A General Protocol for Gene Expression Analysis in Chemically Mutant Coffee Plants in Response to Rust (Hemileia vastatrix Berk & Broome) Infection.

Coffea spp. is the source of one of the most widely consumed beverages in the world. However, the cultivation of this crop is threatened by Hemileia vastatrix Berk & Broome, a fungal disease, which reduces the productivity and can cause significant economic losses. In this protocol, coffee leaf segment derived from a chemical mutagenesis process are inoculated with uredospores of the pathogen. Subsequently, the gene expression changes are analyzed over the time (0, 5, 24, 48, and 120 h) using quantitative real-time polymerase chain reaction (RT-qPCR). The procedures and example data are presented for expression analysis in the CaWRKY1 gene. This procedure can be applied for quantitative analysis of other genes of interest to coffee breeders and scientists for elucidating the molecular mechanisms involved in the interaction between the plant and pathogen, potentially leading to the development of more efficient approaches for managing this disease.

New insight into the biological activity of Salmo salar NK-lysin antimicrobial peptides.

NK-lysin is a potent antimicrobial peptide (AMP) with antimicrobial activity against bacteria, fungi, viruses, and parasites. NK-lysin is a type of granulysin, a member of the saposin-like proteins family first isolated from a pig's small intestine. In previous work, for the first time, we identified four variants of nk-lysin from Atlantic salmon (Salmo salar) using EST sequences. In the present study, we reported and characterized two additional transcripts of NK-lysin from S. salar. Besides, we evaluated the tissue distribution of three NK-lysins from S. salar and assessed the antimicrobial, hemolytic, and immunomodulatory activities and signaling pathways of three NK-lysin-derived peptides. The synthetic peptides displayed antimicrobial activity against Piscirickettsia salmonis (LF-89) and Flavobacterium psychrophilum. These peptides induced the expression of immune genes related to innate and adaptive immune responses in vitro and in vivo. The immunomodulatory activity of the peptides involves the mitogen-activated protein kinases-mediated signaling pathway, including p38, extracellular signal-regulated kinase 1/2, and/or c-Jun N-terminal kinases. Besides, the peptides modulated the immune response induced by pathogen-associated molecular patterns (PAMPs). Our findings show that NK-lysin could be a highly effective immunostimulant or vaccine adjuvant for use in fish aquaculture.

Design and functional characterization of Salmo salar TLR5 agonist peptides derived from high mobility group B1 acidic tail.

Toll-like receptor 5 (TLR5) responds to the monomeric form of flagellin and induces the MyD88-depending signaling pathway, activating proinflammatory transcription factors such as NF-κB and the consequent induction of cytokines. On the other hand, HMGB1 is a highly conserved non-histone chromosomal protein shown to interact with and activate TLR5. The present work aimed to design and characterize TLR5 agonist peptides derived from the acidic tail of Salmo salar HMGB1 based on the structural knowledge of the TLR5 surface using global molecular docking platforms. Peptide binding poses complexed on TLR5 ectodomain model from each algorithm were filtrated based on docking scoring functions and predicted theoretical binding affinity of the complex. Circular dichroism spectra were recorded for each peptide selected for synthesis. Only intrinsically disordered peptides (6W, 11W, and SsOri) were selected for experimental functional assay. The functional characterization of the peptides was performed by NF-κB activation assays, RT-qPCR gene expression assays, and Piscirickettsia salmonis challenge in SHK-1 cells. The 6W and 11W peptides increased the nuclear translation of p65 and phosphorylation. In addition, the peptides induced the expression of genes related to the TLR5 pathway activation, pro- and anti-inflammatory response, and differentiation and activation of T lymphocytes towards phenotypes such as TH1, TH17, and TH2. Finally, it was shown that the 11W peptide protects immune cells against infection with P. salmonis bacteria. Overall, the results indicate the usefulness of novel peptides as potential immunostimulants in salmonids.

Agonistic effect of peptides derived from a truncated HMGB1 acidic tail sequence in TLR5 from Salmo salar.

Based on the structural knowledge of TLR5 surface and using blind docking platforms, peptides derived from a truncated HMGB1 acidic tail from Salmo salar was designed as TLR5 agonistic. Additionally, a template peptide with the native N-terminal of the acidic tail sequence as a reference was included (SsOri). Peptide binding poses complexed on TLR5 ectodomain model from each algorithm were filtrated based on docking scoring functions and predicted theoretical binding affinity of the complex. The best peptides, termed 6WK and 5LWK, were selected for chemical synthesis and experimental functional assay. The agonist activity by immunoblotting and immunocytochemistry was determined following the NF-κBp65 phosphorylation (p-NF-κBp65) and the nuclear translocation of the NF-κBp65 subunit from the cytosol, respectively. HeLa cells stably expressing a S. salar TLR5 chimeric form (TLR5c7) showed increased p-NF-κBp65 levels regarding extracts from flagellin-treated cells. No statistically significant differences (p > 0.05) were found in the detected p-NF-κBp65 levels between cellular extracts treated with peptides or flagellin by one-way ANOVA. The image analysis of NF-κBp65 immunolabeled cells obtained by confocal microscopy showed increased nuclear NF-κBp65 co-localization in cells both 5LWK and flagellin stimulated, while 6WK and SsOri showed less effect on p65 nuclear translocation (p < 0.05). Also, an increased transcript expression profile of proinflammatory cytokines such as TNFα, IL-1ß, and IL-8 in HKL cells isolated from Salmo salar was evidenced in 5LWK - stimulated by RT-PCR analysis. Overall, the result indicates the usefulness of novel peptides as a potential immunostimulant in S. salar.

Isolation, characterization, and immunomodulatory activity evaluation of probiotic strains from colostrum and canine milk.

Background: This study aimed to characterize potential probiotic strains for use in dogs to prevent infectious enteropathies. Lactic acid bacteria (LAB) isolated from canine milk and colostrum were characterized according to their functional properties, including their resistance to gastrointestinal conditions, inhibitory effect against pathogens, and intestinal adhesion. Methods: The immunomodulatory effects of the strains were also analyzed in in vitro and in vivo studies. Among the strains evaluated, two LAB strains (TUCO-16 and TUCO-17) showed remarkable resistance to pH 3.0, bile salts, and pancreatin, as well as inhibitory effects against pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens. Results: The TUCO-16 and TUCO-17 strains induced a significant increase in the expression of TNF-α, IL-8, and TLR2 in canine macrophages. The oral administration of TUCO-16 and TUCO-17 strains to mice significantly augmented their resistance to pathogenic E. coli or Salmonella intestinal infections. Both canine strains reduced intestinal damage and pathogen counts in the liver and spleen and avoided their dissemination into the bloodstream. These protective effects were related to the ability of TUCO-16 and TUCO-17 strains to differentially modulate the production of IFN-γ, IFN-ß, TNF-α, IL-6, KC, MCP-1, and IL-10 in the intestinal mucosa. Conclusion: Both strains, TUCO-16 and TUCO-17, are potential probiotic candidates for improving intestinal health in dogs, particularly for their ability to inhibit the growth of Gram-negative pathogens common in gastrointestinal infections and modulate the animal's immune response. Further studies are required to effectively demonstrate the beneficial effects of TUCO-16 and TUCO-17 strains in dogs.

Immunomodulatory role of vasoactive intestinal peptide and ghrelin in Oncorhynchus mykiss.

Neuropeptides are a group of peptides derived from precursor proteins synthesized in neuronal and nonneuronal cells. The classical functions of neuropeptides have been extensively studied in mammals, including neuromodulation in the central nervous system, molecular signaling in the peripheral nervous system, and immunomodulation associated mainly with anti-inflammatory activity. In contrast, in teleosts, studies of the immunomodulatory function of these neuropeptides are limited. In Oncorhynchus mykiss, vasoactive intestinal peptide (VIP) mRNA sequences have not been cloned, and the role of VIP in modulating the immune system has not been studied. Furthermore, in relation to other neuropeptides with possible immunomodulatory function, such as ghrelin, there are also few studies. Therefore, in this work, we performed molecular cloning, identification, and phylogenetic analysis of three VIP precursor sequences (prepro-VIP1, VIP2 and VIP3) in rainbow trout. In addition, the immunomodulatory function of both neuropeptides was evaluated in an in vitro model using the VIP1 sequence identified in this work and a ghrelin sequence already studied in O. mykiss. The results suggest that the prepro-VIP2 sequence has the lowest percentage of identity with respect to the other homologous sequences and is more closely related to mammalian orthologous sequences. VIP1 induces significant expression of both pro-inflammatory (IFN-γ, IL-1ß) and anti-inflammatory (IL-10 and TGF-ß) cytokines, whereas ghrelin only induces significant expression of proinflammatory cytokines such as IL-6 and TNF-α.

Association between early sexual initiation and sexually transmitted infections among Peruvian reproductive-age women.

Background: Sexually transmitted infections (STIs) are a serious public health problem worldwide, especially among reproductive-age women. The early sexual onset of sexual intercourse (EOSI) has been suggested as a risk factor, although there is no data at the national level. Objective: To evaluate the association between EOSI and STIs in Peruvian women of childbearing age. Methods: Analytical cross-sectional study with secondary data analyzes of the Peruvian Demographic and Family Health Survey 2018. The outcome was the presence of STIs in the last 12 months and the exposure variable was EOSI (age < 15 years at the time of their first sexual experience). To evaluate the association of interest, crude and adjusted prevalence ratios (aPRs) were calculated using generalized linear models with Poisson family and logarithmic link function. Results: We analyzed data from 31,028 women of childbearing age. The 11.3% reported having STIs in the last 12 months and 20.2% of the participants had an EOSI. After adjusting for potential confounders, we found that EOSI was associated with STIs (aPR: 1.27; 95% CI: 1.08-1.50; p = 0.005). When conducting stratified analysis by area of residence and number of sexual partners, this association was maintained in women living in urban areas (aPR: 1.36; 95% CI: 1.11-1.66; p = 0.003) those who did not report having a history of multiple sexual partners (aPR: 1.27; 95% CI: 1.08-1.51; p = 0.005), and those in the middle (aPR: 1.42; 95% CI: 1.03-1.97; p = 0.034) and highest (aPR: 2.12; 95% CI: 1.33-3.39; p = 0.002) wealth quintiles. Conclusion: Among reproductive-age women from Peru, EOSI was associated with STIs, especially in women living in urban areas, with no history of multiple sexual partners, and belonging to the middle to higher wealth index. The implementation of measures to prevent EOSI and fostering appropriate sexual health counseling for women with EOSI is advised.

Chitosan Microparticles Enhance the Intestinal Release and Immune Response of an Immune Stimulant Peptide in Oncorhynchus mykiss.

The aquaculture industry is constantly increasing its fish production to provide enough products to maintain fish consumption worldwide. However, the increased production generates susceptibility to infectious diseases that cause losses of millions of dollars to the industry. Conventional treatments are based on antibiotics and antivirals to reduce the incidence of pathogens, but they have disadvantages, such as antibiotic resistance generation, antibiotic residues in fish, and environmental damage. Instead, functional foods with active compounds, especially antimicrobial peptides that allow the generation of prophylaxis against infections, provide an interesting alternative, but protection against gastric degradation is challenging. In this study, we evaluated a new immunomodulatory recombinant peptide, CATH-FLA, which is encapsulated in chitosan microparticles to avoid gastric degradation. The microparticles were prepared using a spray drying method. The peptide release from the microparticles was evaluated at gastric and intestinal pH, both in vitro and in vivo. Finally, the biological activity of the formulation was evaluated by measuring the expression of il-1ß, il-8, ifn-γ, Ifn-α, and mx1 in the head kidney and intestinal tissues of rainbow trout (Oncorhynchus mykiss). The results showed that the chitosan microparticles protect the CATH-FLA recombinant peptide from gastric degradation, allowing its release in the intestinal portion of rainbow trout. The microparticle-protected CATH-FLA recombinant peptide increased the expression of il-1ß, il-8, ifn-γ, ifn-α, and mx1 in the head kidney and intestine and improved the antiprotease activity in rainbow trout. These results suggest that the chitosan microparticle/CATH-FLA recombinant peptide could be a potential prophylactic alternative to conventional antibiotics for the treatment of infectious diseases in aquaculture.

Methodological quality and recommendations of hemophilia clinical practice guidelines: A scoping review.

Background and Aims: Hemophilia clinical practice guidelines (CPGs) play a vital role in guiding healthcare professionals' decisions. However, the quality and recommendations of CPGs for hemophilia may vary. This study aimed to assess the methodological quality of hemophilia CPGs published between 2017 and 2021 and compare their recommendations for prophylaxis using clotting factor concentrate. Methods: We conducted a comprehensive search for relevant CPGs in PubMed, TripDatabase, Grades of Recommendation, Assessment, Development, and Evaluation (GRADE) International Guidelines Database, Google Scholar, and Google. We used the AGREE-II instrument to assess the methodological quality of each CPG and compared their recommendations for prophylaxis. Results: Of the 11 CPGs that met the inclusion criteria, 5/11 were developed in upper-middle-income countries, and 6/11 used the GRADE methodology. The primary prophylaxis dose recommendations varied among the CPGs, with 4/11 recommending a low dose, 6/11 recommending an intermediate or high dose, and 1/11 not issuing a recommendation. However, only 2/11 CPGs provided justification for their recommendations on initiation and dose, and no economic evaluations were conducted to support these recommendations. Conclusion: The quality of hemophilia CPGs is not optimal, with inconsistent recommendations for prophylaxis and a lack of justification for these recommendations. To ensure evidence-based and trustworthy recommendations, there is a need for transparency and improvement in the decision-making process of hemophilia CPGs.

A minisatellite-based MLVA for deciphering the global epidemiology of the bacterial cassava pathogen Xanthomonas phaseoli pv. manihotis.

Cassava Bacterial Blight (CBB) is a destructive disease widely distributed in the different areas where this crop is grown. Populations studies have been performed at local and national scales revealing a geographical genetic structure with temporal variations. A global epidemiology analysis of its causal agent Xanthomonas phaseoli pv. manihotis (Xpm) is needed to better understand the expansion of the disease for improving the monitoring of CBB. We targeted new tandem repeat (TR) loci with large repeat units, i.e. minisatellites, that we multiplexed in a scheme of Multi-Locus Variable number of TR Analysis (MLVA-8). This genotyping scheme separated 31 multilocus haplotypes in three clusters of single-locus variants and a singleton within a worldwide collection of 93 Xpm strains isolated over a period of fifty years. The major MLVA-8 cluster 1 grouped strains originating from all countries, except the unique Chinese strain. On the contrary, all the Xpm strains genotyped using the previously developed MLVA-14 microsatellite scheme were separated as unique haplotypes. We further propose an MLVA-12 scheme which takes advantage of combining TR loci with different mutation rates: the eight minisatellites and four faster evolving microsatellite markers, for global epidemiological surveillance. This MLVA-12 scheme identified 78 haplotypes and separated most of the strains in groups of double-locus variants (DLV) supporting some phylogenetic relationships. DLV groups were subdivided into closely related clusters of strains most often sharing the same geographical origin and isolated over a short period, supporting epidemiological relationships. The main MLVA-12 DLV group#1 was composed by strains from South America and all the African strains. The MLVA-12 scheme combining both minisatellite and microsatellite loci with different discriminatory power is expected to increase the accuracy of the phylogenetic signal and to minimize the homoplasy effects. Further investigation of the global epidemiology of Xpm will be helpful for a better control of CBB worldwide.

Factors Associated with COVID-19 Death in a High-Altitude Peruvian Setting during the First 14 Months of the Pandemic: A Retrospective Multicenter Cohort Study in Hospitalized Patients.

Risk factors for COVID-19 death in high-altitude populations have been scarcely described. This study aimed to describe risk factors for COVID-19 death in three referral hospitals located at 3399 m in Cusco, Peru, during the first 14 months of the pandemic. A retrospective multicenter cohort study was conducted. A random sample of ~50% (1225/2674) of adult hospitalized patients who died between 1 March 2020 and 30 June 2021 was identified. Of those, 977 individuals met the definition of death by COVID-19. Demographic characteristics, intensive care unit (ICU) admission, invasive respiratory support (IRS), disease severity, comorbidities, and clinical manifestation at hospital admission were assessed as risk factors using Cox proportional-hazard models. In multivariable models adjusted by age, sex, and pandemic periods, critical disease (vs. moderate) was associated with a greater risk of death (aHR: 1.27; 95%CI: 1.14-1.142), whereas ICU admission (aHR: 0.39; 95%CI: 0.27-0.56), IRS (aHR: 0.37; 95%CI: 0.26-0.54), the ratio of oxygen saturation (ROX) index ≥ 5.3 (aHR: 0.87; 95%CI: 0.80-0.94), and the ratio of SatO2/FiO2 ≥ 122.6 (aHR: 0.96; 95%CI: 0.93-0.98) were associated with a lower risk of death. The risk factors described here may be useful in assisting decision making and resource allocation.

Focused Wound Care Handoff Improves Burn Center Physician-Nursing Communication and Wound Care Education.

Burn patients require changing wound care routines dependent on wound characteristics and operative interventions. Order discrepancies on electronic medical systems and poor communication between providers leads to incorrect wound care treatment which can be harmful to the complex burn patient. By dedicating a daily wound care discussion for each patient involving integral components of the team: physician, charge nurse, and wound care technicians, enhanced communication amongst team members and improved patient care was noted. A single-center burn unit conducted pre- and postintervention survey of nursing staff measuring various components of wound care. The time spent on the wound care discussions were measured daily. Additional time required to conduct the rounds were minimal with nurse reported increased clarification in patient care without additional work burden. Thus, focused wound care meetings assist with communication between providers, clarification of wound care needs, and avoidance of errors without increasing strain on the team.

"É para o seu bem": a "violência perfeita" na assistência obstétrica / "It's for your own good": "perfect violence" in obstetric care

Resumo No Brasil, a violência obstétrica vem sendo pesquisada desde os anos 1980. Na década de 90, no entanto, o fenômeno passou a receber maior destaque. A forma de violência analisada neste trabalho refere-se a uma violência velada, chamada de "violência perfeita". Este ensaio reflete sobre a ocorrência da violência perfeita na obstetrícia, especialmente no que concerne às sutilezas do discurso médico, que pode travestir essa agressão numa forma de cuidado. A violência perfeita pode soar como preocupação da parte do médico com a saúde da gestante, que pode se submeter às recomendações médicas de forma passiva, por acreditar que deve ser o melhor para ela ou para o bebê. Ao praticar a violência perfeita, o obstetra pode interferir no desfecho do parto. A "epidemia" de cesarianas no Brasil tem sido justificada pelos médicos como preferência da mulher, mas pesquisas refutam essa hipótese e provocam a reflexão: quem está de fato escolhendo a modalidade de parto? O presente ensaio nos mostra que observar as sutilezas do discurso médico pode ajudar a responder essa pergunta.

Abstract In Brazil obstetric violence has been researched since the 1980s. However, in the nineties the phenomenon started to have a more prominent position. The form of violence analysed in this study refers to a covert violence, called "perfect violence". This essay reflects about the occurrence of perfect violence in obstetrics, especially regarding the subtleties of medical discourse, which can disguise this aggression as a form of care. This type of violence may sound as if it was a concern with the patient's health on behalf of the doctor, but it can make the patient submit herself to the medical recommendations in a passive manner, believing it is the best for her and her baby. In this way, violence is disguised as good practice, allegedly aiming at what is best for the patient. In practicing perfect violence, the doctor can interfere in the delivery outcome. The caesarean "epidemic" in Brazil has been justified by doctors as the woman's preference, but research undermines this hypothesis and raises the reflection: who is in fact choosing the type of childbirth? This essay shows us that observing the subtleties of medical discourse can help answering this question.

[Clinical practice guideline for Diagnosis and management of cholelithiasis, cholecystitis and choledocholithiasis for the Peruvian Social Security (EsSalud)]. / Guía de práctica clínica para el diagnóstico y manejo de la colelitiasis, colecistitis aguda y coledocolitiasis en el Seguro Social del Perú (EsSalud).

INTRODUCTION: This article summarizes the clinical practice guide (CPG) for the diagnosis and management of cholelithiasis, acute cholecystitis and choledocholithiasis in the Peruvian Social Security (EsSalud). OBJECTIVE: To provide clinical recommendations based on evidence for the management of patients with cholelithiasis, acute cholecystitis and choledocholithiasis in EsSalud. METHODS: a guideline task force (GTF) was formed with internists, general surgeons, gastroenterologists, and methodologists. The group proposed 10 clinical questions to be answered in this Clinical practice guideline (CPG). Systematic searches of preview reviews were performed and when it was necessary, primary studies from PubMed and CENTRAL during 2017 were reviewed. The evidence was selected aiming to answer each proposed question. Certainty of evidence was evaluated using Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology. In periodical work sessions, the group used GRADE methodology for reviewing the evidence and formulating recommendations, good clinical practice items and three flowcharts for diagnosis and treatment. Finally, the CPG was approved by Resolution Nº 046-IETSI-ESSALUD-2017. RESULTS: This CPG approached 10 clinical questions divided into two topics: diagnosis and management. Based on these questions; one strong recommendation, five weak recommendations, and 17 good clinical practice items and three flowcharts were formulated. CONCLUSION: This paper abstracts the methodology and evidence-based conclusions of the CPG for diagnosis and management of cholelithiasis, acute cholecystitis and choledocholithiasis in EsSalud.

Association between the Use of Biomass as Fuel for Cooking and Acute Respiratory Infections in Children under 5 Years of Age in Peru: An Analysis of a Population-Based Survey, 2019.

Background: Acute respiratory infections (ARIs) are the most frequent respiratory diseases associated with the use of biomass as fuel within the home. ARIs are the main cause of mortality in children under 5 years of age. We aimed to evaluate the association between the use of biomass as cooking fuel and ARI in children under 5 years of age in Peru in 2019. Methods: A secondary data analysis of the 2019 Peru Demographic and Family Health Survey (ENDES) has been performed. The outcome variable was a history of ARI. The exposure variable was the use of biomass as fuel for cooking food. To evaluate the association of interest, generalized linear models from the Poisson family with logarithmic link function considering complex sampling to estimate crude prevalence ratio (cPR) and adjusted prevalence ratio (aPR) with their respective 95% confidence intervals have been performed. P values <0.05 were considered statistically significant. Results: A total of 16,043 children were included in the analysis. Of the total, biomass was used as fuel to cook food in the homes of 3,479 (20.0%) children. Likewise, 2,185 (14.3%) of the children had a history of ARI. In the adjusted model, it was found that children living in homes in which biomass was used as cooking fuel had a greater probability of presenting ARI (aPR = 1.13; 95% CI: 1.01-1.28). Conclusions: It has been found that biomass was used to cook food in two of every 10 households. Likewise, almost one-seventh of children under 5 years old presented an ARI. The use of biomass as a source of energy for cooking in the home was associated with a higher probability of presenting ARIs.

New strategy for the design, production and pre-purification of chimeric peptide with immunomodulatory activity in Salmosalar.

The intensive salmon farming is associated with massive outbreaks of infections. The use of antibiotics for their prevention and control is related to damage to the environment and human health. Antimicrobial peptides (AMPs) have been proposed as an alternative to the use of antibiotics for their antimicrobial and immunomodulatory activities. However, one of the main challenges for its massive clinical application is the high production cost and the complexity of chemical synthesis. Thus, recombinant DNA technology offers a more sustainable, scalable, and profitable option. In the present study, using an AMPs function prediction methodology, we designed a chimeric peptide consisting of sequences derived from cathelicidin fused with the immunomodulatory peptide derived from flagellin. The designed peptide, CATH-FLA was produced by recombinant expression using an easy pre-purification system. The chimeric peptide was able to induce IL-1ß and IL-8 expression in Salmo salar head kidney leukocytes, and prevented Piscirickettsia salmonis-induced cytotoxicity in SHK-1 cells. These results suggest that pre-purification of a recombinant AMP-based chimeric peptide designed in silico allow obtaining a peptide with immunomodulatory activity in vitro. This could solve the main obstacle of AMPs for massive clinical applications.

Salmonid MyD88 is a key adapter protein that activates innate effector mechanisms through the TLR5M/TLR5S signaling pathway and protects against Piscirickettsia salmonis infection.

The membrane-anchored and soluble Toll-like Receptor 5 -TLR5M and TLR5S, respectively-from teleost recognize bacterial flagellin and induce the pro-inflammatory cytokines expression in a MyD88-dependent manner such as the TLR5 mammalian orthologous receptor. However, it has not been demonstrated whether the induced signaling pathway by these receptors activate innate effector mechanisms MyD88-dependent in salmonids. Therefore, in this work we study the MyD88 dependence on the induction of TLR5M/TLR5S signaling pathway mediated by flagellin as ligand on the activation of some innate effector mechanisms. The intracellular and extracellular Reactive Oxygen Species (ROS) production and conditioned supernatants production were evaluated in RTS11 cells, while the challenge with Piscirickettsia salmonis was evaluated in SHK-1 cells. Our results demonstrate that flagellin directly stimulates ROS production and indirectly stimulates it through the production of conditioned supernatants, both in a MyD88-dependent manner. Additionally, flagellin stimulation prevents the cytotoxicity induced by infection with P. salmonis in a MyD88-dependent manner. In conclusion we demonstrate that MyD88 is an essential adapter protein in the activation of the TLR5M/TLR5S signaling pathway mediated by flagellin in salmonids, which leads downstream to the induction of innate effector mechanisms, promoting immuno-protection against a bacterial challenge with P. salmonis.

Effects of replacement therapies with clotting factors in patients with hemophilia: A systematic review and meta-analysis.

BACKGROUND: Different prophylactic and episodic clotting factor treatments are used in the management of hemophilia. A summarize of the evidence is needed inform decision-making. OBJECTIVE: To compare the effects of factor replacement therapies in patients with hemophilia. METHODS: We performed a systematic search in PubMed, Central Cochrane Library, and Scopus. We included randomized controlled trials (RCTs) published up to December 2020, which compared different factor replacement therapies in patients with hemophilia. Random-effects meta-analyses were performed whenever possible. The certainty of the evidence was assessed using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology. The study protocol was registered in PROSPERO (CRD42021225857). RESULTS: Nine RCTs were included in this review, of which six compared episodic with prophylactic treatment, all of them performed in patients with hemophilia A. Pooled results showed that, compared to the episodic treatment group, the annualized bleeding rate was lower in the low-dose prophylactic group (ratio of means [RM]: 0.27, 95% CI: 0.17 to 0.43), intermediate-dose prophylactic group (RM: 0.15, 95% CI: 0.07 to 0.36), and high-dose prophylactic group (RM: 0.07, 95% CI: 0.04 to 0.13). With significant difference between these subgroups (p = 0.003, I2 = 82.9%). In addition, compared to the episodic treatment group, the annualized joint bleeding rate was lower in the low-dose prophylactic group (RM: 0.17, 95% CI: 0.06 to 0.43), intermediate-dose prophylactic group (RM of 0.14, 95% CI: 0.07 to 0.27), and high-dose prophylactic group (RM of 0.08, 95% CI: 0.04 to 0.16). Without significant subgroup differences. The certainty of the evidence was very low for all outcomes according to GRADE methodology. The other studies compared different types of clotting factor concentrates (CFCs), assessed pharmacokinetic prophylaxis, or compared different frequencies of medication administration. CONCLUSIONS: Our results suggest that prophylactic treatment (at either low, intermediate, or high doses) is superior to episodic treatment for bleeding prevention. In patients with hemophilia A, the bleeding rate seems to have a dose-response effect. However, no study compared different doses of prophylactic treatment, and all results had a very low certainty of the evidence. Thus, future studies are needed to confirm these results and inform decision making.

Guía de práctica clínica para la prevención, diagnóstico y manejo de infecciones asociadas al acceso de hemodiálisis en el Seguro Social de Salud del Perú (EsSalud) / Clinical practice guidelines for the prevention, diagnosis and management of infections related with hemodialysis access of the Peruvian Health Social Security (EsSalud)

Introducción: El presente artículo resume la guía de práctica clínica (GPC) para la prevención, diagnóstico y manejo de infecciones asociadas al acceso de hemodiálisis en el Seguro Social del Perú (EsSalud). Objetivo: Proveer recomendaciones clínicas basadas en evidencia para la prevención, diagnóstico y tratamiento de infecciones asociadas al acceso de hemodiálisis en EsSalud. Material y Métodos: Se conformó un grupo elaborador de la guía (GEG) que incluyó especialistas y metodólogos. El GEG formuló 7 preguntas clínicas. Se realizó búsquedas sistemáticas de revisiones sistemáticas y estudios primarios en PubMed y CENTRAL de Cochrane entre noviembre del 2019 y marzo del 2020. Se seleccionó la evidencia para responder a las preguntas clínicas planteadas. La certeza de la evidencia fue evaluada usando la metodología Grading of Recommendations Assessment, Development, and Evaluation (GRADE). El GEG usó la metodología GRADE para revisar la evidencia y formular recomendaciones, los puntos de buena práctica clínica y el flujograma de tratamiento. Finalmente, la GPC fue aprobada con Resolución N° 116-IETSI-ESSALUD-2020. Resultados: La presente GPC abordó 7 preguntas clínicas, divididas en tres temas: tamizaje, diagnóstico, estadiaje y tratamiento. En base a estas preguntas se formularon 12 recomendaciones (11 fuertes y 1 condicional), 33 puntos de buena práctica clínica y 2 flujogramas de manejo. Conclusión: El presente artículo resume la metodología y las conclusiones basadas en evidencias de la GPC para la prevención, diagnóstico y tratamiento de infecciones asociadas al acceso de hemodiálisis en EsSalud.

Background: This article summarizes the clinical practice guidelines (CPG) for the prevention, diagnosis and management of infections associated with hemodialysis access of the Peruvian Health Social Security (EsSalud). To provide clinical Objective: recommendations based on evidence for the prevention, diagnosis and management of infections associated with hemodialysis access in EsSalud. Aguideline development Material and Methods:group (GDG) was established, including medical specialists and methodologists. The GDG formulated 7 clinical questions to be answered in this CPG. Systematic searches of systematic reviews and primary studies (when pertinent) were conducted in PubMed, and CENTRAL(Cochrane) from November 2019 to March 2020. The evidence was selected to answer each of the clinical questions. The accuracy of the evidence was evaluated using the Grading of Recommendations Assessment, Development, and Evaluation(GRADE) methodology. In periodic work meetings, the GEG used the GRADE methodology to review the evidence and formulate the recommendations, the points of good clinical practice and the treatment flow chart. Finally, the CPG was approved with Resolution No. 116-IETSI-ESSALUD-2020. The present Results:CPG addressed 7 clinical questions of four topics: prevention, diagnosisandtreatment.Basedonthesequestions,12 recommendations (11strongrecommendationsand1weak recommendation), 33 points of good clinical practice, and 2 flowcharts were formulated. This paper summarizes Conclusion:the methodology and evidence-based conclusions from the CPG for the prevention, diagnosis and management of infections associated with hemodialysis access of EsSalud

Guía de práctica clínica para el diagnóstico y manejo de la colelitiasis, colecistitis aguda y coledocolitiasis en el Seguro Social del Perú (EsSalud) / Clinical practice guideline for Diagnosis and management of cholelithiasis, cholecystitis and choledocholithiasis for the Peruvian Social Security (EsSalud)

RESUMEN Introducción: El presente artículo resume la guía de práctica clínica (GPC) para el diagnóstico y manejo de los pacientes con colelitiasis, colecistitis aguda y coledocolitiasis en el Seguro Social del Perú (EsSalud). Objetivo: Proveer recomendaciones clínicas basadas en evidencia para la evaluación y el manejo de pacientes con colelitiasis, colecistitis aguda y coledocolitiasis en EsSalud. Métodos: Se conformó un grupo elaborador de la guía (GEG) que incluyó médicos especialistas en gastroenterología, cirujanos generales y metodólogos. El GEG formuló 10 preguntas clínicas a ser respondidas por la presente GPC. En septiembre del 2017 se realizaron búsquedas de revisiones sistemáticas y -cuando fue considerado pertinente- estudios primarios en PubMed. Se seleccionó la evidencia para responder 10 preguntas clínicas. La certeza de la evidencia fue evaluada usando la metodología Grading of Recommendations Assessment, Development, and Evaluation (GRADE). En reuniones de trabajo periódicas, el GEG usó la metodología GRADE para revisar la evidencia y formular las recomendaciones, los puntos de buenas prácticas clínicas y el flujograma de evaluación y manejo. Finalmente, la GPC fue aprobada con Resolución N° 021-IETSIESSALUD-2018. Resultados: La presente GPC abordó 10 preguntas clínicas sobre el diagnóstico y manejo de colelitiasis, colecistitis aguda y coledocolitiasis. En base a dichas preguntas se formularon 6 recomendaciones (5 recomendaciones condicionales y 1 recomendación fuerte), 17 puntos de buena práctica clínica y 3 flujogramas. Conclusión: El presente artículo resume la metodología y las conclusiones de la GPC basadas en evidencias para el diagnóstico y tratamiento de la colelitiasis, colecistitis aguda y coledocolitiasis de EsSalud.

ABSTRACT Introduction: This article summarizes the clinical practice guide (CPG) for the diagnosis and management of cholelithiasis, acute cholecystitis and choledocholithiasis in the Peruvian Social Security (EsSalud). Objective: To provide clinical recommendations based on evidence for the management of patients with cholelithiasis, acute cholecystitis and choledocholithiasis in EsSalud. Methods: a guideline task force (GTF) was formed with internists, general surgeons, gastroenterologists, and methodologists. The group proposed 10 clinical questions to be answered in this Clinical practice guideline (CPG). Systematic searches of preview reviews were performed and when it was necessary, primary studies from PubMed and CENTRAL during 2017 were reviewed. The evidence was selected aiming to answer each proposed question. Certainty of evidence was evaluated using Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology. In periodical work sessions, the group used GRADE methodology for reviewing the evidence and formulating recommendations, good clinical practice items and three flowcharts for diagnosis and treatment. Finally, the CPG was approved by Resolution Nº 046-IETSI-ESSALUD-2017. Results: This CPG approached 10 clinical questions divided into two topics: diagnosis and management. Based on these questions; one strong recommendation, five weak recommendations, and 17 good clinical practice items and three flowcharts were formulated. Conclusion: This paper abstracts the methodology and evidence-based conclusions of the CPG for diagnosis and management of cholelithiasis, acute cholecystitis and choledocholithiasis in EsSalud.