Identification of early predictors of clinical outcomes of COVID-19 outbreak in an Italian single center using a machine-learning approach.

OBJECTIVE: SARS-CoV-2 disease (COVID-19) has become a pandemic disease, determining a public health emergency. The use of artificial intelligence in identifying easily available biomarkers capable of predicting the risk for severe disease may be helpful in guiding clinical decisions. The aim of the study was to investigate the ability of interleukin (IL)-6, troponin I, and D-dimer to identify patients with COVID-19 at risk for intensive care unit (ICU)-admission and death by using a machine-learning predictive model. PATIENTS AND METHODS: Data on demographic characteristics, underlying comorbidities, symptoms, physical and radiological findings, and laboratory tests have been retrospectively collected from electronic medical records of patients admitted to Policlinico A. Gemelli Foundation from March 1, 2020, to September 15, 2020, by using artificial intelligence techniques. RESULTS: From an initial cohort of 425 patients, 146 met the inclusion criteria and were enrolled in the study. The in-hospital mortality rate was 15%, and the ICU admission rate was 41%. Patients who died had higher troponin I (p-value<0.01) and IL-6 values (p-value=0.04), compared to those who survived. Patients admitted to ICU had higher levels of troponin I (p-value<0.01) and IL-6 (p-value<0.01), compared to those not admitted to ICU. Threshold values to predict in-hospital mortality and ICU admission have been identified. IL-6 levels higher than 15.133 ng/L have been associated with a 22.91% risk of in-hospital mortality, and IL-6 levels higher than 25.65 ng/L have been associated with a 56.16% risk of ICU admission. Troponin I levels higher than 12 ng/L have been associated with a 26.76% risk of in-hospital mortality and troponin I levels higher than 12 ng/L have been associated with a 52.11% risk of ICU admission. CONCLUSIONS: Levels of IL-6 and troponin I are associated with poor COVID-19 outcomes. Cut-off values capable of predicting in-hospital mortality and ICU admission have been identified. Building a predictive model using a machine-learning approach may be helpful in supporting clinical decisions in a more precise and personalized way.

Valoración de la cicatriz postquirúrgica de pacientes operados por quiste pilonidal según técnicas abiertas versus cerradas

Introducción: El quiste pilonidal puede ser abordado según técnicas quirúrgicas abiertas y cerradas, con distintos resultados estéticos. La cicatriz es un factor importante y representa un indicador de satisfacción. Una herramienta para valorar cicatrices es el cuestionario POSAS 2.0, adaptado transculturalmente para Chile. Objetivo: Conocer la valoración subjetiva de los pacientes operados de quiste pilonidal respecto a su cicatriz, según la técnica quirúrgica. Material y Método: Estudio es de carácter retrospectivo, observacional, transversal, analítico y de cohorte. Se aplica la escala de evaluación por el paciente del cuestionario POSAS 2.0, con una calificación desde 1 a 10, con puntaje total entre 7 y 70. Se recolectaron variables clínico-demográficas. Se aplicó estadística analítica, se compararon los resultados obtenidos entre técnicas abiertas y cerradas. Resultados. Se encuestaron 101 pacientes operados de quiste pilonidal entre 2013 y 2019, de los cuales 59 (59,4%) fueron de sexo masculino. La edad promedio fue de 23 años (DS 7,2). Del total, 22 (21,8%) de los pacientes fueron manejados con técnicas cerradas y 79 con técnicas abiertas (78,2%). El promedio del puntaje total de la escala fue 20 (DS 11,4), 22 en técnicas cerradas y 19,6 en técnicas abiertas. No se encontraron diferencias estadísticamente significativas. Conclusión: La cicatriz postoperatoria es valorada positivamente, con un puntaje que denota características similares a la piel normal. No existen diferencias significativas de la percepción de los pacientes respecto a su cicatriz. Esta es la primera comunicación chilena que evalúa la valoración subjetiva de pacientes sobre la cicatriz postoperatoria de quiste pilonidal.

Introduction: The pilonidal cyst can be approached according to open and closed surgical techniques, with different aesthetic results. The scar is an important factor and represents an indicator of satisfaction. A tool to assess scars is the POSAS 2.0 questionnaire, adapted cross-culturally for Chile. Objective: To know the subjective assessment of patients operated on for pilonidal cyst with respect to their scar, according to the surgical technique. Material and Method: This is a retrospective, observational, cross-sectional, analytical, and cohort study. The patient evaluation scale of the POSAS 2.0 questionnaire is applied, with a score from 1 to 10, with a total score between 7 and 70. Clinical-demographic variables were collected. Analytical statistics were applied, the results obtained between open and closed techniques were compared. Results: We surveyed 101 patients operated on for pilonidal cyst between 2013 and 2019, of which 59 (59.4%) were male. The mean age was 23 years (SD 7.2). Of the total, 22 (21.8%) of the patients were managed with closed techniques and 79 with open techniques (78.2%). The mean total score of the scale was 20 (SD 11.4), 22 in closed techniques and 19.6 in open techniques. No statistically significant differences were found. Conclusion: The postoperative scar is positively valued, with a score that denotes similar characteristics to the normal skin. There are no significant differences in patient's perception of their scar. This is the first chilean communication that evaluates the subjective assessment of patients on the postoperative scar of pilonidal cyst.

Antibacterial and anticancer activity of two NK-lysin-derived peptides from the Antarctic teleost Trematomus bernacchii.

The NK-lysin antimicrobial peptide, first identified in mammals, possesses both antibacterial and cytotoxic activity against cancer cell lines. Homologue peptides isolated from different fish species have been examined for their functional characteristics in the last few years. In this study, a NK-lysin transcript was identified in silico from the head kidney transcriptome of the Antarctic teleost Trematomus bernacchii. The corresponding amino acid sequence, slightly longer than NK-lysins of other fish species, contains six cysteine residues that in mammalian counterparts form three disulphide bridges. Real time-PCR analysis indicated its predominant expression in T. bernacchii immune-related organs and tissues, with greatest mRNA abundance detected in gills and spleen. Instead of focusing on the full T. bernacchii derived NK-lysin mature molecule, we selected a 27 amino acid residue peptide (named NKL-WT), corresponding to the potent antibiotic NK-2 sequence found in human NK-lysin. Moreover, we designed a mutant peptide (named NKL-MUT) in which two alanine residues substitute the two cysteines found in the NKL-WT. The two peptides were obtained by solid phase organic synthesis to investigate their functional features. NKL-WT and NKL-MUT displayed antibacterial activity against the human pathogenic bacterium Enterococcus faecalis and the ESKAPE pathogen Acinetobacter baumannii, respectively. Moreover, at the determined Minimum Inhibitory Concentration and Minimum Bactericidal Concentration values against these pathogens, both peptides showed high selectivity as they did not exhibit any haemolytic activity on erythrocytes or cytotoxic activity against mammalian primary cell lines. Finally, the NKL-MUT selectively triggers the killing of the melanoma cell line B16F10 by means of a pro-apoptotic pathway at a concentration range in which no effects were found in normal mammalian cell lines. In conclusion, the two peptides could be considered as promising candidates in the fight against antibiotic resistance and tumour proliferative action, and also be used as innovative adjuvants, either to decrease chemotherapy side effects or to enhance anticancer drug activity.

A New Potential Resource in the Fight against Candida auris: the Cinnamomum zeylanicum Essential Oil in Synergy with Antifungal Drug.

Candida auris is a multidrug-resistant fungus known to be a global public health problem. The skin-based transmission, together with the marked resistance to drugs, resulted in its rapid spread to all continents. The aim of this study was to identify an essential oil (EO) active in the fight against C. auris. A total of 15 EOs were tested against 10 clinical strains of C. auris. Cinnamomum zeylanicum EO (CZ-EO) was the most effective (MIC90 and MFC90 equal to 0.06% vol/vol). Three fractions obtained from CZ-EO, and the cinnamaldehyde (CIN), the major chemical compound, were tested to identify the principal compound effectives against C. auris. All CIN-containing samples showed anti-fungal activity. To study the synergy with fluconazole, CZ-EO, its active fraction (FR2), and CIN were tested in checkerboard tests. Results show that CZ-EO and FR2, but not CIN, synergize with fluconazole. Furthermore, only the copresence of CZ-EO or FR2 synergize with fluconazole at therapeutic concentrations of the drug (0.45 ± 0.32 µg/mL and 0.64 ± 0.67 µg/mL, respectively), while CIN only shows additive activity. In vivo studies conducted on Galleria mellonella larvae show the absence of toxicity of CZ-EO up to concentrations of 16% vol/vol, and the ability of CZ-EO to reactivate the efficacy of fluconazole when formulated at synergic concentrations. Finally, biochemical tests were made to study the mechanism of action of CZ-EO. These studies show that in the presence of both fluconazole and CZ-EO, the activity of fungal ATPases decreases and, at the same time, the amount of intracellular drug increases. IMPORTANCE This study highlights how small doses of CZ-EO are able to inhibit the secretion of fluconazole and promote its accumulation in the fungal cell. In this manner, the drug is able to exert its pharmacological effects bypassing the resistance of the yeast. If further studies will confirm this synergy, it will be possible to develop new therapeutic formulations active in the fight against C. auris resistances.

Frailty in Chile: Development of a Frailty Index Score Using the Chilean National Health Survey 2016-2017.

BACKGROUND: The Frailty Index (FI) is used to quantify and summarize vulnerability status in people. In Chile, no development and assessment of a FI have been explored. OBJECTIVE: To develop and evaluate a FI using representative data from Chilean adults aged 40 years and older stratified by sex. DESIGN: Cross-sectional study. SETTING: National representative data from the Chilean National Health Survey 2016-2017 (CNHS 2016-2017). PARTICIPANTS: 3,036 participants older than 40 years with complete data for all variables. MEASUREMENTS: A 49-item FI was developed and evaluated. This FI included deficits from comorbidities, functional limitations, mental health status, physical activity, anthropometry, medications, and falls. A score between 0 and 1 was calculated for each person. Descriptive statistics and linear regression models were employed to evaluate the FI's performance in the population. Comparative analyses were carried out to evaluate the FI score by age (1<60 and ≥ 60 years). RESULTS: The mean FI score was 0.15 (SD:0.09), with a 99% upper limit of 0.46. Scores were greater in women than men (0.17 [SD:0.09]) vs. 0.12 [0.08]); in people older than 80 years (0.22 [0.11]), and in people with ≤8 years of education (0.18 [0.10]) compared with those with >12 years (0.12 [0.08]). The average age-related increase in the FI was 2.3%. When a cut-off point ≥ 0.25 was applied, the prevalence of frail individuals was 11.8% (95% CI: 10.0 to 13.8) in the general population. The prevalence was higher in women 15.9% [95% CI: 13.3 to 18.9] than men 7.4% [95% CI: 5.3 to 10.1]. In a comparative analysis by age, higher FI mean scores and prevalence of frail were observed in people ≥ 60 than younger than 60. CONCLUSIONS: The mean FI score and frailty prevalence were higher in women than men, in people with fewer years of formal education, and incremented markedly with age. This FI can be used for early detection of frailty status focusing on women and middle-aged people as a strategy to delay or prevent frailty-related consequences.

Fishers' perception of the interaction between the South American sea lions and the Chinook salmon fishery in southern Chile.

We studied how the South American sea lion (SASL, Otaria flavescens) interacts with the operation of an artisanal fishery of Chinook salmon, a non-native species in Chile, using a combination of biological and social approaches, including a valuation by fishers about this interaction. During austral summer of 2019, an observer onboard artisanal fishing boats characterized the attack behavior of SASLs to gillnet-captured Chinook salmon during 33 hauls and analyzed which factors may affect the intensity of attacks. To analyze the relationship between fishers and SASLs, a Likert scale about the perception and views about nature was applied. A total of 23 interviews-including 35 open and 16 closed questions-with fishers were conducted to describe how they perceived the interactions with SASLs. Interactions with SASLs were recorded in 35% of the fishing events and varied depending on both operational factors, such as the number of boats, as well as environmental factors, such as moon's luminosity. Even though SASL interactions resulted in seven fish (~ 70 kg) damaged of a total catch of 2815 kg (2.5%) during the survey, boats with a damaged catch by SASL lost up to 11% of their revenue. This is consistent with 87% of the interviewed fishers who considered that the conflict with the SASL negatively impacts their activity and results in economic losses. A negative perception towards SASLs likely results from personal experience and revenue loss, even though impacts of SASL interactions at the scale of the entire fishery may be less important. While older fishers with less formal education have a productivist and instrumental focus, younger fishers with a more sustainable and conservationist view of fishing offer an opportunity to lead an improved local understanding of the relationship between salmon, SASLs, and humans.

Síndrome de resección anterior baja y su relación con la calidad de vida: aplicación de cuestionario EuroQol-5 / Low anterior resection syndrome and quality of life: application of EuroQol-5 questionnaire

Resumen Introducción: Las cirugías con preservación de esfínter tienen como consecuencia el desarrollo de una disfunción defecatoria con diferentes grados, la cual es conocida como síndrome de resección anterior baja (LARS) y es medida con el cuestionario LARS Score. Objetivo: Determinar la asociación del cuestionario EuroQol-5 (calidad de vida) con los diferentes grados de LARS Score. Materiales y Método: Estudio de tipo transversal, aplicando el cuestionario LARS Score y EuroQol-5 a pacientes operados por cáncer de recto medio y bajo, durante el periodo 2004-2017. Se realiza análisis demográfico y del tipo de cirugía. Para determinar asociaciones entre variables se utilizan diferentes pruebas estadísticas, considerando significativo un valor de p < 0,05. Resultados: Se encuestó a 54 pacientes, 62,16% hombres, promedio de edad 58,44 años, el 37,03% presentó LARS Mayor. Los índices promedio de calidad de vida para pacientes No LARS es 0,75, para LARS Menor es 0,69 y para LARS Mayor es 0,61, la diferencia entre índices presenta un valor p = 0,246. 46,3% presenta problemas en actividades habituales. LARS Mayor presenta un Odd-Ratio de 3,8 y 4,7 para dolor/malestar y angustia/depresión respectivamente. 70% de los pacientes con LARS Mayor presentaron resección total del mesorrecto (TME) y el 45% corresponde a menores de 65 años. Discusión: No existe diferencia estadísticamente significativa entre los índices de calidad de vida según LARS Score. LARS Mayor tiene mayor posibilidad de desarrollar algún grado de dolor/malestar y angustia/depresión. El porcentaje de LARS Mayor es acorde a lo publicado y la TME es uno de los factores de mayor impacto en el desarrollo de LARS. Conclusiones: El LARS Score se relaciona de manera no significativa con el índice de calidad de vida entregado por el cuestionario EuroQol-5D, existiendo una tendencia a disminuir la calidad de vida a medida que empeora el LARS.

Introduction: Sphincter-sparing surgeries result in the development of a defecatory dysfunction with different degrees, which is known as low anterior resection syndrome (LARS) and is measured with the LARS Score questionnaire. Objective: To determine the association of the EuroQol-5 questionnaire with the different degrees of LARS Score. Materials and Method: Crosssectional study, applying the LARS Score and EuroQol-5 questionnaire to patients operated with low and middle rectal cancer, during the period 2004-2017. Demographic analysis and type of surgery are performed. Different statistical tests are used to determine associations between variables, considering a significant p value < 0.05. Results: 54 patients were surveyed, 62.16% men, mean age 58.44 years, 37.03% presented Mayor-LARS. The average quality of life indices for Non-LARS patients is 0.75, for Minor-LARS is 0.69 and for Mayor-LARS is 0.61, the difference between indices presents a p value = 0.246. 46.3% present problems in habitual activities. LARS Mayor presents an Odd-Ratio of 3.8 and 4.7 for pain/discomfort and anguish/depression respectively. 70% of patients with LARS Mayor presented SMT and 45% corresponded to those under 65 years of age. Discussion: There is no statistically significant difference between the quality of life indices according to the LARS Score. Mayor-LARS is more likely to develop some degree of Pain/Discomfort and anguish/depression. The percentage of Mayor-LARS is according to what has been published and the TME is one of the factors with the greatest impact on the development of LARS. Conclusion: The LARS Score is non-significantly related to the quality of life index provided by the EuroQol-5D questionnaire, and there is a tendency to decrease quality of life as the LARS worsens.

Características epidemiológicas y manifestaciones clínicas de pacientes con proctitis infecciosa / Epidemiologic characteristics and clinical manifestations in patients with infectious proctitis

Resumen Objetivo: Describir y caracterizar los aspectos epidemiológicos y clínicos de los pacientes con proctitis infecciosa en nuestro hospital. Materiales y Método: Se trata de un estudio de tipo transversal de pacientes con sintomatología perianal y del canal anal, concordante con proctitis, que consultaron en el policlínico de coloproctología entre enero de 2017 y diciembre de 2018, a quienes se les realizó estudio de secreción anal para confirmar infección de transmisión sexual. Resultados: Son 46 pacientes hombres, 26 años promedio de edad, un 6% heterosexuales. Un 65% había consultado previamente (ninguno a un coloproctólogo). El síntoma más frecuente: ano húmedo (97,8%) y el signo: dermitis perianal (100%). De este grupo, 20 nunca se habían realizado estudio de VIH y 50% resultó ser seropositivo. Presentaban infecciones de transmisión sexual más comunes: Gonorrea (43,4%) y sífilis (31,2%) y en un 32% hubo presencia de más de un germen. Discusión: Los resultados sugieren que la proctitis infecciosa es una patología de pacientes jóvenes quienes tienen relaciones sexuales sin protección, más del 50% no utiliza condón. La mayoría de los pacientes presentan consultas previas con médicos no proctólogos. Los gérmenes con mayor frecuencia que provocan proctitis infecciosa: Neisseria gonorrhoeae, Chlamydia trachomatis, Treponema pallidum. Conclusión: La presencia de una dermitis perianal sin respuesta a tratamiento habitual, muy severa y/o en pacientes de riesgo, debe hacer sospechar proctitis infecciosa.

Objective: Describe and characterize epidemiological and clinical aspects of patients with infectious proctitis at our hospital. Materials and Method: This is a cross-sectional study of patients with perianal and anal canal symptoms consistent with proctitis, who consulted at the Clinical Hospital of The University of Chile coloproctology outpatient clinic between January 2017 and December 2018, with perianal and anal canal symptoms, consistent with proctitis, and with confirmed sexually transmitted disease by anal secretion study. Results: Total of 46 patients, all male with average age of 26 years old. 6% heterosexuals. 65% had previous medical consults (none with a proctologist). The most common symptom was wet anus (97.8%), and the most common physical finding was perianal dermatitis (100%). 20 patients had never been tested for HIV, and 50% were positive for this disease. The most common sexually transmitted diseases were gonorrhea (43.4%) and syphilis (31.2%). In 32% of the patients, the culture informed more than one pathogen. Discussion: The results suggest that infectious proctitis is a disease of young patients who have unprotected sex. More than 50% do not use a condom. Most patients have prior consults, but none with a proctologist. The most frequent pathogen that causes infectious proctitis: Neisseria gonorrhoeae, Chlamydia trachomatis, and Treponema pallidum. Conclusion: The presence of severe perianal dermatitis, lack of response to common treatment, and/or in patients with risk factors, infectious proctitis should be suspected.

Perforación y obstrucción colónica: conducta de los coloproctólogos latinoamericanos / Perforation and obstruction of the colon: behavior of the latin american colon and rectal surgeons

Resumen Objetivo: Conocer las diferentes conductas que realizan los cirujanos coloproctólogos latinoamericanos en relación con las urgencias colónicas. Materiales y Método: Estudio transversal, utilizando encuesta vía web con preguntas de selección múltiple. La encuesta fue enviada a las distintas Sociedades Coloproctológicas Latinoamericanas, así como a la Asociación Latinoamericana de Coloproctología (ALACP), para su distribución. La encuesta fue escrita en español neutro y traducida al portugués. Se utilizó análisis estadísticos descriptivos y analítico. Resultados: 441 encuestas respondidas completamente de 16 países diferentes. El 85% realiza resección y anastomosis sin ostomía de protección en obstrucciones de colon derecho. En las perforaciones del colon izquierdo, se realiza operación de Hartmann en el 63,3% de los casos que presentan peritonitis purulentas y en el 94,5% de las peritonitis fecaloideas. Discusión: En las obstrucciones colónicas, la resección con anastomosis primaria, es una conducta poco discutida en colon derecho, a diferencia de las obstrucciones del lado izquierdo, en donde realizar una operación de Hartmann es una conducta tan válida como la resección y anastomosis. En los cuadros de perforación, la decisión de resección y anastomosis primaria es multifactorial, tomando relevancia la estabilidad hemodinámica del paciente. En estos últimos casos, realizar una resección con ostomía, es la respuesta de gran parte de los encuestados. Conclusiones: Los resultados de cada situación, en su mayoría, presentan una tendencia clara hacia una conducta en particular; solo en el caso de obstrucción de colon izquierdo, se observan dos conductas (operación de Hartmann o anastomosis primaria) ambas validadas por la literatura internacional.

Objective: Learn about the different management options performed by latin american colon and rectal surgeons, in relation to colonic emergencies. Materials and Method: Cross-sectional study, using web survey with multiple-choice questions. The survey was sent to the different Latin America Coloproctological Societies, as well as to ALACP, for distribution. The survey was written in neutral Spanish and translated into Portuguese. Descriptive and analytical statistical analysis was used. Results: 441 complete surveys, from 16 different countries. 85% perform resection and anastomosis without diverting ostomy in obstructions of the right colon. In perforations of the left colon, Hartmann's procedure is performed in 63.3% of case with purulent peritonitis and in 94.5% of fecaloid peritonitis. Discussion: In colonic obstructions, resection with primary anastomosis, is little discussed behavior in the right colon, unlike obstructions on the left side, where performing a Hartmann operation is a behavior as valid as resection and anastomosis. In colonic perforation, the decision of resection and primary anastomosis is multifactorial, taking into account the hemodynamic stability of the patient. In the latter cases, performing an ostomy is the response of a large part of the surveyed. Conclusions: The results in each situation, for the most part, present a clear tendency towards a particular behavior; only in the case of left colon obstruction, two behaviors (Hartmann procedure or primary anastomosis) are both validated by international literature.

Clinical characteristics, management and outcome of patients with invasive candidiasis hospitalized in Internal Medicine Units: findings from a registry by the Italian Scientific Society FADOI.

PURPOSE: Invasive candidiasis (IC) is a challenging clinical condition, burdened by relevant mortality and morbidity. There is limited knowledge on the occurrence and management of IC in Internal Medicine Units (IMUs). Aim of this study was to provide real-world data on this topic. METHODS: Consecutive objectively diagnosed cases of IC were collected in this prospective registry, which involved 18 IMUs in Italy. Patients were followed-up to 90 days from the diagnosis of candidemia. RESULTS: A total of 111 patients were observed (median age 78, IQR 67-83) for an overall incidence of infection of 1.89 cases/1000 hospital admissions. Candida albicans was the most frequent isolated species (62%), followed by Candida parapsilosis (17%) and Candida glabrata (13%). Echinocandins and fluconazole were used as initial therapy in 56.8 and 43.2% of patients, respectively. Antifungal therapy was started within 24 h in 18.9% of patients, in 40.6% in the period 1-3 days, and in 40.5% of patients more than 3 days after blood cultures. Death rate was 19.8% at 30 days and 40.5% at 90 days. At multivariable analysis concomitant bacteremia (i.e. polymicrobial sepsis), and fluconazole as the initial therapy were associated with an increased risk of death at 90 days. CONCLUSIONS: The incidence of IC is not negligible, and our registry confirmed that these patients have a relevant mortality rate at 90 days. Concomitant bacteremia, featuring polymicrobial sepsis, and starting antifungal treatment with fluconazole instead of echinocandins independently increase the risk of death. Efforts are needed to improve the awareness and management of IC in IMUs.

Increased Faecalibacterium abundance is associated with clinical improvement in patients receiving rifaximin treatment.

Compositional and functional alterations of the gut microbiota are involved in the pathogenesis of several gastrointestinal diseases. Rifaximin is often used to induce disease remission due to its eubiotic effects on the gut microbiota. To investigate the correlation between changes in the gut microbiota composition and symptoms improvement in patients who present a clinical response to rifaximin treatment. Patients with ulcerative colitis (UC), Crohn's disease (CD), irritable bowel syndrome (IBS) and diverticular disease (DD) undergoing rifaximin treatment for clinical indication were enrolled in the study. Rifaximin was administered at the dose of 1,200 mg/day for 10 days. Faecal samples were collected at baseline and at the end of treatment; clinical improvement was assessed by Mayo score for UC, CD Activity Index (CDAI) for CD, IBS severity scoring system (IBS-SSS) for IBS and global symptomatic score (GSS) for DD. Twenty-five patients were included in the analysis and a clinical improvement was recorded for 10/25 (40%) of them. Microbial alpha diversity showed a slight increase in clinical responders (P=0.271), while it decreased in patients who did not improved (P=0.05). A significant post-treatment increase in Faecalibacterium abundance was observed in patients with a positive response (log2FC 1.959, P=0.042). Roseburia abundance decreased in both groups, whereas Ruminococcus decreased only in patients who clinically improved. Clinical improvement consequent to rifaximin treatment is associated with an increase in Faecalibacterium abundance. Achieving a positive shift in the gut microbiota composition seems a key event to obtain a clinical benefit from treatment.

Synthetic Oligomers Mimicking Capsular Polysaccharide Diheteroglycan are Potential Vaccine Candidates against Encapsulated Enterococcal Infections.

Infections caused by Enterococcus spp. are a major concern in the clinical setting. In Enterococcus faecalis, the capsular polysaccharide diheteroglycan (DHG), composed of ß-d-galactofuranose-(1 → 3)-ß-d-glucopyranose repeats, has been described as an important virulence factor and as a potential vaccine candidate against encapsulated strains. Synthetic structures emulating immunogenic polysaccharides present many advantages over native polysaccharides for vaccine development. In this work, we described the synthesis of a library of DHG oligomers, differing in length and order of the monosaccharide constituents. Using suitably protected thioglycoside building blocks, oligosaccharides up to 8-mer in length built up from either Galf-Glcp or Glcp-Galf dimers were generated, and we evaluated their immunoreactivity with antibodies raised against DHG. After the screening, we selected two octasaccharides, having either a galactofuranose or glucopyranose terminus, which were conjugated to a carrier protein for the production of polyclonal antibodies. The resulting antibodies were specific toward the synthetic structures and mediated in vitro opsonophagocytic killing of different encapsulated E. feacalis strains. The evaluated oligosaccharides are the first synthetic structures described to elicit antibodies that target encapsulated E. faecalis strains and are, therefore, promising candidates for the development of a well-defined enterococcal glycoconjugate vaccine.

Biomarkers of fungal infection: Expert opinion on the current situation.

The introduction of non-culture-based diagnostic techniques is revolutionizing the world of microbiological diagnosis and infection assessment. Fungi are no exception, and the introduction of biomarkers has opened up enormous expectations for better management of these entities. Biomarkers are diverse, their targets are also diverse and their evaluation has been done preferably in an individualized use and with deficient designs. Less is known about the value of the combined use of biomarkers and the impact of the negativity of two or more biomarkers on antifungal treatment decisions has been poorly studied. Given the paucity of prospective, randomized and definitive studies, we have convened experts from different fields, with an interest in invasive fungal infections, to answer some questions about the current relevant use of fungal biomarkers. This document summarizes the answers of these experts to the different questions.

Laboratory handling practice for faecal microbiota transplantation.

AIMS: Faecal microbiota transplantation (FMT) consists of the infusion of faeces from a healthy donor to the gastrointestinal tract of a recipient patient to treat disease associated with alterations in gut microbiota. The objective of this article was to describe laboratory workflow of an FMT laboratory to provide tips for preparing the faecal suspensions to be infused. METHODS AND RESULTS: Twenty-stool solutions obtained from ten donors were prepared using two different protocols: magnet plate emulsion (MPE) and Seward StomacherTM Emulsion (SSE). We evaluated parameters such as preparation time, handiness, and aerobic and anaerobic microbial count. For three donors, we monitored bacterial counts after defrosting at different time-points. MPE requires more time than SSE. In terms of microbial load, both methods showed similar values, with small and statistically differences (P ≤ 0·05) regarding anaerobes in favour of SSE. Frozen aliquots showed the same bacterial load values after defrosting. CONCLUSION: Although both methods allow an easy and available preparation of a stool suspension, SSE seems more suitable, particularly for stool banking. Aerobic and anaerobic species are preserved with both protocols; and safety for laboratory operators is guaranteed. SIGNIFICANCE AND IMPACT OF THE STUDY: In recent years, FMT has become a fascinating and interesting subject. Nevertheless, there are no real guidelines describing laboratory facilities and procedures. This paper aims to be a useful and simple guide to increase the number FMT centres as much possible.

Rapid molecular tests for detection of antimicrobial resistance determinants in Gram-negative organisms from positive blood cultures: a systematic review and meta-analysis.

BACKGROUND: Timely detection of antimicrobial (cephalosporin/carbapenem) resistance (AMR) determinants is crucial to the clinical management of bloodstream infections caused by Gram-negative bacteria (GNB). OBJECTIVES: To review and meta-analyse the evidence for using commercially available molecular tests for the direct detection of AMR determinants in GNB-positive blood cultures (PBCs). DATA SOURCES: PubMed, Scopus and ISI Web of Knowledge. STUDY ELIGIBILITY CRITERIA: Clinical studies evaluating the performance of two major commercial systems, namely the Verigene® and FilmArray® systems, for rapid testing of GNB-PBCs, in comparison with the phenotypic or genotypic methods performed on GNB-PBC isolates. METHODS: Literature search according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses criteria and, for meta-analysis of sensitivity and specificity of both systems, bivariate random-effects model. RESULTS: Twenty studies were identified (3310 isolates) from 2006 to 2019. Nine studies were conducted in East Asia. In 15 studies using phenotypic comparators (1930 isolates), 1014 (52.5%) isolates were Escherichia coli, and 287 (14.9%) of all the isolates displayed AMR phenotypes. In five studies using genotypic comparators (1380 isolates), 585 (42.4%) were E. coli, and 100 (7.2%) of all the isolates displayed AMR genotypes. Pooled sensitivity and specificity estimates for detection of AMR determinants by the Verigene (i.e. CTX-M, IMP, KPC, NDM, OXA and VIM) and/or FilmArray (i.e. KPC) systems were 85.3% (95% CI 79.9%-89.4%) and 99.1% (95% CI 98.2%-99.5%), respectively, across the 15 studies, and 95.5% (95% CI 89.2%-98.2%) and 99.7% (95% CI 99.1%-99.9%), respectively, across the five studies. CONCLUSIONS: Our findings show that the Verigene and FilmArray systems may be a valid adjunct to the conventional microbiology (phenotypic or genotypic) methods used to identify AMR in GNBs. The FilmArray system detects only one AMR genotype, namely KPC, limiting its use. Both Verigene and FilmArray systems can miss important cephalosporin/carbapenem resistance phenotypes in a minority of cases. However, the sensitivity and specificity of both systems render them valuable clinical tools in timely identification of resistant isolates. Further studies will establish the prominence of such rapid diagnostics as standard of care in individuals with bloodstream infections.

Conjugation of Different Immunogenic Enterococcal Vaccine Target Antigens Leads to Extended Strain Coverage.

Enterococci have emerged as important nosocomial pathogens due to their resistance to the most commonly used antibiotics. Alternative treatments or prevention options are aimed at polysaccharides and surface-related proteins that play important roles in pathogenesis. Previously, we have shown that 2 Enterococcus faecium proteins, the secreted antigen A and the peptidyl-prolyl cis-trans isomerase, as well as the Enterococcus faecalis polysaccharide diheteroglycan, are able to induce opsonic and cross-protective antibodies. Here, we evaluate the use of glycoconjugates consisting of these proteins and an enterococcal polysaccharide to develop a vaccine with broader strain coverage. Diheteroglycan was conjugated to these 2 enterococcal proteins. Rabbit sera raised against these glycoconjugates showed Immunoglobulin G titers against the corresponding conjugate, as well as against the respective protein and carbohydrate antigens. Effective opsonophagocytic killing for the 2 sera was observed against different E. faecalis and E. faecium strains. Enzyme-linked immunosorbent assays against whole bacterial cells showed immune recognition of 22 enterococcal strains by the sera. Moreover, the sera conferred protection against E. faecalis and E. faecium strains in a mouse infection model. Our results suggest that these glycoconjugates are promising candidates for vaccine formulations with a broader coverage against these nosocomial pathogens and that the evaluated proteins are potential carrier proteins.