Fatty Acid Unsaturation Degree of Plasma Exosomes in Colorectal Cancer Patients: A Promising Biomarker.

Even though colorectal cancer (CRC) is one of the most preventable cancers, it is currently one of the deadliest. Worryingly, incidence in people <50 years has increased unexpectedly, and for unknown causes, despite the successful implementation of screening programs in the population aged >50 years. Thus, there is a need to improve early diagnosis detection strategies by identifying more precise biomarkers. In this scenario, the analysis of exosomes is given considerable attention. Previously, we demonstrated the exosome lipidome was able to classify CRC cell lines according to their malignancy. Herein, we investigated the use of the lipidome of plasma extracellular vesicles as a potential source of non-invasive biomarkers for CRC. A plasma exosome-enriched fraction was analyzed from patients undergoing colonoscopic procedure. Patients were divided into a healthy group and four pathological groups (patients with hyperplastic polyps; adenomatous polyps; invasive neoplasia (CRC patients); or hereditary non-polyposis CRC. The results showed a shift from 34:1- to 38:4-containing species in the pathological groups. We demonstrate that the ratio Σ34:1-containing species/Σ38:4-containing species has the potential to discriminate between healthy and pathological patients. Altogether, the results reinforce the utility of plasma exosome lipid fingerprint to provide new non-invasive biomarkers in a clinical context.

A Combination of Apple Vinegar Drink with Bacillus coagulans Ameliorates High Fat Diet-Induced Body Weight Gain, Insulin Resistance and Hepatic Steatosis.

Obesity is a worldwide epidemic characterized by excessive fat accumulation, associated with multiple comorbidities and complications. Emerging evidence points to gut microbiome as a driving force in the pathogenesis of obesity. Vinegar intake, a traditional remedy source of exogenous acetate, has been shown to improve glycemic control and to have anti-obesity effects. New functional foods may be developed by supplementing traditional food with probiotics. B. coagulans is a suitable choice because of its resistance to high temperatures. To analyze the possible synergic effect of Vinegar and B. coagulans against the metabolic alterations induced by a high fat diet (HFD), we fed twelve-week-old C57BL/6 mice with HFD for 5 weeks after 2 weeks of acclimation on a normal diet. Then, food intake, body weight, blood biochemical parameters, histology and liver inflammatory markers were analyzed. Although vinegar drink, either alone or supplemented with B. coagulans, reduced food intake, attenuated body weight gain and enhanced glucose tolerance, only the supplemented drink improved the lipid serum profile and prevented hepatic HFD-induced overexpression of CD36, IL-1ß, IL-6, LXR and SREBP, thus reducing lipid deposition in the liver. The beneficial properties of the B. coagulans-supplemented vinegar appear to be mediated by a reduction in insulin and leptin circulating levels.

Influence of Storage Temperature and Packaging on Bacteria and Yeast Viability in a Plant-Based Fermented Food.

Optimization of food storage has become a central issue for food science and biotechnology, especially in the field of functional foods. The aim of this work was to investigate the influence of different storage strategies in a fermented food product (FFP) and further determine whether the regular storage (room temperature (RT) and standard packaging (SP)) could be refined. Eight experimental conditions (four different temperatures × two packaging) were simulated and changes in FFP's microbial ecology (total bacteria, lactic acid bacteria (LAB), and yeasts) and physicochemical characteristics (pH and moisture content (MC)) were determined following 1, 3, 6, and 12 months. All conditions tested showed a decline in microbial content due to the effect of the temperature, 37 °C being the most detrimental condition, while -20 and 4 °C seemed to be better than RT in some parameters. Vacuum packaging (VP) only had a major effect on MC and we found that VP preserved greater MC values than SP at 3, 6, and 12 months. The correlation analysis revealed that total bacteria, LAB, and yeasts were positively associated, and also both pH and MC showed a correlation. According to our results and with the purpose to maintain the load of viable microorganisms, we observed that the best storage conditions should contemplate SP and freezing or cooling temperature during a period no longer than 3 months.

A Fermented Food Product Containing Lactic Acid Bacteria Protects ZDF Rats from the Development of Type 2 Diabetes.

Type 2 diabetes (T2D) is a complex metabolic disease, which involves a maintained hyperglycemia due to the development of an insulin resistance process. Among multiple risk factors, host intestinal microbiota has received increasing attention in T2D etiology and progression. In the present study, we have explored the effect of long-term supplementation with a non-dairy fermented food product (FFP) in Zucker Diabetic and Fatty (ZDF) rats T2D model. The supplementation with FFP induced an improvement in glucose homeostasis according to the results obtained from fasting blood glucose levels, glucose tolerance test, and pancreatic function. Importantly, a significantly reduced intestinal glucose absorption was found in the FFP-treated rats. Supplemented animals also showed a greater survival suggesting a better health status as a result of the FFP intake. Some dissimilarities have been observed in the gut microbiota population between control and FFP-treated rats, and interestingly a tendency for better cardiometabolic markers values was appreciated in this group. However, no significant differences were observed in body weight, body composition, or food intake between groups. These findings suggest that FFP induced gut microbiota modifications in ZDF rats that improved glucose metabolism and protected from T2D development.

Identifying potential areas of expansion for the endangered brown bear (Ursus arctos) population in the Cantabrian Mountains (NW Spain).

Many large carnivore populations are expanding into human-modified landscapes and the subsequent increase in coexistence between humans and large carnivores may intensify various types of conflicts. A proactive management approach is critical to successful mitigation of such conflicts. The Cantabrian Mountains in Northern Spain are home to the last remaining native brown bear (Ursus arctos) population of the Iberian Peninsula, which is also amongst the most severely threatened European populations, with an important core group residing in the province of Asturias. There are indications that this small population is demographically expanding its range. The identification of the potential areas of brown bear range expansion is crucial to facilitate proactive conservation and management strategies towards promoting a further recovery of this small and isolated population. Here, we used a presence-only based maximum entropy (MaxEnt) approach to model habitat suitability and identify the areas in the Asturian portion of the Cantabrian Mountains that are likely to be occupied in the future by this endangered brown bear population following its range expansion. We used different spatial scales to identify brown bear range suitability according to different environmental, topographic, climatic and human impact variables. Our models mainly show that: (1) 4977 km2 are still available as suitable areas for bear range expansion, which represents nearly half of the territory of Asturias; (2) most of the suitable areas in the western part of the province are already occupied (77% of identified areas, 2820 km2), 41.4% of them occurring inside protected areas, which leaves relatively limited good areas for further expansion in this part of the province, although there might be more suitable areas in surrounding provinces; and (3) in the eastern sector of the Asturian Cantabrian Mountains, 62% (2155 km2) of the land was classified as suitable, and this part of the province hosts 44.3% of the total area identified as suitable areas for range expansion. Our results further highlight the importance of increasing: (a) the connectivity between the currently occupied western part of Asturias and the areas of potential range expansion in the eastern parts of the province; and (b) the protection of the eastern sector of the Cantabrian Mountains, where most of the future population expansion may be expected.

Suitability of a new mixed-strain starter for manufacturing uncooked raw ewe's milk cheeses.

Most raw milk Ossau-Iraty cheeses are currently manufactured on-farm using the same commercial streptococcal-lactococcal starter (S1). One way to enhance the microbial diversity that gives raw milk its advantages for cheese-making is to formulate new starters combining diverse, characterized strains. A new starter (OI) combining 6 raw milk strains of lactococci, recently isolated and characterized, was tested in parallel with the current starter by making 12 Ossau-Iraty raw milk cheeses at 3 farmhouses under the conditions prevailing at each farm. Compliance of the sensory characteristics with those expected by the Ossau-Iraty professionals, physicochemical parameters and coliforms were quantified at key manufacturing steps. The new starter OI gave cheeses having proper compliance but having lower compliance than the S1 cheeses under most manufacturing conditions, while managing coliform levels equally well as starter S1. This lower compliance relied more on the absence of Streptococcus thermophilus in starter OI, than on the nature of the lactoccocal strains present in starter OI. The study also shows that variations in 5 technological parameters during the first day of manufacture, within the range of values applied in the 3 farmhouses, are powerful tools for diversifying the scores for the sensory characteristics investigated.

Psychiatric care at a national mental institution during the Spanish Civil War (1936-39): Santa Isabel de Leganés.

The scanty research available regarding the health of the mentally ill during the Spanish Civil War is largely due to the loss of most documents, and to the difficulty in accessing the existing archives for decades. Up to the present time, historiography has described overcrowded facilities for the mentally disturbed and the fact that old buildings such as convents and spas were turned into establishments for treating patients with mental problems during the Civil War. However, research reviewing the institutional life and conditions of psychiatric patients during this war is still rather scarce.The aim of our article is to discuss the characteristics of the patients at Santa Isabel National Mental Asylum between 1936 and 1939, as well as the functioning of this institution located in Leganés, a city to the south of Madrid (Spain). The method for this study includes a review of the medical records, statistical registers and other documents kept in the institution's Historical Archive. In addition, using documents from other Spanish archives, as well as information obtained from contemporary and secondary sources, we attempt to describe similarities to and differences from other mental institutions.

Monitoring the sensory quality of canned white asparagus through cluster analysis.

BACKGROUND: White asparagus is one of the 30 vegetables most consumed in the world. This paper unifies the stages of their sensory quality control. The aims of this work were to describe the sensory properties of canned white asparagus and their quality control and to evaluate the applicability of agglomerative hierarchical clustering (AHC) for classifying and monitoring the sensory quality of manufacturers. RESULTS: Sixteen sensory descriptors and their evaluation technique were defined. The sensory profile of canned white asparagus was high flavor characteristic, little acidity and bitterness, medium firmness and very light fibrosity, among other characteristics. The dendrogram established groups of manufacturers that had similar scores in the same set of descriptors, and each cluster grouped the manufacturers that had a similar quality profile. CONCLUSION: The sensory profile of canned white asparagus was clearly defined through the intensity evaluation of 16 descriptors, and the sensory quality report provided to the manufacturers is in detail and of easy interpretation. AHC grouped the manufacturers according to the highest quality scores in certain descriptors and is a useful tool because it is very visual. © 2015 Society of Chemical Industry.

Biodiversity and growth dynamics of lactic acid bacteria in artisanal PDO Ossau-Iraty cheeses made from raw ewe's milk with different starters.

The biodiversity and growth dynamics of Lactic Acid Bacteria (LAB) in farm-house Ossau-Iraty cheeses were investigated from vat milk to 180 days of ripening in six independent batches made from six raw ewe's milks using five typical cheese-making methods. Commercial starter S1 was used for three batches, starter S1 combined with S2 for one batch and no starter for two batches. Up to ten LAB species from five genera and up to two strains per species were identified per milk; up to eleven species from five genera and up to three strains per species were identified per cheese. Lactococcus lactis, Lactobacillus paracasei, Enterococcus faecalis, Enterococcus faecium, Enterococcus durans, and Leuconostoc mesenteroides were detected in all cheeses. Lactococci reached the highest counts irrespective of the milk and starter used. Lactococci and enterococci increased during manufacture, and mesophilic lactobacilli increased during ripening. Strain and species numbers, the percentage of isolates originating from the raw milk, maximum counts of each genus/species and time for reaching them, all varied according to whether or not a starter was used and the composition of the starter. The genotypes of strains within species varied according to the raw milk used. This generated distinct LAB microbiotas throughout manufacture and ripening that will certainly impact on the characteristics of the ripened cheeses.

Physicochemical, microbiological and sensory profiles of fermented milk containing probiotic strains isolated from kefir.

A two-strain starter culture containing Lactobacillus plantarum CIDCA 83114, a potential probiotic strain isolated from kefir grains, and Streptococcus thermophilus CIDCA 321 was tested for the preparation of a fermented milk product. Kluyveromyces marxianus CIDCA 8154, a yeast with immunomodulatory properties was included to formulate a three-strain starter culture. Supernatants of enterohaemorragic Escherichia coli, shiga-toxin-producing strain, along with a two-strain or a three-strain starter culture were included in the medium of Vero-cell surface cultures. The results demonstrated that these combinations of microorganisms antagonize the cytopathic action of shiga toxins. The cell concentration of Lb. plantarum did not decrease during fermentation, indicating that the viability of this strain was not affected by low pH, nor did the number of viable bacteria change during 21 days of storage in either fermented products. The number of viable yeasts increases during fermentation and storage. Trained assessors analyzed the general acceptability of fresh fermented milks and considered both acceptable. The milk fermented with the two-strain starter culture was considered acceptable after two week of storage, while the product fermented with the three-strain starter culture remained acceptable for less than one week. The main changes in sensory attributes detected by the trained panel were in sour taste, milky taste and also in fermented attributes. The correlation between different sensory attributes and acceptability indicated that the panel was positively influenced by milky attributes (taste, odour, and flavour) as well as the intensity of flavour. In conclusion, the two-strain starter culture would be the more promising alternative for inclusion of that potential probiotic lactobacillus in a fermented milk product.

Use of wild Lactobacillus strains in an adjunct culture for a Roncal-type cheese.

The effect of an added adjunct culture consisting of facultatively heterofermentative lactobacilli (FHL) on the volatile compounds and sensory characteristics of a Spanish ewes'-milk cheese was examined. Three cheese batches were prepared using a commercial starter, one from raw milk, another from pasteurized milk, and a third from pasteurized milk with an added culture of wild Lactobacillus. paracasei+Lb. plantarum. Analysis of the volatile compounds was carried out by the purge and trap method and gas chromatography with a mass spectrometer and disclosed a total of 86 compounds belonging to the chemical families hydrocarbons, fatty acids, esters, ketones, aldehydes, and alcohols. After ageing for 120 and 240 days, the cheese samples underwent sensory analysis by a panel of expert assessors. The attributes evaluated were characteristic odour and odour intensity and characteristic aroma and aroma intensity. Pasteurization of the milk had an effect on the formation of certain volatile compounds, adversely affecting the characteristic flavour of the cheese. Use of the adjunct culture in addition to the commercial starter improved the flavour of the cheese made from the pasteurized milk, which earned sensory scores similar to those awarded to the cheese made from the raw milk. Use of adjunct cultures consisting of indigenous FHL strains could help to conserve the traditional characteristics of Roncal cheese made from pasteurized milk, although some technical adjustments to the Regulations would be needed.

Poly (acrylic acid) chitosan interpolymer complexes for stomach controlled antibiotic delivery.

The aim of this study was to develop a stomach-specific drug delivery system to increase the efficacy of amoxicillin against Helicobacter pylori. Polyacrylic acid (PAA), chitosan (CS), and amoxicillin (A) were employed to obtain polyionic complexes. The design of the hydrogel delivery system was based on the swellable approach; with a floating feature to prolong the Gastric Residence Time (GRT). The polyionic complex (PAA:CS:A 2.5:5:2) showed a sustained drug release profile in enzyme-free simulated gastric fluid (SGF) and pH 4.0. A pH independent swelling-eroding pattern with adequate maximum swelling ratios of 17.76 and 13.42 was obtained at in SGF and pH 4.0, respectively, with similar eroding profiles in both pH media. This network carrier provides an amoxicillin protective effect towards the hydrolytic degradation in SGF. The in vivo study was performed on healthy volunteers, using the [13C] octanoic acid breath test. The proposed hydrogel showed a prolonged GRT of up to 3 h. The preliminary results from this study suggest that amoxicillin polyionic complexes have potential for improving local antibiotic therapy against H. pylori.

Chitosan-poly(acrylic) acid polyionic complex: in vivo study to demonstrate prolonged gastric retention.

The aim of this study was to develop a chitosan-poly(acrylic) acid based controlled drug release system for gastric antibiotic delivery. Different mixtures of amoxicillin (A), chitosan (CS), and poly(acrylic) acid (PAA) were employed to obtain these polyionic complexes. A non-invasive method was employed for determining the gastric residence time of the formulations. It was studied the swelling behavior and drug release from these complexes. Gastric emptying rate study was performed by means of the [13C]octanoic acid breath test. The gastric emptying rates of two different formulations (conventional and gastric retentive system) were studied. Swelling studies indicated that the extent of swelling was greater in the polyionic complexes than in the single chitosan formulations. The amoxicillin diffusion from the hydrogels was controlled by the polymer/drug interaction. The property of these complexes to control the solute diffusion depends on the network mesh size, which is a significant factor in the overall behavior of the hydrogels. The gastric half-emptying time of the polyionic complex was significantly delayed compared to the reference formulation, showing mean values of 164.32+/-26.72 and 65.06+/-11.50min, respectively (P<0.01). The results of this study suggest that, these polyionic complexes are good systems for specific gastric drug delivery.

PCR and RAPD identification of L. plantarum strains isolated from ovine milk and cheese. Geographical distribution of strains.

Lactobacilli, and specifically Lactobacillus plantarum, are an important group of microorganisms in ovine cheeses, even though they are not ordinarily included in the starter cultures added. The present study effected counts of lactobacilli in Roncal Protected Designation of Origin (PDO) milk and cheese samples and isolated a total of 1026 strains. The strains were identified to species level by polymerase chain reaction (PCR) using L. plantarum-specific oligonucleotide primers, and the strains belonging to this species were then characterized by randomly amplified polymorphic DNA (RAPD). The percentage of L. plantarum present in the cheeses depended on the plant where the cheese was manufactured. Cluster analysis of the RAPD profiles obtained revealed seven main clusters. On comparing the strains, most of the strains present in the cheese were found not to have come from the raw milk.

Interpolymer complexes of poly(acrylic acid) and chitosan: influence of the ionic hydrogel-forming medium.

Non-covalent polyionic complexes were developed for localized antibiotic delivery in the stomach. Freeze-dried interpolymer complexes based on polyacrylic acid (PAA) and chitosan (CS) were prepared in a wide range of copolymer compositions by dissolving both polymers in acidic conditions. The influence of hydrogel-forming medium on the swelling and drug release was evaluated. The properties of these complexes were investigated by using scanning electron microscopy, dynamic swelling/eroding and release experiments in enzyme-free simulated gastric fluid (SGF). The electrostatic polymer/polymer interactions generate polyionic complexes with different porous structures. In a low pH environment, the separation of both polymer chains augmented as the amount of cationic and carboxilic groups increased within the network. However, the presence of higher amount of ions in the hydrogel-forming medium produced a network collapse, decreasing the maximum swelling ratio in SGF. PAA:CS:A (1:2.5:2)-1.75 M complexes released around 54% and 71% of the amoxicillin in 1 and 2 h, respectively, in acidic conditions. A faster drug release from this interpolymer complex was observed when the ionic strength of the hydrogel-forming medium increased. Complexes with a high amount of both polymer chains within the network, PAA:CS:A(2.5:5:2), showed a suitable amoxicillin release without being affected by an increased amount of ions in the hydrogel-forming medium. These freeze-dried interpolymer complexes could serve as potential candidates for amoxicillin delivery in an acidic enviroment.

Release of amoxicillin from polyionic complexes of chitosan and poly(acrylic acid). Study of polymer/polymer and polymer/drug interactions within the network structure.

Polyionic complexes of chitosan (CS) and poly(acrylic acid) (PAA) were prepared in a wide range of copolymer composition and with two kind of drugs. Release of amoxicillin trihydrate and amoxicillin sodium from these different complexes were studied. The swelling behavior of and solute transport in swellable hydrogels were investigated to check the effect of polymer/polymer and polymer/drugs interactions. The electrostatic polymer/polymer interactions take place between the cationic groups from CS and the anionic ones from PAA. The diffusion of amoxicillin trihydrate was controlled only by the swelling/eroding ratio of the polyionic complexes. The swelling degree of amoxicillin sodium hydrogels was more extensive when compared to the swelling degree of amoxicillin trihydrate formulations. It was concluded that the water uptake was mainly governed by the degree of ionization. Restriction of amoxicillin sodium diffusion could be achieved by polymer/ionized-drug interaction that retards the drug release. Freeze-dried polyionic complexes could serve as suitable candidates for amoxicillin site-specific delivery in the stomach.

Formation of Biogenic Amines in Idiazábal Ewe's-Milk Cheese: Effect of Ripening, Pasteurization, and Starter.

The importance of biogenic amines stems from their toxicity and their potential as a cause of food poisoning. After fish, cheese has been the foodstuff most often responsible for cases of food poisoning caused by ingestion of biogenic amines. Changes in biogenic amine (histamine, tyramine, putrescine, cadaverine, tryptamine, isopentylamine, spermidine, and phenylethylamine) content during the ripening of a cheese made from raw ewe's milk were studied, together with the effects of pasteurization and a commonly used commercial starter and indigenous starter cultures. Biogenic amines were determined by high-pressure liquid chromatography following extraction from the cheese and derivatization with dansyl chloride. Levels of histamine, potentially the most toxic amine, in the semihard Idiazábal cheese were very low. Tyramine, putrescine, and cadaverine were the main amines present in this type of cheese. Levels of those amines increased during ripening. Pasteurization of the milk brought about a reduction in the biogenic amine content of the ripened cheese, irrespective of the starter culture employed. The indigenous starter cultures considered in this study produced higher levels of histamine and cadaverine and lower levels of tyramine and putrescine than the commercial starter culture tested.