ABSTRACT Objective This pilot study investigated whether single nucleotide polymorphisms (SNP) in the NOX5 gene (NADPH oxidase 5) are associated with the type 2 diabetes (T2D) risk. Subjects and methods A total of 1579 patients with T2D and 1627 age- and sex-matched healthy subjects were recruited for this study. Genotyping of common SNPs, namely rs35672233, rs3743093, rs2036343, rs311886, and rs438866, was performed using the MassArray-4 system. Results SNP rs35672233 was associated with an increased risk of T2D (OR = 1.67, 95% CI 1.29-2.17, FDR = 0.003). The H3 haplotype (rs35672233T-rs3743093G-rs2036343A-rs311886C-rs438866C) increased T2D risk (OR = 1.65, 95% CI 1.27-2.13, FDR = 0.001). The rs35672233 polymorphism and H3 haplotype were found to have an association with T2D risk only in subjects with a body mass index greater than 25 kg/m2 (FDR < 0.01). Environmental risk factors, such as chronic psycho-emotional stress, sedentary lifestyle, high-calorie diet and SNP rs35672233 were jointly associated with T2D susceptibility. A haplotype comprising the allele rs35672233-C and conferring protection against T2D, was associated with elevated levels of antioxidants such as total glutathione and uric acid, as well as reduced levels of two-hour postprandial glucose in the plasma of patients. The NOX5 polymorphisms showed no associations with diabetic complications. Conclusion The present study is the first to establish associations between polymorphisms in NOX5 and the risk of type 2 diabetes mellitus, and provides a new line of evidence for the crucial role of oxidative stress-related genes in disease susceptibility. (SNP NOX NADPH 5 TD T D (T2D 157 162 age sexmatched sex matched SNPs rs rs3743093 rs2036343 rs311886 rs438866 MassArray4 MassArray 4 MassArray- system rs3567223 OR 167 1 67 1.67 95 1.292.17, 129217 1.29 2.17, 29 17 1.29-2.17 0.003. 0003 0.003 . 0 003 0.003) H rs35672233Trs3743093Grs2036343Ars311886Crs438866C rsTrsGrsArsCrsC rs35672233T rs3743093G rs2036343A rs311886C rs438866C G C (rs35672233T-rs3743093G-rs2036343A-rs311886C-rs438866C 165 65 1.65 1.272.13, 127213 1.27 2.13, 27 13 1.27-2.13 0.001. 0001 0.001 001 0.001) kgm2 kgm kg m2 m kg/m 0.01. 0.01 01 0.01) factors psychoemotional psycho emotional stress lifestyle highcalorie high calorie susceptibility rs35672233C rsC acid twohour two hour complications mellitus stressrelated related 15 16 rs374309 rs203634 rs31188 rs43886 rs356722 6 1.6 9 292 1.292.17 12921 129 1.2 217 2.17 1.29-2.1 000 0.00 00 Trs Grs Ars Crs rsT rsG rsA 272 1.272.13 12721 127 213 2.13 1.27-2.1 0.0 rs37430 rs20363 rs3118 rs4388 rs35672 1. 1.292.1 1292 12 21 2.1 1.29-2. 1.272.1 1272 1.27-2. 0. rs3743 rs2036 rs311 rs438 rs3567 1.292. 2. 1.29-2 1.272. 1.27-2 rs374 rs203 rs31 rs43 rs356 1.292 1.29- 1.272 1.27- rs37 rs20 rs3 rs4 rs35 rs2

ABSTRACT Objective This pilot study investigated whether single nucleotide polymorphisms (SNP) in the NOX5 gene (NADPH oxidase 5) are associated with the type 2 diabetes (T2D) risk. Subjects and methods A total of 1579 patients with T2D and 1627 age- and sex-matched healthy subjects were recruited for this study. Genotyping of common SNPs, namely rs35672233, rs3743093, rs2036343, rs311886, and rs438866, was performed using the MassArray-4 system. Results SNP rs35672233 was associated with an increased risk of T2D (OR = 1.67, 95% CI 1.29-2.17, FDR = 0.003). The H3 haplotype (rs35672233T-rs3743093G-rs2036343A-rs311886C-rs438866C) increased T2D risk (OR = 1.65, 95% CI 1.27-2.13, FDR = 0.001). The rs35672233 polymorphism and H3 haplotype were found to have an association with T2D risk only in subjects with a body mass index greater than 25 kg/m2 (FDR < 0.01). Environmental risk factors, such as chronic psycho-emotional stress, sedentary lifestyle, high-calorie diet and SNP rs35672233 were jointly associated with T2D susceptibility. A haplotype comprising the allele rs35672233-C and conferring protection against T2D, was associated with elevated levels of antioxidants such as total glutathione and uric acid, as well as reduced levels of two-hour postprandial glucose in the plasma of patients. The NOX5 polymorphisms showed no associations with diabetic complications. Conclusion The present study is the first to establish associations between polymorphisms in NOX5 and the risk of type 2 diabetes mellitus, and provides a new line of evidence for the crucial role of oxidative stress-related genes in disease susceptibility. (SNP NOX NADPH 5 TD T D (T2D 157 162 age sexmatched sex matched SNPs rs rs3743093 rs2036343 rs311886 rs438866 MassArray4 MassArray 4 MassArray- system rs3567223 OR 167 1 67 1.67 95 1.292.17, 129217 1.29 2.17, 29 17 1.29-2.17 0.003. 0003 0.003 . 0 003 0.003) H rs35672233Trs3743093Grs2036343Ars311886Crs438866C rsTrsGrsArsCrsC rs35672233T rs3743093G rs2036343A rs311886C rs438866C G C (rs35672233T-rs3743093G-rs2036343A-rs311886C-rs438866C 165 65 1.65 1.272.13, 127213 1.27 2.13, 27 13 1.27-2.13 0.001. 0001 0.001 001 0.001) kgm2 kgm kg m2 m kg/m 0.01. 0.01 01 0.01) factors psychoemotional psycho emotional stress lifestyle highcalorie high calorie susceptibility rs35672233C rsC acid twohour two hour complications mellitus stressrelated related 15 16 rs374309 rs203634 rs31188 rs43886 rs356722 6 1.6 9 292 1.292.17 12921 129 1.2 217 2.17 1.29-2.1 000 0.00 00 Trs Grs Ars Crs rsT rsG rsA 272 1.272.13 12721 127 213 2.13 1.27-2.1 0.0 rs37430 rs20363 rs3118 rs4388 rs35672 1. 1.292.1 1292 12 21 2.1 1.29-2. 1.272.1 1272 1.27-2. 0. rs3743 rs2036 rs311 rs438 rs3567 1.292. 2. 1.29-2 1.272. 1.27-2 rs374 rs203 rs31 rs43 rs356 1.292 1.29- 1.272 1.27- rs37 rs20 rs3 rs4 rs35 rs2

Resumen Introducción. La miel Klanceng (Tetragonula laeviceps) es muy demandada por sus beneficios. No hay información sobre la fecha de caducidad de la miel Klanceng. Objetivo. Predecir la vida útil de la miel Klanceng mediante la evaluación del efecto del calentamiento (40 ºC / 48 h y 70 ºC / 7 h) y el almacenamiento durante dos años. Materiales y métodos. El análisis se llevó a cabo tres veces (2020, 2021 y 2022) en el Laboratorio de la Agencia Nacional de Investigación de Innovación, Yakarta, Indonesia. Se tomaron muestras de miel Klanceng (Tetragonula laeviceps) de una granja de abejas en Magetan, Java Oriental, Indonesia. La muestra (5 kg) se dividió en 3: sin proceso de calentamiento (UT), calentamiento a 40 °C / 48 h (T1) y calentamiento a 70 °C / 7 h (T2). Luego se analizaron HMF, diastasa, invertasa, fosfatasa ácida (AP), glucosa oxidasa (GO), DPPH, absorbancia y actividad fenólica a los 0, 12 y 24 meses. Resultados. Los resultados mostraron que el calentamiento corto a alta temperatura (70 °C / 7 h) tuvo un mayor impacto en la disminución de la actividad enzimática en comparación con el calentamiento prolongado a baja temperatura (40 °C / 48 h). El almacenamiento tuvo un impacto significativo en el aumento de 5-hidroximetil (furano)-2-carbaldehído (HMF) en comparación con el calentamiento. Durante el almacenamiento (24 meses) el valor de HMF superó el límite máximo. Conclusiones. El valor de HMF (55,33 ± 0,57 mg/kg) superó el límite máximo permitido (máximo 40 ppm), esto se debió al efecto significativo del calentamiento en T2 y el proceso de almacenamiento. La vida útil de la miel Klanceng fue de dos años, siempre que no se calentara a altas temperaturas (70 °C).

Abstract Introduction. Klanceng honey (Tetragonula laeviceps) is in great demand because of its benefits. There is no information on the expiration date of the Klanceng honey. Objective. To predict the shelf life of Klanceng honey by evaluating the effect of heating (40 ºC / 48 h and 70 ºC / 7 h) and storage for two years. Materials and methods. The analysis was carried out three times (2020, 2021, and 2022) at the National Innovation Research Agency Laboratory, Jakarta, Indonesia. Samples of Klanceng honey (Tetragonula laeviceps) were taken from a bee farm in Magetan, East Java, Indonesia. The sample (5 kg) was divided into 3: without heating process (UT), heating at 40 ºC / 48 h (T1), and heating at 70 ºC / 7 h (T2). Then analyzed the activity of HMF, diastase, invertase, acid phosphatase (AP), glucose oxidase (GO), DPPH, honey color, and phenolic at 0, 12 and 24-months. Results. The results showed that short heating at high temperature (70 ºC / 7 h) had a greater impact on decreasing enzyme activity compared to prolonged heating at low temperature (40 ºC / 48 h). Storage had a major impact on the increase of 5-hydroxymethyl (furan)-2-carbaldehyde (HMF) compared to the heating process. During storage (24-months) the HMF value exceeded the maximum limit. Conclusions. The HMF value (55.33 ± 0.57 mg/kg) exceeded the maximum allowable limit (max 40 ppm), this was due to the significant effect of heating on T2 and the storage process. The shelf life of Klanceng honey was two years, as long as it was not heated to high temperatures (70 ºC).

Oxidative stress is identified as the common pathogenic factor that leads to insulin resistance in diabetics. Malondialdehyde is a product of lipid peroxidation. Aim The aim of this study was to determine the variation in the Salivary malondialdehyde (MDA) among subjects with and without T2DM in comparison to the fasting blood and Salivary glucose. Methods This study involved 29 healthy participants as Controls (group I) and 29 participants with Type 2 Diabetes Mellitus as Cases (group II). Salivary Glucose was analysed by glucose oxidase end-point assay. Thiobarbituric acid (TBA) assay method was considered for estimation of MDA in fasting saliva. Data was Statistically analysed using SPSS20. Parametric test was performed to analyse the data. Results The correlation calculated between FBG with FSG level was found to be highly significant. A positive correlation between MDA levels with FBG was found. The relationship between FBG and FSG (r = 0.7815, p < 0.05), FBG and MDA (r =0.3678, p < 0.05) and FSG and MDA (r = 0.2869, p < 0.05) were found to be positively significant. Conclusion Saliva as a unique body fluid can serve as a medium for biochemical analysis only in standard settings and with multiple measures to be used as a diagnostic tool in par with the gold standard serum. Salivary MDA levels can be considered as one of the oxidative stress markers in Type 2 Diabetic condition. diabetics peroxidation (MDA TDM T DM group I II. II . II) endpoint end point TBA (TBA saliva SPSS20 SPSS data significant r 07815 0 7815 0.7815 0.05, 005 0.05 , 05 03678 3678 =0.3678 02869 2869 0.2869 serum condition SPSS2 0781 781 0.781 00 0.0 0367 367 =0.367 0286 286 0.286 078 78 0.78 0. 036 36 =0.36 028 28 0.28 07 7 0.7 03 3 =0.3 02 0.2 =0. =0

SUMMARY: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) can protect diabetic kidneys, but the mechanisms are unclear. This work is to investigate the potential mechanisms of C. tinctoria Nutt. in the treatment of diabetic nephropathy based on network pharmacology analysis of its active ingredients. Twelve small molecular compounds of C. tinctoria Nutt. and targets related to diabetic nephropathy were docked by Discovery Studio 3.0. DAVID database was used for GO enrichment and KEGG pathway analysis. Cytoscape 3.6.1 was used to construct active ingredient-target network. Cell viability was detected with MTT. Glucose consumption was analyzed with glucose oxidase method. Protein expression was measured with Western blot and immunofluorescence. Electron microscopy observed autophagosomes. The core active ingredients of C. tinctoria Nutt. included heriguard, flavanomarein, maritimein, and marein. Twenty-one core targets of the 43 potential targets were PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK, and ABL2. These 21 core targets were significantly enriched in 50 signaling pathways. Thirty- four signaling pathways were closely related to diabetic nephropathy, of which the top pathways were PI3K/AKT, insulin, and mTOR, and insulin resistance. The enriched GO terms included biological processes of protein phosphorylation, and the positive regulation of PI3K signaling and cytokine secretion; cellular components of cytosol, extracellular region, and extracellular space; and molecular function of protein kinase activity, ATP binding, and non-membrane spanning protein tyrosine kinase activity. In vitro experiments found that marein increased the expression of phosphorylated AKT/AKT in human renal glomerular endothelial cells of an insulin resistance model induced by high glucose, as well as increased and decreased, respectively, the levels of the microtubule-associated proteins, LC3 and P62. C. tinctoria Nutt. has many active ingredients, with main ingredients of heriguard, flavanomarein, maritimein, and marein, and may exert anti-diabetic nephropathy effect through various signaling pathways and targets.

RESUMEN: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) puede proteger riñones diabéticos, sin embargo los mecanismos son desconocidos. Este trabajo se realizó para investigar los potenciales mecanismos de C. tinctoria Nutt. en el tratamiento de la nefropatía diabética basado en el análisis de farmacología en red de sus principios activos. Doce compuestos moleculares pequeños de C. tinctoria Nutt. y los objetivos relacionados con la nefropatía diabética fueron acoplados por Discovery Studio 3.0. La base de datos DAVID se utilizó para el enriquecimiento GO y el análisis de la vía KEGG. Se usó Cytoscape 3.6.1 para construir una red de ingrediente-objetivo activa. La viabili- dad celular se detectó mediante MTT. El consumo de glucosa se analizó con el método de glucosa oxidasa. La expresión proteica fue determinada mediante Western blot e inmunofluorescencia. En la microscopía electrónica se observó autofagosomas. Los principales ingredientes activos de C. tinctoria Nutt. incluyeron heriguard, flavanomarein, maritimin y marein. Veintiún de los 43 objetivos potenciales fueron PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK y ABL2. Estos 21 objetivos principales se enriquecieron significativamente en 50 vías de señalización. Treinta y cuatro vías de señalización estuvieron estrechamente relacionadas con la nefropatía diabética, de las cuales las principales vías fueron PI3K/ AKT, insulina y mTOR, y resistencia a la insulina. Los términos GO enriquecidos incluyeron procesos biológicos de fosforilación proteica, la regulación positiva de la señalización de PI3K y la secreción de citoquinas; componentes celulares del citosol, región extracelular y espacio extracelular; y la función molecular de la actividad de la proteína quinasa, la unión de ATP y la actividad de la proteína tirosina quinasa que no se extiende por la membrana. Los experimentos in vitro encontraron que la mareína aumentaba la expresión de AKT/AKT fosforilada en células endoteliales glomerulares renales humanas en un modelo de resistencia a la insulina inducida por niveles elevados de glucosa, así como aumentaron y disminuyeron respectivamente, los niveles de las proteínas asociadas a los microtúbulos, LC3 y P62. C. tinctoria Nutt. tiene muchos principios activos, con ingredientes principales de heriguard, flavanomarein, maritimain y marein, y puede ejercer un efecto de nefropatía antidiabética a través de distintass vías de señalización y objetivos.

ABSTRACT The study was conducted to investigate the effect of GOX on performance, egg quality, and nutrient digestibility in laying hens. In total, 432, 50-week-old Hy-Line brown breeder hens were assigned into four treatments, and fed a basal diet with GOX at 0, 100, 200 and 300 units for 10 weeks, respectively. A Quadratic decrease in FI in week 3 (p<0.05) and linear increase in egg production in week 6 to 10 and overall experiment period (p<0.05) and Quadratic increase in egg production in week 7 (p<0.05), a linear decrease in broken egg rate in week 6 (p<0.05) a quadratic increase in egg weight on day 14 (p<0.05), alinear increase in egg weight on day 28 (p<0.05), and linear decrease in yolk color on day 7 (p<0.05), a linear increase in yolk color on day 42 and day 70 (p<0.05), and linear increase in haugh unit on day 28 and 70 (p<0.05), a linear increase albumen height on day 28 and day 56 (p<0.05), and linear decrease in shell color on day 14 (p<0.05) and day 28(p<0.05), a linear and quadratic increases in eggshell strength and eggshell thickness on day 56 (p<0.05), and linear increase in eggshell strength and eggshell thickness on day 70 (P0.05) were observed with the addition of GOX the the diet. Conclusion: This study suggested that the supplementation of GOX may have beneficial effects on feed intake and egg quality in laying hens.

RESUMO CONTEXTO E OBJETIVO: Considerando a associação entre câncer colorretal (CCR), a resistência à insulina, à obesidade e o papel proeminente da grelina nessas doenças metabólicas, foi explorado se os níveis plasmáticos de grelina estavam associados ao CCR. Além disso, nos pacientes com CCR foram pesquisadas as possíveis correlações entre a grelina, insulina, resistência insulínica e índice de massa corporal (IMC) como indicadores de obesidade. MÉTODOS: Foram incluídos neste estudo 170 indivíduos, sendo 82 com CRC e 88 controles. Os níveis plasmáticos de grelina, insulina e glicose foram medidos em todos os sujeitos utilizando métodos ELISA e glicose oxidase. Além disso, a resistência à insulina foi avaliada pelo cálculo do índice HOMA-IR. RESULTADOS: Os pacientes com CRC apresentaram redução dos níveis de grelina (P<0,001) e maior índice HOMA-IR (P<0.001) do que os controles. Curiosamente, quando os pacientes com CRC foram estratificados com base no local do tumor, níveis mais baixos de grelina e maior índice de HOMA-IR foram observados nos indivíduos com câncer de cólon ou retal versus controles também. Além disso, houve uma correlação negativa entre idade e IMC independente entre os níveis de grelina e HOMA-IR (r=-0,365, P<0,05) e uma correlação negativa independente da idade entre os níveis de grelina e IMC (r=-0,335, P<0,05) no subgrupo retal. CONCLUSÃO: Nossos achados apoiam o papel da grelina em relação à resistência à insulina e à obesidade na suscetibilidade do CRC; no entanto, ela precisa ser corroborada por estudos posteriores.

ABSTRACT BACKGROUND AND OBJECTIVE: Considering the association between colorectal cancer (CRC) and both insulin resistance and obesity, and the prominent role of ghrelin in these metabolic disorders, we explored whether plasma levels of ghrelin were associated with CRC. Moreover, in the patients with CRC the possible correlations between ghrelin and insulin, insulin resistance, and body mass index (BMI) as an indicator of obesity were examined. METHODS: A total of 170 subjects, including 82 cases with CRC and 88 controls were enrolled in this study. Plasma levels of ghrelin, insulin, and glucose were measured in all the subjects using ELISA and glucose oxidase methods. Furthermore, insulin resistance was assessed by calculating HOMA-IR index. RESULTS: The cases with CRC had decreased ghrelin levels (P<0.001) and a higher HOMA-IR index (P<0.001) than controls. Interestingly, when CRC patients were stratified based on tumor site, lower ghrelin levels and a higher HOMA-IR index were observed in the patients with either colon or rectal cancer vs. controls too. Additionally, there were an age and BMI-independent negative correlation between ghrelin levels and HOMA-IR (r=-0.365, P<0.05), and an age-independent negative correlation between ghrelin levels and BMI (r=-0.335, P<0.05) in the rectal subgroup. CONCLUSION: Our findings support a role for ghrelin in connection with insulin resistance and obesity in CRC susceptibility; however, it needs to be corroborated by further studies.

BACKGROUND: Hypoglycaemia is a common presenting feature of illness among children admitted with various conditions into the paediatric emergency roomOBJECTIVE: To determine the prevalence of hypoglycaemia among children admitted into a Nigerian teaching hospital, identify some of its associated factors and assess its relationship to patient outcomeMETHODS: In this cross-sectional study, 388 children were consecutively recruited. Their sociodemographic characteristics, clinical features and management outcome were obtained. Plasma glucose was measured using the glucose oxidase method and classified as hypoglycaemia (<3.4 mmol/L), low glycaemia (3.4 - <4.4 mmol/L) , normoglycaemia (4.4 - 7.7 mmol/L) and hyperglycaemia (>7.8 mmol/LRESULTS: Hypoglycaemia occurred in 50 (12.9%) subjects, while 55 (14.2%) had low glycaemia. At bivariate analysis, hypoglycaemia was associated with age (p=0.012), >6 hours from the last meal (p=0.004), medication use before presentation (p=0.014) and some markers of illness severity such as loss of consciousness (p<0.001), respiratory distress (p=0.004), dehydration (p=0.002), shock (p=0.004), and hypoxia (p=0.001). After logistic regression, a time > 6 hours from last meal (adjusted odds ratio (aOR) 2.98; 95% confidence interval (CI) 1.27 - 6.98), hypoxia (aOR 3.10; 95% CI 1.39 - 6.90), and passage of watery stools (aOR 2.92; 95% CI 1.26 - 6.78) were associated with hypoglycaemia. Hypoglycaemic subjects were 2.9 times more likely to die than normoglycaemic ones (p<0.001) but this association was lost at multivariable analysisCONCLUSION: Hypoglycaemia complicates common childhood illnesses and may be associated with poorer outcomes. Plasma glucose measurement and subsequent appropriate action should be core parts of the immediate management of ill children

ABSTRACT Objective Compare the concordance degree between plasma glucose and glucose measurements on Dried Blood Spots (DBS) during pregnancy. Subjects and methods Glucose measurement was performed in pregnant women after a fast of 8-12 hours. Venous blood was collected with sodium fluoride, the plasma was separated, and glucose measured by the enzymatic oxidase glucose method. Capillary blood samples were collected and analyzed by DBS. For statistics, the paired Student’s t test, interclass correlation coefficient (ICC), graphic approach of Altman and Bland, and survival – concordance plot were used. Results 307 pregnant women were evaluated, 88.6% without diabetes and 11.4% with previous diabetes. The glucose ranged from 66 to 190 mg/dL [3.66 to 10.55 mmol/L] in plasma and from 53 to 166 mg/dL [2.94 to 9.21 mmol/L] in DBS. The glucose average values were 88.1 ± 12 mg/dL [4.98 ± 0.67 mmol/L] in plasma and 89.2 ±11,5 mg/dL, [4.95 ± 0.64 mmol/L] in DBS – p-value = 0.084. The ICC value was moderate (0.510), and Pearson’s correlation coefficient was r = 0.507 p < 0.001. Altman and Bland’s graph showed that difference between the values obtained by both methods is -24.62 to 22.3 mg/dL [-1.37 to 1.24 mmol/L]. Significant fixed bias (-1,16 average difference) and proportional bias (r = 0.056; p = 0.33) were not observed. Anemia was associated with differences between plasma glucose and DBS measurements (p = 0.031). Conclusion Capillary glucose in DBS correlates with plasma glucose; however, the methods do not present good concordance. The presence of anemia worsens this result.

Abstract: Fructooligosaccharides (FOS) are fructose polymers, linked by β-glucosidic bonds (2-1), considered low caloric food sweeteners with prebiotic function and can be obtained from sucrose by biosynthesis. The fruits of the carob tree (Prosopis pallida) contain mainly carbohydrates where sucrose stands out. For this reason, the production of FOS from carob extract was optimized using a pectinase from A. aculeatus. The response surface methodology using was applied to optimize the parameters that affected production: time, temperature and carob extract; then they were verified experimentally. The biosynthesis process of FOS was carried out with carob extract at the concentrations of 20, 35 and 50 ° Bx, Pectinase at 10, 35 and 61mg, at 45, 55 and 65 °C for 0,5, 9,5 and 18 h, with stirring at 150 rpm The analysis methods were glucose oxidase and AOAC 999.03, for glucose and FOS respectively and the Minitab 16 program. The optimal reaction conditions for the production of FOS were: carob extract 38 °Bx, enzyme 31 mg, 54 °C, 150 rpm and 12 h. The experimental yield in the transformed extract was FOS 16, 1% quantified by HPLC.

RESUMEN Los fructooligosacáridos (FOS) son polímeros de fructosa, unidos mediante enlaces glucosídicos β(2-1), considerados edulcorantes de bajo aporte calórico con función prebiótica que pueden ser obtenidos a partir de sacarosa por biosíntesis. Los frutos del algarrobo (Prosopis pallida) contienen principalmente carbohidratos y, en particular, alta concentración de sacarosa. Por tal motivo, se optimizó la producción de FOS a partir del extracto de algarrobo usando pectinasa de A. aculeatus. La metodología de superficie de respuesta se aplicó para optimizar los parámetros que afectan la producción como: tiempo, temperatura y extracto de algarrobo; después se verificaron experimentalmente. El proceso de biosíntesis de FOS se realizó con extracto de algarrobo a las concentraciones de 20, 35 y 50 °Bx, pectinasa a 10, 35 y 61 mg, a 45, 55 y 65 °C durante 0,5; 9,5 y 18 h, con agitación a 150 rpm. Los métodos de análisis para glucosa y FOS fueron glucosa oxidasa y AOAC 999,03 respectivamente y el programa Minitab 16. Las condiciones óptimas de reacción para la producción de FOS fueron: extracto de algarrobo 38 °Bx, pectinasa 31 mg, 54 °C, 150 rpm y 12 h. El rendimiento experimental en el extracto transformado fue FOS 16,1 % cuantificados por HPLC.

Abstract Immobilization process is used to facilitate the enzyme recovery and reusability. The aims of this study were to evaluate the effect of different amounts of chitosan on immobilization efficiency of Glucose Oxidase (GOX), pH effect on enzyme activity and changes of kinetic characteristics of enzyme immobilized on Magnetic Chitosan Nanoparticles (MCNP). GOX was immobilized on MCNP with cross bonds and Schiff base covalent connections. The sample with 0.3 g of chitosan had the highest immobilization efficiency (41.30%). Morphology of nanoparticles was investigated by Scanning Electron Microscopy (SEM). Immobilization of GOX on magnetic nanoparticles was verified with Fourier Transform Infrared (FTIR). The effect of pH on enzyme activity was similar on both immobilized enzyme (IE) and free enzyme (FE). The evaluation of kinetic parameters for IE and FE has showed that the effectiveness factor was 0.45. Generally, it could be concluded that magnetic nanoparticles with 0.3 g chitosan could provide a suitable medium for immobilization of GOX.

BACKGROUND. Identifying women with gestational diabetes mellitus (GDM) allows interventions to improve perinatal outcomes. A fasting plasma glucose (FPG) level >5.1 mmol/L is 100% specific for a diagnosis of GDM. The International Association of Diabetes and Pregnancy Study Groups acknowledges that FPG <4.5 mmol/L is associated with a low probability of GDM.OBJECTIVES. The validity of selective screening based on the presence of risk factors was compared with the universal application of FPG >4.5 mmol/L to identify women with GDM. FPG >4.5 mmol/L or the presence of one or more risk factors was assumed to indicate an intermediate to high risk of GDM and therefore the need for an oral glucose tolerance test (OGTT).METHODS. Consecutive black South African (SA) women were recruited to a 2-hour 75 g OGTT at 24 - 28 weeks' gestation in an urban community health clinic. Of 969 women recruited, 666 underwent an OGTT, and of these 589 were eligible for analysis. The glucose oxidase laboratory method was used to measure plasma glucose concentrations. The World Health Organization GDM diagnostic criteria were applied. All participants underwent a risk factor assessment. The χ² test was used to determine associations between risk factors and a positive diagnosis of GDM. The sensitivity and specificity of a positive diagnosis of GDM were calculated for FPG >4.5 mmol/L, FPG >5.1 mmol/L, and the presence of one or more risk factors.RESULTS. The prevalence of overt diabetes mellitus and GDM was 0.5% and 7.0%, respectively. Risk factor-based selective screening indicated that 204/589 (34.6%) of participants needed an OGTT, but 18/41 (43.9%) of positive GDM diagnoses were missed. Universal screening using the FPG threshold of >4.5 mmol/L indicated that 152/589 (25.8%) of participants needed an OGTT, and 1/41 (2.4%) of positive diagnoses were missed. An FPG of >5.1 mmol/L identified 36/41 (87.8%) of GDM-positive participants. The sensitivity and specificity of the presence of one or more risk factors were 56% and 67%, respectively. The sensitivity and specificity of FPG >4.5 mmol/L were 98% and 80%, respectively.CONCLUSIONS. Universal screening using FPG >4.5 mmol/L had greater sensitivity and specificity in identifying GDM-affected women and required fewer women to undergo a resource-intensive diagnostic OGTT than risk factor-based selective screening. A universal screening strategy using FPG >4.5 mmol/L may be more efficient and cost-effective than risk factor-based selective screening for GDM in black SA women.

A mathematical model describing the reduction of Hydrogen peroxide (H2O2) to water in a metal dispersed conducting polymer film is discussed. The model is based on a system of reaction-diffusion equations containing a non-linear term related to Michaelis-Menten kinetics of the enzymatic reaction. The approximate analytical expressions corresponding to the concentration of substrate and product for steady and non-steady state conditions have been obtained using a new approach to homotopy perturbation method (HPM). Approximate analytical expressions of the electrochemical oxidation current are also presented for steady and non-steady state conditions. The numerical simulation (Matlab program) response for concentration profiles was carried out and compared with the analytical results of this work and are found to be in good agreement. The influence of initial substrate concentration, the thickness of the film as well as the diffusion layer and kinetic parameters on the current response were investigated. A graphical procedure for estimating the kinetic parameters from the expression of the current response is also proposed.

We evaluated a hybrid catalytic power source for less invasive internal electroporation with better tissue reach than the widely used and more invasive external electroporation. We modelled how open-circuit voltage optimizes platinum-loading in catalysts to improve the electrochemical activity (ECA) possible from bioelectrogenesis through these systems and address the high costs of nano-drug delivery systems. The effects of the catalysts' convective flux and proton concentration were modelled for an enzyme (glucose oxidase) biofuel cell that was fed glucose substrate at a current rate under isothermal physiological conditions. Glucose concentrations were varied relative to anode catalyst loading models with 0.1-0.5 mg cm-2 platinum and alloyed (Pt-Ru-Ni) with a narrow particle size distribution. Using the free (solvation) electron model, bioelectrochemical activity (BECA) and a high open circuit voltage were generated by 5.5,10 and 20 mM glucose with 20 kU L-1 glucose oxidase at 37 °C. BECA (glucose oxidase), on its own, produced pulses of various intensities for nano-microsecond durations whereas the hybrid BECA-ECA (glucose oxidase and platinum) anode catalyst provided sustainable pulses of microseconds-minute durations. Enhanced catalysis with the hybrid BECA-ECA's open circuit voltage favours compatibility of a hybrid-powered nano-drug delivery system for internal electroporation.

Resumen Objetivo. El diagnóstico de la hipoglucemia en hurones (Mustela putorius furo) se basa en la detección de un nivel de glucosa en la sangre inferior a 60-70 mg / dl. Una concentración de glucosa en sangre por debajo de estos niveles es muy sugestiva de tumor de células beta pancreáticas (insulinoma), que comprende aproximadamente el 25% de todas las neoplasias diagnosticadas en esta especie. En este estudio se pretende comprobar si los bajos valores de glucosa en sangre obtenidos mediante el uso de glucómetros portátiles son fiables como método de diagnóstico de hipoglucemia en esta especie. Material y métodos. En este estudio se analizan 31 muestras de sangre de hurones de distinta edad y sexo, mediante el uso de tres tipos de glucómetros portátiles, comparando dichos resultados con las medidas obtenidas mediante un analizador de bioquímica líquida que usa el método de la glucosa oxidasa, considerado como uno de los métodos estándar de medida. Resultados. Uno de los tres glucómetros utilizado en este estudio subestima los valores de glucosa en sangre en esta especie, mientras que otro los sobreestima, lo que supone un error sistemático (predecible) de los equipos que se podría solucionar calibrándolos para esta especie. El otro glucómetro que presenta mayor concordancia con los valores de referencia presenta un error aleatorio elevado (impredecible) más difícil de corregir. Por tanto ninguno de los tres equipos sería adecuado para realizar un diagnóstico fiable de hipoglucemia en hurones. Conclusiones. Estos resultados indican que las concentraciones de glucosa en sangre en hurones deben confirmarse con un analizador de laboratorio validado para realizar el diagnóstico de hipoglucemia de forma fiable, y los glucómetros usarse sólo para hacer un seguimiento de la enfermedad.

Abstract Aims. The diagnosis of hypoglycemia in ferrets (Mustela putorius furo) is based on the detection of a blood glucose level below 60-70 mg / dl. A blood glucose concentration below these levels is very suggestive of pancreatic beta cell tumor (insulinoma), which comprises of approximately 25% of all neoplasms diagnosed in this species. This study aims to check if the low blood glucose values obtained through the use of portable glucometers are reliable as a method of diagnosing hypoglycemia in this species. Material and methods. In this study, 31 samples of ferrets blood of different ages and sexes are analyzed, using three types of portable glucometers, comparing these results with the measurements obtained by a liquid biochemistry analyzer using the glucose oxidase method, considered as one of the standard measurement methods. Results. One of the three glucometers used in this study underestimates blood glucose values in this species, while another overestimates them, which is a systematic error (predictable) of the equipment that could be solved by calibrating them for this species. The other glucometer is more consistent with the reference values, but has a high random error (unpredictable), that is more difficult to correct. Therefore none of the three teams would be adequate to make a reliable diagnosis of hypoglycemia in ferrets. Conclusions. These results indicate that blood glucose concentrations in ferrets should be confirmed with a validated laboratory analyzer to reliably diagnose hypoglycemia, and blood glucose meters used only to monitor the disease.