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A New Type of Inherited Catalase Deficiencies: Its Characterization and Comparison to the Japanese and Swiss Type of Acatalasemia
Abstract
Thirteen Hungarian families that exhibited inherited catalase deficiencies have been detected. Differences between the deficiencies reported from Hungary and the previously reported Swiss acatalasemia were characterized using biochemical analysis of the catalase proteins. Molecular biological methods were used to compare the previously reported types of catalase deficiencies in Japan and the Hungarian deficiencies. Three mutations (a GA insertion in exon 2, a G insertion in exon 2, and a T to G substitution in intron 7) are responsible for decreased catalase activity in 7 of the 13 Hungarian kindreds; the other 6 families have not yet been characterized. These are not the mutations observed in Japan. Changes in lipid and carbohydrate metabolism and the high incidence (12.7%) of diabetes mellitus in the Hungarian kindreds suggest that individuals with inherited catalase deficiency are at risk of atherosclerosis and diabetes mellitus. The Hungarian subjects were detected during screening of a large population for catalase activity; no overt disease state was associated with the deficiencies. We hypothesize that the increased risk of disease may be due to prolonged exposure to elevated levels of blood hydrogen peroxide due to the lack of normal removal of hydrogen peroxide by blood catalase.
... Vélhetõen azonban a táplálkozási körülményeknek és a szájhigiéniának talán még nagyobb szerepe lehetett az orális gangraena létrejöttében, mint a katalázhiánynak, mivel a késõbb leírt esetekben, amikorra jelentõsen megváltoztak a táplálkozási és a szájhigiéniai szokások, ez az elváltozás már nem alakult ki (17). A késõbi években detektált acatalasaemiás és hypocatalasaemiás egyéneknél (Svájc, 1963; Magyarország, 1992) orális gangraena nem fordult elõ (18,19). ...
... Magyarországon több mint 23 000 egyén vérkatalázaktivitásának vizsgálatával elsõként egy acatalasaemiás és 12 hypocatalasaemiás család volt detektálható (19)(20)(21). A két acatalasaemiás nõvér közül mindketten és a 63 hypocatalasaemiásból heten diabetes mellitusban szenvedtek (kilencük közül nyolcan a 2-es típusban, míg egyikõjük az 1-es típusban). ...
... A világ 11 országában 59 acatalasaemiás családot és 113 acatalasaemiás egyént tart számon az irodalom, de mindössze két olyan nukleotidmutáció (japán A és B) ismert, amely katalázhiányos szindrómát okoz. A Magyarországon detektált, veleszületett katalázhiányos családokban az ismert két japán típusú mutáció nem volt kimutatható, és a svájci típusra jellemzõ, megváltozott elektroforetikus mobilitás sem volt tapasztalható (19). Ezek ismeretében feltételezhetõ, hogy a magyarországi acatalasaemiáért genetikailag egy vagy több új katalázgén-mutáció lehet a felelõs. ...
CLINICAL EXAMINATION OF ENZYMECATALASE AND ITS MUTATIONSIN HUNGARYHydrogen peroxide is formed during physio-logical and pathological processes in the humanbody. Hydrogen peroxide in high concentrationand its more reactive compound (hydroxylradical) can damage cells. Recent findings showthat hydrogen peroxide in low concentration isrequired for many physiological pathways of cellsignaling such as diabetes, platelet activation,inflammation, immune response. In humantissues the steady state concentration of hydro-gen peroxide changes between 0.1 nmol/l-100micromol/l.The main regulator of hydrogen peroxidemetabolism is the enzyme catalase. This enzymehas a fast and high activity on high hydrogenperoxide concentrations and it shows a weakeffect on its physiologic concentration.Acatalasemia, the inherited deficiency of en-zyme catalase seems to be rather a syndromethan a simple enzyme deficiency as it wasthought earlier. It is associated with changes inlipid, carbohydrate and erythrocyte metabo-lisms.Contrary to the the worldwide (11 countries)distribution of acatalasemia only two syndromecausing catalase mutations have been reportedfrom Japan. Recently, four novel catalase muta-tions detected in one acatalasemic and 12hypocatalasemic Hungarian families which mu-tations are responsible for the decreased
(17) A kataláz enzim klinikai vonatkozásai és mutációi Magyarországon. Available from: https://www.researchgate.net/publication/238098991_A_katalaz_enzim_klinikai_vonatkozasai_es_mutacioi_Magyarorszagon [accessed Mar 09 2018].
... Vélhetõen azonban a táplálkozási körülményeknek és a szájhigiéniának talán még nagyobb szerepe lehetett az orális gangraena létrejöttében, mint a katalázhiánynak, mivel a késõbb leírt esetekben, amikorra jelentõsen megváltoztak a táplálkozási és a szájhigiéniai szokások, ez az elváltozás már nem alakult ki (17). A késõbi években detektált acatalasaemiás és hypocatalasaemiás egyéneknél (Svájc, 1963; Magyarország, 1992) orális gangraena nem fordult elõ (18,19). ...
... Magyarországon több mint 23 000 egyén vérkatalázaktivitásának vizsgálatával elsõként egy acatalasaemiás és 12 hypocatalasaemiás család volt detektálható (19)(20)(21). A két acatalasaemiás nõvér közül mindketten és a 63 hypocatalasaemiásból heten diabetes mellitusban szenvedtek (kilencük közül nyolcan a 2-es típusban, míg egyikõjük az 1-es típusban). ...
... A világ 11 országában 59 acatalasaemiás családot és 113 acatalasaemiás egyént tart számon az irodalom, de mindössze két olyan nukleotidmutáció (japán A és B) ismert, amely katalázhiányos szindrómát okoz. A Magyarországon detektált, veleszületett katalázhiányos családokban az ismert két japán típusú mutáció nem volt kimutatható, és a svájci típusra jellemzõ, megváltozott elektroforetikus mobilitás sem volt tapasztalható (19). Ezek ismeretében feltételezhetõ, hogy a magyarországi acatalasaemiáért genetikailag egy vagy több új katalázgén-mutáció lehet a felelõs. ...
Hydrogen peroxide is formed during physiological and pathological processes in the human body. Hydrogen peroxide in high concentration and its more reactive compound (hydroxyl radical) can damage cells. Recent findings show that hydrogen peroxide in low concentration Is required for many physiological pathways of cell signaling such as diabetes, platelet activation, inflammation, immune response. In human tissues the steady state concentration of hydrogen peroxide changes between 0.1 nmol/l-100 μmol/l. The main regulator of hydrogen peroxide metabolism is the enzyme catalase. This enzyme has a fast and high activity on high hydrogen peroxide concentrations and it shows a weak effect on its physiologic concentration. Acatalasemia, the inherited deficiency of enzyme catalase seems to be rather a syndrome than a simple enzyme deficiency as it was thought earlier. It is associated with changes in lipid, carbohydrate and erythrocyte metabolisms. Contrary to the the worldwide (11 countries) distribution of acatalasemia only two syndrome causing catalase mutations have been reported from Japan. Recently, four novel catalase mutations detected in one acatalasemic and 12 hypocatalasemic Hungarian families which mutations are responsible for the decreased catalase activity.
... The clinical, biochemical, and genetic characterization and comparison to the Japanese and Swiss type of the inherited catalase deficiency showed that the Hungarian acatalasemia is a new type of this syndrome [6]. ...
... Ϯ 17.4 MU/l and female 52.0 Ϯ 12.4 MU/l vs. 100.7 Ϯ 8.8 MU/l) may yield a higher hydrogen peroxide concentration. The erythrocyte catalase provides a general protection against intra-and extracellular hydrogen peroxide [1,2,27] and its decreased levels via increased hydrogen peroxide have shown that they may be responsible for the alteration in lipid and carbohydrate metabolism [5,6,10]. ...
Elevated plasma homocysteine can generate oxygen free radicals and hydrogen peroxide. The enzyme catalase is involved in the protection against hydrogen peroxide. We examined the effect of oxidative stress promoted by homocysteine on erythrocyte metabolism (blood hemoglobin, MCV, folate, B12, serum LDH, LDH isoenzymes, haptoglobin) in the oxidative stress sensitive Hungarian patients with inherited catalase deficiency. The plasma homocysteine (HPLC method, Bio-Rad), folate, B12 (capture binding assay, Abbott), blood hemoglobin concentrations, blood catalase activity (spectrophotometric assay of hydrogen peroxide), and MCV values were determined in 7 hypocatalasemic families including hypocatalasemic (male:12, female:18) patients and their results were compared to those of the normocatalasemic (male:17 female: 12) family members. We found decreased (p <.036) folate (ng/ml) concentrations (male hypocatalasemic 5.44 +/- 2.81 vs. normocatalasemic 7.56 +/- 1.97, female 5.01 +/- 1.93 vs. 6.61 +/- 1.91), blood hemoglobin (p <.010, male:140.2 +/- 11.0 vs. 153.6 +/- 11.6 g/l, female: 128.4 +/- 10.9 vs. 139.6 +/- 9.2 g/l). Increased levels of MCV (p <.001) were detected in hypocatalasemic patients (male: 98.6 +/- 3.4 vs. 90.1 +/- 7.5 fl, female: 95.9 +/- 3.9 vs. 90.1 +/- 2.5 fl), plasma homocysteine (p <.049, male: 9.72 +/- 3.61 vs. 7.36 +/- 2.10 umol/l, female: 9.06 +/- 3.10 vs. 6.84 +/- 2.50 umol/l) and not significant (p >.401) plasma B12 (male: 336 +/- 108 vs. 307 +/- 76 pg/ml, female: 373 +/- 180 vs. 342 +/- 75 pg/ml). The serum markers of hemolysis (LDH, LDH isoenzymes, haptoglobin) did not show significant (p >.228) signs of oxidative erythrocyte damage. We report firstly on increased plasma homocysteine concentrations in inherited catalase deficiency. The increased plasma homocysteine and inherited catalase deficiency together could promote oxidative stress via hydrogen peroxide. The patients with inherited catalase deficiency are more sensitive to oxidative stress of hydrogen peroxide than the normocatalasemic family members. This oxidative stress might be responsible for the decreased concentration of the blood hemoglobin via the oxidation sensitive folate and may contribute to the early development of arteriosclerosis and diabetes in these patients.
... Takahara unexpectedly discovered Japanese patients with acatalasemia (also known as Takahara disease) by severe oral gangrene after exposure to H 2 O 2 (34,35). Changes in lipid and carbohydrate metabolism and the high incidence (12.7%) of diabetes mellitus in the Hungarian acatalasemia suggest that this inherited catalase deficiency is a risk factor for atherosclerosis and diabetes mellitus (6,7). Deficiency of catalase may predispose to cumulative oxidant damage in pancreatic beta cells (6,7). ...
... Changes in lipid and carbohydrate metabolism and the high incidence (12.7%) of diabetes mellitus in the Hungarian acatalasemia suggest that this inherited catalase deficiency is a risk factor for atherosclerosis and diabetes mellitus (6,7). Deficiency of catalase may predispose to cumulative oxidant damage in pancreatic beta cells (6,7). However, the effect of acatalasemia on the progression of kidney disease is poorly understood. ...
Tissue homeostasis is determined by the balance between oxidants and antioxidants. Catalase is an important antioxidant enzyme regulating the level of intracellular hydrogen peroxide and hydroxyl radicals. The effect of catalase deficiency on renal tubulointerstitial injury induced by unilateral ureteral obstruction (UUO) has been studied in homozygous acatalasemic mutant mice (C3H/AnLCs(b)Cs(b)) compared with wild-type mice (C3H/AnLCs(a)Cs(a)). Complete UUO caused interstitial cell infiltration, tubular dilation and atrophy, and interstitial fibrosis with accumulation of type IV collagen in obstructed kidneys (OBK) of both mouse groups. However, the degree of injury showed a significant increase in OBK of acatalasemic mice compared with that of wild-type mice until day 7. The deposition of lipid peroxidation products including 4-hydroxy-2-hexenal, malondialdehyde, and 4-hydroxy-2-nonenal was severer in dilated tubules of acatalasemic OBK. Apoptosis in tubular epithelial cells significantly increased in acatalasemic OBK at day 4. Expression of caspase-9, a marker of mitochondrial pathway-derived apoptosis, increased in dilated tubules of acatalasemic mice. The level of catalase activity remained low in acatalasemic OBK until day 7 without compensatory upregulation of glutathione peroxidase activity. The data indicate that acatalasemia exacerbated oxidation of renal tissue and sensitized tubular epithelial cells to apoptosis in OBK of UUO. This study demonstrates that catalase deficiency enhanced tubulointerstitial injury and fibrosis in a murine model of UUO and thus supports the protective role of catalase in this model.
... Being widely expressed, CAT limits the toxic effect of H 2 O 2 by catalyzing its decomposition into water and oxygen in all aerobic cells (3). Several rare CAT gene sequence variations have been identified in association with acatalasemia, which is characterized by severe catalase deficiency (4,5). Reports on the prevalence of possible oxidative stress-related diseases or other diseases in catalase-deficient individuals are scarce. ...
A low level of the central antioxidant enzyme catalase has been suggested to be a risk factor for diseases influenced by oxidative stress. In this study, we investigated the possible association of the catalase -262C/T polymorphism with survival, physical and cognitive functioning, and a number of oxidative stress-mediated disorders. The study population was 2223 Danish individuals, aged 45-93 years, drawn from three population-based surveys. The results suggest that the catalase -262C/T polymorphism is not associated with either survival, or the majority of the age-related phenotypes investigated. However, our data indicate a statistical significant association of TT homozygosity with improved physical functioning as well as a trend of the T allele conferring an improved general cognitive functioning, although these results did not remain significant after correcting for multiple testing. The results raise the hypothesis that the catalase -262T allele serves as protection against neurodegenerative and physical decline, although replication in other studies is warranted for confirmation of these findings.
... Oxidative stress with alterations in profile of antioxidant enzymes in erythrocytes is also related to many others specific pathologies (Matés et al. 1999), e.g. diabetes (Góth andEaton 2000, Sailaja et al. 2003), hypertension (Yuan et al. 1996, Johnson 2002 etc. Changes of erythrocyte CAT activity in dependence on oxygen consumption also occur in patients with hyperthyroidism (Kurasaki et al. 1986, Alicigüzel et al. 2001. ...
The erythrocytes represent an important source of antioxidant capacity of the blood. Catalase (EC 1.11.1.6.) is one of the enzymatic components of their antioxidant defense system. The objective of this study was to follow erythrocyte catalase (CAT) in 7-, 15-, 21-, 35-, 60- and 90-day-old Wistar rats of both sexes in normoxia and after exposure to intensive acute hypobaric hypoxia. During the development CAT activity increases in both sexes, but the rise was usually higher in females. Hypobaric hypoxia increased CAT activity in all studied age groups of both sexes. However, higher CAT activity in females was less affected by hypoxia than the lower activity in males. This was true for nearly all age groups studied. It can be concluded that both ontogenetic aspects and sex differences play a major role in establishing the activity of CAT, which is an important part of the antioxidant defense of the organism.
... Catalase is an antioxidant enzyme in microsomes of liver and plays a major role in detoxification of peroxides and reactive oxygen species (ROS) (Glorieux and Calderon, 2017). Overexpression of catalase protects against inflammation associated injury including atherosclerosis and diabetic complications (Góth, 2001) F7/Coagulation Factor VII −0.68 # Level 1 confirmation. Coagulation factor VII is synthesized in liver and adipose tissue and is part of the coagulation pathway where it binds to Tissue Factor (TF). ...
Metabolic disorders, such as obesity and type 2 diabetes have a large impact on global health, especially in industrialized countries. Tissue-specific chronic low-grade inflammation is a key contributor to complications in metabolic disorders. To support therapeutic approaches to these complications, it is crucial to gain a deeper understanding of the inflammatory dynamics and to monitor them on the individual level. To this end, blood-based biomarkers reflecting the tissue-specific inflammatory dynamics would be of great value. Here, we describe an in silico approach to select candidate biomarkers for tissue-specific inflammation by using a priori mechanistic knowledge from pathways and tissue-derived molecules. The workflow resulted in a list of candidate markers, in part consisting of literature confirmed biomarkers as well as a set of novel, more innovative biomarkers that reflect inflammation in the liver and adipose tissue. The first step of biomarker verification was on murine tissue gene-level by inducing hepatic inflammation and adipose tissue inflammation through a high-fat diet. Our data showed that in silico predicted hepatic markers had a strong correlation to hepatic inflammation in the absence of a relation to adipose tissue inflammation, while others had a strong correlation to adipose tissue inflammation in the absence of a relation to liver inflammation. Secondly, we evaluated the human translational value by performing a curation step in the literature using studies that describe the regulation of the markers in human, which identified 9 hepatic (such as Serum Amyloid A, Haptoglobin, and Interleukin 18 Binding Protein) and 2 adipose (Resistin and MMP-9) inflammatory biomarkers at the highest level of confirmation. Here, we identified and pre-clinically verified a set of in silico predicted biomarkers for liver and adipose tissue inflammation which can be of great value to study future development of therapeutic/lifestyle interventions to combat metabolic inflammatory complications.
... Os polimorfismos de Mn-SOD, CAT, GPX1 e haptoglobina têm sido descritos como interessantes biomarcadores em associação com doenças (Taufer et al., 2005;Ratnasinghe et al., 2000;Wassel, 2000;Góth, 2001;Hu e Diamond, 2003;Góth et al., 2004;Ichimura et al., 2004;Sadrzadeh e Bozorgehr, 2004;Akyol et al., 2005;Oberley, 2005;Ravn-Haren et al., 2006;Carter e Worwood, 2007;Oh et al., 2007). Desta forma, estudos que examinem a distribuição das frequências alélicas e genotípicas de tais marcadores em várias etnias poderão fornecer uma melhor compreensão de seu significado biológico para estudos antropogênicos e de associação com doenças. ...
Tese (doutorado)—Universidade de Brasília, Instituto de Ciências Biológicas, Programa de Pós-Graduação em Biologia Animal, 2009. O consumo diário de antioxidantes naturais protege contra danos oxidativos causados por espécies reativas de oxigênio (ERO) e pode reduzir o risco de câncer, aterosclerose e outras doenças degenerativas. O treinamento físico induz adaptações benéficas, mas exercícios extenuantes ou com frequência de treinamento muito elevada aumentam a geração de ERO, resultando em danos oxidativos no DNA e nos tecidos. O pequi (Caryocar brasiliense Camb.) é uma fruta típica do Cerrado Brasileiro, bem conhecida na culinária regional e usada na medicina popular para tratar várias enfermidades. Sua polpa contém diversos antioxidantes como carotenóides, vitamina C e compostos fenólicos, e sua composição de ácidos graxos é representada principalmente pelos ácidos oléico (51,37 a 55,87%) e palmítico (35,17 a 46,79%). O objetivo deste estudo foi investigar os efeitos antigenotóxicos, antioxidantes e farmacológicos de extratos da polpa do pequi, através de estudos in vivo em camundongos e atletas humanos (corredores de rua). Os testes pré-clínicos e toxicológicos em camundongos mostraram que os extratos não tiveram efeito clastogênico ou genotóxico sobre as células e ambos protegeram os animais contra danos oxidativos no DNA induzidos por bleomicina ou ciclofosfamida, dois fármacos antineoplásicos. Entretanto, o efeito protetor dependeu da dose. O teste de TBARS, usado para avaliar a atividade antioxidante, mostrou que, nas doses testadas, o extrato aquoso elevou a peroxidação lipídica nos camungongos de ambos os sexos, especialmente nos machos; o extrato orgânico aumentou a peroxidação lipídica apenas nos machos, sem efeitos nas fêmeas. O extrato orgânico (óleo de pequi) foi escolhido para investigar os efeitos antioxidantes contra danos oxidativos induzidos pelo exercício em atletas corredores de rua. Para avaliar se os efeitos antioxidantes do óleo de pequi foram influenciados pelos genótipos das enzimas antioxidantes e da haptoglobina (Hp), foram investigados os polimorfismos dos genes Mn-SOD ( Val9Ala), CAT ( 21A/T), GPX1 (Pro198Leu) e Hp. As avaliações foram feitas após corridas ao ar livre em terreno plano, antes e depois da ingestão de 400mg de óleo de pequi em cápsulas durante 14 dias. A pressão arterial dos voluntários foi checada antes das corridas. Após as duas corridas, foram colhidas amostras de sangue e essas foram submetidas ao teste do cometa, teste TBARS, hemograma, lipidograma pós-prandial e análises bioquímicas de creatina-fosfoquinase (CPK), transaminase glutâmica-oxalacética (TGO), transaminase glutâmica-pirúvica (TGP), proteína Creativa ultra-sensível (PCR-US). Os resultados dos testes de TGO, TGP e cometa indicaram influência do sexo, e uma redução significativa de danos após o tratamento com óleo de pequi foi observada no grupo de mulheres para TGO e TGP e em ambos os sexos para o teste do cometa. Para esses marcadores, também houve influência do genótipo Mn-SOD, cuja frequência mais alta de heterozigotos foi relacionada a menores danos no DNA e nos tecidos, além de exibirem melhor resposta ao óleo de pequi. Para CPK e PCR-US, os resultados indicaram que o sexo exerceu efeito significativo, e uma queda não-significativa nos valores foi observada apenas para os homens. No eritrograma, a tendência geral de redução de eritrócitos, hemoglobina e hematócrito após pequi foi particularmente associada a uma maior expansão do plasma, sendo tais resultados influenciados por sexo, idade (eritrócitos, hemoglobina) e genótipos de haptoglobina (eritrócitos, hemoglobina e hematócrito), Mn-SOD (hematócrito), CAT (hemoglobina e hematócrito) e GPX1 (hemoglobina). A redução significativa da anisocitose foi relacionada principalmente ao efeito protetor antioxidante do óleo de pequi. Os resultados do leucograma e da série plaquetária sugeriram que o óleo de pequi foi biologicamente eficiente em reduzir a inflamação provocada pelo exercício agudo. Queda significativa nos valores da pressão arterial, de colesterol total e LDL pósprandiais foram observadas no grupo 45 anos, sugerindo que o óleo de pequi apresentou efeitos cardiovasculares protetores, principalmente para aqueles atletas incluídos na faixa etária de maior risco. Para os homens, o teste TBARS apresentou resultados contrários àqueles obtidos com camundongos e, novamente, nenhum efeito foi observado nas mulheres. Porém, tais resultados foram influenciados por idade, distância percorrida e genótipos Hp e Mn-SOD. Mediante o apresentado, podemos concluir que o óleo de pequi, além de possuir várias propriedades nutricionais, apresentou efeitos antioxidantes e cardiovasculares protetores e, após testes adicionais e com a definição da dose adequada, também poderá ser usado como adjuvante na quimioterapia do câncer, na forma de suplemento na dieta. _______________________________________________________________________________ ABSTRACT The daily consumption of natural antioxidants protects against oxidative damage caused by reactive oxygen species (ROS) and can reduce the risk of cancer, atherosclerosis and other degenerative diseases. Physical training induces beneficial adaptations, but exhausting exercise above habitual intensity or training increases generation of ROS, resulting in oxidative damages in DNA and in tissues. Pequi (Caryocar brasiliense Camb.) is a typical fruit found in the Brazilian Cerrado (a savanna-like biome), well-known in regional cookery and used in folk medicine to treat various illnesses. Its pulp contains several antioxidant such as carotenoids, vitamin C and phenolic compounds, and its fatty acid composition is represented mainly by oleic (51.37 to 55.87%) and palmitic (35.17 to 46.79%) acids. The aim of this study was to investigate the antigenotoxic, antioxidant and pharmacological effects of pequi fruit pulp extracts through in vivo studies in mice and human athletes (street runners). Preclinical and toxicological tests in mice showed that the extracts had no clastogenic or genotoxic effect and both protected mice against oxidative DNA damage induced by bleomycin or cyclophosphamide, two antineoplastic drugs. However, the protective effect depended on the dose. TBARS assay, used to assess the antioxidant activity, showed that, at the tested doses, the aqueous extract enhanced lipid peroxidation in mice of both sexes, especially in males; the organic extract enhanced lipid peroxidation only in males, with no effect in females. Then, we chose the organic extract (pequi oil) to investigate its antioxidant effects against exercise-induced oxidative damages in runners. To estimate if the antioxidant effects of the pequi oil were influenced by antioxidant enzymes and haptoglobin (Hp) genotypes, the Mn-SOD ( Val9Ala), CAT ( 21A/T), GPX1 (Pro198Leu) and Hp genes’ polymorphisms were investigated. Evaluations took place after outdoor races on flat land before and after ingestion of 400mg pequioil capsules for 14 days. The arterial pressure of volunteers was checked before the races. Blood samples were taken after the two races and submitted to comet assay, TBARS assay, blood count (hemogram), postprandial lipid profile dosage and biochemical analyses of creatine phosphokinase (CPK), glutamic-oxalacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), high sensitivity C-reactive protein (hs-CRP). Results of TGO, TGP and comet tests indicated influence of sex, and significant damage decrease after pequi oil treatment was observed in the women group for TGO and TGP tests and in both sexes for the comet test. For these markers, there were also influences of Mn-SOD genotype, whose higher frequency of heterozygous was related to lower DNA damage and tissue injuries, besides presenting a better response to the pequi oil. Regarding CPK and hs-CRP, results indicated that sex exerted significant effect, and a nonsignificant decrease in the values was observed only for men. Results of erythrogram indicated that the general downward trend of erythrocytes, hemoglobin and hematocrit after treatment was particularly associated to a higher plasma expansion, and they were influenced by sex, age (erythrocytes, hemoglobin) and haptoglobin (erythrocytes, hemoglobin and hematocrit), Mn-SOD (hematocrit), CAT (hemoglobin and hematocrit) and GPX1 (hemoglobin) genotypes. The significant decrease in the anisocytosis was related mainly to the protective antioxidant effect of pequi oil. Results of leukogram and platelets parameters (platelet count, plateletocrit, mean platelet volume MPV, platelet deviation weight PDW) suggested that pequi oil was biologically efficient to reduce the inflammation provoked by acute exercise. Significant decrease in the values of arterial pressure, postprandial total cholesterol and LDL-C was observed in age group 45 years, suggesting that pequi oil presented protective cardiovascular effects, mainly for those athletes included in the age group of higher risk. For males, TBARS assay presented results contrary to those obtained with mice and, again, no effect was observed in females. However, these results were influenced by age, distance covered, Hp and Mn-SOD genotypes. By means of our results we can conclude that pequi-oil, as well as possessing many other nutritional properties, showed protective antioxidant and cardiovascular effects and, after more tests and with adequate adjustment of the dose, it also could be a useful dietary supplement as adjuvant to the chemotherapy of cancer.
... Overproduction enzyme activities. and/or insufficient detoxification of hydrogen peroxide may trig-ger a cascade of neurotoxic events contributing to the neural Switzerland, and Hungary have been characterized [76,[89][90][91] using damage characteristic of the disease. [63] clinical, biochemical and molecular genetic methods, while the identification of cases in other countries has been sporadic and In a study of 137 controls and 137 patients with Alzheimers relatively poorly characterized. ...
Enzyme catalase seems to be the main regulator of hydrogen peroxide metabolism. Hydrogen peroxide at high concentrations is a toxic agent, while at low concentrations it appears to modulate some physiological processes such as signaling in cell proliferation, apoptosis, carbohydrate metabolism, and platelet activation. Benign catalase gene mutations of 5′ noncoding region (15) and intron 1 (4) have no effect on catalase activity and are not associated with disease.
Catalase gene mutations have been detected in association with diabetes mellitus, hypertension, and vitiligo. Decreases in catalase activity in patients with tumors is more likely to be due to decreased enzyme synthesis rather than to catalase mutations.
Acatalasemia, the inherited deficiency of catalase has been detected in 11 countries. Its clinical features might be oral gangrene, altered lipid, carbohydrate, homocysteine metabolism and the increased risk of diabetes mellitus. The Japanese, Swiss, and Hungarian types of acatalasemia display differences in biochemical and genetic aspects. However, there are only limited reports on the syndrome causing these mutations.
These data show that acatalasemia may be a syndrome with clinical, biochemical, genetic characteristics rather than just a simple enzyme deficiency.
... [4,5] In 1966, Ogata et al. suggested lack of catalase in blood to be the cause of the disease. [6] A high incidence of diabetes mellitus was reported by Goth et al. in Hungarian acatalasemia patients [7,8] and was associated with catalase enzyme mutations. [9] Recently, Goth et al. and Masuoka et al. reported that the diabetes mellitus was associated with low catalase activity in blood. ...
Catalase is one of the major antioxidant enzymes that catalyzes the hydrolysis of H2O2. The aim of this study was to suggest a new method for the assay of catalase activity. For this purpose, an amperometric biosensor based on glucose oxidase for determination of catalase activity was developed. Immobilization of glucose oxidase was made by a cross-linking method with glutaraldehyde on a Clark-type electrode (dissolved oxygen probe). Optimization and characterization properties of the biosensor were studied and determination of catalase activity in defined conditions was investigated in artificial serum solution. The results were compared with a reference method.
... Like acatalasemia, hypocatalasemia usually does not cause any health problems. [12] ...
Catalase deficiency, also known as acatalasemia, is a disease marked by abnormally low levels of the enzyme catalase. Many individuals with acatalasemia have never had any health issues and are now diagnosed because they have affected family members. Some of the first people diagnosed with acatalasemia developed open sores (ulcers) within their mouths, resulting in soft tissue death (gangrene). Takahara disease is a disorder that occurs when acatalasemia causes mouth ulcers and gangrene. Because of changes in oral hygiene, these complications are rarely seen in more recent cases of acatalasemia. According to studies, people with acatalasemia are more likely to develop type 2 diabetes, which is the most common type of diabetes. Type 2 diabetes affects a higher proportion of people with acatalasemia than the general population, and the disorder strikes at a younger age. Acatalasemia may also be a risk factor for other common, complex diseases, according to researchers.
... Hungarian and Japanese hypocatalsemics have a catalase activity of about 50% compared to normocatalsemics (Ogata et al., 1977a;Goth, 2001) while Swiss hypocatalasemics have almost normal activities (Aebi et al., 1977). Hungarian subjects with catalase deficiency have higher levels of erythrocyte SOD and GPX activitities (Goth and Eaton, 2000). ...
... MnSOD upregulation was shown to mitigate apoptosis in brain [104], diabetic cardiopathy [105], cell signaling death in liver [106], and restored redox balance in the skeletal muscle following exercise [107]. ROS are frequently associated with pathophysiology and an increasing number of diseases are associated with ROS activity as an etiologic agent or contributing factor [108]. Several studies have shown that MnSOD induction can protect against neurotoxic conditions [102], cardiomyopathy [105,109], and diabetic disorders [110,111]. ...
Oxidative stress is characterized by imbalanced reactive oxygen species (ROS) production and antioxidant defenses. Two main antioxidant systems exist. The nonenzymatic system relies on molecules to directly quench ROS and the enzymatic system is composed of specific enzymes that detoxify ROS. Among the latter, the superoxide dismutase (SOD) family is important in oxidative stress modulation. Of these, manganese-dependent SOD (MnSOD) plays a major role due to its mitochondrial location, i.e., the main site of superoxide ( [Formula: see text] ) production. As such, extensive research has focused on its capacity to modulate oxidative stress. Early data demonstrated the relevance of MnSOD as an [Formula: see text] scavenger. More recent research has, however, identified a prominent role for MnSOD in carcinogenesis. In addition, SOD downregulation appears associated with health risk in heart and brain. A single nucleotide polymorphism which alters the mitochondria signaling sequence for the cytosolic MnSOD form has been identified. Transport into the mitochondria was differentially affected by allelic presence and a new chapter in MnSOD research thus begun. As a result, an ever-increasing number of diseases appear associated with this allelic variation including metabolic and cardiovascular disease. Although diet and exercise upregulate MnSOD, the relationship between environmental and genetic factors remains unclear.
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... Lower activities of CAT and SOD were shown in premature infants during first 72 h of their life in comparison with full-term infants and even during aging [33,34] . Less than 10% of normal erythrocyte CAT activity was observed in homozygous carrier of inherited CAT deficiency-acatalasemia [35] , and less than 50% in heterozygous subject's hypocatalasemia [36] . An elevated SOD and CAT activities during aging in human erythrocytes has been reported [33] . ...
Despite more than 300 theories to explain the aging process, oxidative stress theory offers the best mechanism to explain aging and age related disorders. Several studies has shown the importance of oxidative stress during aging. PubMed, Science Direct and Springer online data bases are taken into consideration to write this mini-review. Human erythrocytes are most abundant and specialized cells in the body. Erythrocytes were extensively studied due to their metabolism and gas transport functions. Recent studies on erythrocytes have provided us detailed information of cell membrane and its structural organization that may help in studying the aging and age associated changes. The susceptibility of an organism is associated with the antioxidant potential of the body. Erythrocytes have potent antioxidant protection consisting of enzymatic and non-enzymatic pathways that counteract with reactive oxygen species, thus maintaining the redox regulation in the body. The non-enzymatic and enzymatic antioxidants and other biomarkers associated with erythrocyte membrane transport functions are the main content of this review. Biomarkers of oxidative stress in erythrocytes and its membrane were taken into the consideration during human aging that will be the main subject of this mini- review.
... In addition, CAT reacts with H ϩ donors, such as methanol, ethanol, formic acid, or phenols, with peroxidase activity. Specific mutations/polymorphisms in CAT gene may result in catalase deficiency (also known as acatalasemia), as reported in Japanese, Swiss, and Hungarian individuals (59,60,158). The deficiency or mutation of CAT has been associated with low H 2 O 2 removal and, therefore, oxidative stress leading to various diseases and abnormalities, such atherosclerosis, hyperlipidemia, gangrene, hypertension, neurodegeneration, and diabetes (60). ...
It is now accepted that nutrient abundance in the blood, especially glucose, leads to the generation of reactive oxygen species (ROS), ultimately leading to increased oxidative stress in a variety of tissues. In the absence of an appropriate compensatory response from antioxidant mechanisms the cell, or indeed tissue, becomes overwhelmed by oxidative stress, leading to the activation of intracellular stress associated pathways. Activation of the same or similar pathways also appear to play a role in mediating insulin resistance, impaired insulin secretion and late diabetic complications. The ability of antioxidants to protect against the oxidative stress induced by hyperglycemia and elevated free fatty acid (FFA) levels in vitro, suggests a causative role of oxidative stress in mediating the latter clinical conditions. In this review, we describe common biochemical processes associated with oxidative stress driven by hyperglycemia and/or elevated FFA and the resulting clinical outcomes; beta-cell dysfunction and peripheral tissue insulin resistance.
The enzyme catalase is the main regulator of hydrogen peroxide metabolism. Deficiency of catalase may cause high concentrations of hydrogen peroxide and increase the risk of the development of pathologies for which oxidative stress is a contributing factor, for example, type 2 diabetes mellitus. Catalase deficiency has been reported to be associated with increased frequency of diabetes mellitus in a cohort of patients in Hungary. In this cohort, the majority of mutations in the catalase gene occur in exon 2.
Type 2 diabetic patients (n=308) were evaluated for mutations in intron 1 (81 bp), exon 2 (172 bp) and intron 2 (13 bp) of the catalase gene. Screening for mutations utilized PCR single-strand conformational polymorphism (SSCP) and PCR heteroduplex methods. Verification of detected mutations was by nucleotide sequence analysis.
A total of 11 catalase gene mutations were detected in the 308 subjects (3.57%, p<0.001). Five of the 11 were at two previously reported mutation sites: exon 2 (79) G insertion and (138) GA insertion. Six of the 11 were at five previously unreported catalase mutation sites: intron 1 (60) G-->T; intron 2 (7) G-->A and (5) G-->C; exon 2 (96) T-->A; and exon 2 (135) T-->A. The novel missense mutations on exon 2 (96 and 135) are associated with 59% and 48% decreased catalase activity, respectively; the novel G-->C mutation on intron 2 (5) is associated with a 62% decrease in catalase activity. Mutations detected on intron 1 (60) and intron 2 (7) showed no change in catalase activity. The G-->C mutation on intron 2 (5) might be a splicing mutation. The two missense mutations on exon 2 (96) and (135) cause substitutions of amino acids 53 (Asp-->Glu) and 66 (Glu-->Cys) of the catalase protein. These are close to amino acids that are important for the binding of heme to catalase, 44 (Val) and 72-75 (Arg, Val, Val, His). Changes in heme binding may be responsible for the activity losses.
Mutations that cause decreased catalase activity may contribute to susceptibility to inherited type 2 diabetes mellitus. Exon 2 and neighboring introns of the catalase gene may be minor hot spots for type 2 diabetes mellitus susceptibility mutations.
Pertussis, also called whooping cough, is caused by Bordetella pertussis . The disease may show an atypical course, particularly in neonates and elderly patients. A rapid and safe diagnostic method is thus essential for appropriate treatment and prophylaxis. Culture has been considered the gold standard for detection of B. pertussis , but this method often lacks sensitivity, and a minimum of 4 days may be required to obtain results (1)(2). PCR is a rapid, sensitive, and specific method for the diagnosis of pertussis (3)(4)(5).
In this study, a new molecular assay was established based on real-time PCR and including a homologous internal control (IC). We evaluated the performance of this assay with a commercially available genomic DNA isolate and with clinical samples.
The new molecular assay consisted of a protocol for manual extraction of DNA followed by generation of the amplification product by real-time PCR. The assay was based on the amplification of a 181-bp fragment of the repetitive insertion sequence IS481, which has been described in B. pertussis and Bordetella holmesii and may be present in Bordetella bronchiseptica (6)(7)(8)(9)(10) (see Table 1 in the Data Supplement that accompanies the online version of this Technical Brief at http://www.clinchem.org/content/vol51/issue12). We determined assay linearity and detection limit by analyzing 10-fold dilutions of the ATCC genomic DNA isolate 9797D from B. pertussis . Interassay variation was determined with 7 dilutions of the genomic DNA isolate (5 determinations on 5 different days), whereas intraassay variation was determined with 3 samples (5 determinations within a single assay). All assays for determination of inter- and intraassay variation included negative …
Although oxygen is a prerequisite to life, at concentrations beyond the physiological limits it may be hazardous to the cells. Since the lungs are directly exposed to very high amounts of oxygen, it is imperative for the organ to possess defences against possible oxidative challenge. The lungs are therefore endowed with an armamentarium of a battery of endogenous agents called antioxidants. The antioxidant species help the lungs ward off the deleterious consequences of a wide variety of oxidants/reactive oxygen species such as superoxide anion, hydroxyl radical, hypohalite radical, hydrogen peroxide and reactive nitrogen species such as nitric oxide, peroxynitrite, nitrite produced endogenously and sometimes accessed through exposure to the environment. The major non-enzymatic antioxidants of the lungs are glutathione, vitamins C and E, beta-carotene, uric acid and the enzymatic antioxidants are superoxide dismutases, catalase and peroxidases. These antioxidants are the first lines of defence against the oxidants and usually act at a gross level. Recent insights into cellular redox chemistry have revealed the presence of certain specialized proteins such as peroxiredoxins, thioredoxins, glutaredoxins, heme oxygenases and reductases, which are involved in cellular adaptation and protection against an oxidative assault. These molecules usually exert their action at a more subtle level of cellular signaling processes. Aberrations in oxidant: antioxidant balance can lead to a variety of airway diseases, such as asthma, chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis which is the topic of discussion in this review.
The enzyme catalase is the main regulator of hydrogen peroxide metabolism. Recent findings suggest that a low concentration of hydrogen peroxide may act as a messenger in some signalling pathways whereas high concentrations are toxic for many cells and cell components. Acatalasemia is a genetically heterogeneous condition with a worldwide distribution. Yet only two Japanese and three Hungarian syndrome-causing mutations have been reported. A large-scale (23 130 subjects) catalase screening program in Hungary yielded 12 hypocatalasemic families. The V family with four hypocatalasemics (60.6 +/- 7.6 MU/L) and six normocatalasemic (103.6 +/- 23.5 MU/L) members was examined to define the mutation causing the syndrome. Mutation screening yielded four novel polymorphisms. Of these, three intron sequence variations, namely G-->A at the nucleotide 60 position in intron 1, T-->A at position 11 in intron 2, and G-->T at position 31 in intron 12, are unlikely to be responsible for the decreased blood catalase activity. However, the novel G-->A mutation in exon 9 changes the essential amino acid Arg 354 to Cys 354 and may indeed be responsible for the decreased catalase activity. This inherited catalase deficiency, by inducing an increased hydrogen peroxide steady-state concentration in vivo, may be involved in the early manifestation of type 2 diabetes mellitus for the 35-year old proband.
In the vasculature, reactive oxygen species (ROS) generated by both mitochondrial respiration and enzymatic sources serve as integral components of cellular signaling and homeostatic mechanisms. Because ROS are highly reactive biomolecules, the cellular redox milieu is carefully maintained by small-molecule antioxidants and antioxidant enzymes to prevent the deleterious consequences of ROS excess. When this redox balance is perturbed, because of either increased ROS production or decreased antioxidant capacity, oxidant stress is increased in the vessel wall and, if not offset, vascular dysfunction ensues. A number of heritable polymorphisms of pro-oxidant enzymes, including 5-lipoxygenase, cyclooxygenase-2, nitric oxide synthase-3, and NAD(P)H oxidase, have been identified and found to modulate ROS production and, thereby, the risk of atherothrombotic cardiovascular disease in individuals with these genetic polymorphisms. Similarly, heritable deficiency of the antioxidant enzymes catalase, glutathione peroxidases, glutathione-S-transferases, heme oxygenase, and glucose-6-phosphate dehydrogenase favors ROS accumulation, and has been associated with an increased risk of vascular disease. Individually, each of these polymorphisms imposes a state of uncompensated oxidant stress on the vasculature and collectively comprise the oxidative enzymopathies.
Catalase (EC 1.11.1.6) is an enzyme which is present mainly in the peroxisomes of mammalian cells. It is a tetrameric enzyme consisting of four identical, tetrahedrally arranged subunits of 60 kDa, each containing in its active center a heme group and NADPH. Catalase has two enzymatic activities depending on the concentration of H2O2. If the concentration of H2O2 is high, catalase acts catalytically, i.e. removes H2O2 by forming H2O and O2 (catalatic reaction). However, at a low concentration of H2O2 and in the presence of a suitable hydrogen donor, e.g. ethanol, methanol, phenol, and others, catalase acts peroxidically, removing H2O2, but oxidizing its substrate (peroxidatic reaction). The review article presents current knowledge about the structure, properties, and functions of catalase in living organisms.
The effects of hydrogen peroxide on normal and acatalasemic erythrocytes were examined. Severe hemolysis of acatalasemic erythrocytes and a small tyrosine radical signal (g = 2.005) associated with the formation of ferryl hemoglobin were observed upon the addition of less than 0.25 mM hydrogen peroxide. However, when the concentration of hydrogen peroxide was increased to 0.5 mM, acatalasemic erythrocytes became insoluble in water and increased the tyrosine radical signal. Polymerization of hemoglobin and aggregation of the erythrocytes were observed. On the other hand, normal erythrocytes exhibited only mild hemolysis by the addition of hydrogen peroxide under similar conditions. From these results, the scavenging of hydrogen peroxide by hemoglobin generates the ferryl hemoglobin species (H-Hb-Fe(IV)=O) plus protein-based radicals (*Hb-Fe(IV)=O). These species induce hemolysis of erythrocytes, polymerization of hemoglobin, and aggregation of the acatalasemic erythrocytes. A mechanism for the onset of Takarara disease is proposed.
Patients with low (inherited and acquired) catalase activities who are treated with infusion of uric acid oxidase because they are at risk of tumour lysis syndrome may experience very high concentrations of hydrogen peroxide. They may suffer from methemoglobinaemia and haemolytic anaemia which may be attributed either to deficiency of glucose-6-phosphate dehydrogenase or to other unknown circumstances. Data have not been reported from catalase deficient patients who were treated with uric acid oxidase. It may be hypothesized that their decreased blood catalase could lead to the increased concentration of hydrogen peroxide which may cause haemolysis and formation of methemoglobin. Blood catalase activity should be measured for patients at risk of tumour lysis syndrome prior to uric acid oxidase treatment.
In this study, a specific enzyme catalase was immobilised onto the surface of two different biodegradable materials, starch cellulose acetate (SCA) and starch polycrapolactone (SPCL) blends. This immobilisation was achieved by several different routes, mainly by covalent binding and an adsorption method using as activation agents epichlorohydrin, cyanogen bromide (CNBr), and aminopropyltriethoxysilane. The effect of the coupling pH of the enzyme-support reaction was determined in terms of activity recovery (%). The catalase immobilised on SCA showed higher activity recovery (%) for all the methods used as compared with results obtained with SPCL. The immobilisation process using epichlorohydrin as an activation agent and polyethylenimine as a spacer-arm enhanced the stability and the half-lives at pH 7.0, 30 degrees C, for immobilised catalase on both SCA and SPCL. The half-lives were respectively, 1162 and 870 h compared with other treatments and free enzyme (480 h). The free glycerol present in the immobilisation medium was also a factor that contributed toward the better performance regarding the long-term stability at 30 degrees C and neutral pH. The extension of the morphological modifications on the surface of the materials was observed by scanning electron microscopy. In general, the results indicated that the chemical modification with epichlorohydrin could provide a simple and rather efficient technique to modify the starch-based materials' surface that might be useful in several biomedical applications.
Gestational diabetes occurs with variable severity in 3%–5% of all pregnancies and may be related to oxidative stress and impaired antioxidant defenses (1). Antioxidant enzymes include superoxide dismutase, which produces hydrogen peroxide, and catalase, which consumes hydrogen peroxide. Catalase is the main regulator of hydrogen peroxide metabolism (2), which is associated with diabetes mechanisms such as Glut 4 expression, insulin secretion, insulin signaling, protein tyrosine phosphatase regulation, and glucose transport stimulation (3). Hydrogen peroxide has novel insulin-like effects, e.g., inhibition of lipolysis and reactivation of phosphoenolpyruvate carboxy kinase (4)(5), and insulin moderates hydrogen peroxide generation (6)(7) and catalase synthesis (8). High concentrations of hydrogen peroxide may damage heme proteins, cause cell death, and together with redox active metal ions, produce highly toxic hydroxyl radicals.
High catalase activity in erythrocytes seems to provide antioxidant defense for tissues with low catalase activity, particularly pancreatic beta cells. Catalase is important in antioxidant defense against hydrogen peroxide (9)(10), but there are conflicting reports of decreases (11)(12), increases (13), and no change(14) in catalase activity in diabetes. A high incidence (14%) of diabetes mellitus observed in 63 Hungarian patients with inherited catalase deficiency (1 with type 1 and 7 with type 2 diabetes) could be associated with damage to oxidation-sensitive pancreatic beta cells by exposure to long-term increased hydrogen peroxide concentrations (15), but there have been conflicting reports from small studies of maternal and embryonic catalase in rat (16) and human(17) gestational diabetes.
We compared blood catalase activity in patients with gestational diabetes, pregnant patients without diabetes, and nonpregnant nondiabetic individuals. We examined the catalase gene mutations associated with decreased catalase activities and evaluated the effects of decreased maternal blood catalase activity on complications in newborns.
Study participants included 60 pregnant women with gestational diabetes …
The seafood industry is of considerable importance to both the New Zealand and global economies and therefore tissue preservation technologies that increase product quality and/or prolong shelf life have the potential to add significant value. Technologies for maintaining the viability of isolated tissues also have a wide range of other medical and industrial applications. This thesis examines the relationship between metabolic function, oxidation and cell death and the resulting stability of the non-viable tissues during long term storage in chinook salmon (Oncorhynchus tshawytscha) red and white skeletal muscle tissue. This research also looks at the role of the aquatic anaesthetic AQUI-S™, in which the active ingredient is isoeugenol (a lipid soluble antioxidant), and other antioxidant compounds in preserving metabolic function in viable tissues and tissue stability in nonviable tissues. Perfusion of salmon tails at 15℃ over 5 or 10 hours with oxygen saturated saline resulted in significant increases in protein and lipid oxidation (protein carbonyl and TBARS concentrations respectively) in the red muscle, but not the white muscle. The introduction of ascorbic acid and uric acid into the saline did not reduce the oxidation in the red muscle despite significantly increasing their respective concentrations in the tissue. This indicates the difficulties associated with attempting to extend tissue viability by delivering free oxygen to the tissue and also highlights the difference in susceptibility of the two muscle types to oxidation. Tail fillets from salmon harvested in both rested and exhausted physiological states using AQUI-S™, and fillets from exhausted salmon harvested without AQUI-S™, were exposed to air at 15℃ for up to 96 hours. Protein carbonyls increased in a roughly linear fashion over the entire 96 hours in all three groups. Both lipid peroxides (TBARS) and uric acid concentrations began to increase in the exhausted group after 30 hours. In contrast, no significant increases in lipid peroxides or uric acid was seen in the fillets from either group harvested using AQUI-S™. Vitamin E concentrations reduced slowly but did not change significantly despite the oxidation that was evident in the tissue. These processes also occurred in salmon tail fillets during storage at 6℃. The measurement of ATP related compounds provides an effective indicator of both the metabolic state of the tissue post-harvest and the quality. The breakdown of these compounds is also associated with the production of ammonia and hydrogen peroxide. Fresh rested salmon fillets had high concentrations of ATP and creatine phosphate, which were both depleted after 12 hours storage at 15℃. This indicates that cell viability lasted a number of hours following harvesting. These metabolites were depleted in exhausted fillets and metabolic potential appeared to be immediately compromised. The concentration of the taste enhancing compound IMP was significantly reduced in fresh rested tissue, but increased during storage, and was significantly higher than in exhausted tissues following 12 hours of storage at 15℃. This indicates that some properties of rested tissues may improve with limited storage times. The accumulation of uric acid - the metabolic end point for ATP related compounds - was also significantly reduced in rested tissue and increases in K-value were slowed. AQUI-S™ showed an ability to preserve tissue function through its anaesthetic action allowing tissue to be harvested in a rested state, and to reduce late stage lipid oxidation in stored salmon tail fillets. The antioxidant action of isoeugenol in salmon fillets may be mediated through its ability to chelate transition metals released during tissue degradation. This research shows that during reperfusion and during fillet storage there is a significant level of oxidative stress, which needs to be minimized while maintaining basic tissue metabolism to prolong tissue and cellular viability. The development of future technologies to preserve tissue viability may depend on the development of a synthetic oxygen carrying compound with properties similar to red blood cells. This may allow more control over oxygen delivery, potentially reducing the oxidative stress associated with high concentrations of free oxygen in solution. However, preserving cell viability will also require the maintenance of endogenous antioxidant function and there is also the potential to use iron chelating compounds including plant derived flavonoids to preserve non-viable tissues. Future research in these areas is necessary.
Peroxisomes perform a multitude of biosynthetic and catabolic functions, many of which are related to lipid metabolism. Peroxisomal disorders result either from deficiency of a single peroxisomal enzyme or protein, or from a defect in the complex mechanism of peroxisomal biogenesis, resulting in deficiency of several or multiple peroxisomal functions. These can be assessed by a battery of biochemical assays, enabling a biochemical phenotype to be defined that is specific and diagnostic for each of the peroxisomal disorders. Some peroxisomal disorders have unique and specific clinical phenotypes, which may be diagnostic. Others share patterns of clinical abnormalities (particularly neurological dysfunction, craniofacial dysmorphism, skeletal defects, sensory deafness, retinopathy) consistent with defined clinical phenotypes, but with considerable overlap and heterogeneity. To a certain extent, the clinical features of a particular disorder reflect the accumulation or deficiency of specific metabolites. Thus, the same clinical phenotypes may be caused by both single enzyme defects and PBDs. Furthermore, the same defect may present with different clinical phenotypes. In general, the severity of the clinical phenotype correlates with the degree of biochemical dysfunction. The clinical heterogeneity of peroxisomal disorders constitutes a diagnostic challenge demanding a high index of suspicion on the clinician's part.
Antioxidant enzymes (superoxide dismutases, catalase and glutathione peroxidase) are components of an organism's mechanisms for combating oxidative stress which is generated in normal metabolism and which may also be a reaction in response to external stimuli. This review identifies the general significance of antioxidant enzymes in health and disease, and some of the diseases that are now believed to have oxidative stress as a component. A discussion is then presented of the molecular mechanisms by which antioxidant enzyme expression is controlled at the transcriptional and post-transcriptional levels. The final sections of the review highlight the effects of exercise and hypertension on antioxidant enzyme expression in a number of different tissues, and the possibilities for future studies in these areas are discussed.
The molecular defects in the catalase gene, levels of m-RNA and properties of the residual catalase studied by scientists are reviewed in human (Japanese, Swiss and Hungarian) and non-human (mouse and beagle dog) acatalasemia with reference to the bioinformatics. Japanese acatalasemia-I, the G to A transition at the fifth position of intron 4 of the catalase gene, limited the correct splicing of the mRNA and synthesized trace catalase with normal properties. Hungarian acatalasemia type C showed a splicing mutation. In the Japanese acatalasemia II and the type A and B of Hungarian acatalasemia, the deletion or insertion of nucleotides was observed in the coding regions, and the frame shift altered downstream amino acid sequences and formed truncated proteins. In the Hungarian acatalasemia D, the substitution of a nucleotide in the exon was found. In mouse and beagle dog acatalasemia, the substitution of nucleotides in the coding regions was also observed. Studies of residual catalase in Swiss, mouse and beagle dog acatalasemia showed that aberrant catalase protein degrades more quickly than normal catalase in cells. The experimental research in genetic toxicology concerning the effect of oxidative stressors (nitrogen monoxide, nitrogen dioxide and so on) on Japanese acatalasemic blood and acatalasemic mice is described. The clinical features of Japanese and Hungarian acatalasemic subjects are also described.
Hydrogen peroxide is enzymatically processed by catalase, and catalase deficiency in blood is known as acatalasemia. We examined whether low catalase activity is a risk factor for diabetes mellitus.
Blood glucose, insulin and glucose tolerance test were examined in acatalasemic and normal mice under non-stress and oxidative stress conditions. Alloxan administration was used as oxidative stress.
Alloxan, which was a drug that caused diabetes mellitus, mostly generated hydrogen peroxide by the reaction of alloxan and reduced glutathione, in vitro. Incidence of hyperglycemia in alloxan-untreated acatalasemic mice was as low as that in the normal mice. However, the incidence of acatalasemia mice treated with alloxan was higher than that in normal mice, and the number of pancreatic beta-cells in the acatalasemic mice was less than that in normal mice.
These results indicate that low catalase activity in the blood is associated with the diabetes mellitus caused by alloxan administration.
DNA transformation of forest tree species is now a striking reality and offers the possibility to generate transgenic trees with useful new characteristics. However, it is important to make the proper environmental assessment of these transgenic trees when established in field trials. For instance, the DNA released into the soil by decaying leaves and roots from the transgenic trees may become available for incorporation by soil microbes. The objective of this study was to investigate the persistence of recombinant plant marker genes in decomposing transgenic poplar leaf material. We studied the stability of the DNA encoding the neomycin phosphotransferase II resistance marker used in tree genetic engineering. DNA persistence in the environment was determined by placing transgenic poplar leaves in permeable bags that were located on weeds, on the soil, and below the soil and left under natural conditions on the site of a field trial for up to 12 months. This work is the first quantitative analysis of tree DNA stability in a natural forest environment. Our data indicate that fragments of the genetically modified DNA are not detectable in the field for more than 4 months.
2 Katedra i Zakład Fizjologii Śląskiej AM w Katowicach, 3 Katedra i Klinika Gastroenterologii Śląskiej AM w Katowicach Abstract - Principal physiological role of catalase is to prevent H2O2 toxicity. Genetic poly- morphisms of catalase may be either the source of pathological events or by contrast, yield a protective cellular effect. Another function of catalase is participation in oxidative metabo- lism of ethanol (EtOH), however, in comparison with main enzymatic systems operating within the liver, a role of catalase seems to be marginal. On the other hand, in central nervous system the catalase is first-line EtOH metabolizing enzyme, which has an important influence on alcohol-related psychopharmacological and behavioral effects. Pharmacological manipulation of enzymatic activity opens a way to alleviation or enforcement of brain effects of EtOH.
Abstract The relevance of reactive oxygen species (ROS) production relies on the dual role shown by these molecules in aerobes. ROS are known to modulate several physiological phenomena, such as immune response, and cell growth and differentiation; on the other hand, uncontrolled ROS production may cause important tissue and cell damage, such as deoxyribonucleic acid oxidation, lipid peroxidation, and protein carbonylation. The manganese superoxide dismutase (MnSOD) antioxidant enzyme affords the major defence against ROS within the mitochondria, which is considered the main ROS production locus in aerobes. Structural and/or functional single nucleotide polymorphisms (SNP) within the MnSOD encoding gene may be relevant for ROS detoxification. Specifically, the MnSOD Ala16Val SNP has been shown to alter the enzyme localization and mitochondrial transportation, affecting the redox status balance. Oxidative stress may contribute to the development of type 2 diabetes, cardiovascular disease, various inflammatory conditions or cancer. The Ala16Val MnSOD SNP has been associated with these and other chronic diseases; however inconsistent findings between studies have made difficult drawing definitive conclusions. Environmental factors, such as dietary antioxidant intake and exercise have been shown to affect ROS metabolism through antioxidant enzyme regulation and may contribute to explain inconsistencies in the literature. Nevertheless, whether environmental factors may be associated to Ala16Val genotypes in human diseases still needs to be clarified.
Catalase (CAT) is a well-studied enzyme that plays an important role in protecting cells against the toxic effects of hydrogen peroxide. In human, it has been implicated in different physiological and pathological conditions. This review summarizes the information available on the function and role of CAT polymorphisms in pathogenesis of various pathophysiological states as well as on the regulation of CAT gene expression. Numerous studies have described the CAT polymorphisms and their link with various diseases. Changes in the CAT levels were reported in many different diseases and polymorphisms in the CAT gene were shown to be associated with different pathophysiological states, e.g. hypertension, diabetes mellitus, insulin resistance, dyslipidaemia, asthma, bone metabolism or vitiligo. Regulation of the CAT gene expression plays an important role in the levels of CAT. The catalase gene expression is regulated by various mechanisms involving e.g. peroxisome proliferator-activated receptor γ (PPARγ), tumour necrosis factor α (TNF-α), p53 protein and hypermethylation of CpG islands in the catalase promoter. Transcription of the CAT gene is mainly influenced by the -262 C/T and -844 A/G polymorphisms. A common polymorphism -262 C/T in the promoter region has been found to be associated with altered CAT activities. Apart from genetic factors, the activities of CAT may be affected by age, seasonal variations, physical activity, or a number of chemical compounds. Future investigations are necessary to elucidate the role of CAT in pathogenesis of oxidative stress-related diseases.
1. Overview of Biomarkers for Diagnosis and Monitoring of Celiac Disease
2. Cystatin C: A Kidney Function Biomarker
3. Procalcitonin: Potential Role in Diagnosis and Management
4. Manganese Superoxide Dismutase and Oxidative Stress Modulation
5. Selenium and Selenium-Dependent Antioxidants in Chronic Kidney Disease
6. Lipidomics:New Insight Into Kidney Disease
Gout is one of the most common forms of arthritis and the prevalence is increasing. Management comprises rapid and effective control of the inflammation in acute gout and sustained urate lowering in the long term. Improving the outcomes for cheaper old drugs and for the increasing number of new, more expensive agents is an important clinical goal. The role of pharmacogenetics in predicting response and adverse events to gout therapies is of considerable interest. Currently, prospective screening is employed to detect HLA-B*5801 carriage and glucose-6-phosphate dehydrogenase deficiency, to minimize occurrence of allopurinol hypersensitivity and pegloticase-related hemolytic anemia. In the future it is likely that other genetic markers of drug response will make the transition to clinical practice to further improve the efficacy and safety of gout therapies. In this review, we will examine the potential clinical relevance of specific genetic variants in the management of gout.
Mammalian erythrocytes or red blood cells (RBCs) are anucleate cells that normally circulate for several months in blood despite limited synthetic capacities and repeated exposures to mechanical and metabolic insults. Their primary purpose is to carry hemoglobin (Hb), a heme-containing protein that accounts for 95% of the total protein in RBCs. The benefits of having Hb contained within cells, as opposed to free in plasma, include the much slower turnover in blood, the metabolic capability of RBCs to maintain iron in Hb in the functional ferrous state, and the ability to control Hb oxygen affinity by altering the concentrations of organic phosphates (especially 2,3DPG). In addition, the presence of free Hb in plasma in concentrations normally found in whole blood would exert an osmotic pressure several times greater than that normally exerted by plasma proteins, profoundly affecting the movement of fluid between the vascular system and tissues.
Enzyme catalase seems to be the main regulator of hydrogen peroxide metabolism. Hydrogen peroxide at high concentrations is a toxic agent, while at low concentrations it appears to modulate some physiological processes such as signaling in cell proliferation, apoptosis, carbohydrate metabolism, and platelet activation. Benign catalase gene mutations of 5′ noncoding region (15) and intron 1 (4) have no effect on catalase activity and are not associated with disease.
Excessive hydrogen peroxide is harmful for almost all cell components, so its rapid and efficient removal is of essential importance for aerobically living organisms. Conversely, hydrogen peroxide acts as a second messenger in signal-transduction pathways. H(2)O(2) is degraded by peroxidases and catalases, the latter being able both to reduce H(2)O(2) to water and to oxidize it to molecular oxygen. Nature has evolved three protein families that are able to catalyze this dismutation at reasonable rates. Two of the protein families are heme enzymes: typical catalases and catalase-peroxidases. Typical catalases comprise the most abundant group found in Eubacteria, Archaeabacteria, Protista, Fungi, Plantae, and Animalia, whereas catalase-peroxidases are not found in plants and animals and exhibit both catalatic and peroxidatic activities. The third group is a minor bacterial protein family with a dimanganese active site called manganese catalases. Although catalyzing the same reaction (2 H(2)O(2)--> 2 H(2)O+ O(2)), the three groups differ significantly in their overall and active-site architecture and the mechanism of reaction. Here, we present an overview of the distribution, phylogeny, structure, and function of these enzymes. Additionally, we report about their physiologic role, response to oxidative stress, and about diseases related to catalase deficiency in humans.
Decomposition of hydrogen peroxide (H2O2 ) at physiological levels was studied in human erythrocytes by means of a recently developed sensitive H2O2 assay. The exponential decay of H2O2 in the presence of purified erythrocyte catalase was followed down to 10−9 mol/L H2O2 at pH 7.4. H2O2 decomposition by purified erythrocyte glutathione peroxidase (GPO) could be directly observed down to 10−7 mol/L H2O2 . No enzyme inhibition was observed at these low H2O2 concentrations. Catalase and GPO activities can be determined separately in a titrated mixture of purified enzymes, which simulates the conditions of H2O2 removal by the erythrocyte. Experiments with fresh human hemolysate allowed us to determine H2O2 decomposition by catalase and GPO using these enzymes in their original quantitative ratio. The different kinetics of these enzymes are shown: H2O2 decomposition by catalase depends linearly on H2O2 concentration, whereas that by GPO becomes saturated at concentrations above 10−6 mol/L H2O2 . Even at very low H2O2 concentrations GPO reaches only approximately 8% of the rate at which catalase simultaneously degrades H2O2 . These data indicate an almost exclusive role for catalase in the removal of H2O2 in normal human erythrocytes.
Acatalasemia was detected in 2 sisters of a Hungarian family. The pedigree of the family showed hypocatalasemia in the children of the patients and in 1 of their brothers, while the other members of the family had normal blood catalase activity. The biochemical characterization (catalase activity, electrophoretic migration, isoelectric point and enzyme stability) of the blood as well as tissue catalase of the acatalasemic patients yielded a catalase form which did not differ from normal.
A rapid, cost-efficient, spectrophotometric assay for serum catalase activity was developed. It was a combination of optimized enzymatic conditions and the spectrophotometric assay of hydrogen peroxide based on formation of its stable complex with ammonium molybdate. Lipemic and icteric sera increased the absorbance without influencing the catalase assay. Due to the high catalase activity in erythrocytes artificial hemolysis increased serum catalase activity. The imprecision of the method was CV less than 5.8% within run as well and day-to-day. The catalase assay performed using polarographic and spectrophotometric determination of hydrogen peroxide yielded a good correlation (r = 0.9602, b = 1.011, a = -0.648, n = 440). In 742 healthy individuals the mean and SD values of serum catalase were 50.5 +/- 18.1 kU/l with 17.7% higher activity in males than in females. Between 14-60 yr the serum catalase increased with age.
Superoxide dismutase is considered important in protection of aerobes against oxidant damage, and increased tolerance to oxidant stress is associated with induction of this enzyme. However, the importance of superoxide dismutase in this tolerance is not clear because conditions which promote the synthesis of superoxide dismutase likewise affect other antioxidant enzymes and substances. To clarify the role of superoxide dismutase per se in organismal defense against oxidant-generating drugs, we employed Escherichia coli transformed with multiple copies of the gene for bacterial iron superoxide dismutase. These bacteria have greater than ten times the superoxide dismutase activity of wild-type E. coli but, importantly, are normal in other oxidant defense parameters including catalase, peroxidases, glutathione, and glutathione reductase. High superoxide dismutase and control bacteria were exposed to the O2- -generating drug paraquat and to elevated pO2. We find; high superoxide dismutase E. coli are more readily killed by paraquat under aerobic, but not anaerobic, conditions. During exposure to paraquat, high superoxide dismutase E. coli accumulate more H2O2. Coincidentally, the reduced glutathione content of high superoxide dismutase E. coli declines more than in control E. coli. E. coli with high superoxide dismutase activity are also more readily killed by hyperoxia. Interestingly, the susceptibility of the parental and high superoxide dismutase E. coli to killing by exogenous H2O2 is not significantly different. Thus, under these experimental conditions, greatly enhanced superoxide dismutase activity accelerates H2O2 formation. The increased H2O2 probably accounts for the exaggerated sensitivity of high superoxide dismutase bacteria to oxidant-generating drugs. These results support the concept that the product of superoxide dismutase, H2O2, is at least as hazardous as the substrate, O2-. We conclude that effective organismal defense against reactive oxygen species may require balanced increments in antioxidant enzymes and cannot necessarily be improved by increases in the activity of single enzymes.
An Hinf1 associated restriction length polymorphism pattern is reported for the catalase gene of Hungarian normocatalasemic individuals and acatalasemic patients. The 2.4-kb pCAT 10 probe revealed 9 bands (2.1, 1.5, 1.2, 1.1, 0.9, 0.8, 0.6, 0.5 and 0.4 kb) with 9 distinct patterns for the controls. The same patterns were detected for the Hungarian acatalasemic patients. The examination of the A to T mutation of the Hungarian acatalasemic patients and their relatives at position -21 in the flanking region with Hinf1 polymorphism could not reveal any difference between the acatalasemic and the normocatalasemic catalase gene.
Purified enzymes were mixed to form a cell-free system that simulated the conditions for removal of hydrogen peroxide within human erythrocytes. Human glutathione peroxidase disposed of hydrogen peroxide (H2O2) at a rate that was only 17% of the rate at which human catalase simultaneously removed hydrogen peroxide. The relative rates observed were in agreement with the relative rates predicted from the kinetic constants of the two enzymes. These results confirm two earlier studies on intact erythrocytes, which refuted the notion that glutathione peroxidase is the primary enzyme for removal of hydrogen peroxide within erythrocytes. The present findings differ from the results with intact cells, however, in showing that glutathione peroxidase accounts for even less than 50% of the removal of hydrogen peroxide. A means is proposed for calculating the relative contribution of glutathione peroxidase and catalase in other cells and species. The present results raise the possibility that the major function of glutathione peroxidase may be the disposal of organic peroxides rather than the removal of hydrogen peroxide.
Decomposition of hydrogen peroxide (H2O2 ) at physiological levels was studied in human erythrocytes by means of a recently developed sensitive H2O2 assay. The exponential decay of H2O2 in the presence of purified erythrocyte catalase was followed down to 10(-9) mol/L H2O2 at pH 7.4. H2O2 decomposition by purified erythrocyte glutathione peroxidase (GPO) could be directly observed down to 10(-7) mol/L H2O2 . No enzyme inhibition was observed at these low H2O2 concentrations. Catalase and GPO activities can be determined separately in a titrated mixture of purified enzymes, which simulates the conditions of H2O2 removal by the erythrocyte. Experiments with fresh human hemolysate allowed us to determine H2O2 decomposition by catalase and GPO using these enzymes in their original quantitative ratio. The different kinetics of these enzymes are shown: H2O2 decomposition by catalase depends linearly on H2O2 concentration, whereas that by GPO becomes saturated at concentrations above 10(-6) mol/L H2O2. Even at very low H2O2 concentrations GPO reaches only approximately 8% of the rate at which catalase simultaneously degrades H2O2. These data indicate an almost exclusive role for catalase in the removal of H2O2 in normal human erythrocytes.
Acatalasemia, a deficiency of enzyme catalase, is an autosomal recessive syndrome with an incidence of 5:106 in Hungary. We have examined the first Hungarian acatalasemic family for the disease-causing mutation. All exons of the catalase gene were screened by PCR-SSCP, PCR-heteroduplex, and nucleotide sequence analysis. The heteroduplex formation detected in exon 2 was verified by nucleotide sequence analysis. We found a GA insertion at nucleotide position 138, increasing the GA repeat number from 4 to 5. This GA insertion caused a frameshift in the amino acid sequence from position 68 to 133 and generated a TGA terminating codon at amino acid position 134. This truncated protein lacks the essential amino acid (histidine 74) in the active center. This finding can explain the decreased blood catalase activity in the Hungarian acatalasemic family.
Partial or near-total lack of erythrocyte catalase activity is a rare condition, generally thought to be benign. However, little is known of the frequency of common diseases of adult onset in human beings with catalase deficiency. We report that, in a series of Hungarian patients with catalase deficiency, there is a higher frequency of diabetes than in unaffected first-degree relatives and the general Hungarian population. We speculate that quantitative deficiency of catalase might predispose to cumulative oxidant damage of pancreatic beta-cells and diabetes.
Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) screening was used for searching mutations of the catalase gene in two Hungarian hypocatalasemic families. A syndrome-causing mutation was found in a PCR product containing exon 7 and its boundaries. Nucleotide sequence analyses detected a G to T substitution at position 5 of intron 7. The effect of this splice site mutation was confirmed by Western blot analyses demonstrating a decreased catalase protein level in these patients. These findings represent a novel type (C) of catalase mutations in the Hungarian acatalasemic/hypocatalasemic patients.
The human hereditary disease Acatalasia (AC) is characterized by low or no catalase activity in all body tissues. We have studied the molecular basis of AC. In order to assess their antioxidant defense status we measured the enzyme activities, protein levels and m-RNA concentrations of catalase, superoxide dismutase and glutathione peroxidase in fibroblasts from a Japanese (AC65) and a Swiss (AC64) patient and several normal individuals. Our results point to genetic heterogeneity. While strain AC64 contained normal levels of catalase mRNA and -protein, strain AC65 was completely devoid of both. A structural mutation in the catalase gene is probably responsible for the inactivation of the enzyme in AC64. Since AC65 contains at least a major portion of the catalase gene it may represent a regulatory mutation in which the gene is not transcribed.
1. With a view to grasp more simply and clearly the characteristics of this disease and in order to find a clue for prompt discovery of cases when encountered in future, the authors undertook a statistical study of the cases already reported by various authors. 2. The cases reported so far amount to 17 familial groups which consisted of 38 acatalasemic cases. These groups were distributed widely throughout Japan. The disease seemed to be prevalent in the rural communities where adherence to the custom of consanguineous marriage occurs. As yet, we have not heard of the occurrence of this disease in other countries. 3. The disease has equal distribution in both sexes. About one half of patients showed a peculiar oral gangrene (Takahara's disease). The great majority of these were noted in those less than 10 years of age. 4. The great majority of them were children whose parents were united in consanguineous marriage and have siblings with acatalasemia. 5. As for the treatment of oral lesions in this disease, extraction of tooth at the site of the lesions, removal of the diseased tissues en masse by resection, and penicillin treatment given concomitantly are effective. The course and the length of time required in healing of the wound due to the operation are about the same as in the case of normal persons. 6. The authors wish to call special attention to the phenomenon peculiar to the acatalasemic blood. The blood of acatalasemic individuals changes to brownish-black color in the absence of foaming or bubble formation upon the application of hydrogen peroxide to blood.
Acatalasemia was one of the earliest described genetic enzyme defects. In 1990, a causal point mutation (a splicing mutation) was first reported in a Japanese patient with acatalasemia. In the present study, the polymerase chain reaction and single-strand conformation polymorphism analysis were used to determine whether the same point mutation was present in unrelated Japanese patients. The subjects studied were the previously examined acatalasemic female, her brother, who is hypocatalasemic, and two other unrelated acatalasemic patients. A single G to A point mutation at the fifth position of intron 4, identical to that previously found, was present in all the studied patients. This finding strongly suggests that only a single mutated allele has spread in the Japanese population.
The abnormalities in acatalasemia at the gene level as well as properties of the residual catalase in Japanese acatalasemia are historically reviewed. The replacement of the fifth nucleic acid, guanine, in the fourth intron by adenine in the acatalasemic gene causes a splicing mutation and hence a deficiency of mRNA. The guanine-to-adenine substitution was detected in two Japanese acatalasemic cases from different families. The properties of the residual catalase are similar to those of normal catalase; the exons are identical. The properties of the residual catalase and the molecular defect in the catalase gene are compared among Japanese, Swiss, and mouse acatalasemias. The physiological role of catalase, as judged from human acatalasemic blood and acatalasemic mice, is also described.
To search for the molecular defect of Japanese-type acatalasemia, we cloned the mutant catalase gene from a person with this deficiency. The nucleotide sequence of the mutant gene was determined for all exons, exon/intron junctions, and 5' and 3' flanking regions, and the findings were compared with the sequence from the normal gene. Seven base differences were found between the two genes. Among them, a G to A substitution at the fifth position of intron 4 (a splicing mutation) seemed most likely to be responsible for the defective catalase synthesis in the subject. To obtain suggestive evidence, we constructed chimeric genes that contained a segment of either the normal or mutant catalase gene, encompassing a 3' part of exon 4, the entire intron 4 and a 5' portion of exon 5, within the third exon of the human alpha-globin gene. When this chimeric gene construct was introduced into simian virus 40-transformed simian cells (COS-7), the transcript of the normal catalase/alpha-globin chimeric gene was spliced correctly, as revealed by Northern blotting and RNase mapping techniques. In contrast, the splicing of the mutant chimeric pre-mRNA occurred between the 5' donor site of the preceding intron and the 3' acceptor site of the intron containing the substitution, thereby skipping one entire exon sequence. Thus, the G to A transition at the fifth position of intron 4 of the catalase gene indeed severely limits the correct splicing of the RNA product. The same splice site mutation was found in the genomic DNA of another acatalasemic individual from an unrelated family. We suggest that this base substitution is the causal mutation of these cases of Japanese-type acatalasemia.
Five Swiss subjects with hereditary acatalasemia (4 homo-, 1 heterozygous) were studied by a series of neutrophil function tests. H2O2 was added to a polymorphonuclear neutrophil leukocyte-suspension to produce a metabolic stress; neutrophil functions related to membrane deformation were subsequently found to be depressed, i.e., chemotaxis, membrane potential, and chemiluminescence. This mechanism might be one pathogenetic factor in the formation of mucosal ulcers in acatalasemic individuals.
The subjects of this study were a hereditary typical acatalasemia and a hereditary atypical acatalasemia. In the latter case, the children did not show hypocatala-semia but one of the cousins proved to be hypocatalasemia. The abdominal muscle and the vermiform appendix from the case of typical acatalasemia had only a minute catalase activity. The liver from the case of atypical acatalasemia indicated a moderate catalase activity and catalase protein was observed in it.
Two Hungarian acatalasemic and eight hypocatalasemic patients revealed normal erythropoesis. Contrary to their decreased defence system against the toxic hydrogen peroxide, the biochemical tests (serum catalase, serum hemoglobin, serum lactate dehydrogenase (LDH) ratio of LDH1 and LDH2 isoenzymes and serum haptoglobin) excluded hemolysis. The normal activity of glutathione peroxidase and the decreased catalase activity could prevent the lysis of the erythrocytes. In the presence of extremely high levels of hydrogen peroxide acute hemolysis may not be excluded; therefore, follow-up of these patients is required.
Japanese-type acatalasemia is characterized by the almost total loss of catalase activity in red cells and is often associated with ulcerating oral lesions. A splicing mutation in intron 4 of catalase gene has so far been a sole disease-causing mutation found in Japanese-type acatalasemic patients. We report here a novel single base deletion in the catalase gene causing Japanese-type acatalasemia. The patient was a 72 year-old Japanese male. His maternal grandmother and his father were first cousins. Molecular analysis using non-RI PCR-SSCP analysis combined with direct sequencing revealed a deletion of the 358th thymine in exon 4 of the patient's catalase gene. The proband was a homozygote and his mother and his three children were heterozygotes for this mutation. The frame shift caused by the nucleotide deletion should alter the downstream amino acid sequence and introduce a new termination codon TGA 43 bp 3' to the mutation. Although the truncated peptide chain consisted of 133 amino acid residues might be translated in the patient's tissue, such an aberrant protein is expected to be extremely unstable and have no catalytic function at all. Our results suggest that Japanese-type acatalasemia is heterogeneous.
203 bp long products containing exon 4 and its junctions from the catalase gene were generated by polymerase chain reaction (PCR). These products were analyzed by single strand conformational polymorphism (SSCP), hetero-duplex formation and nucleotide sequencing. No polymorphism was detected when the Hungarian acatalasemic sisters, their family members and normocatalasemic controls were analyzed. Sequence analyses did not show the G to A point mutation at position 5 of intron 4. This splicing mutation characterizes the Japanese-type of acatalasemia.
In 1756 healthy individuals the mean and S.D. values of blood catalase activity were 111.3 +/- 16.5 MU/l with lower blood catalase for females (107.7 +/- 14.4 MU/l, n = 880) than for males (117.9 +/- 16.8 MU/l, n = 876) while the ratios of blood catalase activity to blood hemoglobin concentration were not different (0.841 +/- 0.107 MU/g versus 0.849 +/- 0.119 MU/g). The decrease of blood catalase with age was greater in males (b = -0.084 MU/l year) than in females (b = -0.016 MU/l year). The screening of 3300 healthy citizens for hypocatalasemia yielded six families (0.18%), and three families were identified out of 1630 clinic patients. These nine families revealed 37 hypocatalasemic patients with 57.5 +/- 11.7 MU/l mean and S.D. of blood catalase activity. Similarly to the Japanese and the Hungarian actalasemic patients, the electrophoretic mobilities of catalase in erythrocytes of hypocatalasemic patients were indistinguishable from that of healthy controls.
The amplified fragment length polymorphism of Hinf1 on the promoter region of the catalase gene in Hungarian acatalasemic and hypocatalasemic patients yielded three different patterns with five bands in total. The sequence analyses revealed A-to-T, C-to-A, and C-to-T mutations at positions -21, -20, and -18 upstream of the translational initiation site. The -21 A-to-T mutations were more frequent in acatalasemic and hypocatalasemic patients (36/2) than in controls (18/14). This mutation had been detected in Japanese acatalasemic patients while the other two are novel mutations. Two extra bands in the Hinf1 pattern are due to star-like activity that cleaved a G/ATTT sequence at position -4 to 0 upstream of the initiation site.
A T-deletion at position 10 of exon 4 for catalase gene was reported as a novel mutation, causing a new genetic type of acatalasemia in Japan. This mutation, destroying a Hinf1 recognition site, was searched for in Hungarian acatalasemic (2) and hypocatalasemic (22) patients and in controls (27) by Hinf1 digestion and sequence analyses of a 203 bp polymerase chain reaction (PCR) product containing the entire exon 4. The Hinf1 polymorphism did not reveal any difference between controls and hypocatalasemic as well as acatalasemic patients. These results were confirmed by sequence analyses showing the T nucleotide for the two acatalasemic and for one unrelated hypocatalasemic patient, as well as for two controls. These findings represent further evidence that acatalasemia is heterogeneous at the DNA level.
The 5' uncoding region (165 bp), exon 1 (63 bp) and part of intron 1 (20 bp) of the catalase gene was amplified by PCR in acatalasemic (2), hypocatalasemic (19) patients and healthy individuals (10). The single strand conformational polymorphism of PCR products showed a highly polymorphic pattern. This polymorphism was supported by nucleotide sequence analyses yielding eight mutations. They are A to T, C to A and C to T at positions -21, -20, -18 of the 5' flanking region, T to C at positions 4, 44, 49 of the non-coding region and C to T and C to A at positions 12, 27 of exon 1. Of these nucleotide substitutions, the fourth, fifth, seventh and eighth are novel mutations. The mutations 1, 3, 6, 8 were present at least at heterozygous level in all acatalasemics and hypocatalasemics. None of these mutations may be the causal mutation(s) of acatalasemia as each of these nucleotide substitutions were detected in healthy subjects with normal blood catalase activity.
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