SPME-GC-MS untargeted metabolomics approach to identify potential volatile compounds as markers for fraud detection in roasted and ground coffee.

Roasted ground coffee has been intentionally adulterated for economic revenue. This work aims to use an untargeted strategy to process SPME-GC-MS data coupled with chemometrics to identify volatile compounds (VOCs) as possible markers to discriminate Arabica coffee and its main adulterants (corn, barley, soybean, rice, coffee husks, and Robusta coffee). Principal Component Analysis (PCA) showed the difference between roasted ground coffee and adulterants, while the Hierarchical Clustering of Principal Components (HCPC) and heat map showed a trend of adulterants separation. The partial Least-Squares Discriminant Analysis (PLS-DA) approach confirmed the PCA results. Finally, 24 VOCs were putatively identified, and 11 VOCs are candidates for potential markers to detect coffee fraud, found exclusively in one type of adulterant: coffee husks, soybean, and rice. The results for possible markers may be suitable for evaluating the authenticity of ground-roasted coffee, thus acting as a coffee fraud control and prevention tool.

Influence of the body mass and visceral adiposity on glucose metabolism in obese women with Pro12Pro genotype in PPARgamma2 gene.

INTRODUCTION: Glucose metabolism may be altered in obesity and genotype for PPAR 2 can influence this variable. OBJECTIVE: To evaluate the influence of body mass (BM) and visceral adiposity (VA) in glucose metabolism in morbid obese women with Pro12Pro genotype. METHODS: Were selected 25 morbidly obese women. Groups were formed according to body mass index (BMI) [G1: 40-45 kg/m² (n = 17); G2: > 45 kg/m² (n = 8)]. Anthropometric, glycemia and insulinemia assessments (fasting, 60 and 120 minutes after high polyunsaturated fatty acids meal) were carried out. The insulin resistance (IR) and insulin sensitivity (IS) were assessed by HOMA-IR and QUICKI respectively. RESULTS: G2 had higher BMI and waist circumference, compared to G1, impaired fasting glucose, low IS and higher IR. The postprandial glucose was normal, but there was a higher insulin peak one hour after the meal in G2. CONCLUSION: Increased BM and VA were associated with worse glucose metabolism suggesting metabolic differences between morbid obese with Pro12Pro genotype.

Introducción: El metabolismo de la glucosa puede estar alterado en la obesidad y el genotipo del gene PPAR 2 puede influir en este variable. Objetivo: Evaluar la influencia de la masa corporal (MC) y de la adiposidad visceral (AV) en el metabolismo de la glucosa en mujeres con obesidad de grado 3 con el genotipo Pro12Pro. Métodos: Se seleccionaron 25 mujeres con obesidad de grado 3. Se formaron grupos de acuerdo con el índice de masa corporal (IMC) [G1: 40-45 kg/m2 (n = 17), G2: > 45 kg/m2 (n = 8)]. Fueron hechas evaluaciones antropométricas, de la glucemia y de la insulinemia (en ayunas, 60 y 120 minutos después de la comida rica en ácidos grasos poliinsaturados). La resistencia a la insulina (RI) y sensibilidad a la insulina (SI) fueron evaluados por el HOMA-IR y QUICKI, respectivamente. Resultados: G2 tuvieron mayor índice de masa corporal y circunferencia de la cintura, en comparación con G1, peor glucemia en ayunas, baja SI y alta RI. La glucosa postprandial fue normal, pero hubo un pico de insulina más alto una hora después de la comida en G2. Conclusión: El aumento de la MC y de la AV se asociaron con peor metabolismo de la glucosa lo que sugiere diferencias metabólicas entre obesos de grado 3 con el genotipo Pro12Pro.

Influence of the body mass and visceral a diposity on glucose metabolism in obese women with Pro12Pro genotype in PPAR gamma2 gene / Influencia de la masa corporal y de la adiposidad visceral en el metabolismo de la glucosa en mujeres obesas con el genotipo Pro12Pro en el gene PPARGAMMA2

INTRODUCTION: Glucose metabolism may be altered in obesity and genotype for PPAR 2 can influence this variable. OBJECTIVE: To evaluate the influence of body mass (BM) and visceral adiposity (VA) in glucose metabolism in morbid obese women with Pro12Pro genotype. METHODS: Were selected 25 morbidly obese women. Groups were formed according to body mass index (BMI) [G1: 40-45 kg/m² (n = 17); G2: > 45 kg/m² (n = 8)]. Anthropometric, glycemia and insulinemia assessments (fasting, 60 and 120 minutes after high polyunsaturated fatty acids meal) were carried out. The insulin resistance (IR) and insulin sensitivity (IS) were assessed by HOMA-IR and QUICKI respectively.RESULTS:G2 had higher BMI and waist circumference, compared to G1, impaired fasting glucose, low IS and higher IR. The postprandial glucose was normal, but there was a higher insulin peak one hour after the meal in G2. CONCLUSION: Increased BM and VA were associated with worse glucose metabolism suggesting metabolic differences between morbid obese with Pro12Pro genotype (AU)

Introducción: El metabolismo de la glucosa puede estar alterado en la obesidad y el genotipo del gene PPAR 2 puede influir en este variable. Objetivo: Evaluar la influencia de la masa corporal (MC) y de la adiposidad visceral (AV) en el metabolismo de la glucosa en mujeres con obesidad de grado 3 con el genotipo Pro12Pro. Métodos: Se seleccionaron 25 mujeres con obesidad de grado 3. Se formaron grupos de acuerdo con el índice de masa corporal (IMC) [G1: 40-45 kg/m2 (n = 17), G2: > 45 kg/m2 (n = 8)]. Fueron hechas evaluaciones antropométricas, de la glucemia y de la insulinemia (en ayunas, 60 y 120 minutos después de la comida rica en ácidos grasos poliinsaturados). La resistencia a la insulina (RI) y sensibilidad a la insulina (SI) fueron evaluados por el HOMA-IR y QUICKI, respectivamente. Resultados: G2 tuvieron mayor índice de masa corporal y circunferencia de la cintura, en comparación con G1, peor glucemia en ayunas, baja SI y alta RI. La glucosa postprandial fue normal, pero hubo un pico de insulina más alto una hora después de la comida en G2. Conclusión: El aumento de la MC y de la AV se asociaron con peor metabolismo de la glucosa lo que sugiere diferencias metabólicas entre obesos de grado 3 con el genotipo Pro12Pro (AU)

Qualitative and quantitative assessment of genetically modified soy in enteral nutrition formulas by polymerase chain reaction based methods / Avaliação qualitativa e quantitativa de soja geneticamente modificada em fórmulas de nutrição enteral

OBJECTIVE: The aim of this work was to investigate the occurrence of Roundup Ready soybean in enteral nutrition formulas sold in Brazil. METHODS: A duplex Polymerase Chain Reaction based on the amplification of the lectin gene and the construction of the recombinant deoxyribonucleic acid of transgenic glyphosate-tolerant soybean (35S promoter and chloroplast transit peptide gene) was performed in order to analyze the deoxyribonucleic acid obtained from nine soy protein isolate-containing formulas. RESULTS: Despite the highly processed nature of the food matrices, amplifiable deoxyribonucleic acid templates were obtained from all tested samples, as judged by the amplification of the lectin gene sequence. However, amplicons relative to the presence of Roundup Ready soybean were restricted to one of the nine enteral nutrition formulas analyzed as well as to the soybean reference powder, as expected. Quantitative analysis of the genetically modified formula by real-time Polymerase Chain Reaction showed a content of approximately 0.3 percent (w/w) of recombinant deoxyribonucleic acid from the Roundup Ready soybean. CONCLUSION: The results show that one of the formulas contained genetically modified soy, pointing to the need of regulating the use of transgenic substances and of specific labeling in this product category.

OBJETIVO: Investigar a ocorrência de soja transgênica em fórmulas de suporte nutricional comercializadas no Brasil. MÉTODOS: Foi desenvolvido o método da reação em cadeia da polimerase duplex, com base na amplificação do gene na lectina, e na construção do ácido desoxirribonucléico recombinante da soja transgênica tolerante a glifosato (promotor 35S e gene de peptídeo de trânsito de cloroplasto), a fim de avaliar o ácido desoxirribonucléico extraído a partir das nove fórmulas contendo isolado protéico de soja. RESULTADOS: Apesar do alto grau de processamento aos quais os produtos avaliados foram submetidos, foi possível extrair ácido desoxirribonucléico amplificável a partir de todas as amostras, demonstrado pela amplificação do gene endógeno (lectina). Adicionalmente, o fragmento relativo à modificação genética da soja transgênica foi detectado em uma das nove amostras avaliadas, bem como na amostra relativa ao material de referência contendo 1,0 por cento de organismo geneticamente modificado. As análises quantitativas realizadas a partir da reação em cadeia da polimerase em tempo real revelaram a presença de aproximadamente 0,3 por cento de ácido desoxirribonucléico recombinante derivado de organismo geneticamente modificado na amostra de fórmula que apresentou resultado positivo. CONCLUSÃO: Os resultados demonstram que uma das fórmulas analisadas apresentava ingredientes derivados de soja geneticamente modificada, apontando para a necessidade de regulamentar a utilização de transgênicos, e de rotulagem específica nessa categoria de produtos.

Aplicabilidade da metodologia de reação de polimerase em cadeia em tempo real na determinação do percentual de organismos geneticamente modificados em alimentos / Applicability of the real-time polymerase chain reaction based-methods in quantification of genetically modified organisms in foods

A detecção de organismos geneticamente modificados na cadeia alimentar é um aspecto importante para todos os assuntos envolvidos no controle de matéria-prima, na indústria de alimentos e na distribuição. A rotulagem e a rastreabilidade de organismos geneticamente modificados são questões atuais que são consideradas para o comércio e a regulamentação. Atualmente, a rotulagem de alimentos processados contendo material transgênico detectável é exigida pela legislação brasileira. O governo brasileiro publicou Decreto nº 4.680 em abril de 2003, que exige rotulagem para todos os alimentos ou ingredientes de alimento, com o limite para rotulagem de 1 por cento. Embora a tecnologia de reação em cadeia da polimerase tenha algumas limitações, a alta sensibilidade e especificidade explicam sua escolha por parte dos laboratórios interessados em realizar análises de detecção de organismos geneticamente modificados e seus derivados. Entre os métodos atualmente disponíveis, aqueles baseados na reação em cadeia da polimerase geralmente são aceitos, considerando a sensibilidade e a confiabilidade na detecção de material geneticamente modificado-derivado em análises de rotina. Neste artigo, apresenta-se uma revisão de métodos atualmente disponíveis baseados na reação em cadeia da polimerase para detecção, identificação e quantificação de organismos geneticamente modificados e seus derivados, discutindo sua aplicabilidade e suas limitações.

Detection of genetically modified organisms in the food chain is an important issue for all subjects involved in raw material control, food industry and distribution. Both labeling and traceability of genetically modified organisms are current issues that are considered for trade and regulation. Currently, labeling of genetically modified foods containing detectable transgenic material is required by the Brazilian legislation. The Brazilian government published the Decree nº 4.680 in April 2003, which requires labeling for all foods or food ingredients, with a stricter labeling threshold of 1 percent. Although polymerase chain reaction technology has some limitations, the high sensitivity and specificity explain why it has been the first choice of most analytical laboratories interested in detection of genetically modified organisms and their derived products. Among the currently available methods, polymerase chain reaction-based methods are accepted, considering the sensitivity and reliability for detection of genetically modified-derived material in routine analysis. In this paper, a review of currently available polymerase chain reaction methods for screening and quantifying genetically modified-derived ingredients is presented, discussing their applicability and limitations.

Gln3p and Nil1p regulation of invertase activity and SUC2 expression in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, sensing and signalling pathways regulate gene expression in response to quality of carbon and nitrogen sources. One such system, the target of rapamycin (Tor) proteins, senses nutrients and uses the GATA activators Gln3p and Nil1p to regulate translation in response to low-quality carbon and nitrogen. The signal transduction, triggered in response to nitrogen nutrition that is sensed by the Tor proteins, operates via a regulatory pathway involving the cytoplasmic factor Ure2p. When carbon and nitrogen are abundant, the phosphorylated Ure2p anchors the also phosphorylated Gln3p and Nil1p in the cytoplasm. Upon a shift from high- to low-quality nitrogen or treatment with rapamycin all three proteins are dephosphorylated, causing Gln3p and Nil1p to enter the nucleus and promote transcription. The genes that code for yeast periplasmic enzymes with nutritional roles would be obvious targets for regulation by the sensing and signalling pathways that respond to quality of carbon and nitrogen sources. Indeed, previous results from our laboratory had shown that the GATA factors Gln3p, Nil1p, Dal80p, Nil2p and also the protein Ure2 regulate the expression of asparaginase II, coded by ASP3. We also had observed that the activity levels of the also periplasmic invertase, coded by SUC2, were 6-fold lower in ure2 mutant cells in comparison to wild-type cells collected at stationary phase. These results suggested similarities between the signalling pathways regulating the expression of ASP3 and SUC2. In the present work we showed that invertase levels displayed by the single nil1 and gln3 and by the double gln3nil1 mutant cells, cultivated in a sucrose-ammonium medium and collected at the exponential phase, were 6-, 10- and 60-fold higher, respectively, in comparison to their wild-type counterparts. RT-PCR data of SUC2 expression in the double-mutant cells indicated a 10-fold increase in the mRNA(SUC2) levels.

Production of L-asparaginase by filamentous fungi.

L-asparaginase production was investigated in the filamentous fungi Aspergillus tamarii and Aspergillus terreus. The fungi were cultivated in medium containing different nitrogen sources. A. terreus showed the highest L-asparaginase (activity) production level (58 U/L) when cultivated in a 2% proline medium. Both fungi presented the lowest level of L-asparaginase production in the presence of glutamine and urea as nitrogen sources. These results suggest that L-asparaginase production by of filamentous fungi is under nitrogen regulation.

Production of L-asparaginase by filamentous fungi

L-asparaginase production was investigated in the filamentous fungi Aspergillus tamarii and Aspergillus terreus. The fungi were cultivated in medium containing different nitrogen sources. A. terreus showed the highest L-asparaginase (activity) production level (58 U/L) when cultivated in a 2 percent proline medium. Both fungi presented the lowest level of L-asparaginase production in the presence of glutamine and urea as nitrogen sources. These results suggest that L-asparaginase production by of filamentous fungi is under nitrogen regulation.