Development and Optimization of a Multiplex Real-Time RT-PCR to Detect SARS-CoV-2 in Human Samples.

PCR and its variants (RT-PCR and qRT-PCR) are valuable and innovative molecular techniques for studying nucleic acids. qPCR has proven to be highly sensitive, efficient, and reproducible, generating reliable results that are easy to analyze. During the COVID-19 pandemic, qPCR became the gold standard technique for detecting the SARS-CoV-2 virus that allowed to confirm the infection event, and those asymptomatic ones, and thus save millions of lives. In-house multiplex qPCR tests were developed worldwide to detect different viral targets and ensure results, follow the infections, and favor the containment of a pandemic. Here, we present the detailed fundamentals of the qPCR technique based on fluorogenic probes and processes to develop and optimize a successful multiplex RT-qPCR test for detecting SARS-CoV-2 that could be used to diagnose COVID-19 accurately.

Virome analyses of Amblyomma cajennense and Rhipicephalus microplus ticks collected in Colombia.

Tick-borne viruses (TBV) have gained public health relevance in recent years due to the recognition of human-associated fatal cases and the increase in tick-borne disease and transmission. However, many tick species have not been studied for their potential to transmit pathogenic viruses, especially those found in Latin America. To gain better understanding of the tick virome, we conducted targeted amplification using broadly-reactive consensus-degenerate pan-viral targeting viruses from the genera Flavivirus, Bandavirus, Uukuvirus, and Orthonairovirus genus. Additionally, we conducted unbiased metagenomic analyses to investigate the presence of viral RNA sequences in Amblyomma cajennense, A. patinoi and Rhipicephalus microplus ticks collected from a horse slaughter plant in Medellín, Colombia. While no viral products were detected by PCR, results of the metagenomic analyses revealed the presence of viral genomes belonging to the genera Phlebovirus, Bandavirus, and Uukuvirus, including Lihan Tick Virus (LTV), which was previously reported in Rhipicephalus microplus from Colombia. Overall, the results emphasized the enormous utility of the next-generation sequencing in identifying virus genetic diversity presents in ticks and other species of vectors and reservoirs.

Laparoscopic management of colovesical fistula in different clinical scenarios.

INTRODUCTION: Colovesical fistula (CVF) is a condition with various aetiologies and presentations. Surgical treatment is necessary in most cases. Due to its complexity, open approach is preferred. However, laparoscopic approach is reported in the management of CVF due to diverticular disease. The aim of this study was to analyse the management and outcome of patients with CVF of different aetiologies treated with laparoscopic approach. PATIENTS AND METHODS: This was a retrospective study. We retrospectively reviewed all patients undergoing elective laparoscopic management of CVF from March 2015 to December 2019. STATISTICAL ANALYSIS USED: None. RESULTS: Nine patients underwent laparoscopic management of CVF. There were no intraoperative complications or conversions to open surgery. A sigmoidectomy was performed in eight cases. In one patient, a fistulectomy with sigmoid and bladder defect closure was performed. In two cases of locally advanced colorectal cancer with bladder invasion, a multi-stage procedure with temporary colostomy was chosen. In three cases, with no intraoperative leakage, we did not perform bladder suture. Four Clavien I-II complications were recorded. Two fragile patients died in the post-operative period. No patients required re-operation. At a median follow-up of 21 months (interquartile range: 6-47), none of the patients had recurrence of fistula. CONCLUSIONS: CVF can be managed with laparoscopic approach by skilled laparoscopic surgeons in different clinical scenarios. Bladder suture is not necessary if leakage is absent. Informed counselling to the patient must be guaranteed concerning the risk of major complications and mortality in case of CVF due to malignant disease.

"What Is the Benefit?": Perceptions and Preferences for Long-Acting Injectable Antiretroviral Therapy Among People Living With HIV.

Long-acting injectable antiretroviral therapy (LA-ART) expands treatment options for people living with HIV (PLWH). This qualitative study characterizes LA-ART awareness, perceptions, and preferences among PLWH engaged in HIV care. From 2019 through 2021, we conducted semistructured in-depth interviews with 71 PLWH sampled from three clinics in three U.S. settings (North Carolina, Washington, DC, Massachusetts). Transcripts were analyzed using narrative and thematic techniques. Participant mean age was 46 years (range 24-72); most were cisgender men (55%) and virally suppressed (73%). Most participants had not heard of LA-ART and reacted with a mix of excitement and cautiousness. Potential LA-ART benefits included easier adherence, privacy, and effectiveness; concerns included effectiveness, side effects, costs, and increased clinic visits. Participants appreciated that LA-ART could support achieving and sustaining viral suppression. To inform their decision, participants wanted more information and convenient access and administration. Findings indicated that a shared decision-making approach and economic and logistical support for PLWH could facilitate LA-ART uptake.

Seroprevalence, infection, and personal protective equipment use among Colombian healthcare workers during the COVID-19 pandemic.

Introduction: Healthcare workers (HCWs) are at the forefront of the COVID-19 response and frequently come into close contact with patients and their virus-contaminated body fluids. Recent studies have identified differential risks of infection and the use of personal protective equipment (PPE) among HCWs. However, available data might be interpreted with caution because of differences in the national health systems, local implementation issues, and adherence limitations to guidelines. A comprehensive description of infection, exposure at work, and biosafety habits during the COVID-19 pandemic has not been conducted among the HCW groups in Latin American populations. Objective: To describe SARS-CoV-2 seroprevalence, infections, and extent of PPE use during the COVID-19 pandemic among HCWs at three different times, including dental practitioners (DP), nursing assistants (NA), physicians (P), and respiratory therapists (RT), from Bogotá, Colombia. Methods: After IRB approval, this cross-sectional study included 307 HCWs. Participants provided nasopharyngeal swabs and blood samples to detect viral RNA (RT-qPCR) and IgM/IgG anti-SARS-CoV-2 (ELFA-ELISA) at baseline (BL) and two follow-ups. Infection prevalence was defined as the number of positive-tested participants (RT-qPCR and/or IgM). Data on clinical status and biosafety habits were collected each time. Results: Differential infection prevalence was found among HCWs through the study timeline (BL: RT-qPCR = 2.6%, IgM = 1.6%; follow-up 1 (45 days after BL): RT-qPCR = 4.5%, IgM = 3.9%; follow-up 2 (60 days after BL): RT-qPCR = 3.58%, IgM = 1.3%. Dental practitioners showed a higher infection frequency in BL and follow-up 1. IgG-positive tested HCWs percentage progressively increased from BL to follow-ups among the whole sample while index values decreased. Limitations in N95 availability and a high perception of occupational risk were reported. Conclusion: A low prevalence of active SARS-CoV-2 infections among HCWs groups was found. Over time, there was an increase in participants showing IgG antibodies, although the levels of these antibodies in the blood decreased. Additionally, HCWs reported limitations in the availability of PPE as well as a variation in their safety practices.

Láser de baja frecuencia para aftas mayores recurrentes. Caso Clínico / Low frequency laser for major recurrent aphthous stomatitis. Clinical case

La estomatitis aftosa recurrente (menor, mayor o herpética) es una de las enfermedades bucodentales más frecuentes, de etiología multifactorial y con predisposición genética. Para esta patología su diagnóstico es clínico, y tomando como base la historia clínica se puede discriminar con otras patologías ulcerativas. La terapia con láser de baja potencia (LLLT) se propone como un tratamiento no invasivo, actuando como un fotomodulador celular, por lo que reduce el dolor, la inflamación, y estimula la regeneración tisular, previniendo daños. En el presente artículo se reporta a un niño de 11 años con una herida dolorosa de 4 meses en el borde posterior de la lengua que apareció tras una mordedura, en el que a la exploración clínica se encontró una lesión amarillenta con zonas enrojecidas, rodeada de un halo blanquecino de bordes redondeados. Para el tratamiento se utilizó LLLT, luego de una semana disminuyó el dolor, el paciente comenzó a comer correctamente, a los 15 días se observó una reducción de los bordes rojizos de la lesión, pasado el mes la lesión había remitido casi por completo. La LLLT se describe como un tratamiento eficaz, no invasivo, reduciendo los síntomas, y mejorando la calidad de vida del paciente al promover la curación de las lesiones.

Recurrent aphthous stomatitis (minor, major or herpetic) is one of the most frequent bucodental diseases, of multifactorial etiology and with genetic predisposition. The diagnosis of this pathology is clinical, and based on the clinical history it can be discriminated with other ulcerative pathologies. Low power laser therapy (LLLT) is proposed as a non-invasive treatment, acting as a cellular photo modulator, which reduces pain, inflammation, and stimulating tissue regeneration, preventing damage. This article reports an 11-year-old boy with a 4-month-old painful wound on the posterior edge of the tongue that appeared after a bite, in which clinical examination revealed a yellowish lesion with reddened areas, surrounded by a whitish halo with rounded edges. LLLT was used for the treatment, after one week the pain decreased, the patient started to eat properly, after 15 days a reduction of the reddish edges of the lesion was observed, after one month the lesion had almost completely remitted. LLLT is described as an effective, non-invasive treatment, which reduces symptoms and improves the patient's quality of life promoting the healing of the lesions.

A Comparison of Lymphoid and Myeloid Cells Derived from Human Hematopoietic Stem Cells Xenografted into NOD-Derived Mouse Strains.

Humanized mice are an invaluable tool for investigating human diseases such as cancer, infectious diseases, and graft-versus-host disease (GvHD). However, it is crucial to understand the strengths and limitations of humanized mice and select the most appropriate model. In this study, we describe the development of the human lymphoid and myeloid lineages using a flow cytometric analysis in four humanized mouse models derived from NOD mice xenotransplanted with CD34+ fetal cord blood from a single donor. Our results showed that all murine strains sustained human immune cells within a proinflammatory environment induced by GvHD. However, the Hu-SGM3 model consistently generated higher numbers of human T cells, monocytes, dendritic cells, mast cells, and megakaryocytes, and a low number of circulating platelets showing an activated profile when compared with the other murine strains. The hu-NOG-EXL model had a similar cell development profile but a higher number of circulating platelets with an inactivated state, and the hu-NSG and hu-NCG developed low frequencies of immune cells compared with the other models. Interestingly, only the hu-SGM3 and hu-EXL models developed mast cells. In conclusion, our findings highlight the importance of selecting the appropriate humanized mouse model for specific research questions, considering the strengths and limitations of each model and the immune cell populations of interest.

Changes in Lolium perenne L. rhizosphere microbiome during phytoremediation of Cd- and Hg-contaminated soils.

The contamination of soil and water by metals such as mercury (Hg) and cadmium (Cd) has been increasing in recent years, because of anthropogenic activities such as mining and agriculture, respectively. In this work, the changes in the rhizosphere microbiome of Lolium perenne L. during the phytoremediation of soils contaminated with Hg and Cd were evaluated. For this, two soil types were sampled, one inoculated with mycorrhizae and one without. The soils were contaminated with Hg and Cd, and L. perenne seeds were sown and harvested after 30 days. To assess changes in the microbiome, DNA isolation tests were performed, for which samples were subjected to two-step PCR amplification with specific 16S rDNA V3-V4 primers (337F and 805R). With mycorrhizae, changes had been found in the absorption processes of metals and a new distribution. While with respect to microorganisms, families such as the Enterobacteriaceae have been shown to have biosorption and efflux effects on metals such as Hg and Cd. Mycorrhizae then improve the efficiency of removal and allow the plant to better distribute the absorbed concentrations. Overall, L. perenne is a species with a high potential for phytoremediation of Cd- and Hg-contaminated soils in the tropics. Inoculation with mycorrhizae modifies the phytoremediation mechanisms of the plant and the composition of microorganisms in the rhizosphere. Mycorrhizal inoculation and changes in the microbiome were associated with increased plant tolerance to Cd and Hg. Microorganism-assisted phytoremediation is an appropriate alternative for L. perenne.

Minigene Splicing Assays and Long-Read Sequencing to Unravel Pathogenic Deep-Intronic Variants in PAX6 in Congenital Aniridia.

PAX6 haploinsufficiency causes aniridia, a congenital eye disorder that involves the iris, and foveal hypoplasia. Comprehensive screening of the PAX6 locus, including the non-coding regions, by next-generation sequencing revealed four deep-intronic variants with potential effects on pre-RNA splicing. Nevertheless, without a functional analysis, their pathogenicity could not be established. We aimed to decipher their impact on the canonical PAX6 splicing using in vitro minigene splicing assays and nanopore-based long-read sequencing. Two multi-exonic PAX6 constructs were generated, and minigene assays were carried out. An aberrant splicing pattern was observed for two variants in intron 6, c.357+136G>A and c.357+334G>A. In both cases, several exonization events, such as pseudoexon inclusions and partial intronic retention, were observed due to the creation or activation of new/cryptic non-canonical splicing sites, including a shared intronic donor site. In contrast, two variants identified in intron 11, c.1032+170A>T and c.1033-275A>C, seemed not to affect splicing processes. We confirmed the high complexity of alternative splicing of PAX6 exon 6, which also involves unreported cryptic intronic sites. Our study highlights the importance of integrating functional studies into diagnostic algorithms to decipher the potential implication of non-coding variants, usually classified as variants of unknown significance, thus allowing variant reclassification to achieve a conclusive genetic diagnosis.

Revegetación natural de áreas afectadas por minería de oro en la selva pluvial tropical del Chocó, Colombia / Natural revegetation of areas impacted by gold mining in the tropical rain forest of Chocó, Colombia

Introducción: La minería de oro a cielo abierto ocasiona impactos drásticos sobre los bosques naturales en regiones tropicales de alta biodiversidad, y la eficiencia de "revegetación", un proceso de resembrar y reconstruir el suelo, continúa siendo poco estudiado. Objetivo: Evaluar el efecto del tiempo sucesional y la distancia al bosque de referencia sobre la estructura biológica y composición de especies en minas de oro revegetadas. Métodos: Realizamos inventarios de vegetación en minas abandonadas en el bosque tropical lluvioso de Chocó, Colombia. Las minas contaban con 6, 10, 15, 19 y 24 años de sucesión natural. En cada escenario, establecimos cuatro parcelas de 2×50 m: a 50 m y 100 m de distancia desde el borde bosque-mina (ecotono). Resultados: Se identificaron 300 especies (193 géneros y 75 familias). La riqueza, diversidad y equitatividad cambian poco con el tiempo sucesional en las minas, pero alcanzan valores superiores en el bosque de referencia. La composición de especies es similar entre las minas con distinto tiempo sucesional, pero difiere ampliamente del bosque de referencia (solo 7 % de semejanza). La calidad del sustrato y las estrategias reproductivas de plantas herbáceas (principalmente Cyperaceae y Melastomataceae) y arbóreas (Cespedesia spathulata y Miconia reducens) que crecen espontáneamente en las minas, juegan un papel importante en la revegetación natural temprana. Conclusiones: 24 años de sucesión natural es poco tiempo para apreciar una recuperación sustancial de la vegetación en las minas si se compara con el bosque, y la distancia desde el ecotono parece no tener efecto significativo; sin embargo, la calidad de sustrato y estrategias reproductivas de plantas herbáceas son importantes en la etapa temprana de recuperación de las áreas impactadas por la minería de oro en Chocó.

Introduction: Open pit gold mining causes drastic impacts on natural forests in tropical regions of high biodiversity, and the efficacy of "revegetation", a process of replanting and rebuilding the soil, is still poorly studied. Objective: To evaluate the effect of successional time, and distance to the reference forest, on the biological structure and species composition of revegetated gold mines. Methods: We inventoried the adjacent forest and abandoned gold mines in the tropical rain forest of Chocó, Colombia. The mines had 6, 10, 15, 19 and 24 years of natural succession. In each scenario, we set four 2 × 50 m plots: two 50 m and two 100 m from the forest-mine edge (ecotone). Results: We identified 300 plant species (193 genera, 75 families). The richness, diversity and evenness changed little with successional time in the mines but reached higher values in the forest. The species composition was similar between the mines with different successional times but differed widely from the forest (only 7 % similarity). The substrate quality and reproductive strategies of herbaceous plants (mainly Cyperaceae and Melastomataceae) and trees (Cespedesia spathulata and Miconia reducens) that grow spontaneously in the mines, play an important role on the early natural revegetation. Conclusions: The 24 years of natural succession have been insufficient to reach a community of a complexity similar to that of the forest, and distance from the ecotone has no significant effect; however, the substrate quality and reproductive strategies of herbaceous plants are important in the early stages of mine recovery in the Chocó.

Continuous prolonged prone positioning in COVID-19-related ARDS: a multicenter cohort study from Chile.

BACKGROUND: Prone positioning is currently applied in time-limited daily sessions up to 24 h which determines that most patients require several sessions. Although longer prone sessions have been reported, there is scarce evidence about the feasibility and safety of such approach. We analyzed feasibility and safety of a continuous prolonged prone positioning strategy implemented nationwide, in a large cohort of COVID-19 patients in Chile. METHODS: Retrospective cohort study of mechanically ventilated COVID-19 patients with moderate-to-severe acute respiratory distress syndrome (ARDS), conducted in 15 Intensive Care Units, which adhered to a national protocol of continuous prone sessions ≥ 48 h and until PaO2:FiO2 increased above 200 mm Hg. The number and extension of prone sessions were registered, along with relevant physiologic data and adverse events related to prone positioning. The cohort was stratified according to the first prone session duration: Group A, 2-3 days; Group B, 4-5 days; and Group C, > 5 days. Multivariable regression analyses were performed to assess whether the duration of prone sessions could impact safety. RESULTS: We included 417 patients who required a first prone session of 4 (3-5) days, of whom 318 (76.3%) received only one session. During the first prone session the main adverse event was grade 1-2 pressure sores in 97 (23.9%) patients; severe adverse events were infrequent with 17 non-scheduled extubations (4.2%). 90-day mortality was 36.2%. Ninety-eight patients (24%) were classified as group C; they exhibited a more severe ARDS at baseline, as reflected by lower PaO2:FiO2 ratio and higher ventilatory ratio, and had a higher rate of pressure sores (44%) and higher 90-day mortality (48%). However, after adjustment for severity and several relevant confounders, prone session duration was not associated with mortality or pressure sores. CONCLUSIONS: Nationwide implementation of a continuous prolonged prone positioning strategy for COVID-19 ARDS patients was feasible. Minor pressure sores were frequent but within the ranges previously described, while severe adverse events were infrequent. The duration of prone session did not have an adverse effect on safety.

Endothelial Dysfunction, HMGB1, and Dengue: An Enigma to Solve.

Dengue is a viral infection caused by dengue virus (DENV), which has a significant impact on public health worldwide. Although most infections are asymptomatic, a series of severe clinical manifestations such as hemorrhage and plasma leakage can occur during the severe presentation of the disease. This suggests that the virus or host immune response may affect the protective function of endothelial barriers, ultimately being considered the most relevant event in severe and fatal dengue pathogenesis. The mechanisms that induce these alterations are diverse. It has been suggested that the high mobility group box 1 protein (HMGB1) may be involved in endothelial dysfunction. This non-histone nuclear protein has different immunomodulatory activities and belongs to the alarmin group. High concentrations of HMGB1 have been detected in patients with several infectious diseases, including dengue, and it could be considered as a biomarker for the early diagnosis of dengue and a predictor of complications of the disease. This review summarizes the main features of dengue infection and describes the known causes associated with endothelial dysfunction, highlighting the involvement and possible relationship between HMGB1 and DENV.

Deletion of the Autism-Associated Protein SHANK3 Abolishes Structural Synaptic Plasticity after Brain Trauma.

Autism spectrum disorders (ASDs) are characterized by repetitive behaviors and impairments of sociability and communication. About 1% of ASD cases are caused by mutations of SHANK3, a major scaffolding protein of the postsynaptic density. We studied the role of SHANK3 in plastic changes of excitatory synapses within the central nervous system by employing mild traumatic brain injury (mTBI) in WT and Shank3 knockout mice. In WT mice, mTBI triggered ipsi- and contralateral loss of hippocampal dendritic spines and excitatory synapses with a partial recovery over time. In contrast, no significant synaptic alterations were detected in Shank3∆11-/- mice, which showed fewer dendritic spines and excitatory synapses at baseline. In line, mTBI induced the upregulation of synaptic plasticity-related proteins Arc and p-cofilin only in WT mice. Interestingly, microglia proliferation was observed in WT mice after mTBI but not in Shank3∆11-/- mice. Finally, we detected TBI-induced increased fear memory at the behavioral level, whereas in Shank3∆11-/- animals, the already-enhanced fear memory levels increased only slightly after mTBI. Our data show the lack of structural synaptic plasticity in Shank3 knockout mice that might explain at least in part the rigidity of behaviors, problems in adjusting to new situations and cognitive deficits seen in ASDs.

An updated review and current challenges of Guanarito virus infection, Venezuelan hemorrhagic fever.

Guanarito virus (GTOV) is a member of the family Arenaviridae and has been designated a category A bioterrorism agent by the US Centers for Disease Control and Prevention. It is endemic to Venezuela's western region, and it is the etiological agent of "Venezuelan hemorrhagic fever" (VHF). Similar to other arenaviral hemorrhagic fevers, VHF is characterized by fever, mild hemorrhagic signs, nonspecific symptoms, thrombocytopenia, and leukopenia. Patients with severe disease usually develop signs of internal bleeding. Due to the absence of reference laboratories that can handle GTOV in endemic areas, diagnosis is primarily clinical and epidemiological. No antiviral therapies are available; thus, treatment includes only supportive analgesia and fluids. GTOV is transmitted by contact with the excreta of its rodent reservoir, Zygodontomys brevicauda. The main reasons for the emergence of the disease may be the increase in the human population, migration, and changes in land use patterns in rural areas. Social and environmental changes could make VHF an important cause of underdiagnosed acute febrile illnesses in regions near the endemic areas. Although there is evidence that GTOV circulates among rodents in different Venezuelan states, VHF cases have only been reported in the states of Portuguesa and Barinas. However, due to the increased frequency of invasions by humans into wildlife habitats, it is probable that VHF could become a public health problem in the nearby regions of Colombia and Brazil. The current Venezuelan political crisis is causing an increase in the migration of people and livestock, representing a risk for the redistribution and re-emergence of infectious diseases.

Methylotrophs and Hydrocarbon-Degrading Bacteria Are Key Players in the Microbial Community of an Abandoned Century-Old Oil Exploration Well.

In this work, we studied the microbial community and the physicochemical conditions prevailing in an exploratory oil well, abandoned a century ago, located in the Cahuita National Park (Costa Rica). According to our analysis, Cahuita well is characterized by a continuous efflux of methane and the presence of a mixture of hydrocarbons including phenanthrene/anthracene, fluoranthene, pyrene, dibenzothiophene, tricyclic terpanes, pyrene, sesquiterpenes, sterane, and n-alkanes. Based on the analysis of 16S rRNA gene amplicons, we detected a significant abundance of methylotrophic bacteria such as Methylobacillus (6.3-26.0% of total reads) and Methylococcus (4.1-30.6%) and the presence of common genera associated with hydrocarbon degradation, such as Comamonas (0.8-4.6%), Hydrogenophaga (1.5-3.3%) Rhodobacter (1.0-4.9%), and Flavobacterium (1.1-6.5%). The importance of C1 metabolism in this niche was confirmed by amplifying the methane monooxygenase (MMO)-encoding gene (pmo) from environmental DNA and the isolation of two strains closely related to Methylorubrum rhodesianum and Paracoccus communis with the ability to growth using methanol and formate as sole carbon source respectively. In addition, we were able to isolated 20 bacterial strains from the genera Pseudomonas, Acinetobacter, and Microbacterium which showed the capability to grow using the hydrocarbons detected in the oil well as sole carbon source. This work describes the physicochemical properties and microbiota of an environment exposed to hydrocarbons for 100 years, and it not only represents a contribution to the understanding of microbial communities in environments with permanently high concentrations of these compounds but also has biotechnological implications for bioremediation of petroleum-polluted sites.

Intergenerational Online Health Information Searching and Brokering: Framing Health Literacy as a Family Asset.

Latino populations are disproportionately impacted by health disparities and face both connectivity and health literacy challenges. As evidenced by the current global pandemic, access to reliable online health-related information and the ability to apply that information is critical to achieving health equity. Through a qualitative study on how Latino families collaborate to access online health resources, this work frames health literacy as a family-level mechanism. Interviews with parent-child dyads combined with online search tasks reveal how families integrate their individual skillsets to obtain, process, and understand online information about illnesses, symptoms, and even medical diagnoses. As they engage in intergenerational online health information searching and brokering, families creatively navigate information and communication technologies (ICTs) to address a range of health needs. Bilingual children help immigrant parents obtain urgent and non-urgent health information needed to care for other family members. When children are tasked with addressing a health need critical to their parent's wellbeing, they collaborate with their parents to obtain, interpret, and apply online health information. Intergenerational online health information searching and brokering thus reveals family-level strengths that can be leveraged to promote both health and digital literacy among marginalized populations.

Management of colovesical fistula: a systematic review.

INTRODUCTION: Colovesical fistulas (CVFs) account for approximately 95% enterovesical fistulas (EVFs). About 2/3 CVF cases are diverticular in origin. It mainly presents with urological signs such as pneumaturia and fecaluria. Diagnostic investigations aim at confirming the presence of a fistula. Although conservative management can be chosen for selected individuals, most patients are mainly treated through surgical interventions. CVF represents a challenging condition, which records high rates of morbidity and mortality. Our systematic review aimed at achieving deeper knowledge of both indications, in addition to short- and long-term outcomes related to CVF management. EVIDENCE ACQUISITION: We performed a systematic literature review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyzes (PRISMA) guidelines. Pubmed/MEDLINE, Embase, Scopus, Cochrane Library and Web of Science databases were used to search all related literature. EVIDENCE SYNTHESIS: The 22 included articles covered an approximately 37 years-study period (1982-2019), with a total 1365 patient population. CVF etiology was colonic diverticulitis in most cases (87.9%). Pneumaturia (50.1%), fecaluria (40.9%) and urinary tract infections (46.6%) were the most common symptoms. Abdomen computed tomography (CT) scan (80.5%), colonoscopy (74.5%) and cystoscopy (55.9%) were the most frequently performed diagnostic methods. Most CVF patients underwent surgery (97.1%) with open approach (63.3%). Almost all patients had colorectal resection with primary anastomosis with or without ostomy and 53.2% patients underwent primary repair or partial/total cystectomy. Four percent anastomotic leak, 1.8% bladder leak and 3.1% reoperations rates were identified. In an average 5-68-month follow-up, overall morbidity, overall mortality and recurrences rates recorded were 8-49%, 0-63% and 1.2%, respectively. CONCLUSIONS: CVF mainly affects males and has diverticular origin in almost all cases. Pneumaturia, fecaluria and urinary tract infections are the most characteristic symptoms. Endoscopic tests and imaging are critical tools for diagnostic completion. Management of CVFs depends on the underlying disease. Surgical treatment represents the final approach and consists of resection and reanastomosis of offending intestinal segment, with or without bladder closure. In many cases, a single-stage surgical strategy is selected. Perioperative and long-term outcomes prove good.

Depression Predicts Chronic Pain Interference in Racially Diverse, Income-Disadvantaged Patients.

BACKGROUND: Chronic pain is one of the most common reason adults seek medical care in the United States, with prevalence estimates ranging from 11% to 40%. Mindfulness meditation has been associated with significant improvements in pain, depression, physical and mental health, sleep, and overall quality of life. Group medical visits are increasingly common and are effective at treating myriad illnesses, including chronic pain. Integrative Medical Group Visits (IMGV) combine mindfulness techniques, evidence based integrative medicine, and medical group visits and can be used as adjuncts to medications, particularly in diverse underserved populations with limited access to non-pharmacological therapies. OBJECTIVE AND DESIGN: The objective of the present study was to use a blended analytical approach of machine learning and regression analyses to evaluate the potential relationship between depression and chronic pain in data from a randomized clinical trial of IMGV in diverse, income-disadvantaged patients suffering from chronic pain and depression. METHODS: The analytical approach used machine learning to assess the predictive relationship between depression and pain and identify and select key mediators, which were then assessed with regression analyses. It was hypothesized that depression would predict the pain outcomes of average pain, pain severity, and pain interference. RESULTS: Our analyses identified and characterized a predictive relationship between depression and chronic pain interference. This prediction was mediated by high perceived stress, low pain self-efficacy, and poor sleep quality, potential targets for attenuating the adverse effects of depression on functional outcomes. CONCLUSIONS: In the context of the associated clinical trial and similar interventions, these insights may inform future treatment optimization, targeting, and application efforts in racialized, income-disadvantaged populations, demographics often neglected in studies of chronic pain.

Influencia de un panel molecular de reacción de polimerasa en cadena múltiple en el diagnóstico o descarte de neumonía asociada a ventilación mecánica en una cohorte chilena de pacientes críticos con COVID-19

Resumen Introducción: La neumonía asociada a ventilación mecánica (NAVM) es frecuente en pacientes críticos con COVID-19. Su diagnóstico precoz es fundamental para su pronóstico. Objetivo: Demostrar la utilidad del panel FilmArray Pneumo (PFA-P) en la confirmación o descarte de NAVM en estos pacientes. Métodos: Estudio retrospectivo de 71 pacientes críticos con COVID-19 con sospecha de NAVM en quienes se realizó cultivos y PFA-P para confirmación diagnóstica. Se describen las características clínicas, microbiología y mortalidad. Se define la validez y seguridad de PFA-P. Resultados: El uso de FAP-P y cultivos descartó NAVM en 29 pacientes (40,8%). En 41 pacientes se confirmó NAVM y la mortalidad a 30 días fue 48,8%. Se estudiaron 48 muestras, los cultivos fueron positivos en 30 (62,5%) y se detectaron 33 bacterias, PFA-P detectó 32 de estas 33 bacterias; 37 bacterias fueron detectadas exclusivamente por PFA-P. Las bacterias prevalentes fueron Klebsiella pneumoniae (31,4%) Pseudomonas aeruginosa (21,4%) y Acinetobacter calcoaceticusbaumannii (14,2%). La sensibilidad, especificidad, valor predictor positivo y valor predictor negativo de PFA-P con respecto a cultivos fue 96,9%, 92,5%, 46,4% y 99,8, respectivamente. Un paciente tuvo NAVM por Burkholderia cepacia bacteria no detectada por FAP-P. Conclusiones: La FAP-P es una técnica molecular eficaz para descartar y diagnosticar la NAVM permitiendo una suspensión rápida de los antimicrobianos o un tratamiento dirigido temprano.

Abstract Background: Ventilator-associated pneumonia (VAP) is frequent in critical COVID-19 patients. Its early diagnosis is essential for its prognosis. Aim: To demonstrate the usefulness of the FilmArray Pneumo panel (FAP-P) in confirming or ruling out VAP in these patients. Methods: Retrospective study of 71 critical COVID-19 patients with suspected VAP in whom cultures and FAP-P were performed for diagnostic confirmation. Clinical characteristics, microbiology and mortality are described. The validity and safety of FAP-P is defined. Results: The use of FAP-P and cultures ruled out VAP in 29 patients (40.8%). In 41 patients, VAP was confirmed and the 30-day mortality was 48.8%. Forty-eight samples were studied, the cultures were positive in 30 (62.5%) and 33 bacteria were detected, FAP-P detected 32 of these 33 bacteria; 37 bacteria were exclusively detected by PFA-P. The most prevalent bacteria were Klebsiella pneumoniae (31.4%), Pseudomonas aeruginosa (21.4%) and Acinetobacter calcoaceticus-baumannii (14.2%). The sensitivity, specificity, positive predictive value and negative predictive value of FAP-P with respect to cultures were 96.9%, 92.5%, 46.4% and 99.8%, respectively. One patient had VAP due to Burkholderia cepacia bacteria not detected by FAP-P. Conclusions: FAP-P is an effective molecular technique to rule out and diagnose VAP, allowing rapid suspension of antibiotics or early targeted treatment.