Molecular Docking and In Vitro Studies of Ochratoxin A (OTA) Biodetoxification Testing Three Endopeptidases.

Ochratoxin A (OTA) is considered one of the main mycotoxins responsible for health problems and considerable economic losses in the feed industry. The aim was to study OTA's detoxifying potential of commercial protease enzymes: (i) Ananas comosus bromelain cysteine-protease, (ii) bovine trypsin serine-protease and (iii) Bacillus subtilis neutral metalloendopeptidase. In silico studies were performed with reference ligands and T-2 toxin as control, and in vitro experiments. In silico study results showed that tested toxins interacted near the catalytic triad, similar to how the reference ligands behave in all tested proteases. Likewise, based on the proximity of the amino acids in the most stable poses, the chemical reaction mechanisms for the transformation of OTA were proposed. In vitro experiments showed that while bromelain reduced OTA's concentration in 7.64% at pH 4.6; trypsin at 10.69% and the neutral metalloendopeptidase in 8.2%, 14.44%, 45.26% at pH 4.6, 5 and 7, respectively (p < 0.05). The less harmful α-ochratoxin was confirmed with trypsin and the metalloendopeptidase. This study is the first attempt to demonstrate that: (i) bromelain and trypsin can hydrolyse OTA in acidic pH conditions with low efficiency and (ii) the metalloendopeptidase was an effective OTA bio-detoxifier. This study confirmed α-ochratoxin as a final product of the enzymatic reactions in real-time practical information on OTA degradation rate, since in vitro experiments simulated the time that food spends in poultry intestines, as well as their natural pH and temperature conditions.

Evaluation of trefoil factor 3 as a non-invasive biomarker of gastric intestinal metaplasia and gastric cancer in a high-risk population.

BACKGROUND: Adenocarcinoma is preceded by chronic atrophic gastritis, gastric intestinal metaplasia and dysplasia. Trefoil factor 3 (TFF3) is a peptide secreted by goblet cells, which is abundantly present in intestinal metaplasia. AIM: To evaluate the utility of serum TFF3 as a non-invasive biomarker for the diagnosis of intestinal metaplasia and gastric cancer. METHODS: Single-center, cross-sectional study of 274 patients who consecutively underwent upper gastrointestinal endoscopy with gastric biopsies (updated Sydney system). TFF3 levels were measured in serum by a commercial ELISA kit. Patients with normal histology or chronic atrophic gastritis without intestinal metaplasia comprised the control group. In addition, 14 patients with invasive gastric cancer were included as a reference group. The association between TFF3 levels and intestinal metaplasia was assessed by logistic regression. RESULTS: Patients with intestinal metaplasia (n=110) had a higher median TFF3 level as compared to controls (n=164), 13.1 vs. 11.9ng/mL, respectively (p=0.024). Multivariable logistic regression showed a no significant association between TFF3 levels and intestinal metaplasia (OR=1.20; 95%CI: 0.87-1.65; p-trend=0.273). The gastric cancer group had a median TFF3 level of 20.5ng/mL, and a significant association was found (OR=3.26; 95%CI: 1.29-8.27; p-trend=0.013). CONCLUSION: Serum levels of TFF3 do not discriminate intestinal metaplasia in this high-risk Latin American population. Nevertheless, we confirmed an association between TFF3 levels and invasive gastric cancer.

Uso de endoprótesis para el tratamiento de enfermedades no neoplásicas de la rodilla / Use of endoprosthesis for the treatment of non-neoplastic pathologies of the knee

Introducción: Las endoprótesis son el método de elección para la reconstrucción luego de las resecciones oncológicas. Los avances en los materiales y diseños permitieron expandir las indicaciones a enfermedades no neoplásicas. Su montaje intraoperatorio simple y rápido, y su estabilidad mecánica inmediata permiten una rehabilitación y una recuperación funcional tempranas. Sin embargo, la tasa de fallas es elevada, aunque distinta de la de las enfermedades oncológicas. Las causas predominantes son diferentes. Objetivos: Analizar nuestra experiencia con el uso de endoprótesis de rodilla y compararla con los estudios publicados, evaluando los resultados funcional y radiográfico, la supervivencia del implante y las causas de su eventual falla. Materiales y Métodos: Se seleccionaron pacientes con enfermedad no neoplásica compleja de rodilla que requirieran una reconstrucción con endoprótesis. Para el examen clínico y la evaluación funcional se utilizó el puntaje de la Musculoskeletal Tumor Society, y para las fallas de los implantes, la clasificación de Henderson y cols. modificada. Resultados:Se estudiaron 12 endoprótesis, con un seguimiento promedio de 3.8 años. Se registraron 2 fallas (18%), con un tiempo promedio hasta la falla de 47.5 meses. Una fue tipo 2 (aflojamiento aséptico) y la otra, tipo 4 (infección). No hubo otras complicaciones. En la evaluación funcional, el puntaje final medio fue del 76,6%. Conclusión: Nuestros resultados respaldan el uso de endoprótesis para enfermedades complejas no neoplásicas de rodilla en pacientes cuidadosamente seleccionados, pese a ser un procedimiento quirúrgico complejo y con muchas complicaciones. Nivel de Evidencia: IV

Introduction: Endoprosthesis is the gold standard for reconstruction after oncological resections. The advances regarding its materials and designs allowed for the expansion of the indications to non-neoplastic pathologies. Its simple and fast intraoperative assembly and its immediate mechanical stability allow for early rehabilitation and functional recovery. However, the failure rate is high, although it is different from oncological pathologies. The predominant causes are varied. Objectives: To analyze our experience in the use of knee endoprosthesis and compare it with the literature, evaluating functional outcomes, radiographic outcomes, implant survival and causes of eventual failure. Materials and Methods: Patients with complex non-neoplastic knee pathology that required reconstruction with endoprosthesis were selected. Clinical history, anamnesis, physical examination, and radiographs were reviewed. For clinical examination and functional evaluation, the MusculoSkeletal Tumor Society Score (MSTS Score) was used. For implant failures, the modified Henderson et al. classification was used. Results: 12 endoprostheses were studied, with an average follow-up of 3.8 years. Failures were recorded in 2 (18%), with a mean time to failure of 47.5 months. One type 2 failure (aseptic loosening) and one type 4 failure (infection) were recorded. No other complications were noted. For the functional evaluation, the mean final score was 76.6%. Conclusion: Our results support the use of endoprostheses for complex non-neoplastic knee diseases in carefully selected patients, despite being a complex surgical procedure with many complications.Keywords: Endoprosthesis; knee; infection; complications. Level of Evidence: IV

Detection of SARS-CoV-2 RNA in serum is associated with increased mortality risk in hospitalized COVID-19 patients.

COVID-19 has overloaded national health services worldwide. Thus, early identification of patients at risk of poor outcomes is critical. Our objective was to analyse SARS-CoV-2 RNA detection in serum as a severity biomarker in COVID-19. Retrospective observational study including 193 patients admitted for COVID-19. Detection of SARS-CoV-2 RNA in serum (viremia) was performed with samples collected at 48-72 h of admission by two techniques from Roche and Thermo Fischer Scientific (TFS). Main outcome variables were mortality and need for ICU admission during hospitalization for COVID-19. Viremia was detected in 50-60% of patients depending on technique. The correlation of Ct in serum between both techniques was good (intraclass correlation coefficient: 0.612; p < 0.001). Patients with viremia were older (p = 0.006), had poorer baseline oxygenation (PaO2/FiO2; p < 0.001), more severe lymphopenia (p < 0.001) and higher LDH (p < 0.001), IL-6 (p = 0.021), C-reactive protein (CRP; p = 0.022) and procalcitonin (p = 0.002) serum levels. We defined "relevant viremia" when detection Ct was < 34 with Roche and < 31 for TFS. These thresholds had 95% sensitivity and 35% specificity. Relevant viremia predicted death during hospitalization (OR 9.2 [3.8-22.6] for Roche, OR 10.3 [3.6-29.3] for TFS; p < 0.001). Cox regression models, adjusted by age, sex and Charlson index, identified increased LDH serum levels and relevant viremia (HR = 9.87 [4.13-23.57] for TFS viremia and HR = 7.09 [3.3-14.82] for Roche viremia) as the best markers to predict mortality. Viremia assessment at admission is the most useful biomarker for predicting mortality in COVID-19 patients. Viremia is highly reproducible with two different techniques (TFS and Roche), has a good consistency with other severity biomarkers for COVID-19 and better predictive accuracy.

IL-6 serum levels predict severity and response to tocilizumab in COVID-19: An observational study.

BACKGROUND: Patients with coronavirus disaese 2019 (COVID-19) can develop a cytokine release syndrome that eventually leads to acute respiratory distress syndrome requiring invasive mechanical ventilation (IMV). Because IL-6 is a relevant cytokine in acute respiratory distress syndrome, the blockade of its receptor with tocilizumab (TCZ) could reduce mortality and/or morbidity in severe COVID-19. OBJECTIVE: We sought to determine whether baseline IL-6 serum levels can predict the need for IMV and the response to TCZ. METHODS: A retrospective observational study was performed in hospitalized patients diagnosed with COVID-19. Clinical information and laboratory findings, including IL-6 levels, were collected approximately 3 and 9 days after admission to be matched with preadministration and postadministration of TCZ. Multivariable logistic and linear regressions and survival analysis were performed depending on outcomes: need for IMV, evolution of arterial oxygen tension/fraction of inspired oxygen ratio, or mortality. RESULTS: One hundred forty-six patients were studied, predominantly males (66%); median age was 63 years. Forty-four patients (30%) required IMV, and 58 patients (40%) received treatment with TCZ. IL-6 levels greater than 30 pg/mL was the best predictor for IMV (odds ratio, 7.1; P < .001). Early administration of TCZ was associated with improvement in oxygenation (arterial oxygen tension/fraction of inspired oxygen ratio) in patients with high IL-6 (P = .048). Patients with high IL-6 not treated with TCZ showed high mortality (hazard ratio, 4.6; P = .003), as well as those with low IL-6 treated with TCZ (hazard ratio, 3.6; P = .016). No relevant serious adverse events were observed in TCZ-treated patients. CONCLUSIONS: Baseline IL-6 greater than 30 pg/mL predicts IMV requirement in patients with COVID-19 and contributes to establish an adequate indication for TCZ administration.

Vedolizumab, an option in patients with inflammatory bowel disease intolerant to thiopurines and refractory to biological agents. / Vedolizumab, una opción en pacientes con enfermedad inflamatoria intestinal intolerantes a tiopurinas y refractarios a biológicos.

Vedolizumab (VDZ), a human monoclonal antibody that binds specifically to α4ß7-integrin, and is approved for the treatment of Crohn's disease (CD) and ulcerative colitis (UC), has demonstrated its efficacy in controlled clinical trials. OBJECTIVE: To describe a population treated with VDZ and to evaluate its long-term efficacy and safety in clinical practice. METHODS: An observational and multicentre study was carried out on patients with inflammatory bowel disease treated with VDZ for at least one year. An evaluation was performed on the activity indices, faecal calprotectin and C-reactive protein levels, hospital admissions, surgeries, and adverse events. RESULTS: A total of 73 patients were analysed (43 UC and 30 CD). More than one anti-TNF and more than one immunosuppressive was previously used by 74 and 23%, respectively, of UC patients, and 90 and 37%, respectively of CD patients. VDZ was stopped in 17 (23%) patients, 10 UC and 7 CD, due to a lack or loss of response before the first year, or due to adverse events. An intensification of the dose was required in 26 (63%) UC, and 16 (53%) CD patients. At 6 months, 70 and 42% of UC patients, and 80 and 43% of CD patients achieved a clinical response and remission, respectively. At one year, 58 and 35% of UC patients and 47 and 43% of CD patients, maintained the clinical response and remission, respectively. The C-reactive protein decreased significantly in both CD and UC patients. However, the decrease in faecal calprotectin was only achieved during follow-up in UC, but not in CD patients. Eight patients with CD that had been treated previously with ustekinumab avoided surgery at one year. A colectomy was performed on 8 (18.6%) UC patients, and 4 (13.3%) CD patients needed surgery. Six patients (8%) (5 UC and 1 CD) had adverse events. The concomitant use of corticosteroids or immunomodulators did not increase the efficacy. Those with a higher number of previous anti-TNF treatments showed less remissions in UC and responses in CD. CONCLUSIONS: After one year of VDZ, a clinical response and remission was induced in a considerable percentage of patients refractory to different biological or immunosuppressive therapies. VDZ can be considered as an alternative in those intolerant to immunosuppressives, with few adverse events.

A Kinetic Study of Furan Formation in Wheat Flour-Based Model Systems during Frying.

Furan is a possible human carcinogen, which is formed in worldwide highly consumed fried starchy foods. In order to elucidate the mechanisms responsible for its occurrence in this food category and propose techniques for its mitigation, the kinetics of furan formation, oil absorption, lipid oxidation, and color change were studied in wheat flour-based model systems during frying at 160, 170, 180, and 190 °C up to 13 min and data were fitted to mathematical models. Additionally, an Arrhenius-type dependency with temperature was evaluated for all studied responses. More drastic frying conditions increased significantly (P ≤ 0.05) the furan content of fried samples. Furan formation followed a sigmoid trend with frying time only for frying temperature of 190 °C (RMS190°C : 7.6%). At lower temperatures, furan generation did not reach the asymptotic concentration level. Color change, lipid oxidation, and oil absorption increased with frying temperature and followed asymptotic relationships with frying time. For all evaluated temperatures, color change (RMS: 4.4% to 12.5%) and polar compound generation (RMS: 2.6% to 7.4%) presented good fit to a first-order kinetic model. Oil absorption was successfully fit to a mass balance-based model (RMS: 10.0% to 19.8%). Under the experimental conditions studied, only color change (EA : 15.47 kJ/mol), lipid oxidation (EA : 6.67 kJ/mol), and oil absorption (EA : 76.98 kJ/mol) presented good fit (RMS: 0.7% to 6.3%) to an Arrhenius-type equation. Based on our results, the keeping of frying temperature below 180 °C and the reduction of the frying time would contribute to reduce not only the final furan occurrence in fried foods but also their oil content.

Discontinuation of Infliximab in Patients With Ulcerative Colitis Is Associated With Increased Risk of Relapse: A Multinational Retrospective Cohort Study.

BACKGROUND & AIMS: Infliximab is a safe and effective therapy for ulcerative colitis (UC). We conducted a multicenter retrospective cohort study that included 7 European countries and Israel to examine whether infliximab discontinuation can be considered for patients who achieve sustained remission. METHODS: We performed a retrospective cohort study, collecting medical records from 13 tertiary care referral inflammatory bowel disease centers of all patients with UC treated with infliximab (n = 193). We compared the disease course of patients with at least 12 months of clinical remission who discontinued infliximab (n = 111) with that of patients who continued scheduled treatment (controls, n = 82). We examined the incidence rates of relapse, hospitalization and colectomy, the comparative effectiveness of different therapeutic strategies after discontinuation, and assessed the rates of response, remission, and adverse effects after infliximab re-initiation. Statistical analyses used time-to-event methods. RESULTS: In the entire cohort, 67 patients (34.7%) relapsed during the follow-up period. The incidence rate of relapse was significantly higher after discontinuation (23.3 per 100 person-years) compared with the control group (7.2 per 100 person-years) in univariable analysis (log-rank P < .001; hazard ratio, 3.41; 95% confidence interval, 1.88-6.20) and multivariable analysis (hazard ratio, 3.70; 95% confidence interval, 2.02-6.77). Rates of hospitalization and colectomy did not differ between groups. Thiopurines appeared to be the best treatment option after infliximab discontinuation (incidence of relapse: 15.0 per 100 person-years for thiopurines, 27.4 per 100 person-years for thiopurines plus aminosalicylates, and 31.2 per 100 person-years for aminosalicylates alone; log-rank P = .032). Response was regained in 77.1% of patients and remission in 51.4% of patients who re-initiated infliximab. However, 17.1% had infusion reactions and 17.1% reported other adverse events. CONCLUSIONS: In a multinational retrospective cohort study of patients with UC in sustained clinical remission, we associated discontinuation of infliximab with an increased risk of relapse. Treatment re-initiation is effective and safe.

Development of alginate microspheres containing thyme essential oil using ionic gelation.

Essential oils are a good antimicrobial and antioxidant agent alternative in human or animal feed. However, their direct use has several disadvantages such as volatilization or oxidation. The development of essential oil microspheres may help to avoid these problems. The objective of the present research was to microencapsulate thyme essential oil by generating emulsions with different dispersion degrees. The emulsions were encapsulated in calcium-alginate microspheres by ionic gelation. The microspheres were evaluated regarding size, shape, encapsulation efficiency, loading capacity and antimicrobial properties. The results indicate that encapsulation efficiency and loading capacity are dependent on concentration and degree of dispersion. The best encapsulation conditions were obtained at 2% v/v of thyme essential oil with a high dispersion degree (18,000rpm/5min), which was achieved with an efficiency of 85%. Finally, the microspheres obtained showed significant antimicrobial effect, especially in gram-positive bacteria.

[Recommendations of the Chilean Association for Digestive Endoscopy for the management of gastric pre-malignant lesions]. / Diagnóstico precoz de cáncer gástrico: Propuesta de detección y seguimiento de lesiones premalignas gástricas: protocolo ACHED.

An expert panel analyzed the available evidence and reached a consensus to release 24 recommendations for primary and secondary prevention of gastric cancer (CG) in symptomatic patients, with indication for upper GI endoscopy. The main recommendations include (1) Search for and eradicate H. pylori infection in all cases. (2) Systematic gastric biopsies (Sydney protocol) in all patients over 40 years of age or first grade relatives of patient with CG, to detect gastric atrophy, intestinal metaplasia or dysplasia. (3) Incorporate the OLGA system (Operative Link on Gastritis Assessment) to the pathological report, to categorize the individual risk of CG. (4) Schedule endoscopic follow-up according to the estimated risk of CG, namely annual for OLGA III- IV, every 3 years for OLGA I- II or persistent H. pylori infection, every 5 years for CG relatives without other risk factors and no follow-up for OLGA 0, H. pylori (-). (4) Establish basic human and material resources for endoscopic follow-up programs, including some essential administrative processes, and (5) Suggest the early CG/total CG diagnosis ratio of each institution and the proportion of systematic recording of endoscopic images, as quality indicators. These measures are applicable using currently available resources, they can complement any future screening programs for asymptomatic population and may contribute to improve the prognosis of CG in high-risk populations.

Management of Helicobacter pylori infection in Latin America: a Delphi technique-based consensus.

AIM: To optimize diagnosis and treatment guidelines for this geographic region, a panel of gastroenterologists, epidemiologists, and basic scientists carried out a structured evaluation of available literature. METHODS: Relevant questions were distributed among the experts, who generated draft statements for consideration by the entire panel. A modified three-round Delphi technique method was used to reach consensus. Critical input was also obtained from representatives of the concerned medical community. The quality of the evidence and level of recommendation supporting each statement was graded according to United States Preventive Services Task Force criteria. RESULTS: A group of ten experts was established. The survey included 15 open-ended questions that were distributed among the experts, who assessed the articles associated with each question. The levels of agreement achieved by the panel were 50% in the first round, 73.3% in the second round and 100% in the third round. Main consensus recommendations included: (1) when available, urea breath and stool antigen test (HpSA) should be used for non-invasive diagnosis; (2) detect and eradicate Helicobacter pylori (H. pylori) in all gastroscopy patients to decrease risk of peptic ulcer disease, prevent o retard progression in patients with preneoplastic lesions, and to prevent recurrence in patients treated for gastric cancer; (3) further investigate implementation issues and health outcomes of H. pylori eradication for primary prevention of gastric cancer in high-risk populations; (4) prescribe standard 14-d triple therapy or sequential therapy for first-line treatment; (5) routinely assess eradication success post-treatment in clinical settings; and (6) select second- and third-line therapies according to antibiotic susceptibility testing. CONCLUSION: These achievable steps toward better region-specific management can be expected to improve clinical health outcomes.

Clinical and epidemiological features of 147 Chilean patients with chronic hepatitis C.

Prevalence, modes of transmission, clinical characteristics and outcomes of hepatitis C (HCV) infection vary in different geographical areas. We aim to describe clinical and epidemiological features of Chilean patients infected with hepatitis C virus. An analysis of demographic, epidemiological, clinical and laboratory data of patients referred to a liver clinic and blood donors with chronic hepatitis C was carried out. 147 patients were evaluated, 68 (46%) were male. Median age was 56 years, median infection age was 27 years and median duration of infection was 27 years. 52.5% of the patients were cirrhotic, and estimated risk of progression to cirrhosis was 16% at 20 years from infection. Risk factors for acquisition of the disease among patients were: Blood transfusion 54%, injection drug use 5%, and risky sexual behavior 2%. No factor was identified in 43% of the patients. Twelve of 64 (18.8%) family members tested positive for HCV antibodies. Genotype 1b was predominant (82%), and 52% of patients had high viral load (>850.000 IU/mL). Liver biopsy was available in 50 patients, showing advanced fibrosis in 54%. These patients were in average 10 years older and tended to have longer duration of infection. Hepatocellular carcinoma was present at the moment of enrollment in 7 patients and developed in 4 more patients during follow up (2.4 years). In conclusion, the natural history and clinical characteristics of HCV infection in Chilean patients is similar to that described elsewhere. The main risk factor was blood transfusion. A significant proportion of patients had advanced liver disease or hepatocellular carcinoma at time of diagnosis.