Frequency of CDH1 germline variants and contribution of dietary habits in early age onset gastric cancer patients in Brazil.

INTRODUCTION: The contribution of CDH1 germline variants to gastric cancer burden among young adults is unknown in Brazil. We aimed to evaluate the frequency of CDH1 germline variants and the diet/lifestyle habits in early age onset gastric cancer (EOGC, ≤ 55 years old) patients. METHODOLOGY: From 2013 to 2015, a total of 88 unrelated and consecutive patients diagnosed with EOGC were enrolled. All CDH1 exons and intronic boundaries were sequenced, and large genomic rearrangements were screened by MLPA. CDH1 transcription analysis was performed for variants that could potentially induce an effect on splicing. The diet and lifestyle habits of EOGC patients were compared to Brazilian population diet and lifestyle, obtained from governmental databases. RESULTS: Of 88 patients, the mean age at EOGC diagnosis was 39 years and 55% fulfilled the criteria for hereditary diffuse gastric cancer. The majority of the tumors were diffuse (74%) and poorly differentiated (80%). In total, 4 novel missense variants of uncertain significance (VUS) were identified: c.313T>A, c.387G>T, c.1676G>A, and c.1806C>A. The MLPA results revealed no rearrangements and CDH1 transcription analysis for variants of interest were inconclusive. EOGC patients had a higher red (OR:2.6, 95%CI:1.4-4.9) and processed (OR:3.1, 95%CI:1.6-6.0) meat intake and higher fruit consumption (OR:0.4, 95%IC:0.3-0.7) compared to eating habits of the Brazilian population. CONCLUSIONS: No unequivocal pathogenic germline CDH1 variants were identified in Brazilian EOGC patients. Dietary habits may be associated with the EOGC development.

Selective enumeration and viability of Bifidobacterium animalis subsp. lactis in a new fermented milk product

One of the key focuses of today's dairy industry worldwide is the continued development of new products, especially probiotic-based products. Buttermilk is originally a by-product of butter making fermented by Mesophilic Aromatic Cultures (MAC). It can also be made by fermentation of pasteurized whole milk or skimmed milk. This product is not marketed in Brazil. The objectives of this work were: (1) to develop a selective medium for Bifidobacterium animalis subsp. lactis enumeration and (2) to determine the viability of this microorganism during the shelf life of the buttermilk. Skim milk added with 10 percent sucrose or 0.03 percent sucralose was pasteurized and inoculated with a composite starter culture consisting of 1 percent MAC (containing Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis biovar. diacetylactis and Leuconostoc mesenteroides subsp. cremoris) and 2 percent Bifidobacterium animalis subsp. lactis. To attain selective counts of Bif. animalis subsp. lactis the MRS agar supplemented with 0.5 percent L-cysteine hydrochloride at 10 percent, 1 percent lithium chloride at 10 percent, 0.01 percent aniline blue and 0.5 percent dicloxacillin at 0.1 percent was modified by increasing the antibiotic concentration, addition of NaCl, adjusting pH to 4.8 or increasing the incubation temperature (from 37 to 45°C). Raising the incubation temperature to 45°C was found to be efficient in inhibiting the MAC cultures, even in media not added with dicloxacillin. Bif. animalis subsp. lactis exhibited high viability in the product. The buttermilk product prepared with sucrose and sweetener contained in excess of 10(8) cfu.ml-1 bifidobacteria throughout the shelf life of the product (28 days).

Atualmente, um dos principais focos da indústria de laticínios em todo o mundo é o desenvolvimento de novos produtos, especialmente probióticos. Buttermilk é originalmente um sub-produto do processamento da manteiga fermentado por Culturas Aromáticas Mesofílicas (MAC). Pode também ser feito pela fermentação de leite integral ou desnatado. Este produto não é comercializado no Brasil. Os objetivos deste trabalho foram o desenvolvimento de meio de cultura seletivo para Bifidobacterium animalis subsp. lactis e a determinação da viabilidade deste microrganismo durante a vida de prateleira do buttermilk produzido. Leite desnatado foi adicionado de 10 por cento da sacarose ou 0,03 por cento de sucralose, pasteurizado e inoculado com 1 por cento de MAC composto por Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis biovar. diacetylactis e Leuconostoc mesenteroides subsp. cremoris e por 2 por cento de Bifidobacterium animalis subsp. lactis. Para obter contagens seletivas de Bif. animalis subsp. lactis, o meio MRS ágar suplementado com 0,5 por cento L-cisteína HCl a 10 por cento, 1 por cento cloreto de lítio a 10 por cento, 0,01 por cento azul de anilina e 0,5 por cento dicloxacilina a 0,1 por cento foi modificado pelo aumento da concentração de antibiótico, adição de NaCl, ajuste de pH para 4,8 ou aumento da temperatura de incubação (de 37 para 45°C). A temperatura de incubação de 45°C foi eficiente para inibir as culturas MAC mesmo sem adição de antibiótico ao meio. Bif. animalis subsp. lactis apresentou alta viabilidade no produto. O buttermilk preparado com sacarose e edulcorante, apresentou mais de 10(8) ufc.ml-1 de Bif. animalis subsp. lactis durante a vida-de-prateleira (28 dias).