Optimizing perinatal wellbeing in pregnancy with obesity: a clinical trial with a multi-component nutrition intervention for prevention of gestational diabetes and infant growth and neurodevelopment impairment.

Pregnancy complicated by obesity represents an increased risk of unfavorable perinatal outcomes such as gestational diabetes mellitus (GDM), hypertensive disorders in pregnancy, preterm birth, and impaired fetal growth, among others. Obesity is associated with deficiencies of micronutrients, and pregnant women with obesity may have higher needs. The intrauterine environment in pregnancies complicated with obesity is characterized by inflammation and oxidative stress, where maternal nutrition and metabolic status have significant influence and are critical in maternal health and in fetal programming of health in the offspring later in life. Comprehensive lifestyle interventions, including intensive nutrition care, are associated with a lower risk of adverse perinatal outcomes. Routine supplementation during pregnancy includes folic acid and iron; other nutrient supplementation is recommended for high-risk women or women in low-middle income countries. This study is an open label randomized clinical trial of parallel groups (UMIN Clinical Trials Registry: UMIN000052753, https://center6.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000060194) to evaluate the effect of an intensive nutrition therapy and nutrient supplementation intervention (folic acid, iron, vitamin D, omega 3 fatty acids, myo-inositol and micronutrients) in pregnant women with obesity on the prevention of GDM, other perinatal outcomes, maternal and newborn nutritional status, and infant growth, adiposity, and neurodevelopment compared to usual care. Given the absence of established nutritional guidelines for managing obesity during pregnancy, there is a pressing need to develop and implement new nutritional programs to enhance perinatal outcomes.

Phytic Acid against Clostridium perfringens Type A: A Food Matrix Study.

This study evaluated the inhibitory effect of phytic acid (PA) on the spore germination and vegetative cells growth of Clostridium perfringens type A, as well as its effect in combination with maltodextrin (MD) in cooked sausages. The addition of 1% PA showed a satisfactory inhibition of spores' germination and vegetative cells growth of C. perfringens in BHI media. The inhibitory effect of 1% PA on vegetative cells was similar to the additive sodium sorbate (SS) at 10%. Subsequently, a mixture of PA-MD (1:1; w/w) was evaluated for the inhibition of C. perfringens spores in cooked sausages. The PA-MD 1.5% and 2.5% had a similar performance to SS 10% and a similar or higher performance than 0.015% NO2 (p < 0.05). In an unprecedented way, the present study demonstrated that PA inhibited spore germination and vegetative cells growth of C. perfringens, highlighting its potential use as an alternative and natural preservative for the meat industry.

Phytic acid accumulation in plants: Biosynthesis pathway regulation and role in human diet.

Phytate or phytic acid (PA), is a phosphorus (P) containing compound generated by the stepwise phosphorylation of myo-inositol. It forms complexes with some nutrient cations, such as Ca, Fe and Zn, compromising their absorption and thus acting as an anti-nutrient in the digestive tract of humans and monogastric animals. Conversely, PAs are an important form of P storage in seeds, making up to 90% of total seed P. Phytates also play a role in germination and are related to the synthesis of abscisic acid and gibberellins, the hormones involved in seed germination. Decreasing PA content in plants is desirable for human dietary. Therefore, low phytic acid (lpa) mutants might present some negative pleiotropic effects, which could impair germination and seed viability. In the present study, we review current knowledge of the genes encoding enzymes that function in different stages of PA synthesis, from the first phosphorylation of myo-inositol to PA transport into seed reserve tissues, and the application of this knowledge to reduce PA concentrations in edible crops to enhance human diet. Finally, phylogenetic data for PA concentrations in different plant families and distributed across several countries under different environmental conditions are compiled. The results of the present study help explain the importance of PA accumulation in different plant families and the distribution of PA accumulation in different foods.

Metabolomic analysis and antioxidant activity of wild type and mutant chia (Salvia hispanica L.) stem and flower grown under different irrigation regimes.

BACKGROUND: Chia (Salvia hispanica L.) is a functional food from Central America. Interest in it is growing rapidly due to the many health benefits from the seed. However, when chia is grown at high latitudes, seed yield may be low whereas a high stem biomass and immature inflorescences are produced. Little is known about the chemical composition and the properties of stems and flowers. In this work, the metabolite profile, the antioxidant activity, and the total polyphenol content of stems and inflorescences were evaluated in a factorial experiment with different chia populations (commercial black chia and long-day flowering mutants G3, G8, and G17) and irrigation (100% and 50% of evapotranspiration). RESULTS: The results show the influence of irrigation and seed source on the antioxidant activity and total polyphenol content of chia flower and stem. Inflorescences exhibit higher antioxidant activity, suggesting their potential use as natural antioxidant. The mutants G3 and G8, at 50% irrigation, contained the highest amounts of compounds with nutraceutical value, especially within the flower. The mutant G17 showed lower antioxidant activity and polyphenol content compared to other seed sources but exhibited high omega 3 content in flowers but low in stems. This indicates that chia varieties should be chosen according to the objective of cultivation. CONCLUSION: These findings, indicating a close relation of metabolite content with irrigation and seed source, may provide the basis for the use of chia flower and stem for their nutraceutical value in the food, feed, and supplement industries. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Hydroalcoholic Extract of Solanum lycocarpum A. St. Hil. (Solanaceae) Leaves Improves Alloxan-Induced Diabetes Complications in Mice.

BACKGROUND: Solanum lycocarpum is a medicinal plant used in Brazil with hypoglycemic activity by its fruits use. However, the fruits production is restricted in some periods of the year, differently of leaves. OBJECTIVE: To evaluate the effects of hydroalcoholic extracts of S. lycocarpum leaves in alloxan-induced diabetic mice. METHODS: Hydroalcoholic extract of S. lycocarpum was characterized by phytochemical and GCMS analysis. The Antidiabetic activity was assessed following treatment for 22 days with S. lycocarpum extract at 125, 250, and 500 mg/kg. Bodyweight, water, and food intake, glycemia, biochemical parameters, anatomy-histopathology of the pancreas, liver and kidney, and expression of target genes were analyzed. In addition, oral acute toxicity was evaluated. RESULTS: Animals treated showed a significant reduction (p < 0.05) in glycemia following a dose of 125 mg/kg. Food intake remained similar for all groups. Decreased polydipsia symptoms were observed after treatment with 250 (p < 0.001) and 500 mg/kg (p < 0.01) compared with diabetic control, although normal rates were observed when 125 mg/kg was administered. A protective effect was also observed in the pancreas, liver, and kidneys, through the regeneration of the islets. Hypoglycemic activity can be attributed to myo-inositol, which stimulates insulin secretion, associated with α-tocopherol, which prevents damage from oxidative stress and apoptosis of ß-pancreatic cells by an increased Catalase (CAT) and Glutathione peroxidase 4 (GPX4) mRNA expression. The toxicological test demonstrated safe oral use of the extract under the present conditions. CONCLUSION: Hydroalcoholic extract of S. lycocarpum promotes the regulation of diabetes in the case of moderate glycemic levels, by decreasing glycemia and exerting protective effects on the islets.

Suplementos nutricionales para prevención de diabetes mellitus gestacional: lecciones aprendidas basadas en la evidencia.

INTRODUCTION: Gestational diabetes mellitus (GDM) affects between 5 and 40% of pregnant women. Recently different interventions with nutritional supplements have been evaluated for prevention of GDM. AIM: To perform a synthesis of the evidence on the efficacy of nutritional supplements (myo-inositol, probiotics, and vitamin D) in the prevention of GDM. METHODS: A systematic search in PubMed and Cochrane library was performed, including systematic reviews of randomized clinical trials (RCTs), published in English or Spanish until May 2020, using the keywords: "prevention", "gestational diabetes", "hyperglycemia and pregnancy", "supplementation", "probiotics", "myo-inositol" and "vitamin D". RESULTS: 10 systematic reviews that met the inclusion criteria were analyzed. Myo-inositol supplementation compared to placebo decreased the incidence of GDM (RR: 0.44 [0.27-0.87]; five RCTs), low quality of evidence. Although the supplementation with vitamin D or probiotics during pregnancy could reduce the incidence of GDM the evidence is limited. CONCLUSIONS: Myo-inositol supplementation is effective for prevention of GDM in high-risk women. Supplementation with vitamin D or probiotics probably decreases the incidence of GDM.

INTRODUCTION: La diabetes mellitus gestacional (DMG) afecta a entre el 5 y el 40% de mujeres embarazadas. Recientemente se han evaluado diferentes intervenciones con suplementos nutricionales para prevenir la DMG. OBJETIVO: Realizar una síntesis de la evidencia sobre eficacia de suplementos nutricionales (mioinositol, probióticos y vitamina D) para prevenir DMG. MÉTODO: Se realizó una búsqueda sistemática en PubMed y la biblioteca Cochrane, se incluyeron revisiones sistemáticas de estudios clínicos aleatorizados (ECAs), publicados en idioma inglés o español hasta mayo de 2020; se utilizaron las palabras clave: "prevención", "diabetes gestacional", "hiperglicemia y embarazo", "suplementación", "probióticos", "mio-inositol" y "vitamina D". RESULTADOS: Se analizaron 10 revisiones sistemáticas que cumplieron los criterios de inclusión. La suplementación con mioinositol comparado con placebo disminuyó la incidencia de DMG (RR: 0.44; IC 95%: 0.27-0.87; cinco ECAs). Si bien la suplementación con vitamina D o probióticos durante el embarazo podría disminuir la incidencia de DMG, la evidencia es limitada. CONCLUSIONES: La suplementación con mioinositol es efectiva para prevenir DMG en mujeres de alto riesgo. La suplementación con vitamina D o probióticos probablemente disminuye la incidencia de DMG.

Uso de mioinositol más Bifidobacterium lactis y Lactobacillus rhamnosus para la prevención de diabetes mellitus gestacional en mujeres mexicanas.

AIM: To compare the incidence of gestational diabetes mellitus (GDM) in women with three or more risk factor to developing GDM supplemented with myo-inositol plus probiotics versus women care without supplementation. METHODS: Retrospective cohort study, group 1, women with supplementation (myo-inositol 2g plus Bifidobacterium lactis and Lactobacillus rhamnosus 5x108 UFC, twice per day, from 12-14 to 28 weeks of gestation; group 2, women with prenatal care without supplementation, matched by age and body mass index (BMI). The primary outcome was the incidence of GDM using the International Association of Diabetes and Pregnancy Study Groups criteria. RESULTS: Group 1 n=48, group 2 n=96. There were no significant baseline differences between groups in age, BMI and number of risk factors. The incidence of GDM in group 1 was n=14 (29.2%), and for group 2 n=46 (47.9%); RR: 0.61 (95% CI: 0.37-0.99; p = 0.03). CONCLUSIONS: Supplementation from 12-14 weeks of gestation with myo-inositol plus probiotics decrease the incidence of GDM in Mexican women.

OBJETIVO: Comparar la incidencia de diabetes mellitus gestacional (DMG) en mujeres con tres o más factores de riesgo para desarrollar DMG suplementadas con mioinositol más probióticos versus mujeres sin suplementación.­. MATERIAL Y ­MÉTODOS: Estudio de cohorte retrospectivo, grupo 1, mujeres con suplementación (mioinositol 2 g más Bifidobacterium lactis y Lactobacillus rhamnosus 5x108unidades formadoras de colonias, dos veces al día, de las 12-14 hasta las 28 semanas de gestación); grupo 2, mujeres con control prenatal habitual sin suplementación, pareadas por edad e índice de masa corporal (IMC). El resultado primario fue la incidencia de DMG utilizando los criterios de la Asociación Internacional de Grupos de Estudio de Diabetes y Embarazo. RESULTADOS: Grupo 1, n = 48, y grupo 2 n = 96. No hubo diferencias significativas en características basales como edad, IMC, y numero de factores de riesgo entre los grupos. La incidencia de DMG en el grupo 1 fue n = 14 (29.2%) y en el grupo 2 n = 46 (47.9%); RR: 0.61 (IC 95%: 0.37-0.99; p = 0.03). CONCLUSIONES: La suplementación desde las 12-14 semanas de gestación con mioinositol más probióticos disminuye la incidencia de DMG en mujeres mexicanas.

In vitro digestion effect on mineral bioaccessibility and antioxidant bioactive compounds of plant-based beverages.

Consumption of plant-based beverages (PBB) is a growing trend; and have been used as viable substitutes for dairy based products. To date, no study has comparatively analyzed mineral composition and effect of in vitro digestion on the bioaccessibility of different PBB. The aim of this research was to investigate the content of essential minerals (calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn)) and to estimate the effect of in vitro digestion in plant-based beverages, and their antioxidant bioactive compounds (phenolic compounds and antioxidant capacity). Moreover, the presence of antinutritional factors, such as myo-inositol phosphates fractions, were evaluated. Samples of PBB (rice, cashew nut, almond, peanut, coconut, oat, soy, blended or not with another ingredients, fortified with minerals or naturally present) and milk for comparison were evaluated. TPC ranged from 0.2 mg GAEq/L for coconut to 12.4 mg GAEq/L for rice and, the antioxidant capacity (DPPH) ranged from 3.1 to 306.5 µmol TE/L for samples containing peanut and oat, respectively. Only a few samples presented myo-inositol phosphates fractions in their composition, mostly IP5 and IP6, especially cashew nut beverages. Mineral content showed a wide range for Ca, ranging from 10 to 1697.33 mg/L for rice and coconut, respectively. The Mg content ranged from 6.29 to 251.23-268.43 mg/L for rice and cashew nut beverages, respectively. Fe content ranged from 0.76 mg/L to 12.89 mg/L for the samples of rice. Zinc content ranged from 0.57 mg/L to 8.13 mg/L for samples of oat and soy, respectively. Significant variation was observed for Ca (8.2-306.6 mg/L) and Mg (1.9-107.4 mg/L) dialyzed between the beverages, with lower concentrations of Fe (1.0 mg/L) and Zn (0.5 mg/L) in dialyzed fractions. This study provides at least 975 analytically determined laboratory results, providing important information for characterization and comparison of different plant-based beverages.

Myo-inositol in health and disease: its impact on semen parameters and male fertility.

BACKGROUND: Myo-inositol (cis-1,2,3,5-trans-4,6-cyclohexanehexol; MI) is the most prominent of nine inositol stereoisomers. MI, its phosphate derivatives, and associated lipids are widely found in vegetables and animal tissues and are known to participate in numerous biological processes. OBJECTIVES: To perform a review analysis on MI presence, functions, and impact in male fertility. MATERIALS AND METHODS: A thorough search of listed publications in PubMed on MI and its derivatives was done. RESULTS: Published information was found and compiled on MI identification, natural dietary sources and absorption, biosynthesis, concentrations, as well as MI as its derivatives (PI, PIP, GPI, IPG) roles in several human tissues and body fluids in health and disease. A section was focused on MI presence, biosynthesis, and functions in the mammalian male genital tract and in spermatozoa, and summarized reports describing the impact of in vivo and in vitro MI supplementation on human semen quality and fertility. Studies reported a discrete improvement in sperm motility in fresh and frozen-thawed semen, and a better sperm performance in natural and assisted fertility. DISCUSSION AND CONCLUSION: MI was reported as an effective supplement for sperm quality. In any case, several study designs lack appropriate controls or data analysis to confirm the relevance of the findings. While promising, larger prospective randomized controlled studies will be required to confirm the positive effect of MI supplementation in male infertility management. Moreover, further investigations are encouraged to unravel MI roles in sperm physiology and the underlying molecular mechanisms.

The insulin-sensitizing mechanism of myo-inositol is associated with AMPK activation and GLUT-4 expression in human endometrial cells exposed to a PCOS environment.

Polycystic ovary syndrome (PCOS) is an endocrine-metabolic disorder characterized by hyperandrogenism and ovulatory dysfunction but also obesity and hyperinsulinemia. These characteristics induce an insulin-resistant state in tissues such as the endometrium, affecting its reproductive functions. Myo-inositol (MYO) is an insulin-sensitizing compound used in PCOS patients; however, its insulin-sensitizing mechanism is unclear. To understand the relationship of MYO with insulin action in endometrial cells, sodium/myo-inositol transporter 1 (SMIT-1) (MYO-transporter), and MYO effects on protein levels related to the insulin pathway were evaluated. SMIT-1 was assessed in endometrial tissue from women with normal weight, obesity, insulin resistance, and PCOS; additionally, using an in vitro model of human endometrial cells exposed to an environment resembling hyperinsulinemic-obese-PCOS, MYO effect was evaluated on p-AMPK and GLUT-4 levels and glucose uptake by Western blot, immunocytochemistry, and confocal microscopy, respectively. SMIT-1 was detected in endometrial tissue from all groups and decreased in PCOS and obesity (P < 0.05 vs. normal weight). In the in vitro model, PCOS conditions decreased p-AMPK levels, while they were restored with MYO (P < 0.05). The diminished GLUT-4 protein levels promoted by PCOS environment were restored by MYO through SMIT-1 and p-AMPK-dependent mechanism (P < 0.05). Also, MYO restored glucose uptake in cells under PCOS condition through a p-AMPK-dependent mechanism. Finally, these results were similar to those obtained with metformin treatment in the same in vitro conditions. Consequently, MYO could be a potential insulin sensitizer through its positive effects on insulin-resistant tissues as PCOS-endometrium, acting through SMIT-1, provoking AMPK activation and elevated GLUT-4 levels and, consequently, increase glucose uptake by human endometrial cells. Therefore, MYO may be used as an effective treatment option in insulin-resistant PCOS women.

Phytase Superdosing in the Diet of Lightweight Replacement Pullets: Performance, Organ Biometry and Bone Characteristics

The aim of this study was to evaluate the effect of two types of phytases (the 1st produced from Escherichia coli and the 2nd from Aspergillus oryzae) with different dosages (300 and 900 Unit Phytase or FTU) on performance, organ biometry and bone quality of replacement pullets in the period of 8 to 17 weeks of age. A total of 288 Hy-Line White laying hens were used at 8 weeks of age. A completely randomized design was adopted in a 2 x 2 factorial scheme, totaling 4 treatments with 8 replicates of 9 birds each. Performance, biometric variables and bone characteristics were evaluated. There was no significant interaction between the factors for any of the evaluated variables. The conversion and the accumulated consumption were influenced (p 0.05) by the phytase dosages, with the highest results being observed for the consumption obtained with 300 FTU, and the best results for the feed conversion obtained with the dosage of 900 FTU. The relative weight of the liver was influenced by the treatments (p 0.05), presenting higher values with bacterial phytase and a dosage of 900 FTU. The sternum length and tibial deformity were influenced by fungal phytase (p 0.05). It is concluded that the use of 900 FTU superdosing and fungal phytase improves the performance and bone characteristics of light replacement pullets.

Phytase Superdosing in the Diet of Lightweight Replacement Pullets: Performance, Organ Biometry and Bone Characteristics

The aim of this study was to evaluate the effect of two types of phytases (the 1st produced from Escherichia coli and the 2nd from Aspergillus oryzae) with different dosages (300 and 900 Unit Phytase or FTU) on performance, organ biometry and bone quality of replacement pullets in the period of 8 to 17 weeks of age. A total of 288 Hy-Line White laying hens were used at 8 weeks of age. A completely randomized design was adopted in a 2 x 2 factorial scheme, totaling 4 treatments with 8 replicates of 9 birds each. Performance, biometric variables and bone characteristics were evaluated. There was no significant interaction between the factors for any of the evaluated variables. The conversion and the accumulated consumption were influenced (p 0.05) by the phytase dosages, with the highest results being observed for the consumption obtained with 300 FTU, and the best results for the feed conversion obtained with the dosage of 900 FTU. The relative weight of the liver was influenced by the treatments (p 0.05), presenting higher values with bacterial phytase and a dosage of 900 FTU. The sternum length and tibial deformity were influenced by fungal phytase (p 0.05). It is concluded that the use of 900 FTU superdosing and fungal phytase improves the performance and bone characteristics of light replacement pullets.(AU)

Diet and Nutritional Interventions with the Special Role of Myo-Inositol in Gestational Diabetes Mellitus Management. An Evidence-Based Critical Appraisal.

BACKGROUND: Gestational Diabetes Mellitus (GDM), defined as glucose intolerance with onset or first recognition during pregnancy, represents one of the most common maternal-fetal complications during pregnancy and it is associated with poor perinatal outcomes. To date, GDM is a rising condition over the last decades coinciding with the ongoing epidemic of obesity and Type 2 Diabetes Mellitus (T2DM). OBJECTIVE: The aim of this review is to discuss the role of diet and nutritional interventions in preventing GDM with the explanation of the special role of myo-inositol (MI) in this matter. METHODS: We performed an overview of the most recent literature data on the subject with particular attention to the effectiveness of diet and nutritional interventions in the prevention of GDM with the special role of MI. RESULTS: Nutritional intervention and physical activity before and during pregnancy are mandatory in women affected by GDM. Moreover, the availability of insulin-sensitizers such as different forms of inositol has dramatically changed the scenario, allowing the treatment of several metabolic diseases, such as those related to glucose dysbalance. Although the optimal dose, frequency, and form of MI administration need to be further investigated, diet supplementation with MI appears to be an attractive alternative for the GDM prevention as well as for the reduction of GDM-related complications. CONCLUSIONS: More studies should be conducted to prove the most effective nutritional intervention in GDM. Regarding the potential effectiveness of MI, further evidence in multicenter, randomized controlled trials is needed to draw firm conclusions.

Forced degradation of l-(+)-bornesitol, a bioactive marker of Hancornia speciosa: Development and validation of stability indicating UHPLC-MS method and effect of degraded products on ACE inhibition.

The antihypertensive activity of the medicinal plant Hancornia speciosa has been previously demonstrated by us, being the activity ascribed to polyphenols and cyclitols like l-(+)-bornesitol. We herein evaluated the stability of the bioactive marker bornesitol submitted to forced degradation conditions. Bornesitol employed in the study was isolated from H. speciosa leaves. An UHPLC-ESI-MS/MS method was developed to investigate bornesitol stability based on MRM (Multiple Reaction Monitoring) acquisition mode and negative ionization mode, employing both specific (m/z 193 → 161 Da) and confirmatory (m/z 193 → 175 Da) transitions. A gradient elution of 0.1% formic acid in water and acetonitrile was performed on a HILIC column. The method was validated and showed adequate linearity (r2 > 0.99), selectivity, specificity, accuracy, and precision (RSD < 2.9%). The method was robust for deliberate variations on dessolvation temperature, but not for changes in the flow rate and dessolvation gas. The results from the stability studies allowed us to classify bornesitol as labile for acidic and alkaline hydrolysis, but as very stable for oxidative and neutral hydrolysis exposure. Bornesitol was categorized as practically stable under photolysis degradation, whereas a considerable reduction on its contents was induced by metal ions and thermolysis exposure. Degraded samples from neutral hydrolysis and thermolysis were assayed in vitro for ACE inhibition and showed a substantial decrease in biological activity as compared to intact bornesitol. myo-Inositol was identified as the major degradation products in both matrices. This is the first report on bornesitol stability under different stress conditions and the obtained data are relevant for the development and quality control of standardized products from H. speciosa leaves.

Effect of enzymatic treatment on phytate content and mineral bioacessability in soy drink.

The objective of this study was to evaluate in vitro bioaccessibility of calcium (Ca), iron (Fe) and zinc (Zn) in soy drink after phytase treatment and correlate it with the content of myo-inositol phosphates. Samples of commercial soy drink products and one sample produced in the laboratory by maceration were evaluated. Phytase was applied using 300 U per liter in 60 min considering the phosphate release. The content of myo-inositol tris-, tetrakis-, pentakis and hexakisphosphate was not observed after phytase treatment. The solubility assay showed an increase from 2.0% to 20.8% for Ca, 2.2% to 37.1% for Fe and 38.8% to 67.4% for Zn after phytase treatment with significant differences (p ≤ 0.05) for most samples. Dialysis assay demonstrated 1.0% to 9.5% for Ca after phytase treatment (p ≤ 0.05) except for one commercial sample. The phytase treatment is a valuable alternative process for improving mineral natural availability in soy drink and decreased the use of salts in the fortification.

Molecular cloning of the myo-inositol oxygenase gene from the kidney of baboons.

The enzyme myo-Inositol oxygenase (MIOX) is also termed ALDRL6. It is a kidney-specific member of the aldo-keto reductase family. MIOX catalyzes the first reaction involved in the myo-inositol metabolism signaling pathway and is fully expressed in mammalian tissues. MIOX catalyzes the oxidative cleavage of myo-Inositol and its epimer, D-chiro-Inositol to D-glucuronate. The dioxygen-dependent cleavage of the C6 and C1 bond in myo-Inositol is achieved by utilizing the Fe2+/Fe3+ binuclear iron center of MIOX. This enzyme has also been implicated in the complications of diabetes, including diabetic nephropathy. The MIOX gene was amplified with reverse transcription-polymerase chain reaction from baboon tissue samples, and the product was cloned and sequenced. MIOX expression in the baboon kidney is described in the present study. The percentages of nucleotide and amino acid similarities between baboons and humans were 95 and 96%, respectively. The MIOX protein of the baboon may be structurally identical to that of humans. Furthermore, the evolutionary changes, which have affected these sequences, have resulted from purifying forces.

Knockdown of Myo-Inositol Transporter SMIT1 Normalizes Cholinergic and Glutamatergic Function in an Immortalized Cell Line Established from the Cerebral Cortex of a Trisomy 16 Fetal Mouse, an Animal Model of Human Trisomy 21 (Down Syndrome).

The Na+/myo-inositol cotransporter (SMIT1) is overexpressed in human Down syndrome (DS) and in trisomy 16 fetal mice (Ts16), an animal model of the human condition. SMIT1 overexpression determines increased levels of intracellular myo-inositol, a precursor of phophoinositide synthesis. SMIT1 is overexpressed in CTb cells, an immortalized cell line established from the cerebral cortex of a Ts16 mouse fetus. CTb cells exhibit impaired cytosolic Ca2+ signals in response to glutamatergic and cholinergic stimuli (increased amplitude and delayed time-dependent kinetics in the decay post-stimulation), compared to our CNh cell line, derived from the cerebral cortex of a euploid animal. Considering the role of myo-inositol in intracellular signaling, we normalized SMIT1 expression in CTb cells using specific mRNA antisenses. Forty-eight hours post-transfection, SMIT1 levels in CTb cells reached values comparable to those of CNh cells. At this time, decay kinetics of Ca2+ signals induced by either glutamate, nicotine, or muscarine were accelerated in transfected CTb cells, to values similar to those of CNh cells. The amplitude of glutamate-induced cytosolic Ca2+ signals in CTb cells was also normalized. The results suggest that SMIT1 overexpression contributes to abnormal cholinergic and glutamatergic Ca2+ signals in the trisomic condition, and knockdown of DS-related genes in our Ts16-derived cell line could constitute a relevant tool to study DS-related neuronal dysfunction.

A new natural source for obtainment of inulin and fructo-oligosaccharides from industrial waste of Stevia rebaudiana Bertoni.

Fructan-type inulin and fructo-oligosaccharides (FOS) are reserve polysaccharides that offer an interesting combination of nutritional and technological properties for food industry. Stevia rebaudiana is used commercially in the sweetener industry due to the high content of steviol glycosides in its leaves. With the proposal of using industrial waste, the objective of the present study was to isolate, characterize and evaluate the prebiotic activity of inulin and FOS from S. rebaudiana stems. The chemical characterization of the samples by GC-MS, NMR and off-line ESI-MS showed that it was possible to obtain inulin molecules from the S. rebaudiana stems with a degree of polymerization (DP) of 12, and FOS with a DP<6. The in vitro prebiotic assay of these molecules indicates a strain specificity in fermentation capacity of fructans as substrate. FOS molecules with a low DP are preferably fermented by beneficial microbiota than inulin molecules with higher DP.

Comparison of methodologies to quantify phytate phosphorus in diets containing phytase and excreta from broilers.

The use of a suitable methodology to quantify the phytate phosphorus (Pphy) content in both the feed and the excreta from broilers is required to enable accurate calculation of the catalytic efficiency of the phytase supplemented in the feed. This study was conducted to compare 2 analytical methodologies (colorimetry and also high-performance liquid chromatography with a refractive index detector) in order to calculate the phytase efficiency by utilizing the results from the methodology that was shown to be the most appropriate. One hundred and twenty broilers were distributed in a (4+1)×2 factorial arrangement, corresponding to 4 diets that were equally deficient in P supplemented with increasing levels of phytase (0, 750, 1,500, and 2,250 units of phytase activity - FTU - per kg of feed) plus 1 positive control diet without phytase, supplied to male and female birds. The result indicated that the colorimetric methodology with an extraction ratio of 1:20 (mass of sample in g:volume of the solvent extractor in mL) was shown to be the most adequate. There was no interaction between the phytase level and the sex of the broilers (p>0.05). Males consumed 12% more Pphy than did females (p<0.01), but the sex of the broilers did not affect (p>0.05) the excretion and retention coefficient of Pphy. The increase in the phytase level of the diet reduced (linear, p<0.01) the Pphy excretion. The greatest Pphy retention was estimated at 87.85% when the diet contained 1,950 FTU/kg (p<0.01), indicating that it is possible to reduce the inorganic P in the formulation at an amount equivalent to 87.85% of the Pphy content present in the feed, which, in this research, corresponds to a decrease in 2.86 g of P/kg of the feed.

Production of D-myo-inositol(1,2,4,5,6)pentakisphosphate using alginate-entrapped recombinant Pantoea agglomerans glucose-1-phosphatase

The glucose-1-phosphatase encoding gene (agp) of Pantoea agglomerans was sequenced and heterologously expressed in Escherichia coli. The enzyme showed very high homology to periplasmatic glucose-1-phosphatases of other members of the Enterobacteriaceae family. It was isolated from transformed Escherichia coli cells in a single step in high yields (32.3 ± 1.2 mg per litre of culture) by Ni-NT agarose affinity chromatography to >95 percent purity as calculated from specific activity determinations. The purified glucose-1-phosphatase was entrapped in alginate beads with an entrapment efficiency of >80 percent. Temperature stability was enhanced as a consequence of entrapment, whereas pH dependence of enzyme activity was not affected. Maximum catalytic activity of entrapped glucose-1-phosphatase was found at 70°C, whereas the free enzyme exhibited maximal activity at 60°C. A single pH optimum at pH 4.5 was determined for the free and the entrapped enzyme. Kinetic parameters for the hydrolysis of sodium phytate were found to be affected by entrapment. They were determined to be K M = 0.84 mmol l-1 and k cat = 8 s-1 at pH 4.5 and 37°C for the entrapped glucose-1-phosphatase and K M = 0.35 mmol l-1 and k cat = 20.5 s-1 for the free enzyme. Complete conversion of phytate into one single myo-inositol pentakisphosphate isomer, identified as D-myo-inositol(1,2,4,5,6)pentakis-phosphate, was shown to be feasible by using the enzyme-loaded alginate beads in batch operations. The entrapped enzyme showed a high operational stability by retaining almost full activity even after ten uses.