Gut microbiome and plasma lipidome analysis reveals a specific impact of Clostridioides difficile infection on intestinal bacterial communities and sterol metabolism.

Clostridioides difficile infection (CDI) causes alterations in the intestinal microbiota, frequently associated with changes in the gut metabolism of bile acids and cholesterol. In addition to the impact on microbiome composition and given the metabolic changes occurring during CDI, our work focuses on the importance to know the effects at the local and systemic levels, both during the infection and its treatment, by paying particular attention to plasma lipid metabolism due to its relationship with CDI pathogenesis. Specific changes, characterized by a loss of microbial richness and diversity and related to a reduction in short-chain acid-producing bacteria and an increase in bile salt hydrolase-producing bacteria, were observed in the gut microbiota of CDI patients, especially in those suffering from recurrent CDI (RCDI). However, gut microbiota showed its ability to restore itself after treatment, resembling healthy individuals, in those patients treated by fecal microbiome transfer (FMT), in contrast with those treated with antibiotics, and displaying increased levels of Eubacterium coprostanoligenes, a cholesterol-reducing anaerobe. Interestingly, changes in plasma lipidome revealed a global depletion in circulating lipids in CDI, with the largest impact on cholesteryl esters. CDI patients also showed a specific and consistent decrease in the levels of lipid species containing linoleic acid-an essential fatty acid-which were only partially recovered after antibiotic treatment. Analysis of the plasma lipidome reflects CDI impact on the gut microbiota and its metabolism, evidencing changes in sterol and fatty acid metabolism that are possibly related to specific alterations observed in gut microbial communities of CDI patients. IMPORTANCE: There is increasing evidence about the influence the changes in microbiota and its metabolism has on numerous diseases and infections such as Clostridioides difficile infection (CDI). The knowledge of these changes at local and systemic levels can help us manage this infection to avoid recurrences and apply the best therapies, such as fecal microbiota transfer (FMT). This study shows a better restoration of the gut in FMT-treated patients than in antibiotic-treated patients, resembling healthy controls and showing increased levels of cholesterol-reducing bacteria. Furthermore, it evidences the CDI impact on plasma lipidome. We observed in CDI patients a global depletion in circulating lipids, particularly cholesteryl esters, and a specific decrease in linoleic acid-containing lipids, an essential fatty acid. Our observations could impact CDI management because the lipid content was only partially recovered after treatment, suggesting that continued nutritional support, aiming to restore healthy lipid levels, could be essential for a full recovery.

Tobacco price elasticity by socioeconomic characteristics in Ecuador.

Smoking is a worldwide epidemic and increased prices are one of the most cost-effective measures to reduce tobacco consumption. This article aims to estimate the price and income elasticity of cigarettes for different population groups in Ecuador. The National Survey of Urban and Rural Household Income and Expenditures (ENIGHUR) 2011-2012 was used, which has information on household cigarette consumption and its sociodemographic characteristics. Deaton's Almost Ideal Demand System, which decouples the effect of quality on the price of the good, was applied. The elasticities were calculated for several groups: urban/rural, income levels (tertiles), education level, sex and age ranges of the household head, and frequency of cigarette purchases in households. The estimated price elasticity nationwide is -0.89 and the income elasticity is 0.41, both statistically significant. Households headed by women (-2.22) are more sensitive to an increase in cigarette prices than those headed by men (-0.65) and households headed by people between 20 and 40 years of age (-2.32) have a higher price elasticity compared to country-level estimations. Differences within other groups are not statistically significant.

Closed-loop technique based on gain balancing for real-time Brillouin optical time-domain analysis.

A closed-loop servo control based on balancing the gain of two probing frequencies is proposed for real-time Brillouin optical time-domain analysis (BOTDA) without post-processing. With the most basic BOTDA hardware setup, the system can perform measurement in 150 ms and track a sudden Brillouin frequency shift (BFS) change in excess of 300 MHz (corresponding to a temperature change of more than 250°C) over ∼5 km of fiber with a spatial resolution of 2 m. Moreover, the feedback loop is independent of the loss experienced by the probe and pump, with no requirement on the BFS uniformity along the fiber. All these advantages make the proposed system suitable for field applications in harsh environments.

Complicaciones de la nefrolitotomía percutánea como tratamiento de la litiasis renal coraliforme

RESUMEN Introducción: La nefrolitotomía percutánea es la primera opción terapéutica para la litiasis renal coraliforme. Objetivo: Caracterizar a los pacientes con complicaciones de la nefrolitotomía percutánea para el tratamiento de la litiasis renal coraliforme. Método: Se estudió una serie de 191 pacientes, operados mediante nefrolitotomía percutánea. Variables estudiadas: tipo de litiasis coraliforme, posición para la técnica, condición de libre de litiasis después de la operación, presencia de complicaciones, momento, tipo y grado según clasificación de Clavien-Dindo. Se hallaron frecuencias absolutas, relativas y se utilizó el test de ji cuadrado para determinar asociación entre variables. Resultados: El 86,9 % tenía menos de 60 años, 67,0 % eran masculinos, 61,7 % presentaba comorbilidades. La litiasis coraliforme era parcial o total (30,3 % y 46,5 %, respectivamente). En 60,2 % afectaba el riñón izquierdo; 58,1 % se operaron en supino y 70,2 % quedaron libre de litiasis con la nefrolitotomía percutánea monoterapéutica. Ocurrieron complicaciones en 19,9 %; 16,2 % fueron postoperatorias, 14,1 % infecciosas, 7,8 % Clavien-Dindo I y 5,2 % IIIb. El tipo de litiasis y la posición de la nefrolitotomía percutánea no se asociaron con las complicaciones (p> 0,05). El grado de la complicación no se relacionó con el tipo de litiasis (p> 0,05). Conclusiones: Las complicaciones postoperatorias más frecuentes son las relacionadas con la infección y el sangrado; predominan ligeramente en los pacientes con litiasis coraliformes parcial, total y en los operados en supino; el grado Clavien-Dindo de las complicaciones, es mayor en las litiasis coraliformes más complejas.

ABSTRACT Introduction: Percutaneous nephrolithotomy is the first therapeutic option for staghorn kidney stones. Objective: To characterize patients with complications of percutaneous nephrolithotomy for the treatment of staghorn renal lithiasis. Method: A series of 191 patients, operated by percutaneous nephrolithotomy, was studied. Variables studied: type of staghorn lithiasis, position for the technique, stone-free condition after the operation, presence of complications, time, type and grade according to the Clavien-Dindo classification. Absolute and relative frequencies were found and the chi-square test was used to determine the association between variables. Results: 86.9 % were less than 60 years old, 67,0 % were male, 61,7 % had comorbidities. The staghorn lithiasis was partial or total (30,3 % and 46,5 %, respectively). In 60,2 % it affected the left kidney; 58.1 % underwent supine surgery and 70,2 % were stone free with monotherapeutic percutaneous nephrolithotomy. Complications occurred in 19,9 %; 16,2 % were postoperative, 14,1 % infectious, 7,8 % Clavien-Dindo I, and 5,2 % IIIb. The type of lithiasis and the position of the percutaneous nephrolithotomy were not associated with complications (p> 0,05). The degree of complication was not related to the type of lithiasis (p> 0,05). Conclusions: The most frequent postoperative complications are those related to infection and bleeding; they slightly predominate in patients with partial and total staghorn stones and in those operated on in the supine position; the Clavien-Dindo grade of complications is higher in the more complex staghorn stones.

Involving society in science: Reflections on meaningful and impactful stakeholder engagement in fundamental research.

Open Science calls for transparent science and involvement of various stakeholders. Here are examples of and advice for meaningful stakeholder engagement.

Multifunctional Amyloids in the Biology of Gram-Positive Bacteria.

Since they were discovered, amyloids have proven to be versatile proteins able to participate in a variety of cellular functions across all kingdoms of life. This multitask trait seems to reside in their ability to coexist as monomers, aggregates or fibrillar entities, with morphological and biochemical peculiarities. It is precisely this common molecular behaviour that allows amyloids to cross react with one another, triggering heterologous aggregation. In bacteria, many of these functional amyloids are devoted to the assembly of biofilms by organizing the matrix scaffold that keeps cells together. However, consistent with their notion of multifunctional proteins, functional amyloids participate in other biological roles within the same organisms, and emerging unprecedented functions are being discovered. In this review, we focus on functional amyloids reported in gram-positive bacteria, which are diverse in their assembly mechanisms and remarkably specific in their biological functions that they perform. Finally, we consider cross-seeding between functional amyloids as an emerging theme in interspecies interactions that contributes to the diversification of bacterial biology.

Imported cysticercosis in Spain: A retrospective case series from the +REDIVI Collaborative Network.

BACKGROUND: Neurocysticercosis (NCC) is the most common parasitic neurological disease worldwide and a major cause of epilepsy. Spain is the country reporting the highest number of NCC imported cases in Europe. METHODOLOGY: Retrospective case series of NCC patients registered in the +REDIVI Network from October 1, 2009 to July 2018. A specific questionnaire, including clinical and diagnostic characteristics, was created and sent to the collaborator centers. RESULTS: 46 cases were included in the analysis. 55% were male, mean age of 40 years. 95.6% were migrants. The median duration since migration from an endemic area was 10 years. Predominant nationalities were Ecuadorians (50%) and Bolivians (30.4%). Frequent locations were parenchymal (87%), subarachnoid (26.1%) and intraventricular cysts (10.9%). Serological analysis was performed in 91.3%, being 54.8% positive. Most prevalent clinical manifestations were persistent headache (60.9%), epilepsy (43.5%) and visual changes (13%). Patients were mainly treated with albendazole (76.1%), corticosteroids (67.4%), and anticonvulsionants (52.2%). 82.5% had a favorable clinical outcome. CONCLUSIONS: Most NCC cases were long-standing migrants. Few clinical differences were observed depending on the cysticerci location. The treatment was often not according to current recommendations, and no uniform criteria were followed when it came to the therapeutic regimen. NCC case management in Spain (including clinician awareness and laboratory capacity improvements) needs to be strengthened.

Two genomic regions encoding exopolysaccharide production systems have complementary functions in B. cereus multicellularity and host interaction.

Bacterial physiology and adaptation are influenced by the exopolysaccharides (EPS) they produce. These polymers are indispensable for the assembly of the biofilm extracellular matrix in multiple bacterial species. In a previous study, we described the profound gene expression changes leading to biofilm assembly in B. cereus ATCC14579 (CECT148). We found that a genomic region putatively dedicated to the synthesis of a capsular polysaccharide (eps2) was overexpressed in a biofilm cell population compared to in a planktonic population, while we detected no change in the transcript abundance from another genomic region (eps1) also likely to be involved in polysaccharide production. Preliminary biofilm assays suggested a mild role for the products of the eps2 region in biofilm formation and no function for the products of the eps1 region. The aim of this work was to better define the roles of these two regions in B. cereus multicellularity. We demonstrate that the eps2 region is indeed involved in bacterial adhesion to surfaces, cell-to-cell interaction, cellular aggregation and biofilm formation, while the eps1 region appears to be involved in a kind of social bacterial motility. Consistent with these results, we further demonstrate using bacterial-host cell interaction experiments that the eps2 region is more relevant to the adhesion to human epithelial cells and the zebrafish intestine, suggesting that this region encodes a bacterial factor that may potentiate gut colonization and enhance pathogenicity against humans.

Biofilm formation displays intrinsic offensive and defensive features of Bacillus cereus.

Biofilm formation is a strategy of many bacterial species to adapt to a variety of stresses and has become a part of infections, contaminations, or beneficial interactions. In this study, we demonstrate that profound physiological changes permit Bacillus cereus to switch from a floating to a sessile lifestyle, to undergo further maturation of the biofilm and to differentiate into the offensive or defensive features. We report that floating and biofilm cells are populations that differentiate metabolically, with members of each subpopulation developing different branches of certain metabolic pathways. Secondly, biofilm populations rearrange nucleotides, sugars, amino acids, and energy metabolism. Thirdly, this metabolic rearrangement coexists with: the synthesis of the extracellular matrix, sporulation, reinforcement of the cell wall, activation of the ROS detoxification machinery and production of secondary metabolites. This strategy contributes to defend biofilm cells from competitors. However, floating cells maintain a fermentative metabolic status that ensures a higher aggressiveness against hosts, evidenced by the production of toxins. The maintenance of the two distinct subpopulations is an effective strategy to face different environmental conditions found in the life styles of B. cereus.

Molecular architecture of bacterial amyloids in Bacillus biofilms.

The formation of biofilms provides structural and adaptive bacterial response to the environment. In Bacillus species, the biofilm extracellular matrix is composed of exopolysaccharides, hydrophobins, and several functional amyloid proteins. We report, using multiscale approaches such as solid-state NMR (SSNMR), electron microscopy, X-ray diffraction, dynamic light scattering, attenuated total reflection Fourier transform infrared (FTIR), and immune-gold labeling, the molecular architecture of B. subtilis and pathogenic B. cereus functional amyloids. SSNMR data reveal that the major amyloid component TasA in its fibrillar amyloid form contain ß-sheet and α-helical secondary structure, suggesting a nontypical amyloid architecture in B. subtilis. Proteinase K digestion experiments indicate the amyloid moiety is ∼100 aa long, and subsequent SSNMR and FTIR signatures for B. subtilis and B. cereus TasA filaments highlight a conserved amyloid fold, albeit with substantial differences in structural polymorphism and secondary structure composition. Structural analysis and coassembly data on the accessory protein TapA in B. subtilis and its counterpart camelysin in B. cereus reveal a catalyzing effect between the functional amyloid proteins and a common structural architecture, suggesting a coassembly in the context of biofilm formation. Our findings highlight nontypical amyloid behavior of these bacterial functional amyloids, underlining structural variations between biofilms even in closely related bacterial species.-El Mammeri, N., Hierrezuelo, J., Tolchard, J., Cámara-Almirón, J., Caro-Astorga, J., Álvarez-Mena, A., Dutour, A., Berbon, M., Shenoy, J., Morvan, E., Grélard, A., Kauffmann, B., Lecomte, S., de Vicente, A., Habenstein, B., Romero, D., Loquet, A. Molecular architecture of bacterial amyloids in Bacillus biofilms.

Increasing prevalence of the epidemic ribotype 106 in healthcare facility-associated and community-associated Clostridioides difficile infection.

Clostridioides difficile is the leading cause of nosocomial diarrhea and antibiotics associated diarrhea, but it is also an increasingly common cause of community diarrhea. In recent years we have observed a progressive increase in the incidence of C. difficile infection (CDI) both at the hospital and community setting that could be explained by the dynamic epidemiology of C. difficile. The present study analyzes changes in the epidemiology of CDI for two years comparing healthcare facility-associated (HCFA) and community-associated (CA) CDI epidemiology, observed in a single laboratory setting. All new episodes of CDI diagnosed during the years 2015-2016 were included in the study and classified as HFCA-, CA- or indeterminate CDI. Isolates were characterized by ribotyping and antimicrobial susceptibility was also determined. A total of 272 primary episodes of different patients were included in the study and classified 55.5% as CA-, 32% as HO-HCFA, 6.25% as CO-HCFA and 6.25% as indeterminate CDI. Overall, ribotype 106 was the most prevalent and also, many patients who suffered recurrent episodes were associated with this ribotype (29%). In fact, ribotype 106 showed a significantly higher recurrence rate than other ribotypes (26% vs 11%, p = 0.03). Moreover, 46% of the moxifloxacin resistant isolates were ribotype 106. No significant differences of antimicrobial resistance were observed between HCFA- and CA-CDI isolates, although fluoroquinolone resistance rates were slightly higher in HCFA-CDI isolates (25% vs 18.5%), and fluoroquinolone resistant ribotypes 106 and 126 were more frequently associated to CA-CDI and ribotype 078 to HCFA-CDI. The increasing incidence of CDI in our health care area is partially explained by the growing prevalence of the epidemic ribotype 106, both in HFCA- and CA-CDI, probably favored by the higher resistance and recurrence rate associated to ribotype 106 isolates.

The Use of Insecticide-Treated Curtains for Control of Aedes aegypti and Dengue Virus Transmission in "Fraccionamiento" Style Houses in México.

Dengue, chikungunya, yellow fever, and Zika viruses transmitted by Aedes aegypti mosquitoes are major public health threats in the tropical and subtropical world. In México, construction of large tracts of "fraccionamientos" high density housing to accommodate population growth and urbanization has provided fertile ground for Ae. aegypti-transmitted viruses. We investigated the utility of pyrethroid-treated window curtains to reduce both the abundance of Ae. aegypti and to prevent dengue virus (DENV) transmission in fraccionamiento housing. Windows and doors of fraccionamiento homes in urban/suburban areas, where Ae. aegypti pyrethroid resistance associated with the Ile1016 knock down resistance (kdr) mutation in the voltage gated sodium channel gene was high, and in rural areas, where kdr resistance was low, were fitted with either insecticide-treated curtains (ITCs) or non-treated curtains (NTCs). The homes were monitored for mosquito abundance and DENV infection. ITCs reduced the indoor abundance of Ae. aegypti and the number of DENV-infected mosquitoes in homes in rural but not in urban/suburban study sites. The presence of non-treated screens also was associated with reduced numbers of mosquitoes in homes. "Super-infested" homes, yielding more than 50 mosquitoes, including DENV-infected mosquitoes, provide a significant public health risk to occupants, visitors, and people in neighboring homes.

[Payment mechanisms and financial resources management for consolidation of Ecuador's health system]. / Mecanismos de pago y gestión de recursos financieros para la consolidación del Sistema de Salud de Ecuador.

OBJECTIVE: Analyze the proposal by the Ministry of Public Health to reform the public financing model in Ecuador with regard to pooling of funds and payment mechanisms. METHOD: A literature review was done of the financing model, the current legal framework, and the budgetary bases in Pubmed, SciELO, LILACS Ecuador, and regional LILACS using the key words health financing, health financing systems, capitation, pooling of funds, health system reform Ecuador, health system Ecuador, and health payment mechanisms. Books and other documents suggested by health systems experts were also included. RESULTS: Review of the financing model enabled identifying the historical segmentation of Ecuador's health system; out of this, the Ministry of Public Health conceived its proposal to reform the financing model. The Ministry's proposed solutions are pooling of funds and payment of services at the first level of care through payment per capita adjusted for socioeconomic and demographic risks. Progress made in reforming the financing model includes design of the proposals and their implementation mechanisms, and discussions with stakeholders. CONCLUSIONS: Implementation of these changes may produce improvements for the health system in efficiency, spreading of risks, incentives for meeting health objectives, as well as contribute to its sustainability and advance toward universal health coverage. Nevertheless, legal, political, and operational constraints are hampering their implementation.

Compound heterozygous KCNQ1 mutations (A300T/P535T) in a child with sudden unexplained death: Insights into possible molecular mechanisms based on protein modeling.

Sudden death in a child is a devastating event with important medical implications for surviving relatives. Because it may be the first manifestation of unknown inherited cardiac disease, molecular autopsy can be helpful to determine the cause of death and identify at risk family members. The aim of the study was to perform a molecular autopsy in a seven year-old girl with sudden unexplained death, to find evidence supporting the possible pathogenicity of mutations identified in inherited cardiac disease genes, and to clinically and genetically assess first-degree relatives. DNA from the index case was extracted from umbilical cord cells stored at birth, and DNA of first-degree relatives from blood samples. Targeted sequencing was performed using a Haloplex design including 81 cardiogenes. Possible functional consequences of the mutations were analyzed using protein modeling and structural mobility analyses. The child was compound heterozygous for KCNQ1 variants p.Ala300Thr and p.Pro535Thr. Ala300Thr is known to cause long QT syndrome in the homozygous state, while Pro535Thr is novel and of unknown clinical significance. The father and sibling were Ala300Thr heterozygous, and had normal QTc intervals at rest and during exercise. The asymptomatic mother was heterozygous for Pro535Thr, and showed borderline QTc at rest, but prolonged QTc during exercise. Protein modeling predicted that Ala300Thr alters the mobility profile of the Kv7.1 tetramer and Thr535 disrupts a calmodulin-binding site, probably causing co-assembly or trafficking defects of the mutant monomer. Altogether, the evidence strongly suggests that this child was affected with a recessive form of Romano Ward syndrome.

Influence of Core Affect in the Differential Efficacy of a Personality Disorder Intervention Program.

The usual emotional experience of the person (affective style) is an influential factor in therapeutic assimilation. Based on a dynamic model of affect shaped dimensionally by the valence and arousal axes (core affect) that fluctuate over time according to the specific context of the individual, its relationship with different variables was investigated and the changes after a 6-month intervention in a specialized hospital unit (N = 103) were observed. The orthogonal structure of core-affect was confirmed. Emotional valence appeared to be positively related to social skills (r = .375; p < .01) and self-esteem (r = .491; p < .01) and negatively to depressive symptoms (r = -.631; p < .01), general disturbance (r = -.395; p < .01) and suicidality (r = -.490; p < .01). Emotional arousal is associated with impulsivity (r = .345; p < .01). The group of patients with an affective style characterized by negative valence and low arousal core-affect gained less therapeutic benefit compared to those with positive valence core-affect (p < .05). Throughout the treatment, valence became more positive (d = .26; IC 95%: 1.9 - 7.2; p = .001), arousal increased (d = .23; IC 95%: 0.2 - 1.7; p = .015) and variability decreased (d = -.44; IC 95%: (-2.9) - (-1.1); p = .001). Changes in the core-affect are related to therapeutic improvement. Adjusting expectations of change can reduce therapeutic frustration, which is as common as it is harmful in the treatment of severe personality disorders.

Household use of insecticide consumer products in a dengue-endemic area in México.

OBJECTIVES: To evaluate the household use of insecticide consumer products to kill mosquitoes and other insect pests, as well as the expenditures for using these products, in a dengue-endemic area of México. METHODS: A questionnaire was administered to 441 households in Mérida City and other communities in Yucatán to assess household use of insecticide consumer products. RESULTS: A total of 86.6% of surveyed households took action to kill insect pests with consumer products. The most commonly used product types were insecticide aerosol spray cans (73.6%), electric plug-in insecticide emitters (37.4%) and mosquito coils (28.3%). Mosquitoes were targeted by 89.7% of households using insecticide aerosol spray cans and >99% of households using electric plug-in insecticide emitters or mosquito coils. Products were used daily or every 2 days in most of the households for insecticide aerosol spray cans (61.4%), electric plug-in insecticide emitters (76.2%) and mosquito coils (82.1%). For all products used to kill insect pests, the median annual estimated expenditure per household that took action was 408 Mexican pesos ($MXN), which corresponded to approximately 31 $US. These numbers are suggestive of an annual market in excess of 75 million $MXN (>5.7 million $US) for Mérida City alone. CONCLUSION: Mosquitoes threaten human health and are major nuisances in homes in the study area in México. Households were found to have taken vigorous action to kill mosquitoes and other insect pests and spent substantial amounts of money on insecticide consumer products.

Molecular epidemiology and resistance profiles of Clostridium difficile in a tertiary care hospital in Spain.

Epidemiological surveillance of Clostridium difficile infection has gained importance in recent years as a result of the rapid spread of epidemic strains, including hypervirulent strains and strains with reduced susceptibility to antimicrobials. The molecular epidemiology and antimicrobial susceptibility of C. difficile in the reference hospital of the Balearic Islands (Spain) is reported in this study. One hundred isolates of toxigenic C. difficile from different patients were selected using rapid dual EIA screening test. All isolates were characterized through toxin profile, PCR ribotyping and, in addition, multi-locus sequence typing (MLST) was performed on fifty selected strains. MICs to metronidazole, vancomycin, erythromycin and moxifloxacin were also determined. A total of 43 different ribotypes were distinguished, with higher prevalence of ribotype 014 (34%). Twenty one per cent of the isolates expressed binary toxin and it is noteworthy that 62% of these were identified as the hypervirulent ribotype 078, the second most prevalent ribotype found in our hospital (13%). A total of 20 different sequence types (STs) were found, including a new described allele and ST. MLST data showed a clear concordance between some ribotypes and STs, mainly represented by ribotype 014/ST-2, ribotype 078/ST-11 and ribotype 001/ST-3. Phylogenetic analysis also revealed that most of the isolates were genetically related, forming a large clonal complex. Finally, ribotypes 078 (ST-11) and 001 (ST-3) were associated with higher resistance to erythromycin and to erythromycin and moxifloxacin, respectively. All these data suggest that the combination of ribotyping and MLST is a good tool for the surveillance of the changing epidemiology of C. difficile. A wide dissemination of clones has been observed in our setting, ribotype 014 (ST-2) being the most prevalent followed by the hypervirulent ribotype 078 (ST-11) and ribotype 001 (ST-3), their spread in our setting probably influenced by their higher resistance.