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The present study evaluated the capacity of three Bacillus species to improve health status and growth performance of Nile Tilapia fed with high levels of soybean meal and challenged with Aeromonas hydrophila. In vitro experiments showed that ß-hemolysin and metalloprotease enzymes were produced by A. hydrophila throughout the exponential growth phase. In vivo experiments showed that 107 colony-forming units (CFUs)/ml of this pathogen killed 50% of control group fishes in 13 days. To evaluate the influence of Bacillus strains on health status and growth performance in Nile Tilapia, 180 fishes (33.44 + 0.05 g) were distributed in 12 tanks of 200 L each, and animals were fed twice per day until satiety. 1) Control group without Bacillus, 2) Bacillus sp1, 3) Bacillus sp2, and 4) Bacillus sp3 groups were formulated containing 106 CFU/g. After 40 days of feeding, the fishes were intraperitoneally injected with 1 ml of A. hydrophila at 2 × 107 CFU/ml, and mortality was recorded. The results showed that cumulative mortality rate was significantly (p< 0.05) lower in the Bacillus sp1 (25%), sp2 (5%), and sp3 (15%) groups, than the control group (50%). Weight gain was also significantly better (p< 0.05) in the Bacillus sp1 (36%), sp2 (67%), and sp3 (55%) groups with respect to the control group (30%). In conclusion, functional diet formulated with high levels of soybean meal and supplemented with Bacillus sp2 could be an alternative to protect Nile tilapia cultures from A. hydrophila infections and improve fish growth performance.

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Some species of Dunaliella produce high levels of ß-carotene, which is the largest natural Vitamin A source. ß-carotene production in Dunaliella is expensive due to low yields obtained under culture conditions used. Since three decades ago synthetic ß-carotene has dominated 98% of world market, even when synthetic is less bioactive than Dunaliella ß-carotene molecule. In vitro experiments have been performed to increase ß-carotene productivity in Dunaliella and some useful results have been obtained. In this review, culture conditions used in those experiments were compared to identify parameters that improved ß-carotene productivity in Dunaliella. Comparison results show that ß-carotene in vitro production can be increased regulating some specific culture conditions.

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BACKGROUND: Diagnosing gastroesophageal reflux disease (GERD) can be challenging given varying symptom presentations, and complex multifactorial pathophysiology. The gold standard for GERD diagnosis is esophageal acid exposure time (AET) measured by pH-metry. A variety of additional diagnostic tools are available. The goal of this consensus was to assess the individual merits of GERD diagnostic tools based on current evidence, and provide consensus recommendations following discussion and voting by experts. METHODS: This consensus was developed by 15 experts from nine countries, based on a systematic search of the literature, using GRADE (grading of recommendations, assessment, development and evaluation) methodology to assess the quality and strength of the evidence, and provide recommendations regarding the diagnostic utility of different GERD diagnosis tools, using AET as the reference standard. KEY RESULTS: A proton pump inhibitor (PPI) trial is appropriate for patients with heartburn and no alarm symptoms, but nor for patients with regurgitation, chest pain, or extraesophageal presentations. Severe erosive esophagitis and abnormal reflux monitoring off PPI are clearly indicative of GERD. Esophagram, esophageal biopsies, laryngoscopy, and pharyngeal pH monitoring are not recommended to diagnose GERD. Patients with PPI-refractory symptoms and normal endoscopy require reflux monitoring by pH or pH-impedance to confirm or exclude GERD, and identify treatment failure mechanisms. GERD confounders need to be considered in some patients, pH-impedance can identify supragrastric belching, impedance-manometry can diagnose rumination. CONCLUSIONS: Erosive esophagitis on endoscopy and abnormal pH or pH-impedance monitoring are the most appropriate methods to establish a diagnosis of GERD. Other tools may add useful complementary information.

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T. macdonaldi is a carnivorous species endemic to the Gulf of California. Indiscriminate exploitation has put totoaba at risk, inducing the development of aquaculture procedures to grow it without affecting the wild population. However, aquafeeds increasing cost and low yields obtained with commercial feeds have motivated researchers to look for more nutritious and cheaper alternatives. Soybean (SB) is the most popular alternative to fishmeal (FM); however, antinutritional factors limit its use in carnivorous species. In this study, we analyzed B. subtilis 9b probiotic capacity to improve growth performance and health status of T. macdonaldi fed with formulations containing 30% and 60% substitution of fish meal with soy protein concentrate (SPC). In addition, we investigated its effect on internal organs condition, their capacity to modulate the intestinal microbiota, and to boost the immunological response of T. macdonaldi against V. harveyi infections. In this sense, we found that T. macdonaldi fed with SPC30Pro diet supplemented with B. subtilis 9b strain and 30% SPC produced better results than SPC30C control diet without B. subtilis and DCML commercial diet. Additionally, animals fed with SPC60Pro diet supplemented with B. subtilis 9b strain and 60% SPC doubled their weight and produced 20% more survival than SPC60C control diet without B. subtilis. Thus, B. subtilis 9b improved T. macdonaldi growth performance, health status, modulated intestinal microbiota, and increased animal's resistance to V. harveyi infections, placing this bacterium as an excellent candidate to produce functional feeds with high levels of SPC.

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Antibiotic resistance is a problem that has been increasing lately; therefore, it is important to find new alternatives to treat infections induced by pathogens that cannot be eliminated with available products. Small antimicrobial peptides (AMPs) known as bacteriocin could be an alternative to antibiotics because they have shown to be effective against a great number of multidrug-resistant microbes. In addition to its high specificity against microbial pathogens and its low cytotoxicity against human cells, most bacteriocin present tolerance to enzyme degradation and stability to temperature and pH alterations. Bacteriocins are small peptides with a great diversity of structures and functions; however, their mechanisms of action are still not well understood. In this review, bacteriocin produced by Bacillus species will be described, especially its mechanisms of action, culture conditions used to improve its production and state-of-the-art methodologies applied to identify them. Bacteriocin utilization as food preservatives and as new molecules to treat cancer also will be discussed.

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BACKGROUND AND AIMS: Irritable bowel syndrome (IBS) is one of the most frequent disorders in clinical practice, with a mean 7.6-10.8% worldwide prevalence. A study showed that 6.1% of patients with diarrhea-predominant IBS (IBS-D) had severe exocrine pancreatic insufficiency (EPI). We aimed to identify the prevalence of EPI based on fecal elastase stool testing (Fel-1) in IBS-D and the clinical characteristics that may predict the diagnosis of EPI. METHODS: Patients aged > 18 years presenting to tertiary hospital outpatient clinics with IBS-D completed validated questionnaires and gave a stool sample where Fel-1 concentration was measured. Patients with Fel-1 < 100 µg/g represented EPI and > 100 to < 200 µg/g underwent testing for pancreatic pathology with laboratory and endoscopic ultrasound (EUS) evaluation. RESULTS: One hundred forty patients (mean age 60 years, females 75.7%) were studied. EPI was found in 5% (95% CI 2.2-10.4), and pancreatic steatosis was the main EUS finding (71%). Dyspepsia was an independent factor associated with EPI (OR 34.7; 95% CI 4.95-366.37, p = 0.0007). After pancreatic enzyme replacement therapy (PERT), patients showed a significant improvement in the Bristol stool scale (p < 0.0001), bowel movements per day (p < 0.005), distension score (0.0009), pain score (0.0277) and IBS severity (0.0034). CONCLUSION: EPI is present in 5% of patients who fulfill Rome IV criteria for D-IBS, and dyspepsia was an independent symptom strongly associated with EPI. Pancreatic steatosis was the main endoscopic ultrasound finding. After PERT therapy, patients had significantly improved stool frequency, stool consistency, abdominal pain, distension and IBS severity score.

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The high mortality rate of breast cancer is mainly caused by the metastatic ability of cancer cells, resistance to chemotherapy and radiotherapy, and tumor regression capacity. In recent years, it has been shown that the presence of breast cancer stem cells is closely associated with the migration and metastatic ability of cancer cells, as well as with their resistance to chemotherapy and radiotherapy. The tumor microenvironment is one of the main molecular factors involved in cancer and metastatic processes development, in this sense it is interesting to study the role of platelets, one of the main communicator cells in the human body which are activated by the signals they receive from the microenvironment and can generate more than one response. Platelets can ingest and release RNA, proteins, cytokines and growth factors. After the platelets interact with the tumor microenvironment, they are called "tumor-educated platelets." Tumor-educated platelets transport material from the tumor microenvironment to sites adjacent to the tumor, thus helping to create microenvironments conducive for the development of primary and metastatic tumors. It has been observed that the clone capable of carrying out the metastatic process is a cancer cell with stem cell characteristics. Cancer stem cells go through a series of processes, including epithelial-mesenchymal transition, intravasation into blood vessels, movement through blood vessels, extravasation at the site of the establishment of a metastatic focus, and site colonization. Tumor-educated platelets support all these processes.

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BACKGROUND: Virtual reality simulation in gastrointestinal endoscopy is an educational tool that allows repetitive instruction in a non-patient care environment. AIM: To determine the impact of a virtual endoscopy training curriculum applying an objective pre- and post-training analysis on trainee endoscopists. METHODS: A before-after training study was carried out. Subjects were first year fellows of gastroenterology, who completed a questionnaire and then performed two pre-training simulated cases. The virtual endoscopy training curriculum consisted of an 8-h workday utilizing two GI MENTOR™ in a specialized clinical simulation center. After the training, all subjects completed the same two cases they did in the pre-training. Pre- and post-training results' comparisons were made by paired t test. RESULTS: Totally, 126 subjects were included (mean age 30 years, 61% female). A significant improvement from pre- to post-training was observed in psychomotor skills (total time, percentage, and number of balloons exploded) and endoscopic skills (cecal intubation time, percentage of examined mucosa, and efficacy of screening). There was also an improvement in the quality of the endoscopic study; percentage of examined mucosa over 85% showed a significant improvement post-training with an adjusted OR of 2.72 (95% CI 1.51-4.89, p = 0.001). CONCLUSIONS: Virtual endoscopy training curriculum produces a significant improvement in the trainee endoscopists performance and their psychomotor skills and introduces the concept of a quality endoscopic study in a non-patient, risk-free environment.

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Beneficial microorganisms maintain the ecosystems, plants, animals and humans working in healthy conditions. In nature, around 95% of all microorganisms produce beneficial effects by increasing nutrients digestion and assimilation, preventing pathogens development and by improving environmental parameters. However, increase in human population and indiscriminate uses of antibiotics have been exerting a great pressure on agriculture, livestock, aquaculture, and also to the environment. This pressure has induced the decomposition of environmental parameters and the development of pathogenic strains resistant to most antibiotics. Therefore, all antibiotics have been restricted by corresponding authorities; hence, new and healthy alternatives to prevent or eliminate these pathogens need to be identified. Thus, probiotic bacteria utilization in aquaculture systems has emerged as a solution to prevent pathogens development, to enhance nutrients assimilation and to improve environmental parameters. In this sense, B. subtilis is an ideal multifunctional probiotic bacterium, with the capacity to solve these problems and also to increase aquaculture profitability.

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Cervical cancer (CC) is one of the leading causes of cancer-associated mortalities in women from developing countries. Similar to other types of cancer, CC is considered to be a multifactorial disease, involving socioeconomic, cultural, immunological and epigenetic factors, as well as persistent human papilloma virus (HPV) infection. It has been well established that cancer stem cells (CSCs) play an important role in defining tumor size, the speed of development and the level of regression following treatment; therefore, CSCs are associated with a poor prognosis. CSCs have been detected in many types of cancer, including leukemia, pancreatic, colon, esophagus, liver, prostate, breast, gastric and lung cancer. In cervical cancer, CSCs have been associated with resistance to normally used drugs such as cisplatin. The present review summarizes the strategies that high-risk HPV viruses (HPV-16 and HPV-18) have developed to transform normal epithelial cells into cancer cells, as well as the cellular pathways and studies associated with the identification of cervical cancer stem cell biomarkers. In this sense, the present review provides state of the art information regarding CC development.

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BACKGROUND: Bacillus thuringiensis toxins are effective against multiple biological targets such as insects, nematodes, mites, protozoa, and importantly, human cancer cells. One of the main mechanisms by which Cry toxins to trigger cell death is the specific recognition of cadherin-like membrane cell receptors. OBJECTIVE: This work aimed to assess the cytotoxicity of the Cry1Ab and Cry1Ac toxins from Bacillus thuringiensis in HeLa, cervical cancer cell line, as well as their antitumor activity in mouse models. METHODS: We analyzed several biological targets of Cry1Ab and Cry1Ac including erythrocytes, insect larvae, as well as cancer and non-cancer cell lines. The viability of HeLa, SiHa, MCF7 and HaCat cells was assessed by MTT 24 h after the administration of Cry toxins. We also studied apoptosis as a possible cytotoxicity mechanism in HeLa. The capacity of Cry toxins to eliminate tumors in xenograft mouse models was also analyzed. RESULTS: Both toxins, Cry1Ab and Cry1Ac, showed specific cytotoxic activity in HeLa (HPV18+) cervical cancer cell line, with a Cry1Ab LC50 of 2.5 µg/ml, and of 0.5 µg/ml for Cry1Ac. Apoptosis was differentially induced in HeLa cells using the same concentration of Cry1Ab and Cry1Ac toxins. Cry1Ac eliminated 50% of the tumors at 10 µg/ml, and eliminate 100% of the tumors at 30 and 50 µg/ml. CONCLUSION: Bacillus thuringiensis Cry1A toxins show dual cytotoxic activity, in insects as well as in HeLa cancer cell line.

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PURPOSE: To compare fiber posts of several calibers and trademarks to their corresponding root canal preparation drills. METHODS: Three widely used endodontic post brands and their drills were evaluated: Exacto, ParaPost Taper Lux, and Macro-Lock Illusion X-RO. Fiber posts and drills were microphotographed with a scanning electron microscope and images were analyzed using ImageJ image processing software. Fiber post diameter on apical extreme (Pd0), fiber post diameter at 5 mm from the apical extreme (Pd5), drill diameter on apical extreme (Dd0) and drill diameter at 5 mm from the apical extreme (Dd5) were analyzed. The data were statistically analyzed using student t-test. RESULTS: Exacto posts 0.5 showed larger dimensions than their corresponding drills (P< 0.05) at Pd0. Macro-Lock posts showed no significant differences vs. their drills at Pd0 in any of the studied groups. ParaPost drills 4.5, 5 and 5.5 were statistically significantly larger than their posts at Dd0 (P< 0.05). Exacto posts 0.5 and 1 showed larger dimensions than their drills measured at Pd5 (P< 0.05). Exacto posts number 2 showed smaller calibers than their corresponding drills at Pd5 (P< 0.05). Macro-Lock drills number 4 and ParaPost drills number 5 were larger than their posts at Dd5 (P< 0.05). CLINICAL SIGNIFICANCE: Poor spatial correspondence between post and drill dimensions can adversely affect the film thickness of the resin cement, diminishing bond strength due to polymerization shrinkage. The lack of correspondence in size between posts and drills may lead to the formation of empty chambers between the post and endodontic obturation with excessive luting cement thickness, thus inducing critical C-Factor stresses.

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GERD is a highly prevalent disease in our country. It has a deep impact in patient´s quality of life, representing extremely high costs regarding health. The correct understanding of its pathophysiology is crucial for the rational use of diagnoses methods and the implementation of appropriate treatment adjusted to each individual case. In this review we evaluate this disorder based on the best available evidence, focusing in pathophysiological mechanisms, its epidemiology, modern diagnosis methods and current management standards.

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Introducción: La incontinencia fecal mayor es un trastorno que modifica significativamente la calidad devida. Un grupo particularmente afectado son las mujeres con antecedentes de trauma obstétrico. Dentrode ellos, los más graves son los de cuarto grado que involucran la totalidad de las capas del tabique rectovaginal, produciendo una comunicación completa entre la luz rectal y la vagina, generando una cloaca. Sibien la incidencia de éstas, es de alrededor del 0,3% de los partos, el efecto que tiene sobre las pacienteses devastador. El único tratamiento efectivo para este tipo de lesiones es la reparación quirúrgica.Objetivo: Evaluar el impacto que presenta la corrección quirúrgica de la cloaca por trauma obstétrico en lacalidad de vida de las afectadas.Material y Métodos: Análisis prospectivo secundario y ampliado de una serie consecutiva de pacientestratadas por desgarro perineal completo tipo cloaca durante el año 2013. Se evaluó la demografía de lamuestra, la paridad de las pacientes, el tiempo medio hasta la consulta desde el último parto, la manometríapre y postoperatoria, la evaluación de incontinencia fecal pre y postoperatorio. Para valorar la severidad dela incontinencia fecal se utilizó el índice CCF-FIS y el índice de severidad de incontinencia fecal (FISI). Parala evaluación de calidad de vida se utilizó la encuesta FIQLS de la Sociedad Americana de Cirujanos delColon y Recto (ASCRS).Resultados: Tres pacientes fueron intervenidas entre enero de 2013 y diciembre de 2013. En el examenfísico, el 100% de las pacientes presentaron una cloaca perineal. El score CCF-FIS preoperatorio fue del16,7 (16 a 18 puntos). El puntaje de FISI pre-operatorio fue de 54,3 (52 a 57). Las tres pacientes refirieronalteraciones en su actividad social y sexual. Se reevaluaron las pacientes al tercer mes de postoperatorio yluego del cierre de la colostomía...

Introduction: The major fecal incontinence is a disorder that significantly change the quality of life. Aparticularly affected group are women with a history of obstetric trauma and presenting demonstrationsimmediately. Among them, the most serious are the fourth degree involving all the layers of the rectovaginalseptum, producing a complete communication between the rectal lumen and vagina, creating a sewer.Although their incidence is about 0.3% of births, the effect on patients is devastating.Objective: To evaluate the impact making the surgical correction of the cloaca by obstetrical trauma in thequality of life of those affected.Material and Methods: Secondary and expanded Prospective analysis of a consecutive series of patientstreated by complete perineal tear type cloaca in 2013. The demographics of the sample was evaluated theparity of the patients, the median time to the query from the last delivery, pre and postoperative manometry,assessment of pre-and postoperative fecal incontinence. To assess the severity of fecal incontinence CCFFISindex and the severity of fecal incontinence (FISI) was used. The FIQLS survey by the American Societyof Colon and Rectal Surgeons (ASCRS) was used for the evaluation of quality of lifeResults: Three patients were operated between January 2013 and December 2013 on physical examination, 100% of patients had a perineal cloaca. The CCF-FIS preoperative score was 16.67 (16-18 points). Thescore FISI pre-surgery was 54.33 (52-57). The three patients reported changes in their social and sexualactivity. Patients at the third month after surgery and after colostomy closure were reassessed...

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Scleroderma is a chronic autoimmune disease of unknown cause characterized by fibrotic skin and multiple organs involvement, including the gastrointestinal tract. It occurs mainly in women between 35 and 65 years of age. It is classified as limited or diffuse based on the extent of skin involvement. Gastrointestinal dysmotility is observed in up to 90% of patients with a diffuse and limited scleroderma. It may involve any segment of the gastrointestinal tract from the esophagus to the anus and is related to collagen deposition at the level of enteric and vascular smooth muscle. Gastroparesis is a condition characterized by abnormal gastric motility, delay gastric emptying, in the absence of a mechanical obstruction to outflow. Gastric scintigraphy with radiolabeled solid food is the gold standard for the diagnosis of gastroparesis. Two cases of patients with systemic scleroderma and severe gastroparesis are presented in order to discuss the diagnostic and therapeutic approach, emphasizing the utility of gastric emptying scintigraphy.

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Twenty eight Bacillus thuringiensis strains isolated from the Tijuana-Ensenada region of northwestern Mexico were analyzed to determine the distribution of cry and cyt genes. Crystal production by the strains was examined by scanning electron microscopy, which showed the predominance of cubic crystals. Alkaline-dissolved and trypsin activated crystals were also analyzed by SDS-PAGE, yielding bands of 40-200 kDa. The cry1 and cry2 genes were molecularly characterized using general and newly designed specific primers in addition to other oligonucleotides (cry3, cry4, cry8, cry9, cry11, Nem, cry25, cry29 and cyt), resulting in the identification of novel gene combinations. The use of specific primers for cry1A, cry1B, cry1C, cry1D, cry1E, cry1F and cry2Aa, cry2Ab, cry2Ac, cry2Ad showed differences in the distribution of cry1 (36%), cry2 (71%), and cyt (40%) in strains from Tijuana-Ensenada compared to other previously studied regions. Bioassays were conducted on Manduca sexta larvae to analyze the Cry insecticidal capacity of the isolated strains. The hemolytic activity of the Cyt toxin from the same strains was assessed in human erythrocytes.

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Functional feed supplemented with alternative-economic nutrient sources (protein, carbohydrates, lipids) and probiotics are being considered in shrimp/fish aquaculture production systems as an option to increase yield and profits and to reduce water pollution. In this study the probiotic potential to formulate functional feeds have been evaluated using four dietary treatments: Treatment 1 (B + Bs); Bacillus subtilis potential probiotic strain was supplemented to a soybeanmeal (SBM)-carbohydrates (CHO) basal feed. Treatment 2 (B + Bm); Bacillus megaterium potential probiotic strain was supplemented to the same SBM-CHO basal feed. In Treatment 3 (B); SBM-CHO basal feed was not supplemented with probiotic strains. Treatment 4 (C); fishmeal commercial feed (FM) was utilized as positive control. Feeding trials evaluated the survival, growth, and food conversion ratio and stress tolerance of juvenile Litopenaeus vannamei (Boone) Pacific white shrimp. Best overall shrimp performance was observed for animals fed with Treatment 1 (B+Bs); additionally, stress tolerance and hemolymph metabolites also showed the best performance in this treatment. SBM-CHO basal feed not supplemented with probiotic strains (B) presented smaller growth and lower feed conversion ratio (FCR). Shrimps fed with the fishmeal commercial feed (C) presented the lowest stress tolerance to high ammonia and low oxygen levels. Specifically selected B. subtilis strains are recommended to formulate functional and economical feeds containing high levels of vegetable; protein and carbohydrates as main dietary sources in L. vannamei cultures.

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PURPOSE: Angiodysplasia is a frequent cause of colonic bleeding. Argon plasma coagulation was reported to be useful in endoscopic hemostasis. However, the long-term outcomes have been poorly evaluated. This study was designed to assess the long-term outcomes of bleeding patients with colonic angiodysplasia treated by argon plasma coagulation. METHODS: A cohort of 100 patients with gastrointestinal bleeding caused by colonic angiodysplasia was studied prospectively. The endoscopic intervention was considered successful if there was no further overt bleeding and hemoglobin concentration was stabilized (primary end point). Secondary end points were the requirement of blood transfusions, the need for surgery, bleeding-related mortality, and overall mortality. RESULTS: Overt bleeding resolved and hemoglobin levels were stabilized without transfusions or iron therapy in 85 of 100 patients (85 percent) after a median follow-up of 20 (range, 6-62) months. Transfusion requirements ceased in 90 percent of patients and only one required surgery. No patient died because of hemorrhage. In the subgroup of patients with anemia, mean hemoglobin levels increased from 9.3 (range, 5.5-12.2) g/dl before treatment to 12.6 (range, 7.4-16.7) g/dl after treatment (P < 0.01). The probability of remaining free of rebleeding at one and two year follow-up was 98 percent (95 percent confidence interval, 96-100) and 90 percent (95 percent confidence interval, 83-97), respectively. Among 118 procedures, only two complications were observed (1.7 percent). CONCLUSIONS: Endoscopic argon plasma ablation therapy is useful in the management of bleeding from colonic angiodysplasia.

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We sought to assess the prevalence, severity of symptoms, and risk factors of uninvestigated dyspepsia in a population-based study in Argentina. Eight hundred thirty-nine valid questionnaires were evaluated. Dyspepsia was present in 367 subjects (43.2%; 95% confidence interval [CI], 39.8-46.6); 110 (13.6%) had overlap with gastroesophageal reflux disease (GERD). The group with dyspepsia without GERD consisted of 257 subjects (29.6%; 95% CI, 26.5-32.7), 183 (71.1%) had ulcer-like dyspepsia, and 74 (28.9%) had dysmotility-like dyspepsia. Symptoms were considered very severe in 1.9%, severe in 14.0%, moderate in 59.5%, and mild in 24.5% of the subjects. Dyspepsia was associated with a score >14 on the psychosomatic symptom scale (PSC) (OR, 2.52; 95% CI, 1.75-3.61), a family history of diseases of the esophagus or stomach (OR, 1.73; 95% CI, 1.19-2.52) and an educational level >12 years (OR, 1.55; 95% CI, 1.05-2.29). Dyspepsia is especially prevalent in Argentina. In a significant proportion of dyspeptic subjects, the severity of symptoms interferes with daily activities. A higher PSC, positive family history, and a higher educational level are risk factors for dyspepsia.

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