Characterization of Systemic Disease Development and Paw Inflammation in a Susceptible Mouse Model of Mayaro Virus Infection and Validation Using X-ray Synchrotron Microtomography

Mayaro virus (MAYV) is an emerging arthropod-borne virus endemic in Latin America and the causative agent of arthritogenic febrile disease. Mayaro fever is poorly understood; thus, we established an in vivo model of infection in susceptible type-I interferon receptor-deficient mice (IFNAR−/−) to characterize the disease. MAYV inoculations in the hind paws of IFNAR−/− mice result in visible paw inflammation, evolve into a disseminated infection and involve the activation of immune responses and inflammation. The histological analysis of inflamed paws indicated edema at the dermis and between muscle fibers and ligaments. Paw edema affected multiple tissues and was associated with MAYV replication, the local production of CXCL1 and the recruitment of granulocytes and mononuclear leukocytes to muscle. We developed a semi-automated X-ray microtomography method to visualize both soft tissue and bone, allowing for the quantification of MAYV-induced paw edema in 3D with a voxel size of 69 µm3. The results confirmed early edema onset and spreading through multiple tissues in inoculated paws. In conclusion, we detailed features of MAYV-induced systemic disease and the manifestation of paw edema in a mouse model extensively used to study infection with alphaviruses. The participation of lymphocytes and neutrophils and expression of CXCL1 are key features in both systemic and local manifestations of MAYV disease

Virus Identification in Unknown Tropical Febrile Illness Cases Using Deep Sequencing

Dengue virus is an emerging infectious agent that infects an estimated 50–100 million people annually worldwide, yet current diagnostic practices cannot detect an etiologic pathogen in ∼40% of dengue-like illnesses. Metagenomic approaches to pathogen detection, such as viral microarrays and deep sequencing, are promising tools to address emerging and non-diagnosable disease challenges. In this study, we used the Virochip microarray and deep sequencing to characterize the spectrum of viruses present in human sera from 123 Nicaraguan patients presenting with dengue-like symptoms but testing negative for dengue virus. We utilized a barcoding strategy to simultaneously deep sequence multiple serum specimens, generating on average over 1 million reads per sample. We then implemented a stepwise bioinformatic filtering pipeline to remove the majority of human and low-quality sequences to improve the speed and accuracy of subsequent unbiased database searches. By deep sequencing, we were able to detect virus sequence in 37% (45/123) of previously negative cases. These included 13 cases with Human Herpesvirus 6 sequences. Other samples contained sequences with similarity to sequences from viruses in the Herpesviridae, Flaviviridae, Circoviridae, Anelloviridae, Asfarviridae, and Parvoviridae families. In some cases, the putative viral sequences were virtually identical to known viruses, and in others they diverged, suggesting that they may derive from novel viruses. These results demonstrate the utility of unbiased metagenomic approaches in the detection of known and divergent viruses in the study of tropical febrile illness

Goodbye Hartmann trial: a prospective, international, multicenter, observational study on the current use of a surgical procedure developed a century ago

Background: Literature suggests colonic resection and primary anastomosis (RPA) instead of Hartmann's procedure (HP) for the treatment of left-sided colonic emergencies. We aim to evaluate the surgical options globally used to treat patients with acute left-sided colonic emergencies and the factors that leading to the choice of treatment, comparing HP and RPA. Methods: This is a prospective, international, multicenter, observational study registered on ClinicalTrials.gov. A total 1215 patients with left-sided colonic emergencies who required surgery were included from 204 centers during the period of March 1, 2020, to May 31, 2020. with a 1-year follow-up. Results: 564 patients (43.1%) were females. The mean age was 65.9 ± 15.6 years. HP was performed in 697 (57.3%) patients and RPA in 384 (31.6%) cases. Complicated acute diverticulitis was the most common cause of left-sided colonic emergencies (40.2%), followed by colorectal malignancy (36.6%). Severe complications (Clavien-Dindo ≥ 3b) were higher in the HP group (P < 0.001). 30-day mortality was higher in HP patients (13.7%), especially in case of bowel perforation and diffused peritonitis. 1-year follow-up showed no differences on ostomy reversal rate between HP and RPA. (P = 0.127). A backward likelihood logistic regression model showed that RPA was preferred in younger patients, having low ASA score (≤ 3), in case of large bowel obstruction, absence of colonic ischemia, longer time from admission to surgery, operating early at the day working hours, by a surgeon who performed more than 50 colorectal resections. Conclusions: After 100 years since the first Hartmann's procedure, HP remains the most common treatment for left-sided colorectal emergencies. Treatment's choice depends on patient characteristics, the time of surgery and the experience of the surgeon. RPA should be considered as the gold standard for surgery, with HP being an exception

Grau de exposição a praguicidas organoclorados em moradores de aterro a céu aberto Levels of exposure to organochlorine pesticides in open-air dump dwellers

OBJETIVO: Estimar o grau de exposição interna aos praguicidas organoclorados (POP) dos moradores de um atêrro a céu aberto. MÉTODOS: Estudo observacional, de aferição simultânea e controlado, cujo fator de estudo foi residir em área contaminada por POP e o efeito foi a concentração sanguínea dessas substâncias. A população de estudo foi de 238 pessoas residentes em um aterro a céu aberto (Pilões), no município de Cubatão, SP; e a população de controle ficou dimensionada em 258 pessoas, também residentes no mesmo município (Cota 200). Foram analisados os praguicidas HCB, p-p'DDT, p-p'DDE, p-p'DDD, o-p'DDT, a HCH; b HCH; g HCH; Aldrin; Dieldrin; Endrin; Heptaclor; Heptaclor-epoxi e Mirex. RESULTADOS: Os teores médios sangüíneos de HCB em Pilões foram de 4,66 µg/L, 155 vezes maior que a média na Cota 200 (0,03 µg/L). Em Pilões, os teores médios de DDT total foram de 3,71 µg/L, duas vezes maior do que na Cota 200 (1,85 µg/L) e o HCH total apresentou concentrações sangüíneas seis vezes maiores em Pilões, 0,84 µg/L, contra 0,13 µg/L. CONCLUSÕES: Evidenciou-se associação positiva entre residir em Pilões e apresentar teores sangüíneos de POP, com risco muitas vezes maior quando comparado a localidades sem a presença desses contaminantes.<br>OBJECTIVE: To estimate the degree of internal exposure to organochlorine pesticides (persistent organic pollutants, POP) among inhabitants of an open-air dump. METHODS: This was an observational study with simultaneous measurement and control, in which the criterion for subjects was that they dwelled in an area contaminated by POP and the effect of such contamination was the concentration of such substances into the blood. The study population consisted of 238 people living on an open-air dump in Pilões, in the municipality of Cubatão, SP, Brazil, and the control population was formed by 258 people, also resident in that municipality, in the Cota 200 district. The following pesticides were analysed: hexachlorobenzene (HCB), p-p'DDT, p-p'DDE, p-p'DDD, o-p'DDT, a HCH (hexachlorocyclohexane); b HCH; g HCH; aldrin; dieldrin; endrin; heptachlor; heptachlor-epoxy and mirex. RESULTS: The average blood level of HCB in Pilões was 4.66 µg/l, which was 155 times higher than the average for Cota 200 (0.03 µg/l). In Pilões the average blood concentration of total DDT was 3.71 µg/l, which was twice the level found in Cota 200 (1.85 µg/l). Total HCH presented blood concentrations that were six times greater in Pilões (0.84 µg/l versus 0.13 µg/l). CONCLUSIONS: A positive association was shown between dwelling in Pilões and the presence of POP in the blood, with much higher risks than in localities without the presence of these contaminants