RESUMO
This article refers to the paper "Assessment of table olive fermentation by functional data analysis" (Ruiz-Bellido et al., 2016) [1]. The dataset include pH, titratable acidity, yeast count and area values obtained during fermentation process (380 days) of Aloreña de Málaga olives subjected to five different fermentation systems: i) control of acidified cured olives, ii) highly acidified cured olives, iii) intermediate acidified cured olives, iv) control of traditional cracked olives, and v) traditional olives cracked after 72 h of exposure to air. Many of the Tables and Figures shown in this paper were deduced after application of Functional Data Analysis to raw data using a routine executed under R software for comparison among treatments by the transformation of raw data into smooth curves and the application of a new battery of statistical tools (functional pointwise estimation of the averages and standard deviations, maximum, minimum, first and second derivatives, functional regression, and functional F and t-tests).
RESUMO
For the first time, functional data analysis (FDA) was used to assess the effects of different treatments on Protection Denomination of Origin Aloreña de Málaga table olive fermentations, focusing on the evolution of yeast population. The analysis of fermentation by a conventional approach led to scarce information. However, the transformation of microbial (and also physicochemical) data into smooth curves allowed the application of a new battery of statistical tools for the analysis of fermentations (functional pointwise estimation of the averages and standard deviations, maximum, minimum, first and second derivatives, functional regression, and functional F and t-tests). FDA showed that all the treatments assayed led to similar trends in yeast population while changes in pH and titratable acidity profiles led to several significant differences. Therefore, FDA represents a promising and valuable tool for studying table olive fermentations and for food microbiology in general.
Assuntos
Fermentação , Microbiologia de Alimentos/métodos , Olea/microbiologia , Leveduras/metabolismo , Interpretação Estatística de Dados , Concentração de Íons de Hidrogênio , Leveduras/crescimento & desenvolvimentoRESUMO
This study uses an "omics" approach to evaluate the bacterial biodiversity changes during fermentation process of natural green cracked Aloreña de Málaga table olives, from raw material to fermented fruit. For this purpose, two industries separated by almost 20km in Guadalhorce Valley (Málaga, Spain) were analysed for obtaining both brines and fruit samples at different moments of fermentation (0, 7, 30 and 120days). Physicochemical and microbial counts during fermentation showed the typical evolution of this type of processes, apparently dominated by yeasts. However, high-throughput barcoded pyrosequencing analysis of V2-V3 hypervariable region of the bacterial 16S rRNA gene showed at 97% identity the presence of 131 bacterial genera included in 357 operational taxonomic units, not detected by the conventional approach. The bacterial biodiversity was clearly higher in the olives at the moment of reception in the industry and during the first days of fermentation, while decreased considerably as elapse the fermentation process. The presence of Enterobacteriaceae and Lactobacillaceae species was scarce during the four months of study. On the contrary, the most important genus at the end of fermentation was Celerinatantimonas in both brine (95.3% of frequency) and fruit (89.4%) samples, while the presence of well-known spoilage microorganisms (Pseudomonas and Propionibacterium) and halophilic bacteria (Modestobacter, Rhodovibrio, Salinibacter) was also common during the course of fermentation. Among the most important bacterial pathogens related to food, only Staphylococcus genus was found at low frequencies (<0.02% of total sequences). Results show the need of this type of studies to enhance our knowledge of the microbiology of table olive fermentations. It is also necessary to determine the role played by these species not previously detected in table olives on the quality and safety of this fermented vegetable.
Assuntos
Bactérias/genética , Microbiologia de Alimentos , Olea/microbiologia , Leveduras/genética , Biodiversidade , Fermentação , Humanos , Metagenoma , RNA Ribossômico 16S , Sais , EspanhaRESUMO
Glutaminase (EC 3.5.1.2) is a key enzyme in rapidly proliferating cells. Using anti-sense technology, an Ehrlich ascites tumor cell line (0.28AS-2) with reduced glutaminase activity has been obtained. We investigated the in vivo growth characteristics of the 0.28AS-2 cells. When injected i.p. into normal Swiss albino mice, the 0.28AS-2 cells were unable to grow. On the contrary, when injected into nude mice, they developed into solid tumors. Mice inoculated with 0.28AS-2 cells kept immunologic memory and rejected a second inoculation with parental Ehrlich ascites tumor cells. Expression of both polymorphic epithelial mucin-1 (MUC-1) and the enzyme N-acetyl-alpha-D-galactosaminidase, proteins implicated in host immune system escape, were markedly diminished in 0.28AS-2 cells. Study of the immune system response in mice inoculated with 0.28AS-2 cells revealed an increase in splenic CD18 cells and the presence of a large number of activated F4/80+ macrophages in the ascites cavity. These features, not observed in mice inoculated with parental Ehrlich ascites tumor cells, indicate that a distinctive, strong immune response occurred in animals inoculated with 0.28AS-2 cells. Our results suggest that inhibition of glutaminase expression using anti-sense technology induces phenotypic changes in Ehrlich ascites tumor cells that allow the development of an effective anti-tumor immune response, which makes the cells unable to develop in vivo tumors.
Assuntos
Carcinoma de Ehrlich/enzimologia , Glutaminase/metabolismo , Hexosaminidases/metabolismo , Proteínas de Neoplasias/metabolismo , RNA Antissenso/metabolismo , Animais , Carcinoma de Ehrlich/imunologia , Carcinoma de Ehrlich/patologia , Feminino , Glutaminase/antagonistas & inibidores , Glutaminase/imunologia , Hexosaminidases/imunologia , Imunidade Celular , Memória Imunológica , Camundongos , Camundongos Nus , Mucina-1/metabolismo , Proteínas de Neoplasias/imunologia , Fator de Crescimento Transformador beta/metabolismo , Células Tumorais Cultivadas , alfa-N-AcetilgalactosaminidaseRESUMO
Phosphate-activated glutaminase has a critical role in tumours and rapidly dividing cells and its activity is correlated with malignancy. Ehrlich ascites tumour cells transfected with the pcDNA3 vector containing an antisense segment (0.28 kb) of rat kidney glutaminase showed impairment in the growth rate and plating efficiency, as well as a shortage in the glutaminase protein and activity. The C-terminal segment used is well conserved in all glutaminase sequences known. The transfected cells, named 0.28AS-2, displayed remarkable changes in their morphology compared with the parental cell line. The 0.28AS-2 cells also lost their tumourigenic capacity in vivo. Control mice developed an ascitic tumour, with a lifespan of 16+/-1 days, when inoculated with 10(7) cells/mouse; on the contrary, animals inoculated with transfected cells up to 2.5 times the cell numbers of control mice did not develop tumours and behaved as healthy animals. The ability to revert the transformed phenotype of antisense-transfected cells confirms the relevance of glutaminase in the transformation process and could provide new ways for the study of gene therapy.