In vitro digestibility and angiotensin converting enzyme (ACE) inhibitory activity of solid-state fermented fava beans (Vicia faba L.).

Reducing the content of quickly absorbed carbohydrates and saturated fats in snack formulations while increasing the consumption of high-quality proteins are effective strategies to prevent obesity in childhood. Thus, the nutritional value, digestibility, and functionality of fava beans (Vicia faba L.) fermented with Pleurotus ostreatus were examined as potential ingredients for food design. Solid-state fermentation enhanced the protein content by 16% with a rise in essential (25%) and non-essential (15%) amino acids while decreasing total carbohydrate content and tannin levels. Moreover, fermentation modified the amino acid profile released during digestion, increasing amino acids such as valine, isoleucine, and threonine, which are vital for health and development in childhood. Furthermore, the bioaccessible fraction of the fermented bean showed a 60% of ACE inhibition and improved magnesium bioaccessibility. Consequently, fava beans fermented with Pleurotus ostreatus emerged as a new ingredient in the development of new protein-rich snacks tailored for children and adolescents.

Faecal lipid profile as a new marker of fat maldigestion, malabsorption and microbiota.

BACKGROUND: Fat malabsorption in children with cystic fibrosis (CF) leads to poor nutritional status and altered colonic microbiota. This study aimed at establishing the faecal lipid profile in children with CF, and exploring associations between the faecal lipidome and microbiota. METHODS: Cross-sectional observational study with children with CF and an age-matched control group. Faecal lipidome was analysed by UHLC-HRMS and microbiota profiling by 16S rRNA amplicon sequencing. RESULTS: Among 234 identified lipid species, five lipidome clusters (LC) were obtained with significant differences in triacylglycerols (TG), diacylglycerols (DG), monoacylglycerols (MG) and fatty-acids (FA): LC1 subjects with good digestion and absorption: low TG and low MG and FA; LC2 good digestion and poor absorption: low TG and high MG and FA; LC3 Mild digestion and poor absorption: intermediate TG and high MG and FA; LC4 poor digestion and absorption: high TG and high MG and FA; LC5 outliers. Bacteroidota and Verrucomicrobiota decreased over LC1-LC4, while Proteobacteria increased. Nutritional status indicators were significantly higher in LC1 and decreased over LC2-LC4. CONCLUSION: Assessing faecal lipidome may be relevant to determine how dietary lipids are digested and absorbed. This new evidence might be a method to support targeted nutritional interventions towards reverting fat maldigestion or malabsorption. IMPACT: Lipidomic analysis enabled the identification of the lipid species related to maldigestion (triglycerides) or malabsorption (monoglycerides and fatty acids). Children with cystic fibrosis can be grouped depending on the faecal lipidome profile related to dietary fat maldigestion or malabsorption. The lipidome profile in faeces is related to the composition of microbiota and nutritional status indicators.

In vitro screening of the impact of dietary prebiotic components, probiotic strains, and their symbiotic combinations on colonic microbiota in children with cystic fibrosis.

Children with Cystic Fibrosis (CF) are more likely to have intestinal dysbiosis due to recurrent antibiotic therapy and the conventional hypercaloric diet administered to them. This study aimed at evaluating the effect of isolated prebiotic components and probiotic strains, and their combinations as potential synbiotics, on the intestinal microbiota of CF children. A static in vitro colonic fermentation model was used by colonizing vials with faecal inoculum, a culture medium, and the substrates to be tested. Post treatment, aliquots were taken to determine ammonium, lactate, and short-chain fatty acids production and to profile the microbiota composition by 16s rRNA sequencing. At genus level, Escherichia-Shigella decreased (15.8%) with the treatment pectin + L. rhamnosus, followed by the beta-glucan + L. salivarius (15.5%). Inversely, the most increase in Bacteroides (44%) was obtained by the treatment with Pectin + L. reuteri. Lactate and acetic acid production was significantly increased with prebiotics and their combinations with L. rhamnosus and L. salivarius. In conclusion, the use of beta-glucan and pectin in combination with probiotic strains from the Lactobacillaceae family suggest potential to modulate dysbiosis and metabolic activity on CF colonic microbiota, encouraging further studies in animal studies or clinical settings to confirm the findings in vivo.

Protein digestibility and ACE inhibitory activity of fermented flours in older adults and standard gastrointestinal simulation.

Consumption of essential amino acids responsible for muscle protein synthesis is important in preventing sarcopenia among older individuals. This population may experience gastrointestinal disorders that inhibit protein digestibility, making it crucial to address. Therefore, solid-state fermentation (SSF) using Pleurotus ostreatus and air drying has been suggested as a means of improving the protein digestibility of lentils and quinoa. SSF combined with air drying at 70 °C resulted in a slight increase in protein hydrolysis compared to unfermented samples. SSF was found to boost the proportion of small peptides to 35 %. Following digestion, SSF and drying yielded bioactive peptides of 1400 and 450 Da, with a range of 11 % to 28 %, respectively, and peptides < 190 Da making up 60 % of the total. SSF promoted valine, leucine, and isoleucine generation; however, hot air drying reduced free amino acids due to the amino acid-reducing sugar bonding but was never lower than the initial content of its unfermented counterpart. Furthermore, SSF and drying at 70 °C improved the release of hydrophobic amino acids (>70 mg/g dry basis) and negatively charged amino acids (>20 mg/g dry basis) in lentils during digestion. The SSF samples exhibited lower angiotensin converting enzyme (ACE) inhibitory activity, ≤35 %, compared to unfermented flours after digestion. However, the ACE inhibitory activity increased in SSF-dried samples, in part because of melanoidins generated during drying. Finally, lower values of protein digestibility and thus smaller peptides, amino acid profile, and ACE inhibitory activity of fermented flours were found in the older adult digestion model.

Effect of beta-glucan supplementation on cystic fibrosis colonic microbiota: an in vitro study.

BACKGROUND: Children with cystic fibrosis (CF) present with gut dysbiosis, and current evidence impedes robust recommendations on the use of prebiotics. This study aimed at establishing the prebiotic potential of a commercial beta-glucan on the in vitro colonic microbiota of a child with CF compared to a healthy counterpart (H). METHODS: A dynamic simulator of colonic fermentation (twin-SHIME® model) was set up including the simulation of the proximal (PC) and distal colon (DC) of the CF and the H subjects by colonizing the bioreactors with faecal microbiota. During two weeks the system was supplied with the beta-glucan. At baseline, during treatment and post-treatment, microbiota composition was profiled by 16 S rRNA and short-chain fatty acids (SCFA) production was determined by GS-MS. RESULTS: At baseline, Faecalibacterium, was higher in CF' DC than in the H, along higher Acidaminococcus and less Megasphaera and Sutterella. Beta-glucan supplementation induced increased microbiota richness and diversity in both subjects during the treatment. At genus level, Pseudomonas and Veillonella decreased, while Akkermansia and Faecalibacterium increased significantly in CF. CONCLUSION: The supplementation with beta-glucan suggests positive results on CF colonic microbiota in the in vitro context, encouraging further research in the in vivo setting. IMPACT: Current evidence supports assessing the effect of prebiotics on modifying cystic fibrosis microbiota. The effect of beta-glucan supplementation was evaluated in a controlled dynamic in vitro colonic ecosystem. Beta-glucan supplement improved diversity in cystic fibrosis colonic microbiota. The treatment showed increased abundance of Faecalibacterium and Akkermansia in cystic fibrosis. New evidence supports the use of prebiotics in future clinical studies.

Association between Dietary Intake and Faecal Microbiota in Children with Cystic Fibrosis.

A "high-fat, high-energy diet" is commonly recommended for children with cystic fibrosis (CF), leading to negative consequences on dietary patterns that could contribute to altered colonic microbiota. The aim of this study was to assess dietary intake and to identify possible associations with the composition of faecal microbiota in a cohort of children with CF. A cross-sectional observational study was conducted, including a 3-day food record simultaneously with the collection of faecal samples. The results showed a high fat intake (43.9% of total energy intake) and a mean dietary fibre intake of 10.6 g/day. The faecal microbiota was characterised at the phylum level as 54.5% Firmicutes and revealed an altered proportion between Proteobacteria (32%) and Bacteroidota (2.2%). Significant associations were found, including a negative association between protein, meat, and fish intake and Bifidobacterium, a positive association between lipids and Escherichia/Shigella and Streptococcus, a negative association between carbohydrates and Veillonella and Klebsiella, and a positive association between total dietary fibre and Bacteroides and Roseburia. The results reveal that a "high-fat, high-energy" diet does not satisfy dietary fibre intake from healthy food sources in children with CF. Further interventional studies are encouraged to explore the potential of shifting to a high-fibre or standard healthy diet to improve colonic microbiota.

In Vitro Digestion Assessment (Standard vs. Older Adult Model) on Antioxidant Properties and Mineral Bioaccessibility of Fermented Dried Lentils and Quinoa.

The growing number of older adults necessitates tailored food options that accommodate the specific diseases and nutritional deficiencies linked with ageing. This study aims to investigate the influence of age-related digestive conditions in vitro on the phenolic profile, antioxidant activity, and bioaccessibility of minerals (Ca, Fe, and Mg) in two types of unfermented, fermented, and fermented dried quinoa and lentils. Solid-state fermentation, combined with drying at 70 °C, significantly boosted the total phenolic content in Castellana and Pardina lentils from 5.05 and 6.6 to 10.5 and 7.5 mg gallic acid/g dry weight, respectively, in the bioaccessible fraction following the standard digestion model, compared to the unfermented samples. The phenolic profile post-digestion revealed elevated levels of vanillic and caffeic acids in Castellana lentils, and vanillic acid in Pardina lentils, while caffeic acids in Castellana lentils were not detected in the bioaccessible fraction. The highest antioxidant potency composite index was observed in digested fermented dried Castellana lentils, with white quinoa samples exhibiting potency above 80%. Mineral bioaccessibility was greater in fermented and fermented dried samples compared to unfermented ones. Finally, the digestive changes that occur with ageing did not significantly affect mineral bioaccessibility, but compromised the phenolic profile and antioxidant activity.

Effect of Lactobacillaceae Probiotics on Colonic Microbiota and Metabolite Production in Cystic Fibrosis: A Comparative In Vitro Study.

Cystic Fibrosis-related gut dysbiosis (CFRGD) has become a recognised complication in children with this condition, and current evidence remains insufficient to guide the selection of probiotic strains for supplementation treatments. The aim of this study was to characterise the effect of three probiotic strains on CFRGD by means of a dynamic in vitro simulation of the colonic fermentation (SHIME®). The configuration of the system included three bioreactors colonised with the faecal inoculum of a child with cystic fibrosis. For 20 days, each bioreactor was supplied daily with either Lacticaseibacillus rhamnosus GG (ATCC 53103 TM), Limosilactobacillus reuteri (DSM 17938) or Lactiplantibacillus plantarum (DSM 22266). The baseline microbiota was characterised by a high abundance of Prevotella, Faecalibacterium and Acidaminococcus genera. After 20 days of supplementation, L. rhamnosus and L. plantarum reduced Prevotella significantly, and the three strains led to increased Faecalibacterium and Bifidobacterium and decreased Acidaminococcus, with some of these changes being maintained 10 days after ceasing supplementation. The metabolic activity remained unaltered in terms of short-chain fatty acids, but branched-chain fatty acids showed a significant decrease, especially with L. plantarum. Additionally, ammonia decreased at 20 days of supplementation, and lactate continuously increased with the three strains. The effects on colonic microbiota of L. rhamnosus, L. reuteri or L. plantarum were established, including increased beneficial bacteria, such as Faecalibacterium, and beneficial metabolites such as lactate; and on the other hand, a reduction in pathogenic genera, including Prevotella or Acidaminococcus and branched-chain fatty acids, overall supported their use as probiotics in the context of CFRGD.

Development of Optical Label-Free Biosensor Method in Detection of Listeria monocytogenes from Food.

The present work describes an alternative method for detecting and identifying Listeria monocytogenes in food samples by developing a nanophotonic biosensor containing bioreceptors and optical transducers. The development of photonic sensors for the detection of pathogens in the food industry involves the implementation of procedures for selecting probes against the antigens of interest and the functionalization of the sensor surfaces on which the said bioreceptors are located. As a previous step to functionalizing the biosensor, an immobilization control of these antibodies on silicon nitride surfaces was carried out to check the effectiveness of in plane immobilization. On the one hand, it was observed that a Listeria monocytogenes-specific polyclonal antibody has a greater binding capacity to the antigen at a wide range of concentrations. A Listeria monocytogenes monoclonal antibody is more specific and has a greater binding capacity only at low concentrations. An assay for evaluating selected antibodies against particular antigens of Listeria monocytogenes bacteria was designed to determine the binding specificity of each probe using the indirect ELISA detection technique. In addition, a validation method was established against the reference method for many replicates belonging to different batches of meat-detectable samples, with a medium and pre-enrichment time that allowed optimal recovery of the target microorganism. Moreover, no cross-reactivity with other nontarget bacteria was observed. Thus, this system is a simple, highly sensitive, and accurate platform for L. monocytogenes detection.

Impact of Air-Drying Temperature on Antioxidant Properties and ACE-Inhibiting Activity of Fungal Fermented Lentil Flour.

Solid-state fermentation (SSF) with Pleurotus ostreatus enhances the nutritional value of legumes. However, drying can cause significant changes in physical and nutritional properties of the final products. Thus, this work studies the impact of air-drying temperature (50, 60, and 70 °C) on relevant properties (antioxidant properties, ACE-inhibitory capacity, phytic acid, colour, and particle size) of two fermented lentils flour (Pardina and Castellana) using freeze-drying as a reference method. Castellana variety is a better substrate for Pleurotus, generating four times more biomass. In addition, an almost total reduction of phytic acid from 7.3 to 0.9 mg/g db is achieved in this variety. Air-drying significantly decreased the particle size and the final colour with ΔE > 20; nonetheless, the temperature does not play a crucial role. SSF decreased the total phenolic content and the antioxidant capacity regardless of the variety, however, drying at 70 °C increased total phenolic content (186%) in fermented Castellana flour. Comparing drying methods, freeze-drying implied a higher decrease in those parameters, reducing the TPC from 2.4 to 1.6 and from 7.7 to 3.4 mg gallic acid/g db in Pardina and Castellana dried flours. Finally, the flours inhibit the angiotensin I-converting-enzyme, and fermentation and drying increased their potential cardiovascular benefits.

Prevalence and Characterization of Beta-Lactam and Carbapenem-Resistant Bacteria Isolated from Organic Fresh Produce Retailed in Eastern Spain.

Fresh fruits and vegetables are potential reservoirs for antimicrobial resistance determinants, but few studies have focused specifically on organic vegetables. The present study aimed to determine the presence of third-generation cephalosporin (3GC)- and carbapenem-resistant Gram-negative bacteria on fresh organic vegetables produced in the city of Valencia (Spain). Main expanded spectrum beta-lactamase (ESBL)- and carbapenemase-encoding genes were also detected in the isolates. One hundred and fifteen samples were analyzed using selective media supplemented with cefotaxime and meropenem. Resistance assays for twelve relevant antibiotics in medical use were performed using a disc diffusion test. A total of 161 isolates were tested. Overall, 33.5% presented multidrug resistance and 16.8% were resistant to all ß-lactam antibiotics tested. Imipenem resistance was observed in 18% of isolates, and low resistance levels were found to ceftazidime and meropenem. Opportunistic pathogens such as Acinetobacter baumannii, Enterobacter spp., Raoultella sp., and Stenotrophomonas maltophilia were detected, all presenting high rates of resistance. PCR assays revealed blaVIM to be the most frequently isolated ESBL-encoding gene, followed by blaTEM and blaOXA-48. These results confirm the potential of fresh vegetables to act as reservoirs for 3GC- and carbapenem-producing ARB. Further studies must be carried out to determine the impact of raw organic food on the spread of AMRs into the community.

Exploring the Impact of Solid-State Fermentation on Macronutrient Profile and Digestibility in Chia (Salvia hispanica) and Sesame (Sesamum Indicum) Seeds.

Fermentation of plant-based substrates with edible fungi enhances the nutrient profile and digestibility, but it has been scarcely applied to edible seeds, which are rich in healthy lipids. In this study, chia and sesame seeds were solid-state fermented with Pleurotus ostreatus, followed by drying and milling. Fermentation led to increased content of lipid and protein in both seeds' products, and a change in fatty acid profile in favor of increased polyunsaturated fatty acids. Then, the samples were subjected to in vitro digestion. Lipolysis, determined by nuclear magnetic resonance, was higher in sesame than in chia products, and the fermented counterparts had increased values compared to the controls. In terms of physical properties, fermentation showed reduced particle size and increased matrix degradation and decreased viscosity of the digestion medium, which were related to increased lipolysis. In conclusion, applying solid-state fermentation on chia and sesame seeds could be a recommendable approach.

Non-dipper effect in hypertense patients after renal transplantation by 24-hour ambulatory blood pressure monitoring.

OBJECTIVE: Determinate instant and after 1-month non-dipper effect in hypertense patients after renal transplant by 24-hour ambulatory blood pressure monitoring in Hospital General de Zona No. 50, San Luis Potosí, Mexico. METHOD: Descriptive, longitudinal and prospective cohort study of a non-probability convenience sampling in post-transplant patients with hypertension. We collected data from MAPA and includes age, sex, cardiovascular risk factors in variables. Use of central tendency and dispersion measures for descriptive analysis and t Student for inferential analysis. RESULTS: 19 patients were included, 11 male (57.9%) and 8 females (42.1%), with age range 20 to 49 years (median of 30.2 years ± 7.7). Where the non-dipper effect in the first take was 89.5% and in the second take 84.2%. CONCLUSIONS: There is a high frequency of the non-dipper pattern in patients at one month of kidney transplant, the persistence of this hypertension may be, among others, by the use of immunosuppressants. A new category for non-dipper classification is described.

OBJETIVO: Determinar el efecto non-dipper inmediato y posterior a 1 mes en pacientes adultos hipertensos postrasplante renal con monitoreo continuo de la presión arterial de 24 horas, en el Hospital General de Zona No. 50 de San Luis Potosí, México. MÉTODO: Estudio de tipo cohorte, longitudinal, prospectivo, con muestreo no probabilístico por conveniencia de casos consecutivos en pacientes receptores de trasplante renal con hipertensión arterial. Se recogieron los siguientes datos: edad, sexo, factores de riesgo cardiovascular, uso de antihipertensivos o inmunosupresores, y monitoreo ambulatorio de la presión arterial de 24 horas. Se aplicaron medidas de tendencia central y de dispersión para análisis descriptivo, y prueba t de Student para análisis inferencial. RESULTADOS: Se incluyeron 11 hombres (57.9%) y 8 mujeres (42.1%), con una edad de 20 a 49 años (media 30.2 ± 7.7), en los que el efecto non-dipper inmediato fue del 89.5% y posterior a 1 mes fue del 84.2%. CONCLUSIONES: Existe una alta frecuencia del patrón non-dipper en pacientes a 1 mes del trasplante renal. La persistencia de la hipertensión puede ser, entre otras causas, por el uso de inmunosupresores. Se describe una nueva categoría para la clasificación non-dipper.

Antioxidants Bioaccessibility and Lactobacillus salivarius (CECT 4063) Survival Following the In Vitro Digestion of Vacuum Impregnated Apple Slices: Effect of the Drying Technique, the Addition of Trehalose, and High-Pressure Homogenization.

To benefit the health of consumers, bioactive compounds must reach an adequate concentration at the end of the digestive process. This involves both an effective release from the food matrix where they are contained and a high resistance to exposure to gastrointestinal conditions. Accordingly, this study evaluates the impact of trehalose addition (10% w/w) and homogenization (100 MPa), together with the structural changes induced in vacuum impregnated apple slices (VI) by air-drying (AD) and freeze-drying (FD), on Lactobacillus salivarius spp. salivarius (CECT 4063) survival and the bioaccessibility of antioxidants during in vitro digestion. Vacuum impregnated apple slices conferred maximum protection to the lactobacillus strain during its passage through the gastrointestinal tract, whereas drying with air reduced the final content of the living cells to values below 10 cfu/g. The bioaccessibility of antioxidants also reached the highest values in the VI samples, in which the release of both the total phenols and total flavonoids to the liquid phase increased with in vitro digestion. The addition of trehalose and homogenization at 100 MPa increased the total bioaccessibility of antioxidants in FD and AD apples and the total bioaccessibility of flavonoids in the VI samples. Homogenizing at 100 MPa also increased the survival of L. salivarius during in vitro digestion in FD samples.

[Clinical and electromyographic findings in patients with diabetic polyneuropathy]. / Manifestaciones clínicas y alteraciones electroneuromiográficas en pacientes con diabetes tipo 2 y polineuropatía.

BACKGROUND: Diabetic neuropathy represents a polyneuropathy with electrophiosiological alterations. Electroneuromyography (NMD) plays an important role in the evaluation of patient with diabetes mellitus type 2 (DM2) and doubtful neuropathy. OBJECTIVE: To determine clinical manifestations in patients with distal symmetrical sensory polyneuropathy (DSSP) and to correlate them with electromyographic alterations. MATERIAL AND METHODS: Transversal, analytical study. 138 patients over 18 years old, with DM2 and PSSD were selected. They underwent physical examination, laboratory studies and electromyography (EM) with 4-channel Nicolet electromyograph. Measures of central tendency and their dispersion were analyzed; data normality with Kolmogorov-Smirnov; Student's t test and Spearman's correlation. RESULTS: Thalar hyperkeratosis was the most frequent clinical finding in 103 (74%) patients. The most frequent symptoms were paresthesia in 132 (95.7%) patients and tingling in 93 (67.4%) patients. Exploration of superficial sensitivity determined neuropathy in 42 (30.4%) patients finding greater insensitivity in the medial plantar nerve territory. In the EM, the nerve with the greatest absent response was the left lateral plantar nerve in 51 (59%) patients. A significant correlation (p < 0.05) was found between the variables of EM with age, years of evolution and levels of glycated hemoglobin A1c. CONCLUSIONS: The higher the lack of glycemic control, the chronobiology of the patient and the time of illness, the greater the electromyographic affection.

INTRODUCCIÓN: la neuropatía diabética representa una polineuropatía con alteración electrofiosiológica. La electroneuromiografía (ENM) desempeña un papel importante en la evaluación del paciente con diabetes mellitus tipo 2 (DM2) y neuropatía dudosa. OBJETIVO: determinar las manifestaciones clínicas en pacientes con polineuropatía sensitiva simétrica distal (PSSD) y correlacionarlas con alteraciones electroneuromiográficas. MATERIAL Y MÉTODOS: estudio transversal, analítico. Se seleccionaron 138 pacientes mayores de 18 años, con DM2 y PSSD. Se les realizó exploración física, estudios de laboratorio y ENM con electromiógrafo Nicolet de 4 canales. Se analizaron las medidas de tendencia central y su dispersión; normalidad de datos con Kolmogorov-Smirnov; t de Student y correlación de Spearman. RESULTADOS: la hiperqueratosis talar fue el hallazgo clínico más frecuente en 74% de los pacientes. Los síntomas más frecuentes fueron parestesias en 95.7% y hormigueo en 67.4%. La exploración de sensibilidad superficial determinó neuropatía en 30.4%, encontrando mayor insensibilidad en el territorio del nervio plantar medial. En la ENM, el nervio con mayor respuesta ausente fue el plantar lateral izquierdo en 59%. Se encontró correlación significativa (p < 0.05) entre las variables electroneuromiograficas con la edad, años de evolución y niveles de hemoglobina glucosilada A1c (HbA1c). CONCLUSIONES: a mayor descontrol glucémico, cronobiología del paciente y tiempo del padecimiento, mayor es la afección electroneuromiográfica.

Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels.

Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 µg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.

Optimization of pre- treatments with Propidium Monoazide and PEMAX™ before real-time quantitative PCR for detection and quantification of viable Helicobacter pylori cells.

Accurate detection of H. pylori in different environmental and clinical samples is essential for public health strtdudies. Now, a big effort is being made to design PCR methodologies that allow for the detection of viable and viable but non-culturable (VBNC) H. pylori cells, by achieving complete exclusion of dead cells amplification signals. The use of DNA intercalating dyes has been proposed. However, its efficacy is still not well determined. In this study, we aimed to test the suitability of PMA and PEMAX™ dyes used prior to qPCR for only detecting viable cells of H. pylori. Their efficiency was evaluated with cells submitted to different disinfection treatments and confirmed by the absence of growth on culture media and by LIVE/DEAD counts. Our results indicated that an incubation period of 5 min for both, PMA and PEMAX™, did not affect viable cells. Our study also demonstrated that results obtained by using intercalating dyes may vary depending on the cell stress conditions. In all dead cell's samples, both PMA and PEMAX™ pre-qPCR treatments decreased the amplification signal (>103 Genomic Units (GU)), although none of them allowed for its disappearance confirming that intercalating dyes, although useful for screening purposes, cannot be considered as universal viability markers. To investigate the applicability of the method specifically to detect H. pylori cells in environmental samples, PMA-qPCR was performed on samples containing the different morphological and viability states that H. pylori can acquire in environment. The optimized PMA-qPCR methodology showed to be useful to detect mostly (but not only) viable forms, regardless the morphological state of the cell.

A Primary Prevention Clinical Risk Score Model for Patients With Brugada Syndrome (BRUGADA-RISK).

OBJECTIVES: The goal of this study was to develop a risk score model for patients with Brugada syndrome (BrS). BACKGROUND: Risk stratification in BrS is a significant challenge due to the low event rates and conflicting evidence. METHODS: A multicenter international cohort of patients with BrS and no previous cardiac arrest was used to evaluate the role of 16 proposed clinical or electrocardiogram (ECG) markers in predicting ventricular arrhythmias (VAs)/sudden cardiac death (SCD) during follow-up. Predictive markers were incorporated into a risk score model, and this model was validated by using out-of-sample cross-validation. RESULTS: A total of 1,110 patients with BrS from 16 centers in 8 countries were included (mean age 51.8 ± 13.6 years; 71.8% male). Median follow-up was 5.33 years; 114 patients had VA/SCD (10.3%) with an annual event rate of 1.5%. Of the 16 proposed risk factors, probable arrhythmia-related syncope (hazard ratio [HR]: 3.71; p < 0.001), spontaneous type 1 ECG (HR: 3.80; p < 0.001), early repolarization (HR: 3.42; p < 0.001), and a type 1 Brugada ECG pattern in peripheral leads (HR: 2.33; p < 0.001) were associated with a higher risk of VA/SCD. A risk score model incorporating these factors revealed a sensitivity of 71.2% (95% confidence interval: 61.5% to 84.6%) and a specificity of 80.2% (95% confidence interval: 75.7% to 82.3%) in predicting VA/SCD at 5 years. Calibration plots showed a mean prediction error of 1.2%. The model was effectively validated by using out-of-sample cross-validation according to country. CONCLUSIONS: This multicenter study identified 4 risk factors for VA/SCD in a primary prevention BrS population. A risk score model was generated to quantify risk of VA/SCD in BrS and inform implantable cardioverter-defibrillator prescription.

Novel Insights Into the Effects of Interleukin 6 Antagonism in Non-ST-Segment-Elevation Myocardial Infarction Employing the SOMAscan Proteomics Platform.

Background Interleukin 6 concentration is associated with myocardial injury, heart failure, and mortality after myocardial infarction. In the Norwegian tocilizumab non-ST-segment-elevation myocardial infarction trial, the first randomized trial of interleukin 6 blockade in myocardial infarction, concentration of both C-reactive protein and troponin T were reduced in the active treatment arm. In this follow-up study, an aptamer-based proteomic approach was employed to discover additional plasma proteins modulated by tocilizumab treatment to gain novel insights into the effects of this therapeutic approach. Methods and Results Plasma from percutaneous coronary intervention-treated patients, 24 in the active intervention and 24 in the placebo-control arm, drawn 48 hours postrandomization were randomly selected for analysis with the SOMAscan assay. Employing slow off-rate aptamers, the relative abundance of 1074 circulating proteins was measured. Proteins identified as being significantly different between groups were subsequently measured by enzyme immunoassay in the whole trial cohort (117 patients) at all time points (days 1-3 [7 time points] and 3 and 6 months). Five proteins identified by the SOMAscan assay, and subsequently confirmed by enzyme immunoassay, were significantly altered by tocilizumab administration. The acute-phase proteins lipopolysaccharide-binding protein, hepcidin, and insulin-like growth factor-binding protein 4 were all reduced during the hospitalization phase, as was the monocyte chemoattractant C-C motif chemokine ligand 23. Proteinase 3, released primarily from neutrophils, was significantly elevated. Conclusions Employing the SOMAscan aptamer-based proteomics platform, 5 proteins were newly identified that are modulated by interleukin 6 antagonism and may mediate the therapeutic effects of tocilizumab in non-ST-segment-elevation myocardial infarction.

The relationship between sleep quality and all-cause, CVD and cancer mortality: the Southall and Brent REvisited study (SABRE).

BACKGROUND: Both long and short sleep duration increase risk of mortality. Most previous studies have been performed in Europeans and have focused on sleep duration. Thus, we aimed to investigate the association between sleep quality and mortality across three different ethnic groups. METHODS: We used data from the Southall and Brent REvisited Study (SABRE) cohort, which comprises first generation migrant South Asian and African Caribbean men and women, aged 40-69 years, recruited between 1988 and 1991. In sum, 4399 participants provided complete data at baseline and follow-up. Of those, 1656 died by December 2017. Our exposures (eg, difficulty falling asleep, early morning waking and waking up tired in the morning) were self-reported and our primary outcome was mortality. We used Cox proportional hazards models to analyse our data, adjusting for baseline-measured confounders. RESULTS: None of the sleep measures were strongly associated with all-cause mortality in Europeans or African Caribbeans, whilst in South Asians difficulty falling asleep was related to an increased risk of all-cause mortality (HR = 1.28, 95%CI = 1.01; 1.61). In Europeans, early morning waking was associated with a moderately increased risk of cardiovascular death (HR = 1.31, 95%CI = 1.05; 1.63); alternately, this association was not as strong in the other groups. CONCLUSION: Our findings suggest that the relationship between sleep quality and mortality may differ by ethnic group, but formal heterogeneity tests indicated that the strongest difference in HRs was observed for early morning waking and cardiovascular disease (CVD) mortality across the three groups (Cochran's Q test p = 0.036). As such, these results are novel and provide support for ethnic differences in sleep quality and mortality, and may have implications for precision medicine.