviors / 대한간학회지

BACKGROUND/AIMS: Genetic variations of ethanol-metabolizing enzymes can affect alcohol drinking behavior. The aims of this study were to investigate and compare the distributions of these genetic polymorphisms between a healthy control group and a heavy drinker group which included an alcoholic liver cirrhosis group. METHODS: Genotypes of ADH2, ALDH2, CYP2E1, and catalase were identified by polymerase chain reaction and restriction fragment length polymorphism. Genomic DNA was extracted from peripheral leukocytes in 42 healthy controls, 12 heavy drinkers, and 30 alcoholic liver cirrhosis patients. RESULTS: 1) The genotype frequencies of ALDH2 (1*1), ADH2 (1*1), CYP2E1 (c1c1), and catalase1 (TT) were 69%, 55%, 38%, and 12%, respectively in healthy Korean males. 2) There was a significant difference in the distribution of the genetic polymorphism of ALDH2 between the control group and heavy drinker group (12 heavy drinkers and 30 alcoholic liver cirrhosis patients). The genotype frequency of ALDH2 mutant, ALDH2 (1*2) and ALDH2 (2*2) in the heavy drinker group (12%) was significantly lower than that in the control group (30%). 3) We didn't find anyone with ALDH2 homozygote mutant (DD) in the heavy drinker group. 4) There was no significant difference in the distribution of genetic polymorphisms in ADH2, CYP2E1 and catalase1 between the two groups. CONCLUSIONS: These results suggest that the absence of ALDH2 mutant genotype is strongly related to heavy drinking behavior. We can not prove, however, any evidence that the polymorphisms of other ethanol-metabolizing enzymes are associated with the determination of alcohol-drinking behavior.

Association Between Polymorphisms of Ethanol-Metabolizing Enzymes and Susceptibility to Alcoholic Cirrhosis in a Korean Male Population

Alcohol is oxidized to acetaldehyde by alcohol dehydrogenase (ADH) and cytochrome P-4502E1 (CYP2E1), and then to acetate by aldehyde dehydrogenase (ALDH). Polymorphisms of these ethanol-metabolizing enzymes may be associated with inter-individual difference in alcohol metabolism and susceptibility to alcoholic liver disease. We determined genotype and allele frequencies of ALDH2, CYP2E1, ADH2, and ADH3 in male Korean patients with alcoholic cirrhosis (n=56), alcoholics without evidence of liver disease (n=52), and nondrinkers (n=64) by using PCR or PCR-directed mutagenesis followed by restriction enzyme digestion. The prevalences of heterozygous ALDH2*1/*2 plus homozygous ALDH2*2/*2 in patients with alcoholic cirrhosis (7.1%) and alcoholics without evidence of liver disease (3.8%) were significantly lower than that in nondrinkers (45.3%). The c2 allele frequencies of the CYP2E1 in alcoholic cirrhosis, alcoholics without evidence of liver disease, and nondrinkers were 0.21, 0.20, and 0.20, respectively. Allele frequencies of ADH2*2 in the three groups were 0.78, 0.74, and 0.77 and those of ADH3*1 were 0.94, 0.98, and 0.95. Therefore, we confirmed the observation that the ALDH2*2 gene protects against the development of alcoholism. However, the development of cirrhosis in Korean alcoholic patients was not associated with polymorphisms of ethanol-metabolizing enzymes.

Some biochemical changes in heme synthesis in iron deficiency.

Some enzymes and intermediates of heme synthesis were determined in blood and urine of 26 women with severe iron deficiency anemia (IDA). Erythrocyte free protoporphyrin was almost doubled and delta-aminolevulinate dehydrase significantly raised. But urinary excretion of delta-aminolevulinic acid and reticulocyte ferrochelatase were significantly reduced in iron deficiency anemia. Hence these could serve as useful indices of iron deficiency and consequent anemia.

Effect of alcohol dependence on the levels of duodenal disaccharidases in human subjects.

BACKGROUND: The aim of the study was to detect the duodenal enzyme activity in patients of alcohol dependence and to compare with non-alcoholic patients of non-ulcer dyspepsia. METHODS: Disaccharidases (lactase, sucrase, maltase) were estimated in 20 non alcoholic patients of non-ulcer dyspepsia and 20 alcoholics admitted to the drug de-addiction and treatment centre of PGIMER, Chandigarh, India. RESULTS: No significant influence of alcohol on enzyme levels in patients of alcohol dependence when compared to patients of non-ulcer dyspepsia was observed. However, a significant decrease in lactase level was noted in patients consuming more than 125 gm/day of alcohol. CONCLUSION: Amount of consumption of alcohol showed decrease in lactase enzyme, but not in maltase and sucrase. There was no effect of duration of alcohol consumption on dissacharidases in the two groups.

Indicators of inflammation and cellular damage in chronic asymptomatic or oligosymptomatic alcoholics: correlation with alteration of bilirubin and hepatic and pancreatic enzymes

Indicadores bioquimicos e hematimetricos de inflamacao e lesao celular foram correlacionados com bilirrubina e enzimas hepaticas e pancreatica em 30 alcoolistas cronicos do sexo masculino internados em hospital psiquiatrico para desintoxicacao e tratamento do alcoolismo. A aspartato aminotransferase, alanino aminotransferase, gamaglumiltransferase, fosfatase alcalina e bilirrubina total estavam alteradas em 90(por cento), 63(por cento), 87(por cento), 23(por cento) e 23(por cento) dos casos, respectivamente. Entre os indicadores de inflamacao (desidrogenase latica, alterada em 16 dos casos; alfa-1 globulina, 24(por cento), alfa-2 globulina, 88(por cento); contagem de leucocitos, 28(por cento) nenhum estava correlacionado com as alteracoes da bilirrubina e enzimas hepaticas...

Effect of chronic ethanol feeding on intestinal alkaline phosphatase activity in rats.

The effect of chronic ethanol feeding has been studied on intestinal alkaline phosphatase (IAP) activity. The enzyme was assayed using p-nitrophenyl phosphate (PNPP), phenylphosphate (PhP) and beta-glycerophosphate (beta GP) as the substrates. Feeding of ethanol for 10 days did not effect the enzyme activity but it was markedly elevated in animals fed ethanol for 20 or 30 days. However, ethanol administration to rats, for over 6 wk exhibited a decline in IAP activity, both in the soluble and membrane fractions of intestine. Kinetic studies revealed an enhancement in Vmax with no change in apparent Km after 30 days of ethanol feeding and a decrease in Vmax after 6 wk of ethanol ingestion. These results were confirmed by assaying the enzyme activity in non-denatured polyacrylamide gels using 5-bromo-4-chloro-3-indoly1 phosphate (BCIP) as the substrate. These findings suggest differential changes in IAP activity in response to ethanol feeding in rats.

Modificaciones de los niveles de acido delta aminolevulico dehidrasa en pacientes alcoholicos cronicos / Changes in the levels of delta-aminolevulinic acid dehydratase in chronic alcoholic patients

Estudios clínicos y bioquímicos fueron realizados sobre 58 pacientes con alcoholismo crónico a su vez divididos en 3 grupos: A (hepatopatia alcohólica), B (hepatopatía y neuropatía alcohólica) y C (neuropatía alcohólica). Simultáneamente se investigaron varios parámetros relacionados con la biosíntesis del hemo. Se comprobó que el ácido delta-aminolevúlico (ALA), porfobilinógeno (PBG) urinarios y las porfirinas urinarias y fecales, no presentaron diferencias significativas entre los grupos estudiados. También se determinaron en eritrocitos las actividades de ALA-dehidrasa (ALA-D), uroporfirinógeno-I-sintetasa y uroporfirinógeno-III-sintetasa. De todos los datos obtenidos se comprobó que solamente el ALA-dehidrasa en los grupos B y C se hallaba significativamente disminuida (P < 0,002). La disminución de los niveles de ALA-D se correspondió con la severidad de la neuropatía.

Efeitos intermediários da sobrecarga alcoólica moderada sobre os níveis séricos da gamaglutamiltransplanspeptidase, transminase glutâmico-oxalacética e transaminase glutâmico-pivúrica em adultos jovens saudáveis / Intermediate effects of a moderate alcohol uptake on serum gamma-glutamyltranspeptidase, glutamic-oxaloacetic transminase, and glutamic-pyruvic transaminase levels in healthy yong adults

Os autores estudaram o efeito de uma sobrecarga alcoólica de 0,75g/kg de peso nos níveis séricos de gama-glutamiltranspeptidase (GGT), transaminase glutâmico-oxalacética (TGO) e transaminase glutâmico-pirúvica (TGP) em 14 voluntários jovens saudáveis em um período de 72 horas. Nao houve alteraçöes significativas nas enzimas medidas. Concluem que uma sobrecarga alcoólica moderada em indivíduos jovens saudáveis nao altera GGT, TGO e TGP.

Valor da enzima gama-glutamil transferase (gama GT) sérica no diagnóstico do alcoolismo / Value of serum gamma-glutamyltransferase in the diagnosis of alcoholism

Faz-se uma revisäo e discute-se a utilidade da dosagem da gama-glutamil transferase sérica (gama-GT) no diagnóstico do alcoolismo. Segundo dados da literatura, a sua sensibilidade varia de 26-95% e a sua especificidade de 85-94%. Alguns fatores, tais como o tipo de populaçäo estudada, diferentes critérios de normalidade empregados, uso concomitante de outras drogas, patologias de origem näo alcoólica, contribuem para a discrepância dos resultados encontrados. Embora útil no seguimento de pacientes, para monitorizaçäo do tratamento, o emprego da gama-GT para fins de detecçäo de alcoolismo em grandes populaçöes tem limitaçöes, dependendo da prevalência de alcoolismo. Do ponto de vista prático, a presença de níveis séricos elevados da gama-GT pode ser útil identificar consumo excessivo de álcool desde que associada a informaçöes obtidas por questionários padronizados para a detecçäo do alcoolismo e levando-se em consideraçäo dados clínicos

Niveles de transaminasas y fosfatasa alcalina en sujetos con dosaje etílico positivo / Levels of transaminases and alkaline phosphatase in subjects with ethyl positive dosage

El presente trabajo se realizó en el Laboratorio Central del HCSFP y del Centro de Reconocimiento Médico SFP. con la finalidad de investigar los efectos del alcohol sobre los valores de las enzimas transaminasas y fosfatasa alcalina. Se evaluó a personas que por accidentes de tránsito fueron conducidos al Centro de Reconocimiento Médico por personal G.C. y que presentaron prueba cualitativa de alcoholemia positiva. Previo descarte de sueros hemolizados y lechosos, seleccionamos 90 muestras con alcoholemia a partir de 0.61 gr/l y comprendidos entre los 20 y 62 años. Los sujetos se agruparon de acuerdo a tres niveles de alcoholemia y tres grupos etarios. Para el dosaje de las transaminasas se usó el método de Reitman y Frankel modificado y la hidrolisis del paranitrofenol para la fosfatasa alcalina, así como el método de Sheftel modificado para el dosaje de alcohol etílico. Se evaluán resultados, no observándose variaciones anormales en las cifras de las enzimas estudiadas (transaminasas y fosfatasa alcalina) ni con el grado de alcoholemia ni con la edad