Estudio de la bioaccesibilidad in vitro de zinc en arroz (oryza sativa) y su relación con el contenido de ácido fitico / In vitro bioaccesibility of zinc in rice (oryza sativa) and its relationship with phytic acid content

La bioaccesibilidad de un nutriente en un alimento sirve para determinar la calidad nutricional de éste para el consumo humano. El arroz es uno de los alimentos más importantes en la dieta por su gran aporte calórico y nutricional. El objetivo de este estudio fue analizar la bioaccesibilidad in vitro del zinc en arroz mediante espectrofotometría ultravioleta-visible y espectrometría de absorción atómica de llama y su relación con el contenido de ácido fítico. El porcentaje de bioaccesibilidad del zinc, respecto al porcentaje de ácido fítico, presentó una relación logarítmica inversamente proporcional (r= −0,669; p<0,05). Los valores porcentuales de bioaccesibilidad del zinc y ácido fítico en las líneas de arroz evaluadas se hallaron en un rango de 1,98,7% y 0,039-0,946% respectivamente. Se encontró que el ácido fítico afecta la bioaccesibilidad del zinc y que ésta no estuvo ligada a la concentración total del zinc presente en las líneas de arroz evaluadas. Las técnicas implementadas para cuantificar el zinc dializado presentaron diferencias significativas y se mostró que la técnica ultravioleta-visible no fue apta para este tipo de ensayos.

The bioavailability of a nutrient in a food serves to determine the nutritional quality for human consumption. Rice is one of the most important foods in diet due to its caloric and nutritional contribution. The objective of this study was to analyze the in vitro bioavailability of zinc in rice by ultraviolet-visible spectrophotometry and flame atomic absorption spectrometry and its relationship with phytic acid content. The percentage of zinc bioaccessibility with respect to phytic acid percentage, showed an inverse proportional logarithmic relationship (r= −0.669; p<0.05). The percentage values of zinc bioavailability and phytic acid in the evaluated rice varieties had a range of 1.9-8.7% and 0.039-0.946%, respectively. Phytic acid affected the bioaccessibility of zinc and was not linked to the total concentration of zinc present in the rice lines evaluated. The techniques implemented to quantify zinc dialyzed presented significant differences. It was shown that the ultraviolet-visible technique was not suitable for this type of assay.

Feasibility of sentinel lymph node dissection using Tc-99m phytate in papillary thyroid carcinoma

PURPOSE: Various methods of sentinel lymph node (SLN) biopsy in thyroid cancer have been introduced. Tc-99m phytate as a radiotracer has been successfully utilized for SLN biopsy in breast, cervix, and endometrial cancer. We assessed the feasibility of SLN dissection using Tc-99m phytate in papillary thyroid carcinoma (PTC). METHODS: Seventeen patients with PTC were prospectively enrolled. Ultrasound-guided peritumoral injection of 55.5 MBq Tc-99m phytate in 0.25-mL normal saline was performed. Preoperative single-photon emission-computed tomography (SPECT) and intraoperative gamma-probe were used for SLN detection during operation. RESULTS: Identification rate of SLNs was 70.6% (12 of 17) with SPECT, and 88.2% (15 of 17) with gamma-probe. Combined SPECT and gamma-probe had identification rates of 88.2% (15 of 17). Identification rates of SLNs in central LN compartments were 82.4% (14 of 17) and 41.2% (7 of 17) in lateral LN compartments. Overall sensitivity, specificity, positive predictive value, and negative predictive value of the results of SLN biopsy were 91.6%, 100%, 88.4%, and 100%, respectively. Eight patients (47.1%) had metastasis in SLNs; all patients had SLN metastasis in the central compartment and 2 patients had SLN metastasis in both the central and lateral compartments. CONCLUSION: Combined SPECT and gamma-probe could detect SLNs with an 88.2% identification rate in PTC. SLN biopsy using Tc-99m phytate is technically feasible. Further investigation is warranted for clinical application of Tc-99m phytate in PTC.

Antimutagenic and Antioxidant Activities of Thai Rice Brans

BACKGROUND: Rice bran is the outer layer of the rice grain, and contains high amounts of bioactive phytochemicals. Here, we investigated and compared chemopreventive properties of purple and white rice bran extracts. METHODS: Rice bran was extracted with dichloromethane and methanol. Chemical constituents in the extracts were analyzed by colorimetric assay and high performance liquid chromatography. The mutagenicity and antimutagenicity of the extracts were determined via the Salmonella mutation assay. The anticarcinogenic enzyme induction and antioxidant activities of the extracts were examined using Hepa1c1c7 cells and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, respectively. RESULTS: The methanol extracts of rice bran contained high amounts of phenolic acids, flavonoids, anthocyanins, and phytic acid, whereas large amounts of γ-oryzanol and vitamin E were presented in the dichloromethane extract. None of the extracts were mutagenic to Salmonella typhimurium. All rice bran extracts had strong antimutagenic effects against aflatoxin B1- and 2-amino-3,4-dimethylimidazo [4,5-f]quinoline-induced mutagenesis. The inhibitory effect against 2-aminofluorene-induced mutagenesis was found in the dichloromethane extract, while only the methanol extract of purple rice bran exhibited antimutagenic effects against benzo(a)pyrene. None of the extracts induced quinone reductase activity in Hepa1c1c7 cells. Additionally, the greatest antioxidant capacity was found in the methanol extract of purple rice bran. CONCLUSIONS: The methanol extract of purple rice bran containing high amount of phenolic acids, flavonoids, anthocyanins, and phytic acid showed the most effective antioxidant and antimutagenic activities by inhibiting mutagenic metabolizing enzymes and/or scavenging free radicals. These results demonstrate the nutritional and medical value of Thai rice for cancer prevention.

Phytase production through response surface methodology and molecular characterization of Aspergillus fumigatus NF191.

Phytase play an important role in phytic acid catalysis that act as a food inhibitor in cereals. Here, we isolated high phytase producing isolates NF191 closely related to Aspergillus fumigatus sp. from piggery soil. DNA was isolated from the fungal culture and amplified the ITS region using ITS1 and ITS4 primer using PCR. The 400-900 bp amplicon was gel eluted and subjected to sequencing. The sequencing results were assembled and compared with NCBI data base which showed the 99% identity of Aspergilllus fumigatus. Different carbon sources viz., fructose, galactose, lactose, dextrose, sucrose, maltose and different nitrogen sources (organic & inorganic) NH4Cl, NH4NO3, (NH4)2SO4, KNO3, NaNO3, urea, yeast extract, peptone, beef extract were tested for optimal production. The 0.3% dextrose, 0.5% NH4NO3 and 96 h incubation time showed the best production and enzyme activity at 45 ºC incubation temperature. The selected parameters, dextrose, ammonium sulphate and incubation time, when employed with statistical optimization approach involving response surface optimization using Box Behnken Design, gave a 1.3 fold increase in phytase production compared to unoptimized condition.

Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The KM for sodium phytate hydrolysis was 30.9 mM, while the kcat and kcat/KM were 1.46 ×105 s−1 and 4.7 × 106 s−1.M−1, respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg2+, Cd2+, K+ and Ca2+, and it was drastically inhibited by F−. The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.

Dietas com fitase para frangos de corte alojados em ambientes com diferentes sistemas de climatização / Diets with phytase for broilers housed in environments with differenta air-conditioning systems

O experimento foi conduzido para avaliar o efeito de diferentes níveis de fitase nas dietas de frangos de corte sobre as variáveis de desempenho, rendimento de carcaça e cortes, incorporação de proteína bruta, cálcio e fósforo na carcaça e teores de nitrogênio, cálcio e fósforo na cama das aves aos 41 dias de idade. Seiscentos e quarenta pintos de um dia foram distribuídos em boxes em delineamento inteiramente casualizado, esquema fatorial 5 x 2, com cinco níveis de fitase: 0, 1000, 2000, 3000 e 4000 FTU por kg de ração, com quatro repetições e 16 aves na unidade experimental, associados a dois ambientes (galpão com ventilação e nebulização e galpão com ventilação sem nebulização). As aves receberam dietas isonutritivas à base de milho e farelo de soja, formuladas para atender as exigências nutricionais, exceto para fósforo disponível, que foi reduzido em 20% da exigência. O ambiente com ventilação e nebulização reduziu a temperatura média da pele, melhorou a viabilidade da criação e as variáveis de desempenho de frangos de corte. Além disso, aumentou a incorporação de proteína bruta e fósforo na carcaça das aves, bem como elevou os teores de nitrogênio, cálcio e fósforo na cama, em relação ao ambiente com apenas ventilação. A suplementação com fitase até 4000 FTU por kg da dieta aumentou o rendimento de peito.

The experiment was conducted to evaluate effect of different levels of phytase in broiler diets on performance variables, carcass yield and cuts, incorporation of crude protein, calcium and phosphorus in housing and levels of nitrogen, calcium and phosphorus in bed birds at 41 days of age. Six hundred and forty day-old chicks were distributed in boxes in a randomized design, factorial scheme 5 x 2, with five levels of phytase: 0, 1000, 2000, 3000 and 4000 FTU per kg diet, with four replications and 16 birds in experimental unit associated with two rooms (shed with ventilation and fogging and shed with ventilation without fogging). Birds were fed diets isonutrient maize and soybean meal, formulated to meet nutritional requirements, except for available phosphorus, which was reduced in 20% of requirement. The room with ventilation and fogging reduced skin temperature, improved feasibility and performance variables of broilers. It also increased incorporation of crude protein and phosphorus in poultry housing and elevated levels of nitrogen, calcium and phosphorus in bed, in relation to environment with only ventilation. Phytase supplementation to 4000 FTU per kg diet increased breast meat yield.

Minerales, ácido fítico y γ-oryzanol en harinas integrales y salvados de tres variedades de arroz (oryza sativa l) cultivadas en venezuela / Minerals, phytic acid and γ-oryzanol in whole wheat flour and bran of three varieties of rice (oryza sativa l) grown in venezuela

El propósito de esta investigación fue determinar el contenido de algunos minerales, de ácido fítico y de γ-oryzanol en harinas integrales y salvados de tres cultivares (Cimarrón, Fonaiap 1 y Zeta 15) de arroz (Oryza sativa L) cosechados en Venezuela. Los minerales (Ca, Mg, Fe y Zn) fueron cuantificados por espectrofotometría de absorción atómica, mientras que el fósforo (P) y el ácido fítico fueron estimados a través de un método colorimétrico. El γ-oryzanol se cuantificó por HPLC. En general, el contenido de minerales fue mayor en la harina y en el salvado de la variedad Zeta 15. El contenido de P (400,0 ± 5,00 mg/100 g) y Mg (60,0 ± 0,00 mg/100 g) fue menor en la harina de la variedad Cimarrón, mientras que en los salvados de las distintas variedades se observaron valores significativamente altos de P (1860 ± 2,00 - 2185 ± 0,58 mg/100 g) y Ca (66,7 ± 1,15 ­ 73,3 ± 1,53 mg/100 g). Las tres variedades evaluadas representan importantes fuentes de ácido fítico y γ-oryzanol; sin embargo, la variedad Cimarrón fue la que exhibió las menores concentraciones de estos compuestos bioactivos. Basado en los resultados obtenidos, las harinas integrales y los salvados representan una fuente importante de minerales, de ácido fítico y de γ-oryzanol. La concentración de todos los microcomponentes es significativamente superior en los salvados con respecto a las harinas, lo que los convierte en una atractiva alternativa para la obtención de compuestos con actividad biológica de interés para la salud humana(AU)

The objective of this study was to determine the content of some minerals, phytic acid and γ-oryzanol in whole-grain flour and bran in three cultivars (Cimarrón, Fonaiap 1 y Zeta 15) of rice (Oryza sativa L) in Venezuela. The minerals were (Ca, Mg, Fe y Zn) quantified by Atomic Absorption Spectrophotometry while that the phosphorus (P) and phytic acid were determined using a colorimetric method. The γ-oryzanol was quantified by HPLC. In general, the mineral contents were higher in flour than bran in the cultivar Zeta 15. The P (400,0 ± 5,00 mg/100 g) and Mg (60,0 ± 0,00 mg/100 g) contents were lower in flour of the cultivar Cimarrón, while that were observed higher values of P (1860 ± 2,00 - 2185 ± 0,58 mg/100 g) and Ca (66,7 ± 1,15 ­ 73,3 ± 1,53 mg/100 g) in the bran of different cultivars. The three cultivars evaluated are an important source of phytic acid and γ-oryzanol; however, the Cimarrón cultivar exhibited the lowest concentrations of these bioactive compounds. Based on these results, whole-grain flours and brans may offer a good source of minerals, phytic acid and γ-oryzanol. The concentrations of all micro compounds evaluated were higher in the brans than the flours, this become the bran an attractive alternative to obtain compounds with biological activity of interest for human health(AU)

Inappropriate intracervical injection of radiotracer for sentinel lymph node mapping in a uterine cervix cancer patient: importance of lymphoscintigraphy and blue dye injection

Herein, we report a case of sentinel lymph node mapping in a uterine cervix cancer patient, referring to the nuclear medicine department of our institute. Lymphoscintigraphy images showed inappropriate intra-cervical injection of radiotracer. Blue dye technique was applied for sentinel lymph node mapping, using intra-cervical injection of methylene blue. Two blue/cold sentinel lymph nodes, with no pathological involvement, were intra-operatively identified, and the patient was spared pelvic lymph node dissection. The present case underscores the importance of lymphoscintigraphy imaging in sentinel lymph node mapping and demonstrates the added value of blue dye injection in selected patients. It is suggested that preoperative lymphoscintigraphy imaging be considered as an integral part of sentinel lymph node mapping in surgical oncology. Detailed results of lymphoscintigraphy images should be provided for surgeons prior to surgery, and in case the sentinel lymph nodes are not visualized, use of blue dye for sentinel node mapping should be encouraged

Inhibitory effect of inositol hexaphosphate on proliferation of LNCaP cells and its relation to IGFBP 3 expression / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of inositol hexaphosphate (IP6) on proliferation of human prostate carcinoma LNCaP cells and its relation to insulin-like growth factors binding protein-3 (IGFBP-3) expression.</p><p><b>METHODS</b>The siRNA technology was used to silence the IGFBP-3 gene in LNCaP cells. LNCaP cells and IGFBP-3 gene silenced LNCaP cells were exposed to IP6 for 24 h. Cell viability was measured by MTT assay; cell cycle arrest and cell apoptosis were detected by flow cytometry. The expression levels of IGFBP-3 and Bcl-2 mRNA and protein were analyzed by real-time quantitative RT-PCR and Western blotting, respectively.</p><p><b>RESULTS</b>The proliferation of LNCaP cells was be inhibited by IP6 in a dose dependent manner. After exposure to IP6 for 24 h, the cell viability in LNCaP cells and siRNA-treated LNCaP cells was 53.2%±11.6% and 82.3%±10.9%, respectively (P<0.05). After treatment of 1.5 mmol IP6，the apoptosis rate of LNCAP cells and siRNA-treated LNCAP cells was 40.48%±13.21% and 30.43%±10.65%, respectively (P<0.05). The proportion of G1 and G2 phase in LNCAP cells was 70.58%±8.25% and 5.64%±1.23%，after IP6 treatment the percentage of G1 phase cells decreased to 48.66%±11.23% and G2 phase cells increased to 31.11%±9.68%. However, for siRNA treated LNCAP cells, the proportion of G1 phase cells was 58.25%±12.36% and G2 phase cells was 23.85%±12.45%. Higher expression of IGFBP-3 and lower expression of Bcl-2 in LNCaP cells treated with IP6 were found at both mRNA and protein levels. IP6 treatment enhanced IGFBP-3 mRNA expression by 2.21±0.15 folds. In the contrast, expression of Bcl-2 mRNA decreased by 0.69±0.03 folds. Meanwhile, after IGFBP- gene silence Bcl-2 expression was not decreased.</p><p><b>CONCLUSION</b>IP6 can inhibit the proliferation of LNCaP cells, which may be associated with the changes of IGFBP-3 level through Bcl-2 expression.</p>

The ileum as a determinant organ of the functional liver cell mass in rats

PURPOSE: To evaluate if the ileum resection changes the functioning liver cell mass, the hepatic metabolism and the biodistribution of radiopharmaceutical in rats. METHODS: Twelve Wistar rats weighing 285g±34g were randomly divided into the ileum resection group (n = 6) and sham group rats (n = 6). After 30 days, they were anesthetized and 0.1mL of 99m-Tc-phytate (0.66MBq) was injected via femoral vein. After 30 minutes, blood samples were collected for red blood cells radioactive labeling and serum ALT, AST and gammaGT. Liver samples were used for 99m-Tc-phytate percentage of radioactivity/gram of tissue and histopathology. Student 's t test was used with significance 0.05. RESULTS: There was a higher uptake of 99m-Tc-phytate in the liver of sham rats, compared to the ileum resection group (p<0.05). GammaGT, ALT and AST were increased in ileum resection rats compared to sham (p<0.05). The he patocytes count was significantly lower in ileum resection group than in sham (p<0.05). Liver: body mass ratio was lower in experimental animals than in sham group (p<0.05). CONCLUSION: These data support that the ileum has important role in liver function and liver mass regulation, and they have potential clinical implications regarding the pathogenesis of liver injury following lower bowel resection.

Isolation, morphological and molecular characterization of phytate-hydrolysing fungi by 18S rDNA sequence analysis

Phytate is the primary storage form of phosphate in plants. Monogastric animals like poultry, pigs and fishes have very low or no phytase activities in their digestive tracts therefore, are incapable to efficiently utilize phytate phosphorus from the feed. Phytase from microbial sources are supplemented to feedstuff of these to increase the uptake of phytate phosphorus. In the present work efforts were made to isolate and characterize proficient phytase producing fungi from soil. Phytase producing fungi were isolated using phytate specific medium. Fungal isolates were selected according to their higher phytase activities. These isolates were further characterized and identified by morphological and microscopic analysis and confirmed by amplification of 18S rRNA gene, using specific primers. This gene was subsequently sequenced and phylogenetic affiliations were assigned. Fungal isolates were identified as various species of Aspergillus. Phytases from these fungi could be utilized as a feed additive in poultry and swine industries.

Study of thermal behavior of phytic acid

Phytic acid is a natural compound widely used as depigmenting agent in galenic cosmetic emulsions. However, we have observed experimentally that phytic acid, when heated to 150 ºC for around one hour, shows evidence of thermal decomposition. Few studies investigating this substance alone with regard to its stability are available in the literature. This fact prompted the present study to characterize this species and its thermal behavior using thermal analysis (TG/DTG and DSC) and to associate the results of these techniques with those obtained by elemental analysis (EA) and absorption spectroscopy in the infrared region. The TG/DTG and DSC curves allowed evaluation of the thermal behavior of the sample of phytic acid and enabled use of the non-isothermal thermogravimetric method to study the kinetics of the three main mass-loss events: dehydration I, dehydration II and thermal decomposition. The combination of infrared absorption spectroscopy and elemental analysis techniques allowed evaluation of the intermediate products of the thermal decomposition of phytic acid. The infrared spectra of samples taken during the heating process revealed a reduction in the intensity of the absorption band related to O-H stretching as a result of the dehydration process. Furthermore, elemental analysis results showed an increase in the carbon content and a decrease in the hydrogen content at temperatures of 95, 150, 263 and 380 °C. Visually, darkening of the material was observed at 150 °C, indicating that the thermal decomposition of the material started at this temperature. At a temperature of 380 °C, thermal decomposition progressed, leading to a decrease in carbon and hydrogen. The results of thermogravimetry coupled with those of elemental analysis allow us to conclude that there was agreement between the percentages of phytic acid found in aqueous solution. The kinetic study by the non-isothermal thermogravimetric method showed that the dehydration process occurred in two stages. Dehydration step I promoted a process of vaporization of water (reaction order of zero), whereas dehydration step II showed an order of reaction equal to five. This change in reaction order was attributed to loss of chemically bonded water molecules of phytic acid or to the presence of volatile substances. Finally, the thermal decomposition step revealed an order of reaction equal to one. It was not possible to perform the kinetic study for other stages of mass loss.

O ácido fítico é um composto natural muito utilizado como despigmentante em emulsões cosméticas magistrais. No entanto, observou-se experimentalmente que o ácido fítico, quando aquecido a 150 °C durante cerca de uma hora, mostra evidências de decomposição térmica e que poucos estudos envolvendo essa espécie, isoladamente quanto a sua estabilidade, estão disponíveis na literatura. Esse fato motivou o estudo de caracterização e de comportamento térmico dessa espécie, empregando a análise térmica (TG/DTG e DSC) e associando os resultados obtidos com aqueles de análise elementar (AE) e espectroscopia de absorção na região do infravermelho. As curvas TG/DTG e DSC permitiram avaliar o comportamento térmico da amostra de ácido fítico e, com isso, foi possível estudar, pelo método termogravimétrico não isotérmico, a cinética dos três principais eventos de perda de massa, o de desidratação I, desidratação II e de decomposição térmica. A associação das técnicas de espectroscopia de absorção no infravermelho e análise elementar permitiu avaliar os produtos intermediários da decomposição térmica do ácido fítico. Os espectros no infravermelho de amostras isoladas durante o aquecimento evidenciaram a diminuição de intensidade da banda de absorção relativa ao estiramento do grupo O-H como consequência do processo de desidratação. Também, os resultados de análise elementar indicaram que nas temperaturas de 95, 150 e 263 ºC houve aumento no teor de carbono e diminuição do teor de hidrogênio. Visualmente, observou-se o escurecimento do material a 150 ºC, indicando que a decomposição térmica do material iniciou-se nessa temperatura. Na temperatura de 380 ºC, a diminuição do teor de carbono e hidrogênio foi devido ao avanço do processo de decomposição térmica. Os resultados da termogravimetria juntamente com aqueles da análise elementar permitiram concluir que há concordância entre as percentagens de ácido fítico encontrado na solução aquosa. O estudo cinético pelo método termogravimétrico não isotérmico mostrou que o processo de desidratação ocorreu em duas etapas. Na etapa I da desidratação ocorreu um processo de vaporização de água (ordem de reação igual a zero). Na etapa II da desidratação foi encontrada uma ordem de reação igual a cinco. Essa mudança na ordem de reação foi atribuída à perda de moléculas de água quimicamente ligada, ao ácido fítico ou à presença de substâncias voláteis. Finalmente, na etapa de decomposição térmica foi observado que a ordem da reação foi igual a um. Para as outras etapas de perda de massa não foi possível a realização do estudo cinético.

The usefulness and accuracy of sentinel lymph node biopsy using single photon emission computed tomography/computed tomography with 99mTc phytate to detect locoregional lymph node metastases in patients with papillary thyroid carcinoma / 대한외과학회지

PURPOSE: Regional lymph node (LN) metastases are detected in 57-85% of patients with papillary thyroid carcinoma (PTC) and are associated with increased tumor recurrence. However, the management of lymphatic disease in patients with PTC has been ongoing source of debate. We have prospectively assessed the usefulness and accuracy of sentinel LN (SLN) biopsy for the detection of LN metastases in patients with PTC on preoperative imaging using single photon emission computed tomography/computed tomography (SPECT/CT) and 99mTc phytate. METHODS: We prospectively assessed 39 patients with PTC who had risk factors for recurrence or with the necessity of intraoperative LN sampling for suspicious LN metastases on preoperative imaging from August 2010 to March 2011. The patients underwent preoperative lymphoscintigraphy and SPETC/CT and intraoperative SLN biopsy (SLNB). RESULTS: 99mTc lymphoscintigraphy and SPECT/CT localized SLN in 38 patients (97.4%), with the gamma probe identifying 2.15 mean SLNs in the lateral neck of the 39 patients. Skip metastasis was found in one patient, and lateral compartment LN metastasis in 17 (43.5%). The sensitivity, specificity, and accuracy of SLNB for lateral compartment LN metastasis were 88.2%, 100%, and 94.8%, respectively. SLNB was more accurate and useful for lateral than for central compartment LN metastasis. CONCLUSION: SPECT/CT improved SLN detection and anatomical localization compared with lymphoscintigraphy. SLNB in patients with risk factors for recurrence or the necessity of intraoperative LN sampling for suspected LN metastases on preoperative imaging was accurate in detecting LN metastases and may help in deciding whether to perform lateral compartment dissection in patients with PTC.

Fósforo fítico y actividad de fitasa en fórmulas infantiles basadas en cereales

El ácido fítico es un ácido orgánico presente en granos de cereales. El fósforo contenido en su molécula (PFi) no es absorbible por una baja solubilidad, aunque su biodisponibilidad puede incrementarse por la actividad de enzimas fitasas (AFi). Con el objetivo de cuantificar el contenido de PFi y AFi en suplementos elaborados a partir de cereales y destinados a infantes lactantes, se seleccionaron cinco fórmulas de amplia distribución en mercados locales,identificadas de acuerdo a los cereales base de su composición como: A (arroz), T (trigo), ATS (arroz, trigo y aislado de soya), ATM (arroz, trigo y maíz) y ATMS (arroz, trigo, maíz y aislado de soya). Se tomaron cinco muestras por fórmula, cada una correspondiente a un envase (400 a 500 g) elaborado en lotes diferentes y con fecha de vencimiento posterior al muestreo. La proteína cruda varió de 16,8 a 7,2%, con el mayor valor para ATS, y los menores para T y ATM (P< 0,01). El extracto etéreo mostró un rango de 0,31 a 0,75%, mientras la relación calcio:fósforo entre 1,6 en ATM y 1,1 para el resto. El PFi representó el 61,5% del fósforo total en T, con diferencias (P< 0,01) respecto al resto (39,9 ± 6,8%). Sólo se detectó AFi en ATM, T y ATS (570, 300 y 152 U/kg, respectivamente). Se concluye que la composición química se ajustó a lo reportado por los fabricantes, con un elevado contenido de PFi y una AFi fundamentalmente asociados a la presencia de trigo en las fórmulas.

Phytic phosphorus and phytase activity in cerealbased infant formulas. Phytic acid is an organic acid present in cereal grains. The phosphorus inside its molecule (PPhy) is not available because of its low solubility, though the bioavailability could be increased by the activity of phytase enzymes (PhyA). With the purpose of quantifying the PPhy and PhyA in supplements manufactured from cereals and intended for lactating infants, five formulas of wide distribution in local markets were selected and identified depending on the main vegetable ingredients as A (rice), T (wheat), ATS (rice, wheat and soy isolate), ATM (rice, wheat and maize) and ATMS (rice, wheat, maize and soy isolate). Five samples were taken from each formula, each one corresponded to a commercial brand (400 to 500 g), coming from different batches and before their expiration date. The crude protein ranged from 7.2 to 16.8%, with de highest value for ATS and the lowest for T and ATM (P< 0.01). Ether extract varied from 0.31 to 0.75%, while the calcium: phosphorus ratio from 1.6 for ATM, to 1.1 for the rest. The PPhy was 61.5% of the total phosphorus in T, with differences (P<0.01) compared to the other formulas (39.9 ± 6.8%).PhyA was only detected in ATS, T and ATM (152,300 and 570 U/kg, respectively). The chemical composition complied with the manufacturer report, with a high content of PPhy and PhyA associated to the wheat presence in the formula.

Purification, characterization and properties of phytase from Shigella sp. CD2.

Phytases catalyze the release of phosphate from phytic acid. In this study, a phytase producing bacterial strain Shigella sp. CD2 was isolated from the wheat rhizosphere. Phytase production started from the exponential phase of bacterial growth, showing the highest activity during the stationary phase. The enzyme activity was detected in both periplasmic and intracellular fractions. The enzyme was purified by about 133-fold with specific activity 780 U mg-1 protein. The optimum pH and temperature of the enzyme was 5.5 and 60oC, respectively. The enzyme was thermostable and retained 100% and 75% of its activity on pre-incubation at 70o and 80oC for 30 min, respectively. The Km value for the substrate sodium phytate was 0.25 mM. The enzyme was highly specific to substrate phytate, and no activity was detected in presence of other phosphorylated substrates, such as ATP, ADP, glucose 6-phosphate, fructose 6-phosphate and p-nirophenyl phosphate. The activity declined dramatically in presence of Cu2+, Zn2+ and Fe2+ and SDS, whereas Mg2+ and Co2+ slightly enhanced the enzyme activity. The addition of other metal ions or chemicals had little or no effect on phytase activity. The enzyme was resistant to both pepsin and trypsin. Due to high specific activity, substrate specificity, good pH profile, protease insensitivity and thermostability, phytase encoding gene from Shigella sp. CD2 could be an interesting candidate for industrial applications. Further studies on cloning and expression of Shigella phytase gene are currently in progress.

Iron bioavailability of different maize genotypes developed in a breeding program: in vitro and in vivo studies

The objective of this study was to evaluate iron bioavailability of maize genotypes, and analyze the correlation between in vitro and in vivo methods. Dialysable iron was analyzed in 13 genotypes from which 5 were selected for the biological assay. Mean iron content of the genotypes (n=13) was 17.93±2.93 mg kg-1. Phytate varied from 0.77% to 1.03%; phytate: iron molar ratio from 30.64 to 55.41; and soluble iron from 13.17 to 39.63%. The highest value for dialysable iron was 19.14%. In the biological assay, the control group, that received ferrous sulphate, did not present significant difference between the genotypes for Hb gain, Hb gain per gram of iron consumed and HRE. Hb gain did not present a significant correlation with in vitro assay. However, there were positive correlations varying from 0.653 to 0.809. The maize genotypes evaluated presented a good bioavailability since the genotypes showed the same result in hemoglobin gain than control group.

Biodisponibilidade de ferro de diferentes genótipos de milho desenvolvidos em programa de melhoramento genético: estudos in vitro e in vivo. O objetivo deste estudo foi avaliar a biodisponibilidade do ferro de genótipos de milho e analisar a correlação entre métodos in vitro e in vivo. Ferro dialisável foi analisado em13 genótipos, a partir do qual 5 foram selecionados para o ensaio biológico. A média de teor de ferro dos genótipos (n= 13) foi 17,93 ± 2,93 mg kg-1. O teor de fitato variou de 0,77% a 1,03%; razão molar fitato:ferro de 30,64 a 55,41; e ferro solúvel de 13,17 a 39,63%.O valor mais alto para o ferro dialisável foi 19,14%. No ensaio biológico, o grupo controle, que recebeu sulfato ferrso, não apresentou diferença significativa entre os genótipos no ganho Hb, ganho de Hb por grama de ferro consumido e HRE. Ganho de Hb não apresentou correlação significativa com o ensaio in vitro. No entanto, houve correlações positivas variando de 0,653 a 0,809. Os genótipos de milho avaliados apresentaram uma boa biodisponibilidade uma vez que os genótipos apresentaram o mesmo resultado quanto ao ganho de hemoglobina em relação ao grupo controle.

Assessment of iron bioavailability in ten kinds of Chinese wheat flours using an in vitro digestion/Caco-2 cell model / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To compare iron bioavailability (Fe BV) from ten selected kinds of Chinese wheat flours in order to provide scientific basis for further human trials and enable plant breeding programs to screen biofortified wheat cultivars.</p><p><b>METHODS</b>An in vitro digestion/Caco-2 cell model was used to assess Fe BV of ten flour samples from six leading Chinese wheat cultivars and the stability of Fe BV in one cultivar was studied across three growing environments.</p><p><b>RESULTS</b>Significant differences were observed in both Fe BV and Fe bioavailability per gram of food (Fe BVPG) among cultivars (P<0.01) grown at the same location with the same flour extraction rate. Zhongyou 9507 and Jingdong 8 had Fe BV 37%-54% and Fe BVPG 103%-154% higher than the reference control. In the Anyang environment, Zhongyou 9507 had a higher wheat flour-Fe level and Fe BVPG. Differences in Fe BV were detected in cultivars with different flour extraction rates.</p><p><b>CONCLUSION</b>Zhongyou 9507 and Jingdong 8 were identified as the most promising cultivars for further evaluation of efficacy by using human subjects. The growing environments had no effect on Fe BV, but did have a significant effect on Fe BVPG. Fe bioavailabilities in low-extraction (40%) flours were higher than those in high-extraction (78%) flours.</p>

Heat-treated hull flour does not affect iron bioavailability in rats / Farinha de soja com casca tratada termicamente

In this study the chemical composition and iron bioavailability of hull and hull-less soybean flour from the new cultivar UFVTN 105AP was evaluated. The hemoglobin depletion-repletion method was used in Wistar rats. Soybean hull flour presented 37% more total dietary fiber and higher content of iron than hull-less soybean flour. The phytate:iron molar ratio, however, was 2-fold lower in the soybean hull flour in compared to the hull-less soybean flour. Animals fed soybean hull flour presented hemoglobin gains similar to those of the control diet group (p>0.05). The Relative Biological Values of hull and hull-less soybean flour were 68.5% and 67.1%, respectively, compared to the control group. Heat-treated soybean hull flour (150ºC/30 minutes) showed high content of iron and low phytate, which favors the iron bioavailability. Thus, the soybean hull flour is a better source of dietary fiber and iron than hull-less soybean flour at comparable bioavailabilities.

Neste estudo foram avaliadas a composição química e a biodisponibilidade de ferro das farinhas de soja com e sem casca do novo cultivar UFVTN 105AP. O método usado foi de depleção-repleção de hemoglobina em ratos Wistar. A farinha de soja com casca apresentou 37% a mais de fibra alimentar e maior teor de ferro (p 0,05). O Valor Relativo de Biodisponibilidade (VRB) da farinha de soja com casca e sem casca foram 68,5% e 67,1%, respectivamente, em relação ao sulfato ferroso (100%). A farinha de soja com casca submetida ao tratamento térmico (150ºC/30 minutos) apresentou maior teor de ferro e reduzido conteúdo de fitato, favorecendo a biodisponibilidade de ferro. Assim, a farinha de soja com casca apresentou melhor fonte de fibra alimentar e ferro do que a farinha de soja sem casca de comparável biodisponibilidade.