Distinct pattern of one-carbon metabolism, a nutrient-sensitive pathway, in invasive breast cancer: A metabolomic study.

Altered cell metabolism is a hallmark of cancer and critical for its development. Particularly, activation of one-carbon metabolism in tumor cells can sustain oncogenesis while contributing to epigenetic changes and metabolic adaptation during tumor progression. We assessed whether increased one-carbon metabolism activity is a metabolic feature of invasive ductal carcinoma (IDC). Differences in the metabolic profile between biopsies from IDC (n = 47) and its adjacent tissue (n = 43) and between biopsies from different breast cancer subtypes were assessed by gas spectrometry in targeted (Biocrates Life Science ® ) and untargeted approaches, respectively. The metabolomics data were statistically treated using MetaboAnalyst 4.0, SIMCA P+ (version 12.01), Statistica 10 software and t test with p < 0.05. The Cancer Genome Atlas breast cancer dataset was also assessed to validate the metabolomic profile of IDC. Our targeted metabolomics analysis showed distinct metabolomics profiles between IDC and adjacent tissue, where IDC displayed a comparative enrichment of metabolites involved in one-carbon metabolism (serine, glycine, threonine, and methionine) and a predicted increase in the activity of pathways that receive and donate carbon units (i.e., folate, methionine, and homocysteine). In addition, the targeted and untargeted metabolomics analyses showed similar metabolomics profiles between breast cancer subtypes. The gene set enrichment analysis identified different transcription-related functions between IDC and non-tumor tissues that involved one-carbon metabolism. Our data suggest that one-carbon metabolism may be a central pathway in IDC and even in general breast tumors, representing a potential target for its treatment and prevention.

Potential premalignant status of gastric portion excluded after Roux en-Y gastric bypass in obese women: A pilot study.

We evaluated whether the excluded stomach (ES) after Roux-en-Y gastric bypass (RYGB) can represent a premalignant environment. Twenty obese women were prospectively submitted to double-balloon enteroscopy (DBE) with gastric juice and biopsy collection, before and 3 months after RYGB. We then evaluated morphological and molecular changes by combining endoscopic and histopathological analyses with an integrated untargeted metabolomics and transcriptomics multiplatform. Preoperatively, 16 women already presented with gastric histopathological alterations and an increased pH (≥4.0). These gastric abnormalities worsened after RYGB. A 90-fold increase in the concentration of bile acids was found in ES fluid, which also contained other metabolites commonly found in the intestinal environment, urine, and faeces. In addition, 135 genes were differentially expressed in ES tissue. Combined analysis of metabolic and gene expression data suggested that RYGB promoted activation of biological processes involved in local inflammation, bacteria overgrowth, and cell proliferation sustained by genes involved in carcinogenesis. Accumulated fluid in the ES appears to behave as a potential premalignant environment due to worsening inflammation and changing gene expression patterns that are favorable to the development of cancer. Considering that ES may remain for the rest of the patient's life, long-term ES monitoring is therefore recommended for patients undergoing RYGB.

Docosahexaenoic Acid Modulates a HER2-Associated Lipogenic Phenotype, Induces Apoptosis, and Increases Trastuzumab Action in HER2-Overexpressing Breast Carcinoma Cells.

In breast cancer, lipid metabolic alterations have been recognized as potential oncogenic stimuli that may promote malignancy. To investigate whether the oncogenic nature of lipogenesis closely depends on the overexpression of HER2 protooncogene, the normal breast cell line, HB4a, was transfected with HER2 cDNA to obtain HER2-overexpressing HB4aC5.2 cells. Both cell lines were treated with trastuzumab and docosahexaenoic acid. HER2 overexpression was accompanied by an increase in the expression of lipogenic genes involved in uptake (CD36), transport (FABP4), and storage (DGAT) of exogenous fatty acids (FA), as well as increased activation of "de novo" FA synthesis (FASN). We further investigate whether this lipogenesis reprogramming might be regulated by mTOR/PPARγ pathway. Inhibition of the mTORC1 pathway markers, p70S6 K1, SREBP1, and LIPIN1, as well as an increase in DEPTOR expression (the main inhibitor of the mTOR) was detected in HB4aC5.2. Based on these results, a PPARγ selective antagonist, GW9662, was used to treat both cells lines, and the lipogenic genes remained overexpressed in the HB4aC5.2 but not HB4a cells. DHA treatment inhibited all lipogenic genes (except for FABP4) in both cell lines yet only induced death in the HB4aC5.2 cells, mainly when associated with trastuzumab. Neither trastuzumab nor GW9662 alone was able to induce cell death. In conclusion, oncogenic transformation of breast cells by HER2 overexpression may require a reprogramming of lipogenic genetic that is independent of mTORC1 pathway and PPARγ activity. This reprogramming was inhibited by DHA.

Comparison of Virtual Nutri Plus® and Dietpro 5i® software systems for the assessment of nutrient intake before and after Roux-en-Y gastric bypass.

OBJECTIVES: The assessment of nutritional intake before and after bariatric surgery assists in identifying eating disorders, nutritional deficiencies and weight loss/maintenance. The 7-day record is the gold standard for such an assessment and is interpreted using specialized software. This study sought to compare the Virtual Nutri Plus® and Dietpro 5i® software systems in assessing nutrient intake in obese patients with type 2 diabetes mellitus who underwent a Roux-en-Y gastric bypass. METHODS: Nutritional intake was assessed in 10 obese women with type 2 diabetes mellitus before and 3 months after Roux-en-Y gastric bypass. The 7-day record was used to assess food intake and then, the Virtual Nutri Plus® and Dietpro 5i® software systems were used to calculate calorie, macronutrient and micronutrient intake based on validated food chemical composition databases. Clinicaltrials.gov: NCT01251016. RESULTS: During the preoperative period, deficits in the ingestion of total fiber and 15 out of 22 estimated micronutrients were observed when using the Virtual Nutri Plus®, compared to deficiencies in total fiber and 4 micronutrients when using the Dietpro 5i®. During the postoperative period, both the Virtual Nutri Plus® and Dietpro 5i® systems detected deficits in the ingestion of total fiber, carbohydrates and 19 micronutrients, but only the Virtual Nutri Plus® detected deficits in complex B vitamins (except B12) and minerals. CONCLUSION: Virtual Nutri Plus® was more sensitive than Dietpro 5i® for the identification of deficits in nutrient intake in obese, type 2 diabetes mellitus patients undergoing Roux-en-Y gastric bypass.

Efficiency of the 24-hour food recall instrument for assessing nutrient intake before and after Roux-en-Y gastric bypass.

BACKGROUND: Nutritional and food intake assessments before and after bariatric surgery may be important to correct eating habits and nutritional deficiencies. AIM: To assess the efficiency of the 24-hour food recall (24hR) form as a fast method for assessing nutrient intake before and after a Roux-en-Y gastric bypass (RYGB). METHODS: This study analyzed data from 10 obese patients with type 2 diabetes mellitus (T2DM). Food intake (calories, macronutrients, and micronutrients) before and 3 months after RYGB were assessed with the 24hR as well as the seven-day food record (7dR) as the gold standard reference. Virtual Nutri Plus® software was used to quantify nutrients. RESULTS: The 7dR data revealed deficits in the estimated intake of total fiber and 14 out of 22 micronutrients pre- and postoperatively, combined; the 24hR failed to detect intake deficits in only two of these micronutrients (vitamins A and B3). Other postoperative deficits included carbohydrates, vitamin B1, copper, and iron, on which the 24hR was sensitive to only the iron deficit. In our pre- versus post-operative comparison analyses, the 7dR revealed decreases in total calories, carbohydrates, total and all subclasses of fat and fiber, and 12 micronutrients; in the analogous comparison analyses, the 24hR failed to detect decreases in the ingestion of monounsaturated and saturated fats and in six of these micronutrients. CONCLUSIONS: In obese T2DM patients, the 24hR performed reasonably well for probing nutrient intake before and after RYGB, but is not recommended for tracking changes over time, including pre- versus postoperative deficits.

Introducción: Las evaluaciones nutricionales y de ingesta de comida antes y después de cirugía bariátrica puede ser importante para corregir hábitos de alimentación y deficiencias nutricionales. Objetivo: Evaluar la eficacia del formulario de recordatorio de alimento de 24 horas (24hR) como método rápido de evaluación de ingesta de nutrientes antes y después de bypass gástrico en Y de Roux (RYGB). Métodos: Este estudio analizó datos de 10 pacientes obesos con diabetes mellitus de tipo 2 (T2DM). Se evaluó la ingesta de alimentos (calorías, macronutrientes y micronutrientes) antes y 3 meses después de RYGB con el 24hR además del registro de alimentos de siete días (7dR) como patrón de referencia estándar. Se utilizó el software Virtual Nutri Plus® para cuantificar los nutrientes. Resultados: Los datos del 7dR revelaron deficiencias en la ingesta estimada de fibra total y 14 de 22 micronutrientes pre- y postoperación, combinados; el 24hR no pudo detectar deficiencias de ingesta en solo dos de estos micronutrientes (vitaminas A y B3). Otras deficiencias postoperatorias incluyeron carbohidratos, vitamina B1, cobre y hierro, sobre las que el 24hR detectó solo la deficiencia de hierro. En nuestros análisis comparativos pre- y post-operación, el 7dR reveló descensos en calorías totales, carbohidratos, grasas totales y todas las subclasses de grasa y fibra, y 12 micronutrientes; en los análisis comparativos análogos, el 24hR no logró detectar descensos en la ingesta de grasas monoinsaturadas y saturadas y en seis de estos micronutrientes. Conclusiones: En pacientes obesos con T2DM, el 24hR funcionó razonablemente bien demostrando la ingesta de nutrientes antes y después de RYGB, pero no se recomienda para registrar cambios a lo largo del tiempo, incluyendo deficiencias pre- y postoperación.

Comparison of Virtual Nutri Plus® and Dietpro 5i® software systems for the assessment of nutrient intake before and after Roux-en-Y gastric bypass

OBJECTIVES: The assessment of nutritional intake before and after bariatric surgery assists in identifying eating disorders, nutritional deficiencies and weight loss/maintenance. The 7-day record is the gold standard for such an assessment and is interpreted using specialized software. This study sought to compare the Virtual Nutri Plus® and Dietpro 5i® software systems in assessing nutrient intake in obese patients with type 2 diabetes mellitus who underwent a Roux-en-Y gastric bypass. METHODS: Nutritional intake was assessed in 10 obese women with type 2 diabetes mellitus before and 3 months after Roux-en-Y gastric bypass. The 7-day record was used to assess food intake and then, the Virtual Nutri Plus® and Dietpro 5i® software systems were used to calculate calorie, macronutrient and micronutrient intake based on validated food chemical composition databases. Clinicaltrials.gov: NCT01251016. RESULTS: During the preoperative period, deficits in the ingestion of total fiber and 15 out of 22 estimated micronutrients were observed when using the Virtual Nutri Plus®, compared to deficiencies in total fiber and 4 micronutrients when using the Dietpro 5i®. During the postoperative period, both the Virtual Nutri Plus® and Dietpro 5i® systems detected deficits in the ingestion of total fiber, carbohydrates and 19 micronutrients, but only the Virtual Nutri Plus® detected deficits in complex B vitamins (except B12) and minerals. CONCLUSION: Virtual Nutri Plus® was more sensitive than Dietpro 5i® for the identification of deficits in nutrient intake in obese, type 2 diabetes mellitus patients undergoing Roux-en-Y gastric bypass. .

Lipid raft disruption by docosahexaenoic acid induces apoptosis in transformed human mammary luminal epithelial cells harboring HER-2 overexpression.

In HER-2-overexpressing breast cells, HER-2 receptors exist on the cell surface as monomers, homodimers and heterodimers. For signal activation and transduction to occur, HER-2 must be localized to lipid rafts. Therefore, we hypothesized that the amount of lipid rafts on the cell membrane would be a factor in HER-2 signaling. To test this, we used HB4a (an untransformed human mammary epithelial cell line) and HB4aC5.2 cells. HB4aC5.2 cells are HB4a derivatives that have been transfected with five copies of pJ5E.c-ErbB-2 and express approximately 900 times more HER-2 than HB4a cells. In these cells, HER-2 overexpression was accompanied by increased lipid rafts in cell membranes, a hyperactivation of downstream Akt and ERK1/2 proteins, and an increased rate of cell growth compared to HB4a. In addition, HER-2 overexpression was associated with an increased activation of FASN, a key enzyme involved in cellular lipogenesis. Its final product, palmitate, is frequently used to synthesize lipid rafts. We further hypothesized that treatment with docosahexaenoic acid (DHA), an omega-3 fatty acid, would disrupt the lipid rafts and lead to a growth arrest. In HB4aC5.2 cells, but not HB4a cells, we found that DHA treatment disrupted lipid raft; inhibited HER-2 signaling by decreasing activation of Akt, ERK1/2 and FASN proteins; and induced apoptosis. Although little is known about lipid rafts, our data support the idea that disturbances in these microdomains induced by DHA may represent a useful tool for controlling the signaling initiated by HER-2 receptors and its therapeutic potential in the treatment of HER-2 positive breast cancer.

Complementarity of Subjective Global Assessment (SGA) and Nutritional Risk Screening 2002 (NRS 2002) for predicting poor clinical outcomes in hospitalized patients.

BACKGROUND & AIMS: We evaluated the ability of Nutritional Risk Screening 2002 (NRS 2002) and Subjective Global Assessment (SGA) to predict malnutrition related to poor clinical outcomes. METHODS: We assessed 705 patients at a public university hospital within 48 h of admission. Logistic regression and number needed to screen (NNS) were calculated to test the complementarity between the tools and their ability to predict very long length of hospital stay (VLLOS), complications, and death. RESULTS: Of the patients screened, 27.9% were at nutritional risk (NRS+) and 38.9% were malnourished (SGA B or C). Compared to those patients not at nutritional risk, NRS+, SGA B or C patients were at increased risk for complications (p=0.03, 0.02, and 0.003, respectively). NRS+ patients had an increased risk of death (p=0.03), and SGA B and C patients had an increased likelihood of VLLOS (p=0.008 and p<0.0001, respectively). Patients who were both NRS+ and SGA C had lower estimates of NNS than patients who were NRS+ or SGA C only, though their confidence intervals did overlap. CONCLUSIONS: The concurrent application of SGA in NRS+ patients might enhance the ability to predict poor clinical outcomes in hospitalized patients in Brazil.

Efeitos de diferentes emulsões lipídicas sobre a expressão de moléculas de superfície envolvidas no processo de apresentação de antígenos em células mononucleares humanas in vitro / Effects of different lipid emulsions over the expression of surface molecules involved in the antigen presenting process of in vitro human mononuclear cells

Emulsões lipídicas parenterais (EL) podem modular funções leucocitárias, influenciando a resposta imune. O processo de apresentação de antígenos por leucócitos ocorre pela expressão de moléculas de superfície específicas e constitui o ponto-chave para o desencadeamento de resposta imune eficiente / Parenteral lipid emulsion (LE) can modulate leukocytes function and modify immune response. The antigen presenting process by leucocytes occurs through the expression of specific surface molecules and constitutes the key point for the start of an efficient immune response...

Influencia da nutriçäo parenteral total em regime glicidico e lipidico sobre a fagocitose de macrófagos de ratos / Fat and non-fat total parenteral nutrition effect on rat's macrophage phagocytosis

O uso de emulsöes lipídicas em nutriçäo parenteral total tem sido associado a alteraçöes do sistema mononuclear fagocitário. O objetivo do presente estudo foi avaliar a influência da nutriçäo parenteral contendo diferentes emulsöes lipídicassobre a fagocitose de macrófagos (Mf). Setenta ratos Wistar com a veia jugular externa cateterizada receberam, por via intrravenosa, diferentes regimes de nutriçäo parenteral total, isocalóricos, isonitrogenados e isogordurosos ou dieta oral e foram separados em sete grupos: 1) Grupo OS: dieta oral com infusäo intravenosa de soro fisiológico a 9 porcento; 2) Grupo GLU: NPT sem lípides; 3) GrupoLCT: NPT contendo emulsäo lipídica a 10 porcento de trtiglicérides de cadeia longa (TCL); 4) Grupo LCTFO1: NPT contendo TCL e emulsäo lipídica de óleo de peixe(FO) a 10 por cento; 5) Grupo LCTFO2: contendo TCL e FO; 6) Grupo MCT: NPT contendo emulsäo lipídicaa 10 porcento com 50 porcento de triglicérides de cadeia média (TCM) e 50 porcento de TCL; 7) Grupo MCTFO1: NPT contendo TCM, TCL e FO. Após 96 horas de ministraçäo de NPT ou SF, foi injetado, pela veia caudal do rato, 1,0 Ml/kg de peso corpóreo de carväo coloidal. Após três horas os animais foram sacrificados. Fígado, baço e pulmöes forampesados e estudados por imuno-histoquímica, com o método avidina-biotina utilizando o anticorpo anti-MfHAM-56. Verificou-se microscopicamente o número total de Mf (MT) e Mf que haviam fagocitado (MF) o carväo coloidal e estabeleceu-se o índice de fagocitose. O grupo que recebeu dieta oral foi o único a ganhar peso. Em nenhum grupo observou-se diferença no peso dos órgäos. A NPT em regime glicídico e lipídico com TCL inibiu a fagocitose de Mf no fígado,baço e pulmöes. A adiçäo de emulsäo lipidica de óleo de peixe às fórmulas de NPT contendo emulsöes lipídicas de TCM e TCL aumentou o número e a fagocitose de Mf no fígado e pulmöes. Perspectivas apontam a necessidade de estudos com diferentes concentraçöes de lípides em NPT valorizando outras funçöes dos macrófagos.(au)

Influencia da nutriçäo parenteral total em regime glicidico e lipidico sobre a fagocitose de macrofagos de ratos / Effect of fat and fat free total parenteral nutrition on macrophage phagocytosis in rats

O uso de nutricao parenteral total (NPT) tem sido associado a alteracoes anatomicas e funcionais do sistema mononuclear fagocitario. Os efeitos das emulsoes lipidicas parenterais sobre a fagocitose do sistema mononuclear fagocitario ainda sao controversos. O objetivo do presente estudo foi avaliar a influencia da nutricao parenteral total sem lipides e contendo diferentes emulsoes lipidicas sobre a fagocitose de macrofagos. Setenta ratos Wistar com a veia jugular externa cateterizada receberam, por via intravenosa central, diferentes regimes de nutricao parenteral total, isocaloricos (1,16 kcal/mL), isonitrogenados (1,5 g/mL), e isogordurosos (30 a 32 por cento do valor calorico nao proteico) ou dieta oral e foram separados em sete grupos...