The effect of Nigella sativa on the measures of liver and kidney parameters: A systematic review and meta-analysis of randomized-controlled trials.

The aim of this systematic review and dose-response meta-analysis was to determine the effect of Nigella sativa (N.S) supplementation on liver and kidney parameters. We searched PubMed, Scopus, ISI Web of Science, Cochrane central register for controlled trials and Google Scholar from database inception to April 2019 for relevant controlled trials. Mean differences and standard deviations for each outcome were pooled using a random-effects model and a dose-response analysis was performed using a fractional polynomial model. Quality of evidence was evaluated using Cochrane Collaboration Risk of Bias tool and the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology. Nineteen trials (n = 1295 participants) were included in the meta-analysis. We observed that N.S supplements had significant reducing effects on alkaline-phosphatase (ALP) [9 trials, n = 710 participants, weighted mean difference (WMD)= -10.825; 95 %CI: -19.658, -1.992 U/L; P = 0.016; I2 = 75.7 %; P-heterogeneity = 0.000) and blood urea nitrogen (BUN) (12 trials, n = 821 participants, WMD= -1.016; 95 % CI: -1.760, -0.273 U/L; P = 0.007; I2 = 87.7 %; P-heterogeneity = 0.000) concentrations. Subgroup analysis showed that, an intervention of more than 12 weeks was found to have a reducing effect on aspartate- aminotransferase (AST) measures (2 trials, n = 201 participants, WMD= -11.317; 95 % CI: -15.007, -7.626; P = 0.000; I2 = 0.0 %; P-heterogeneity = 0.977). Creatinine levels increased significantly in studies that considered adjusted analysis based on covariates (3 trials, n = 152 participants, WMD = 0.070; 95 % CI: 0.027, 0.112 U/L; P = 0.001; I2 = 0.0 %; P-heterogeneity = 0.788). A daily dose of 1100-1500 mg of N.S supplements was observed to have a substantial reducing effect on ALP levels (5 trials, n = 340 participants, WMD= -11.323; 95 % CI: -21.418, -1.229 U/L; P = 0.028; I2 = 0.00 %; P-heterogeneity = 0.686), while a dosage of more than 2000 mg per day led to a significant increase in BUN concentrations (2 trials, n = 101 participants, WMD= -1.016; 95 % CI: -1.760, -0.273 U/L; P = 0.007; I2 = 87.7 %; P-heterogeneity = 0.000). Our data suggested that N.S supplementation had significant impacts on liver and kidney parameters leading to a decrease in ALP and BUN levels. Longer duration of intervention and normal daily dosages of N.S supplements led to significant reductions in ALP and AST concentrations, respectively, while higher daily dosages increased BUN levels. Hence, in spite of favorable impacts of N.S supplements on liver and kidney parameters, due to the herbal nature of N.S, more studies with high-quality, large-scale, long-term intervention and precise baseline characteristics are needed to assess the exact effective dose, duration and efficacy of N.S supplementation on kidney and liver parameters.

Drug Induced Liver Injury at a Tertiary Hospital in India: Etiology, Clinical Features and Predictors of Mortality.

INTRODUCTION AND AIMS: Drug-induced liver injury (DILI) is rare; however, it is one of the important causes of acute liver failure which results in significant morbidity or mortality. MATERIAL AND METHODS: Patients with suspected DILI were enrolled based on predefined criteria and followed up for at least 6 months or until normalization of liver tests. Causality assessment was done by applying the Roussel Uclaf Causality Assessment Method model. RESULTS: We collected data from 82 individuals diagnosed with DILI at our hospital from 2014 through 2015 (41 men; median age, 38 years). The most commonly implicated drugs were antitubercular therapy (ATT) (49%), antiepileptic drugs (12%), complementary and alternative medicine (CAM) in 10%, antiretroviral drugs (9%) and non-steroidal anti-inflammatory drugs (6%). 8 out of 13 deaths were liver related. Also, liver related mortality was significantly higher for ATT DILI (17.5%) vs. those without (2.4%) (P = 0.02). There was no significant difference in overall as well as liver related mortality in hepatocellular, cholestatic or mixed pattern of injury. Laboratory parameters at one week after discontinuation of drug predicted mortality better than those at the time of DILI recognition. On multivariate logistic regression analysis, jaundice, encephalopathy, MELD (Model for end stage liver disease) score and alkaline phosphatase at one week, independently predicted mortality. CONCLUSION: DILI results in significant overall mortality (15.85%). ATT, anti-epileptic drugs, CAM and antiretroviral drugs are leading causes of DILI in India. Presence of jaundice, encephalopathy, MELD score and alkaline phosphatase at one week are independent predictors of mortality.

Moderate doses of commercial preparations of Ginkgo biloba do not alter markers of liver function but moderate alcohol intake does: A new approach to identify and quantify biomarkers of 'adverse effects' of dietary supplements.

It is difficult to determine if certain dietary supplements are safe for human consumption. Extracts of leaves of Ginkgo biloba trees are dietary supplements used for various purported therapeutic benefits. However, recent studies reported they increased risk of liver cancer in rodents. Therefore, this study assessed the association between ginkgo consumption and liver function using NHANES 2001-2012 data (N = 29,684). Since alcohol is known to adversely affect liver function, association of its consumption with liver function was also assessed. Alcohol and ginkgo extract intake of adult consumers and clinical markers of liver function (alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, gamma glutamyl transferase, lactate dehydrogenase, bilirubin) were examined. Moderate consumers of alcohol (0.80 ± 0.02 drinks/day) had higher levels of aspartate aminotransferase and gamma glutamyl transferase than non-consumers (P < 0.001). There was no difference (P > 0.01) in levels of markers of liver function in 616 ginkgo consumers (65.1 ± 4.4 mg/day intake) compared to non-consumers. While moderate alcohol consumption was associated with changes in markers of liver function, ginkgo intake as typically consumed by U.S. adults was not associated with these markers. Biomarkers measured by NHANES may be useful to examine potential adverse effects of dietary supplements for which insufficient human adverse event and toxicity data are available. TRIAL REGISTRATION NUMBER: Not applicable, as this is secondary analysis of publicly released observational data (NHANES 2001-2012).

Strong Negative Interference by Calcium Dobesilate in Sarcosine Oxidase Assays for Serum Creatinine Involving the Trinder Reaction.

The vasoprotective drug calcium dobesilate is known to interfere with creatinine (Cr) quantifications in sarcosine oxidase enzymatic (SOE) assays. The aim of this study was to investigate this interference in 8 different commercially available assays and to determine its clinical significance. In in vitro experiments, interference was evaluated at 3 Cr levels. For this, Cr was quantified by SOE assays in pooled serum supplemented with calcium dobesilate at final concentrations of 0, 2, 4, 8, 16, 32, and 64 µg/mL. Percent bias was calculated relative to the drug-free specimen. For in vivo analyses, changes in serum concentrations of Cr, cystatin C (CysC; a renal function marker), and calcium dobesilate were monitored in healthy participants of group I before and after oral calcium dobesilate administration. In addition, variations in interference were also examined among different SOE assays using serum obtained from healthy participants of group II. Lastly, Cr levels from the 10 patients treated with calcium dobesilate were measured using 4 SOE assays and liquid chromatography-isotope dilution tandem mass spectrometry (LC-IDMS/MS) for comparison. Our in vitro analyses indicated that the presence of 8 µg/mL calcium dobesilate resulted in a -4.4% to -36.3% reduction in Cr serum concentration compared to drug-free serum for 8 SOE assays examined. In vivo, Cr values decreased relative to the baseline level with increasing drug concentration, with the lowest Cr levels obtained at 2 or 3 hours after drug administration in participants of group I. The observed Cr concentrations for participants in group II were reduced by -28.5% to -3.1% and -60.5% to -11.6% at 0 and 2 hours after administration related to baseline levels. The Cr values of 10 patients measured by Roche, Beckman, Maker, and Merit Choice SOE assays showed an average deviation of -20.0%, -22.4%, -14.2%, and -29.6%, respectively, compared to values obtained by LC-IDMS/MS. These results revealed a clinically significant negative interference with calcium dobesilate in all sarcosine oxidase-based Cr assays, but the degree of interference varied greatly among the assays examined. Thus, extra care should be taken in evaluating Cr quantification obtained by SOE assays in patients undergoing calcium dobesilate therapy.

Impacts of common factors of life style on serum liver enzymes.

AIM: To investigate the impacts of gender, age and factors of life style (alcohol, overweight, coffee and smoking) on serum liver enzymes. METHODS: Serum alanine aminotransferase (ALT) and gamma-glutamyltransferase (GGT) were measured from 6269 apparently healthy individuals (2851 men, 3418 women, mean age 45 ± 12 years, range 25-74 years) in a national cross-sectional health survey. All subjects underwent detailed clinical examinations and interviews including the amount and pattern of alcohol use, coffee consumption and smoking habits. RESULTS: In this population with a mean ± SD alcohol consumption of 65 ± 105 g/wk and body mass index (BMI) of 26.1 ± 4.3 kg/m(2), both ALT and GGT were significantly influenced by alcohol use (P < 0.001) and BMI (P < 0.001), whereas smoking increased only GGT (P < 0.001). A significant effect of age on ALT was seen in men (P < 0.001) whereas not in women. Significant two-factor interactions of alcohol use in men were observed with age (ALT: P < 0.01; GGT: P < 0.001) and BMI (GGT: P < 0.05). For ALT, a significant interaction also occurred between BMI and age (P < 0.005). In contrast, women showed significant interactions of alcohol use with BMI (GGT: P < 0.05), smoking (GGT: P < 0.001), and coffee consumption (GGT: P < 0.001). CONCLUSION: Life-style associated changes in liver enzymes may reflect health risks, which should be considered in the definition of normal limits for liver enzymes.

Analysis of early biochemical markers and regulation by tin protoporphyrin IX in a model of spontaneous osteoarthritis.

Age-related changes in joint tissues lead to osteoarthritis (OA). Detection of early changes in OA patients may help to initiate treatments before the establishment of irreversible joint destruction. STR/ort mice develop with age a severe degenerative joint disease that resembles human OA thus allowing the investigation of biochemical markers as well as new treatments in an accelerated time frame. We have analyzed the changes in serum levels of different mediators during the early phases of idiopathic OA in STR/ort mice. Serum levels of matrix metalloproteinase-3 (MMP-3) but not those of tumor necrosis factor-α, interleukin(IL)-1ß, IL-17 or prostaglandin E(2) correlated with histopathological changes in knees of STR/ort mice at 9 weeks. Treatment of animals with tin protoporphyrin IX (SnPP, 12 mg/kg/dayi.p.) for 4 weeks significantly reduced the progression of OA. Our data suggest that MMP-3 is a sensitive biomarker to detect early OA alterations and that SnPP could be a protective agent in OA.

Determination of trace elements in patients with chronic hepatitis B.

Chronic hepatitis B virus (HBV) infection is a major liver disease worldwide and its clinical manifestations are linked to immune response. The purpose of this study was to evaluate the relationship between selenium, copper, and zinc in comparison with transaminase level in chronic HBV patients. Serum samples of the HBV infected patients were obtained from Tooba medical center, Sari, Iran. Sixty patients were enrolled in this study (36 men and 24 women), mean age: 39.6 ± 12.2 years. The concentration of zinc, selenium, copper and transaminases were determined using an autoanalyzer system. Concentrations of selenium (0.273 ± 0.056 µg/dl) and zinc (2.1 ± 0.037) was elevated in patients with low transaminase levels as were significantly different in comparison with patients with high transaminase level (P<0.05). Serum copper concentration was similar in two groups of patients. Elevated levels of transaminase concentrations were independently associated with low zinc and selenium concentrations in chronic HBV patients. It is concluded that serum zinc and selenium levels are associated with less hepatic damage in chronic HBV patients and might have a protective role during liver injury.

Ação de fatores enzimáticos na obtenção de hidrolisados protéicos de farinha de trigo com baixo teor de fenilalanina / Action of enzymatic factors in obtaining protein hydrolysates of wheat flour, low-phenylalanine

Visando ao preparo de farinha de trigo com baixo teor de fenilalanina (Phe), extraiu-se, enzimaticamente as proteínas, empregando-se uma protease alcalina de Bacillus licheniformis. Em seguida, os extratos protéicos foram hidrolisados pela ação de enzimas comerciais (pancreatina e bromelina) e de extratos enzimáticos obtidos da casca de abacaxi (bruto e purificado), avaliando-se alguns parâmetros enzimáticos, tais como tipo de enzima, tipo de ação enzimática, tipo de associação enzimática e ordem de ação enzimática. O carvão ativado (CA) foi empregado como meio adsorvente e a eficiência da remoção de Phe foi avaliada por espectrofotometria derivada segunda, determinando-se o teor de Phe na farinha de trigo e em seus hidrolisados, após tratamento com CA. O melhor resultado foi encontrado ao se empregar a associação sucessiva do extrato bruto seguida da pancreatina, tendo atingido 66,28 por cento de remoção e o teor final de Phe de 522,44 mg/100 g de hidrolisado.

With the aim of producing wheat flour with low phenylalanine (Phe) content to be introduced in phenylketonuric's diet, the proteins were enzymaticaly extracted, using an alkaline protease from Bacillus licheniformis. Then, the protein extracts were hydrolyzed by the action of commercial enzymes (pancreatin and bromelain) and of enzymatic extracts obtained from pineapple peel (crude and purified). Some enzymatic parameters were evaluated, such as type of enzyme, type of enzyme action, type of enzymatic association and order of enzyme action. The activated carbon (AC) was used as adsorbent and the efficiency of Phe removal was evaluated by second derivative spectrophotometry measuring the Phe content in wheat flour and in their hydrolysates after AC treatment. The best result was found for the successive association of crude extract followed by pancreatin obtaining 66.28 percent of removal and a final Phe content of 522.44 mg/100 g of hydrolysate.

Interaction of glutathione peroxidase-1 and selenium in endemic dilated cardiomyopathy.

BACKGROUND: Keshan disease (KD) is a fatal dilated cardiomyopathy with unknown etiology. We studied the gene-environment interaction in the pathogenesis of KD by assessing the association of low blood selenium and polymorphisms in glutathione peroxidase-1 (GPx-1) gene. METHODS: The concentration of blood selenium and the activity and polymorphisms of GPx-1 in 71KD patients and 290 controls were measured. The functions of rat neonatal cardiomyocytes resulting from overexpression of 2 variants of GPx-1 were studied. RESULTS: Blood concentration of selenium and GPx-1 activity were lower in patients than in controls. Genetic analysis revealed a single nucleotide polymorphism (Pro198Leu) in GPx-1 gene associated with selenium deficiency as well as impaired GPx-1 activity. Gene-environment interaction analysis revealed a synergistic-multiplicative interaction between polymorphism of GPx-1 and selenium deficiency. Overexpression of the GPx-1 leucine-containing allele in cultured cardiomyocytes caused a 30% reduction in selenium-induced GPx-1 activity and increased serum starvation induced apoptosis as compared with that of the wild-type variant 198Pro. CONCLUSION: Selenium deficiency in carriers with the GPx-1 leucine-containing allele is associated with low GPx-1 enzyme activity, which may, in turn, increase the incidence of KD. Results from this unique disease may have broad implications for a gene-environment reaction in the etiology of other diseases.

Ensaios clínicos com as folhas de Cissus sicyoides I. (vitaceae) em pacientes intolerantes à glicose e em diabéticas tipo 2 / Clinical trials with the leaves of Cissus sicyoides I. (vitaceae) in glucose-intolerant and in type 2

A Cissus sicyoides é popularmente empregada para o tratamento do diabetes e é conhecida como insulina vegetal. Realizaram-se ensaios clínicos de fase II com o infuso das folhas da referida planta, para investigar a eficácia terapêutica, desse vegetal, em voluntárias intolerantes à glicose (GIG n=14) e diabéticas tipo 2 (GD n=12). Preparou-se o chá com 1g do pó das folhas secas, diluído em 150 mL de água quente por 10 minutos (uso popular), dose única. A atividade foi avaliada nos tempos basal, 30° e 60° dias. Aplicou-se questionário para reações adversas e realizaram-se exames clínicos e laboratoriais no Hospital Universitário Lauro Wanderley/UFPB. Os dados foram analisados estatisticamente e o nível de significância foi de 5,0 %. Observou-se que: as voluntárias não exerciam atividade física, possuíam parentes com diabetes, eram hipertensas e obesas; o IMC do GD reduziu significativamente com 60 dias, entre os grupos. Concluiu-se que, nesse estudo, não foi evidenciado efeito hipoglicemiante significativo, nas voluntárias, e que nas doses utilizadas a Cissus sicyoides não causou alterações clínicas e laboratoriais confirmando a segurança da utilização da mesma, como alimento, pela população.

Protective effect of pinitol against D-galactosamine-induced hepatotoxicity in rats fed on a high-fat diet.

The protective effect of pinitol against D-galactosamine (GalN)-induced liver damage was examined. Forty male Sprague-Dawley rats were divided into normal control, GalN control, and pinitol groups (0.5%, 1%, and 2%). After 8 weeks of feeding, a single dose of GalN (650 mg/kg) was administered 24 h before their sacrifice. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and tumor necrosis factor-alpha (TNF-alpha) levels were significantly increased after an injection with GalN (P<0.05), but pinitol supplementation at the level of 0.5% reversed these changes to normal levels. Significant decreases in serum triglyceride and cholesterol and increases in hepatic cholesterol were observed in GalN-intoxicated rats. However, supplementation with pinitol significantly attenuated these trends. In addition, pinitol elevated the Mn-superoxide dismutase, glutathione reductase, and catalase activities, prevented hepatic lipid peroxidation, and restored the hepatic GSH levels and cytochrome P450 2E1 function. Thus, 0.5% pinitol supplementation protected the rats from the hepatotoxicity induced by GalN, at least part of its effect being attributable to attenuation of the oxidative stress and inflammatory process promoted by GalN.

Cuticle-degrading proteases produced by the entomopathogenic fungus Beauveria bassiana in the presence ofcoffee berry borer cuticle / Proteases degradadoras de cutícula produzidas pelo fungo entomopatogênico Beauveria bassiana em presença de cutícula de broca do café

A Brazilian isolate of Beauveria bassiana (CG425) that shows high virulence against the coffee berry borer (CBB) was examined for the production of subtilisin-like (Pr1) and trypsin-like (Pr2) cuticle-degrading proteases. Fungal growth was either in nitrate-medium or in CBB cuticle-containing medium under both buffered and unbuffered conditions. In unbuffered medium supplemented with cuticle, the pH of cultures dropped and Pr1 and Pr2 activities were detected in high amounts only at a pH of 5.5 or higher. In buffered cultures, Pr1 and Pr2 activities were higher in medium supplemented with cuticle compared to activities with nitrate-medium. The Pr1 and Pr2 activities detected were mostly in the culture supernatant. These data suggest that Pr1 and Pr2 proteases produced by strain CG425 are induced by components of CBB cuticle, and that the culture pH influences the expression of these proteases, indicating the occurrence of an efficient mechanism of protein secretion in this fungus. The results obtained in this study extend the knowledge about protease production in B. bassiana CG425, opening new avenues for studying the role of secreted proteases in virulence against the coffee berry borer during the infection process.

O isolado brasileiro de Beauveria bassiana (CG425) que apresenta alta virulência contra a broca do café (CBB) foi analisado quanto à produção de proteases degradadoras de cutícula, tipo-subtilisina (Pr1) e tipo-tripsina (Pr2). O crescimento fúngico foi realizado em meio contendo nitrato e em meio contendo cutícula da broca em condições de pH tamponado e não tamponado. Em meio não tamponado, suplementado com cutícula, o pH da cultura caiu e as atividades de Pr1 e Pr2 foram detectadas somente em valores de pH igual ou superior a 5,5. Em culturas tamponadas, as atividades Pr1 e Pr2 foram superiores em meio suplementado com cutícula, comparativamente as atividades em meio contendo nitrato. As atividades Pr1 e Pr2 ocorreram predominantemente no sobrenadante de cultivo. Os dados obtidos sugerem que Pr1 e Pr2 produzidas pelo isolado CG425 são induzidas por componentes da cutícula da broca do café (CBB), e que o pH da cultura influencia a expressão destas proteases, indicando a ocorrência de um mecanismo eficiente de secreção por este fungo. Os resultados obtidos neste estudo aumentam o conhecimento a respeito da produção de proteases por B. bassiana CG425, abrindo novos caminhos para o estudo do papel de proteases na virulência contra a broca do café durante o processo de infecção.

Characterization of a highly effective protein substrate for analysis of JAK2(V617F) Activity.

OBJECTIVE: Identification of JAK2V617F in myeloproliferative disorders makes JAK2 an important marker for disease diagnosis and a highly attractive target for therapeutic drug development. This study is intended to identify a sensitive and specific substrate for assays of the JAK2 enzymatic activity. METHODS: We expressed a glutathione S-transferase (GST) fusion protein designated GST-JAKS, which carries a peptide sequence derived from the autophosphorylation sites of human JAK2. The protein was purified from Escherichia coli cells and was used to analyze to tyrosine kinase activities of purified enzymes and crude cell extracts from cells, including mononuclear cells of JAK2V617F -positive polycythemia vera blood. It was also used to perform JAK2 kinase assays to screen inhibitors of JAK2. RESULTS: GST-JAKS is strongly phosphorylated by activated forms of JAK2 including JAK2V617F and recombinant protein containing its catalytic domain alone. It showed minimal responses to wild-type JAK2 and was not phosphorylated by the epidermal growth receptor and the insulin receptor tyrosine kinases. Kinase assays with GST-JAKS provide a sharp contrast between wild-type and mutant JAK2,V617F and are sensitive enough to detect minute amounts of JAK2V617F found in crude cell extracts. Assays can be scaled up to screen for inhibitors of JAK2 in a dot blot format. CONCLUSION: GST-JAKS is sensitive and specific protein substrate for JAK2 assays. It may have clinical applications in diagnosis of diseases related to abnormal JAK2 activity. It is also an excellent substrate for development of large scale assays to screen JAK2 inhibitors.

A novel therapeutic drug (copper nicotinic acid complex) for non-alcoholic fatty liver.

BACKGROUND: Fatty liver is the accumulation of fat in liver cells, which leads to disruption of the normal liver structure and function. METHODS: A non-alcoholic fatty liver rat model received copper (Cu) (I)-nicotinate complex [CuCl(HNA)2] for 4 weeks. RESULTS: Clinical signs and histopathological examinations showed obvious improvements in rats that received Cu complex who were continuously on an (HCFF) diet than those returned to standard diet with Cu complex. The improvement was matched in total lipids in sera and hepatic tissue, with disappearance of fat droplets from liver sections. Furthermore, the gain in body weight and the corresponding decrease in liver weight, decreased liver transaminases and alkaline phosphatase were prominent. The oxidative stress markers such as nitric oxide, lipid peroxides, glutathione and superoxide dismutase were obviously changed to healthy normal levels. CONCLUSION: The Cu complex may serve as a novel chemical restoring agent in fatty degenerated liver cells and for renewal of their structure and functions. However, clinical trials are required for more evaluation of the Cu complex in humans.

Protective effects of mycelia of Antrodia camphorata and Armillariella tabescens in submerged culture against ethanol-induced hepatic toxicity in rats.

The hepatoprotective effects of the mycelia of Antrodia camphorata and Armillariella tabescens were evaluated in vivo using acute ethanol-intoxicated rats as an experimental model. Animals were orally treated with Antrodia camphorata (0.5 or 1.0 g/kg b.w.) or Armillariella tabescens (0.5 or 1.0 g/kg b.w.) for 10 days whereas controls received vehicle only. At the end of the experimental 10-day period, the animals were administered by gavage with an acute ethanol dose of 5.0 g/kg b.w. diluted in deionized water (6:4, v/v) and sacrificed at 18 h after ethanol administration. The degree of protection was measured by using biochemical parameters like serum transaminases (AST and ALT), alkaline phosphatase (ALP), bilirubin. Meanwhile, the histopathological studies were carried out to support the above parameters. Administration of Antrodia camphorata or Armillariella tabescens markedly prevented ethanol-induced elevation of levels of serum AST, ALT, ALP, and bilirubin comparable with standard drug silymarin.

Actividad de la glutatión reductasa en el embarazo diabético / Glutathione reductase activity in diabetic pregnancy

ANTECEDENTES: El predominio de la diabetes gestacional está aumentando en nuestra población y sus efectos en el metabolismo celular y estatus oxidativo están siendo estudiados. OBJETIVO: Determinar si existe una relación entre la actividad de la glutatión reductasa eritrocitaria, evaluada a través del test CAGRE y la diabetes gestacional. MÉTODO: Estudio transversal de casos y controles, incluyó a 30 embarazadas con diagnóstico de diabetes gestacional y 30 sin patologías asociadas, pertenecientes a la Maternidad del Hospital Barros Luco-Trudeau, Santiago de Chile. La actividad de la glutatión reductasa de eritrocitos fue determinada espectrofotométricamente a través del test CAGRE, la que fue relacionada con variables maternas como: edad, hematocrito, presencia de antecedentes de enfermedades familiares, estado nutricional materno e índice de masa corporal. Para comparar las variables entre ambos grupos, se realizaron medidas de disparidad, posición y análisis de la correlación. RESULTADOS: El grupo de madres diabéticas presentó edad materna, índice de masa de corporal y antecedentes de diabetes gestacional mayores que el grupo control, aunque sin diferencia estadísticamente significativa. El predominio de anemia y la respuesta al suplemento del cofactor FAD en la actividad de la glutatión reductasa eritrocitaria fue similar entre los grupos. La mayor incidencia de diabetes familiar en el grupo control fue estadísticamente significativa. La distribución de los valores de CAGRE, utilizado también como un indicador de los niveles de riboflavina, mostró en el grupo de casos riesgo medio a alto de malnutrición, mientras que en el grupo control la tendencia fue normal o de riesgo bajo. CONCLUSIÓN: Las embarazadas diabéticas gestacionales, presentaron mal nutrición y un estrés oxidativo mayor que el grupo control, evidenciado por el test de CAGRE.

BACKGROUND: The prevalence of the gestational diabetes is increasing in our population and its effects in the cellular metabolism and oxidative status had been studied. OBJECTIVE: Determine if exists a relationship between the erythrocyte glutathione reductase activity evaluated by EGRAC test and the gestational diabetes. METHODS: This traversal study of cases and controls, included at 30 pregnant with diagnostic of gestational diabetes and 30 without associate pathologies, belonging to the Maternity of the Hospital Barros Luco-Trudeau, Santiago-Chile. The activity of the glutathione reductase was determined by spectrophotometric assay through the EGRAC test, and their values were related with maternal variables as: age, hematocrite, presence of antecedents of family illnesses, maternal nutritional status and the body mass index. To compare the variables between both groups, they were carried out measures of disparity, position and analysis of the correlation. RESULTS: We determine that the group of diabetic mothers was older, with higher body mass index and a bigger frequency of antecedents of gestational diabetes that the control group, although without significant difference. The prevalence of anemia and the answer to the supplement with FAD in the activity of the glutathione reductase was similar among the groups. On the other hand, the incidence of familiar diabetes in the group control was bigger. The distribution of the values of EGRAC, also used as an indicator of the riboflavin levels, showed in the group of cases half risk to high of malnutrition, while in the group control the tendency was normal or low risk. CONCLUSION: We can conclude that the gestational diabetics pregnant presented malnutrition and higher oxidative stress that the control group, evidenced by means of the EGRAC test.

Media components and amino acid supplements influencing the production of fruity aroma by Geotrichum candidum

The ability of Geotrichum candidum to produce fruity aroma in food grade sucrose, molasses, corn steep liquor and peptone based culture media was tested by sensory evaluation and analyzed by gas chromatography mass spectrometry. A strong and sweet fruity aroma was produced from molasses, with peptone or corn steep liquor stimulating aroma production. Molasses with peptone supplemented with leucine, valine, or alanine yielded better fruity aroma production and the presence of many esters was consistent with the fruity aroma production.

Geotrichum candidum foi cultivado em diversos meios de cultura contendo sacarose ou melaço e milhocina ou peptona e a produção de aroma frutal foi verificada através de avaliação sensorial e cromatografia gasosa acoplada a espectrometria de massas. Os meios contendo melaço, peptona e leucina, valina ou alanina apresentaram os melhores resultados e a presença de diversos ésteres foi consistente com a formação de aroma frutal.

Cyclodextrin glycosyltransferase from Bacillus licheniformis: optimization of production and its properties

Cyclodextrin glycosyltransferase (EC 2.4.1.19) is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. An alkalophilic Bacillus strain, isolated from cassava peels, was identified as Bacillus licheniformis. CGTase production by this strain was better when potato starch was used as carbon source, followed by cassava starch and amylopectin. Glucose and amylose, on the other hand, acted as synthesis repressors. When the cultivation was supplemented with sodium ions and had the pH adjusted between 6.0 and 9.0, the microorganism maintained the growth and enzyme production capacity. This data is interesting because it contradicts the concept that alkalophilic microorganisms do not grow in this pH range. After ultrafiltration-centrifugation, one protein of 85.2 kDa with CGTase activity was isolated. This protein was identified in plates with starch and phenolphthalein. Determination of the optimum temperature showed higher activities at 25°C and 55°C, indicating the possible presence of more than one CGTase in the culture filtrate. Km and Vmax values were 1.77 mg/mL and 0.0263 U/mg protein, respectively, using potato starch as substrate.

Ciclodextrina glicosiltransferase (EC 2.4.1.19) é uma enzima que produz ciclodextrinas a partir de amido via transglicosilação intramolecular. Uma cepa de Bacillus alcalofílico, isolada de cascas de mandioca, foi identificada como Bacillus licheniformis. A produção de CGTase por esta cepa foi melhor quando amido de batata foi utilizado como fonte de carbono, seguido por amido de mandioca e amilopectina. Glicose e amilose, por outro lado, atuaram como repressor de síntese desta enzima. Quando o cultivo foi suplementado com íons sódio e teve o pH ajustado entre 6,0 e 9,0, o microrganismo manteve a capacidade de crescimento e de produção da enzima. Este dado é interessante pois contraria o conceito de que microrganismos alcalofílicos não apresentam crescimento nesta faixa de pH. Após ultrafiltração-centrifugação, uma proteína de 85,2 kDa com atividade de CGTase foi isolada. Esta proteína foi identificada em placas contendo amido e fenolftaleína. A determinação da temperatura ótima mostrou atividades mais elevadas em 25°C e 55°C, indicando a possível presença de mais de uma CGTase no filtrado de cultura. Valores de Km e Vmax foram 1,77 mg/mL e 0,0263 U/mg proteína, respectivamente, usando amido de batata como substrato.

Differentiation of long-chain fatty acid oxidation disorders using alternative precursors and acylcarnitine profiling in fibroblasts.

The differentiation of carnitine-acylcarnitine translocase deficiency (CACT) from carnitine palmitoyltransferase type II deficiency (CPT-II) and long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency from mitochondrial trifunctional protein deficiency (MTP) continues to be ambiguous using current acylcarnitine profiling techniques either from plasma or blood spots, or in the intact cell system (fibroblasts/amniocytes). Currently, enzyme assays are required to unequivocally differentiate CACT from CPT-II, and LCHAD from MTP. Over the years we have studied the responses of numerous FOD deficient cell lines to both even and odd numbered fatty acids of various chain lengths as well as branched-chain amino acids. In doing so, we discovered diagnostic elevations of unlabeled butyrylcarnitine detected only in CACT deficient cell lines when incubated with a shorter chain fatty acid, [7-2H3]heptanoate plus l-carnitine compared to the routinely used long-chain fatty acid, [16-2H3]palmitate. In monitoring the unlabeled C4/C5 acylcarnitine ratio, further differentiation from ETF/ETF-DH is also achieved. Similarly, incubating LCHAD and MTP deficient cell lines with the long-chain branched fatty acid, pristanic acid, and monitoring the C11/C9 acylcarnitine ratio has allowed differentiation between these disorders. These methods may be considered useful alternatives to specific enzyme assays for differentiation between these long-chain fatty acid oxidation disorders, as well as provide insight into new treatment strategies.