High SARS-CoV-2 tropism and activation of immune cells in the testes of non-vaccinated deceased COVID-19 patients.

BACKGROUND: Cellular entry of SARS-CoV-2 has been shown to rely on angiotensin-converting enzyme 2 (ACE2) receptors, whose expression in the testis is among the highest in the body. Additionally, the risk of mortality seems higher among male COVID-19 patients, and though much has been published since the first cases of COVID-19, there remain unanswered questions regarding SARS-CoV-2 impact on testes and potential consequences for reproductive health. We investigated testicular alterations in non-vaccinated deceased COVID-19-patients, the precise location of the virus, its replicative activity, and the immune, vascular, and molecular fluctuations involved in the pathogenesis. RESULTS: We found that SARS-CoV-2 testicular tropism is higher than previously thought and that reliable viral detection in the testis requires sensitive nanosensors or RT-qPCR using a specific methodology. Through an in vitro experiment exposing VERO cells to testicular macerates, we observed viral content in all samples, and the subgenomic RNA's presence reinforced the replicative activity of SARS-CoV-2 in testes of the severe COVID-19 patients. The cellular structures and viral particles, observed by transmission electron microscopy, indicated that macrophages and spermatogonial cells are the main SARS-CoV-2 lodging sites, where new virions form inside the endoplasmic reticulum Golgi intermediate complex. Moreover, we showed infiltrative infected monocytes migrating into the testicular parenchyma. SARS-CoV-2 maintains its replicative and infective abilities long after the patient's infection. Further, we demonstrated high levels of angiotensin II and activated immune cells in the testes of deceased patients. The infected testes show thickening of the tunica propria, germ cell apoptosis, Sertoli cell barrier loss, evident hemorrhage, angiogenesis, Leydig cell inhibition, inflammation, and fibrosis. CONCLUSIONS: Our findings indicate that high angiotensin II levels and activation of mast cells and macrophages may be critical for testicular pathogenesis. Importantly, our findings suggest that patients who become critically ill may exhibit severe alterations and harbor the active virus in the testes.

Detection of DENV-2 and ZIKV coinfection in southeastern Brazil by serum and urine testing.

PURPOSE: Aedes aegypti mosquito-borne diseases have a significant impact on public health in Brazil. In this study, we investigated the presence of the Zika virus (ZIKV) and dengue virus (DENV) in serum and urine samples from symptomatic participants who attended an Emergency Care Unit located in a city in the northwestern region of São Paulo between February 2018 and April 2019. METHODS: Serum and urine samples were collected from participants suspected of having arbovirus infection. After the extraction of viral RNA, viral detection was performed by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) (One-Step RT-qPCR). RESULTS: A total of 305 participants participated in this study. A total of 283 blood and 270 urine samples were collected. Of 305 patients, 36.4% (111/305) were positive for ZIKV, 43.3% (132/305) for DENV2, and 0.3% (1/305) for DENV1. Coinfection with ZIKV/DENV2 was observed in 13.1% of participants. If only serum samples were used, ZIKV detection would have decreased to 23.3% (71/305). Of all the participants included in the study, only one was suspected of having ZIKV infection based on clinical diagnosis, and the remaining participants were suspected of having DENV. CONCLUSION: By testing serum and urine samples, we increased the detection of both viruses and detected considerable levels of ZIKV and DENV-2 coinfection when compared to other studies. Additionally, we detected an unnoticed ZIKV outbreak in the city. These findings highlight the importance of the molecular diagnosis of arboviruses to aid public health surveillance and management strategies.

Antiviral Evaluation of New Synthetic Bioconjugates Based on GA-Hecate: A New Class of Antivirals Targeting Different Steps of Zika Virus Replication.

Re-emerging arboviruses represent a serious health problem due to their rapid vector-mediated spread, mainly in urban tropical areas. The 2013-2015 Zika virus (ZIKV) outbreak in South and Central America has been associated with cases of microcephaly in newborns and Guillain-Barret syndrome. We previously showed that the conjugate gallic acid-Hecate (GA-FALALKALKKALKKLKKALKKAL-CONH2)-is an efficient inhibitor of the hepatitis C virus. Here, we show that the Hecate peptide is degraded in human blood serum into three major metabolites. These metabolites conjugated with gallic acid were synthesized and their effect on ZIKV replication in cultured cells was evaluated. The GA-metabolite 5 (GA-FALALKALKKALKKL-COOH) was the most efficient in inhibiting two ZIKV strains of African and Asian lineage at the stage of both virus entry (virucidal and protective) and replication (post-entry). We also demonstrate that GA-metabolite 5 does not affect cell growth after 7 days of continuous treatment. Thus, this study identifies a new synthetic antiviral compound targeting different steps of ZIKV replication in vitro and with the potential for broad reactivity against other flaviviruses. Our work highlights a promising strategy for the development of new antivirals based on peptide metabolism and bioconjugation.

Detection of Zika virus in urine from randomly tested individuals in Mirassol, Brazil.

PURPOSE: Studies show that around 80% of Zika virus (ZIKV) infections are asymptomatic. The present study tested urine samples from volunteers, unsuspected of arboviral infection, which attended an emergency care unit (ECU) in Mirassol, Brazil, from March 2018 to April 2019. METHODS: The volunteers were divided into two groups. The first group was composed of outpatients who were not suspected to have an arbovirus infection. This first group was subdivided into two subgroups: outpatients with and without arbovirus-like symptoms. The second group consisted of companions of outpatients treated at the ECU. The second group was also subdivided into two subgroups: totally asymptomatic individuals and those who had arbovirus-like symptoms. RNA was extracted from urine samples, followed by RT-qPCR for ZIKV. RESULTS: We found that 11% (79/697) of the samples tested positive for ZIKV-RNA. Among the ZIKV-RNA-positive individuals, 16.5% (13/79) were companions, of which 61.5% (8/13) were totally asymptomatic and 38.5% (5/13) reported symptoms that could be suggestive of arbovirus infection. In addition, 83.5% (66/79) of the ZIKV-RNA-positive individuals were outpatients without a clinical diagnosis of arbovirus. Of these undiagnosed ZIKV-RNA-positive outpatients, 47% (31/66) had no arbovirus-related symptoms. CONCLUSION: Our study shows the effectiveness of urine as a non-invasive sample to detect the incidence of ZIKV infection. We also highlight the importance of ZIKV molecular diagnosis to aid public health surveillance and prevention of congenital Zika syndrome and other ZIKV-associated diseases.

A novel substitution in NS5A enhances the resistance of hepatitis C virus genotype 3 to daclatasvir.

Hepatitis C virus (HCV) genotype 3 presents a high level of both baseline and acquired resistance to direct-acting antivirals (DAAs), particularly those targeting the NS5A protein. To understand this resistance we studied a cohort of Brazilian patients treated with the NS5A DAA, daclatasvir and the nucleoside analogue, sofosbuvir. We observed a novel substitution at NS5A amino acid residue 98 [serine to glycine (S98G)] in patients who relapsed post-treatment. The effect of this substitution on both replication fitness and resistance to DAAs was evaluated using two genotype 3 subgenomic replicons. S98G had a modest effect on replication, but in combination with the previously characterized resistance-associated substitution (RAS), Y93H, resulted in a significant increase in daclatasvir resistance. This result suggests that combinations of substitutions may drive a high level of DAA resistance and provide some clues to the mechanism of action of the NS5A-targeting DAAs.

Identification of differentially expressed miRNAs in human cells infected with different Zika virus strains.

Infection with distinct Zika virus (ZIKV) strains in in vitro and in vivo models has demonstrated that the host's response to infection is strain-dependent. There has been no analysis of the impact of infection with different ZIKV strains on miRNA expression in human cells. We investigated miRNA expression in PNT1A cells upon infection with an African ZIKV strain (MR766) and a Brazilian ZIKV strain (ZIKVBR) using PCR array. Sixteen miRNAs were modulated in PNT1A cells: six miRNAs were modulated by both strains, while a set of ten miRNAs were modulated exclusively by ZIKVBR infection. In silico analysis showed that nine significant KEGG pathways and eight significant GO terms were predicted to be enriched upon ZIKVBR infection, and these pathways were related to cancer, environmental information processing, metabolism, and extracellular matrix. Differential modulation of miRNA expression suggests that distinct strains of ZIKV can differentially modulate the host response through the action of miRNAs.

Alphacoronavirus Detection in Lungs, Liver, and Intestines of Bats from Brazil.

Bats are flying mammals distributed worldwide known to host several types of Coronavirus (CoV). Since they were reported as the probable source of spillover of highly pathogenic CoV into the human population, investigating the circulation of this virus in bats around the world became of great importance. We analyzed samples from 103 bats from two distinct regions in Brazil. Coronavirus from the Alphacoronavirus genus was detected in 12 animals, 11 from São José do Rio Preto-SP region and 1 from Barreiras-BA region, resulting in a prevalence of 17.18% and 2.56% respectively. The virus was detected not only in intestines but also in lungs and liver. Phylogenetic analysis based on nsP12 genomic region suggests that the sequences group according to host family and sampling location. Studies on the circulation of these viruses in bats remain important to understand the ecology and evolutionary relationship of these pathogens.

Hepatitis C virus in vitro replication is efficiently inhibited by acridone Fac4.

Hepatitis C virus (HCV) affects about 170 million people worldwide. The current treatment has a high cost and variable response rates according to the virus genotype. Acridones, a group of compounds extracted from natural sources, showed potential antiviral actions against HCV. Thus, this study aimed to evaluate the effect of a panel of 14 synthetic acridones on the HCV life cycle. The compounds were screened using an Huh7.5 cell line stably harbouring the HCV genotype 2a subgenomic replicon SGR-Feo-JFH-1. Cells were incubated in the presence or absence of compounds for 72 h and cell viability and replication levels were assessed by MTT and luciferase assays, respectively. At a concentration of 5 µM the acridone Fac4 exhibited a >90 % inhibition of HCV replication with no effect on cell viability. The effects of Fac4 on virus replication, entry and release steps were evaluated in Huh7.5 cells infected with the JFH-1 isolate of HCV (HCVcc). Fac4 inhibited JFH-1 replication to approximately 70 %, while no effect was observed on virus entry. The antiviral activity of Fac4 was also observed on viral release, with almost 80 % of inhibition. No inhibitory effect was observed against genotype 3 replication. Fac4 was able to intercalate into dsRNA, however did not inhibit NS5B polymerase activity or translation driven by the HCV IRES. Although its mode of action is partly understood, Fac4 presents significant inhibition of HCV replication and can therefore be considered as a candidate for the development of a future anti-HCV treatment.

Genome sequencing and genetic characterization of Culex Flavirirus (CxFV) provides new information about its genotypes.

BACKGROUND: Culex Flavivirus (CxFV) is an insect-specific virus that is widely distributed and primarily infects mosquito species from the genus Culex. Its hosts include Culex tritaeniorhynchus, Culex quinquefasciatus, and Anopheles sinensis mosquitoes. Since its original identification, CxFV has been reported in several countries. Despite the increasing number of reports on CxFV, little is known about its genomic characteristics. It is unclear whether the phylogenetic relationships between the strains are influenced by host species and geographic location. RESULTS: We characterized the Brazilian CxFV strain and performed a comprehensive genetic and phylogenetic characterization of CxFV based on all ORF sequences described so far. Our results revealed that the Brazilian strain is in a monophyletic clade with the Mexican strain. Overall, selective pressure indicates that the ORF is undergoing purifying selection. CONCLUSIONS: The phylogenetic analysis revealed a strong association between climate and CxFV ancestry. Also, based on phylogeny and the genetic distance between the main branches of the tree, we propose the classification of the available sequences into two different genotypes. We also suggest the existence of two different subtypes within Genotype 1.

CLINICAL AND FUNCTIONAL EVALUATION OF WRISTS AND HANDS OF SPINAL CORD INJURED PATIENTS.

Introduction: The inability of the spinal cord to propagate sensory and motor stimuli as a result of the disruption of the nerve tracts is called spinal cord injury. Objective: This study analyzes clinically and radiologically the hands and wrists of spinal cord injured patients, evaluating their motor and sensitive functionality, in order to determine if these patients are more likely to develop degenerative alterations. Methods: 14 patients (8 paraplegics and 6 tetraplegics) were evaluated, undergoing anamnesis and clinical examination - a scale of muscular strength (MRC - Medical Research Council) and the amplitude measurement of the movement with a manual goniometer (ROM), were used for objective evaluation - and x-ray exams. The results were compared with pre-existing data from other studies. Results: When asked, only one of the 14 observed patients complained about constant wrist pain, described as level 3 (weak to moderate), based on the visual analog scale (VAS). The motor evaluation, MRC and ROM divided the group of patients into two subgroups: paraplegic and tetraplegic patients. The x-ray analysis showed, based on Kellgren and Lawrence classification, that all exam images fit grades 1 or 2 of osteoarthritis and osteoarthrosis. Conclusion: In conclusion, spinal cord injured patients showed none or minimal clinical and radiological signs of osteoarthritis on hands or wrists. Overall, the hands and wrists of spinal cord-injured patients behave similarly to noninjured patients. Level of Evidence III; Retrospective Comparative Study.

Introdução: A incapacidade da medula espinhal de propagar estímulos sensoriais e motores como resultado do rompimento das vias nervosas é chamada de lesão da medula espinhal. Objetivo: Este estudo analisa clínica e radiograficamente mão e punhos de pacientes lesados medulares, avaliando função motora e sensitiva, a fim de determinar se estes pacientes estariam mais propensos a alterações degenerativas. Métodos: 14 pacientes (8 paraplégicos e 6 tetraplégicos) foram avaliados, passando por anamnese e exame clínico - sendo escala de força muscular (MRC - Medical Research Council) e a medição da amplitude de movimento com um goniômetro manual (ROM) foram utilizados para análise objetiva - e radiografias. Os dados obtidos foram comparados com literatura preexistente. Resultados: Quando questionados, apenas um dos 14 pacientes observados referiu dor crônica nos punhos, descrita como nível 3 (fraca a moderada), baseada na escala visual analógica. A avaliação motora, MRC e ROM dividiram os pacientes em 2 subgrupos: pacientes paraplégicos e tetraplégicos. A análise radiográfica mostrou, baseada na classificação de Kellgren e Lawrence, que todas as imagens se encaixam nos graus 1 ou 2 de osteoartrite e osteoartrose. Conclusão: Conclui-se, então, que pacientes lesados medulares apresentam nenhuma ou mínimas alterações clínicas e radiológicas para osteoartrite ou osteoartrose de punhos ou mãos. Ou seja, no geral, mãos e punhos de pacientes lesados medulares comportam-se como os pacientes sem lesão medular. Nível de Evidência III; Estudo Retrospectivo Comparativo.

Cost and time of hospitalization for elderly people with bone fractures in a reference hospital.

OBJECTIVE: To describe and analyze the aspects regarding the cost and length of stay for elderly patients with bone fractures in a tertiary reference hospital. METHODS: A cross-sectional retrospective study using data obtained from medical records between January and December 2020. For statistical analysis, exploratory analyses, Shapiro-Wilk test, χ2 test, and Spearman correlation were used. RESULTS: During the study period, 156 elderly patients (62.2% women) with bone fractures were treated. The main trauma mechanism was a fall from a standing height (76.9%). The most common type of fracture in this sample was a transtrochanteric fracture of the femur, accounting for 40.4% of cases. The mean length of stay was 5.25 days. The total cost varied between R$2,006.53 and R$106,912.74 (average of R$15,695.76) (updated values). The mean daily cost of hospitalization was R$4,478.64. A positive correlation was found between the length of stay and total cost. No significant difference in cost was observed between the two main types of treated fractures. CONCLUSION: Fractures in the elderly are frequent, resulting in significant costs. The longer the hospital stay for treatment, the higher the total cost. No correlation was found between total cost and number of comorbidities, number of medications used, and the comparison between the treatment of transtrochanteric and femoral neck fractures.

The GA-Hecate Peptide inhibits the ZIKV Replicative Cycle in Different Steps and can Inhibit the Flavivirus NS2B-NS3 Protease after Cell Infection.

BACKGROUND: Peptide drugs are advantageous because they are subject to rational design and exhibit highly diverse structures and broad biological activities. The NS2B-NS3 protein is a particularly promising flavivirus therapeutic target, with extensive research on the development of inhibitors as therapeutic candidates, and was used as a model in this work to determine the mechanism by which GA-Hecate inhibits ZIKV replication. OBJECTIVE: The present study aimed to evaluate the potential of GA-Hecate, a new antiviral developed by our group, against the Brazilian Zika virus and to evaluate the mechanism of action of this compound on the flavivirus NS2B-NS3 protein. METHODS: Solid-phase peptide Synthesis, High-Performance Liquid Chromatography, and Mass Spectrometry were used to obtain, purify, and characterize the synthesized compound. Real-time and enzymatic assays were used to determine the antiviral potential of GA-Hecate against ZIKV. RESULTS: The RT-qPCR results showed that GA-Hecate decreased the number of ZIKV RNA copies in the virucidal, pre-treatment, and post-entry assays, with 5- to 6-fold fewer RNA copies at the higher nontoxic concentration in Vero cells (HNTC: 10 µM) than in the control cells. Enzymatic and kinetic assays indicated that GA-Hecate acts as a competitive ZIKV NS2B-NS3 protease inhibitor with an IC50 of 32 nM and has activity against the yellow fever virus protease. CONCLUSION: The results highlight the antiviral potential of the GA-Hecate bioconjugate and open the door for the development of new antivirals.

Transcription of HIV-1 at sites of intact latent provirus integration.

HIV-1 anti-retroviral therapy is highly effective but fails to eliminate a reservoir of latent proviruses leading to a requirement for life-long treatment. How the site of integration of authentic intact latent proviruses might impact their own or neighboring gene expression or reservoir dynamics is poorly understood. Here we report on proviral and neighboring gene transcription at sites of intact latent HIV-1 integration in cultured T cells obtained directly from people living with HIV, as well as engineered primary T cells and cell lines. Proviral gene expression was correlated to the level of endogenous gene expression under resting but not activated conditions. Notably, latent proviral promoters were 10010,000X less active than in productively infected cells and had little or no measurable impact on neighboring gene expression under resting or activated conditions. Thus, the site of integration has a dominant effect on the transcriptional activity of intact HIV-1 proviruses in the latent reservoir thereby influencing cytopathic effects and proviral immune evasion.

Cryptic circulation of chikungunya virus in São Jose do Rio Preto, Brazil, 2015-2019.

BACKGROUND: Chikungunya virus (CHIKV) has spread across Brazil with varying incidence rates depending on the affected areas. Due to cocirculation of arboviruses and overlapping disease symptoms, CHIKV infection may be underdiagnosed. To understand the lack of CHIKV epidemics in São José do Rio Preto (SJdRP), São Paulo (SP), Brazil, we evaluated viral circulation by investigating anti-CHIKV IgG seroconversion in a prospective study of asymptomatic individuals and detecting anti-CHIKV IgM in individuals suspected of dengue infection, as well as CHIKV presence in Aedes mosquitoes. The opportunity to assess two different groups (symptomatic and asymptomatic) exposed at the same geographic region aimed to broaden the possibility of identifying the viral circulation, which had been previously considered absent. METHODOLOGY/PRINCIPAL FINDINGS: Based on a prospective population study model and demographic characteristics (sex and age), we analyzed the anti-CHIKV IgG seroconversion rate in 341 subjects by ELISA over four years. The seroprevalence increased from 0.35% in the first year to 2.3% after 3 years of follow-up. Additionally, we investigated 497 samples from a blood panel collected from dengue-suspected individuals during the 2019 dengue outbreak in SJdRP. In total, 4.4% were positive for anti-CHIKV IgM, and 8.6% were positive for IgG. To exclude alphavirus cross-reactivity, we evaluated the presence of anti-Mayaro virus (MAYV) IgG by ELISA, and the positivity rate was 0.3% in the population study and 0.8% in the blood panel samples. In CHIKV and MAYV plaque reduction neutralization tests (PRNTs), the positivity rate for CHIKV-neutralizing antibodies in these ELISA-positive samples was 46.7%, while no MAYV-neutralizing antibodies were detected. Genomic sequencing and phylogenetic analysis revealed CHIKV genotype ECSA in São José do Rio Preto, SP. Finally, mosquitoes collected to complement human surveillance revealed CHIKV positivity of 2.76% of A. aegypti and 9.09% of A. albopictus (although it was far less abundant than A. aegypti) by RT-qPCR. CONCLUSIONS/SIGNIFICANCE: Our data suggest cryptic CHIKV circulation in SJdRP detected by continual active surveillance. These low levels, but increasing, of viral circulation highlight the possibility of CHIKV outbreaks, as there is a large naïve population. Improved knowledge of the epidemiological situation might aid in outbreaks prevention.

Analysis of HCV quasispecies dynamic under selective pressure of combined therapy.

BACKGROUND: The quasispecies composition of Hepatitis C virus (HCV) could have important implications with regard to viral persistence and response to interferon-based therapy. The complete NS5A was analyzed to evaluate whether the composition of NS5A quasispecies of HCV 1a/1b is related to responsiveness to combined interferon pegylated (PEG-IFN) and ribavirin therapy. METHODS: Viral RNA was isolated from serum samples collected before, during and after treatment from virological sustained responder (SVR), non-responder (NR) and the end-of-treatment responder patients (ETR). NS5A region was amplified, cloned and sequenced. Six hundred and ninety full-length NS5A sequences were analyzed. RESULTS: This study provides evidence that lower nucleotide diversity of the NS5A region pre-therapy is associated with viral clearance. Analysis of samples of NRs and the ETRs time points showed that genetic diversity of populations tend to decrease over time. Post-therapy population of ETRs presented higher genetic distance from baseline probably due to the bottleneck phenomenon observed for those patients in the end of treatment. The viral effective population of those patients also showed a strong decrease after therapy. Otherwise, NRs demonstrated a continuous variation or stability of effective populations and genetic diversity over time that did not seem to be related to therapy. Phylogenetic relationships concerning complete NS5A sequences obtained from patients did not demonstrate clustering associated with specific response patterns. However, distinctive clustering of pre/post-therapy sequences was observed. In addition, the evolution of quasispecies over time was subjected to purifying or relaxed purifying selection. Codons 157 (P03), 182 and 440 (P42), 62 and 404 (P44) were found to be under positive selective pressure but it failed to be related to the therapy. CONCLUSION: These results confirm the hypothesis that a relationship exists between NS5A heterogeneity and response to therapy in patients infected with chronic hepatitis C.

Kinetic and kinematic evaluation of the ankle joint after achilles tendon reconstruction with free semitendinosus tendon graft: preliminary results.

Chronic rupture of the Achilles tendon (AT) is a surgical challenge and has effects on the gait. The purpose of this study was to evaluate the kinetic and kinematic parameters of the ankle joint in patients with AT rupture operated using a free semitendinosus tendon graft. Thirteen patients were analyzed 6 and 12 months after surgery in a force platform, while the movements were recorded by six infrared cameras. The kinematic variables analyzed included speed, cadence, step length, percentage of stance phase, and range of movement (ROM) of the ankle joint in the sagittal and frontal planes. Kinetic data were obtained by joint movement in different phases of the gait cycle. Functional assessment was performed using the American Orthopaedic Foot and Ankle Society (AOFAS) score. The patients showed a significant increase (P = 0.0215) in AOFAS from 68.5 (±18.7) to 85.2 (±18.0). Speed, cadence, and length of step of the four groups (1A, 1B, 2A, 2B) were lower than the control group (group 3), and the percentage in stance phase was higher for the nonoperated foot 6-month group (1B) compared to the control group (group 3). For the kinematic data, the ROM of the ankle in stance phase increased from 6 to 12 months showing an effect of time between four groups (1A, 1B, 2A, 2B). During swing phase, the ankle ROM was lower in the operated side (effect of side, P = 0.0255) and groups 1A and 2A demonstrated statistical differences when compared with the control group (group 3) (P = 0.0240 and P = 0.0414, respectively). ROM of inversion and eversion presented effect of time among the same groups (P = 0.0059) cited before. There were no differences in kinetic data between groups. This study showed close proximity between the control group and the operated group. Furthermore, improvement was shown when comparing the 6 and 12 months postsurgery periods. The surgical procedure is therefore helpful for the patient and few changes were present in gait and ankle biomechanics.

Tibiotalar arthrodesis in posttraumatic arthritis using the tension band technique.

BACKGROUND: Although a variety of surgical techniques are available for the treatment of ankle degenerative disease, arthrodesis is a common treatment especially for unilateral posttraumatic arthritis in young patients. A clear trend toward internal fixation for ankle arthrodesis is evident, but fusion site compression is known to play an integral role in primary bone healing. METHODS: Between September 2001 and October 2009, 17 adults with painful end-stage posttraumatic ankle arthritis were treated with the tension band technique to achieve tibiotalar arthrodesis. RESULTS: Tibiotalar fusion was obtained in all patients at a mean of 16.6 (range, 13 to 28) weeks. No patient had pain at the last postoperative follow-up examination. AOFAS and Mazur scores were preoperatively 29 and 23.1, respectively, and postoperatively 71.1 and 70.7, respectively. CONCLUSION: Arthrodesis with a tension band was a safe and effective surgical treatment option for posttraumatic ankle arthritis in patients without infection and good bone density, as it was simple, less invasive, and inexpensive and allowed early full weight-bearing. LEVEL OF EVIDENCE: Level IV, retrospective case series.

African ZIKV lineage fails to sustain infectivity in an in vitro mimetic urban cycle.

Zika virus (ZIKV) is an arbovirus maintained in nature in two distinct cycles of transmission: urban and sylvatic. Each cycle includes specific vertebrate and invertebrate hosts, and through alternate infections, a conserved consensus sequence is maintained that might vary depending on the cycle. The current study aimed to investigate the ability of ZIKVAF and ZIKVBR to maintain an infectious cycle by alternating passages in cells mimicking the urban (UC) and semi-sylvatic (SC) cycles. The complete genome of the original inoculum and the last passages for each cycle were sequenced by Sanger. Ten passages were performed, as planned, for ZIKVBR UC, ZIKVAF SC, and ZIKVBR SC. ZIKVBR SC showed significant variation in viral titers along the passages, suggesting that the virus is not well adapted to the non-human primate host. ZIKVAF passage in UC was abrogated in the third passage, showing the inability of the African lineage to sustain cycles in human cells, suggesting a low capacity to establish an urban cycle. Several mutations were found in both strains along the passages, but not occurring at equivalent positions. Further studies are needed to elucidate whether any of these specific mutations affect viral fitness. ZIKV strains behave differently in artificial transmission cycles in vitro: Brazilian ZIKV was able to establish urban and semi-sylvatic cycles in vitro. African ZIKV proved unable to cycle among human and mosquito cells and is compatible only with the semi-sylvatic cycle. The main mutations arose in the NS2A region after artificial transmission cycles for both ZIKV strains but not at equivalent positions.

COST ANALYSIS OF MOTORCYCLE ACCIDENT VICTIMS AT A UNIVERSITY HOSPITAL: PERSPECTIVES FROM 2017 AND 2020.

Introduction: Motorcycle accidents constitute a public health problem that affects public and private health services due to the expenses of the victim's treatment and rehabilitation. Objective: Evaluate the impact of motorcycle accident costs in a university hospital in 2020. Method: Comparative analysis of the costs of motorcycle accident patients in 2020 and 2017. Results: Among 151 patients included in the study, the average cost was U$3,083.54, and the average days of hospitalization were 5.3 days. The patient with the highest cost to the hospital spent U$22,504.05, and the patient with the lowest cost spent U$356.72. The longest stay among these patients was 41 days, and the shortest was one day. The average cost per patient per day for the entire sample was U$581.80. Conclusion: The formulation and application of strategies that promote the reduction of motorcycle accidents in the city of Campinas are necessary. Level of evidence II, Retrospective study.

Introdução: Acidentes motociclísticos constituem um problema de saúde pública que atinge os serviços públicos e privados de saúde, em função dos gastos com o tratamento e com o processo de reabilitação da vítima. Objetivo: Avaliar o impacto dos custos dos acidentes motociclísticos em um hospital universitário em 2020. Método: Análise comparativa dos custos dos pacientes vítimas de acidente motociclístico no ano de 2020 e 2017. Resultados: Dentre 151 pacientes incluídos no estudo, o custo médio foi de U$3.083,54 e a média de dias de internação foi de 5,3 dias. O paciente que apresentou maior custo para o hospital, teve um gasto de U$22.504,05 e o que teve o menor custo, gastou U$356,72. O maior tempo de internação, entre estes pacientes, foi de 41 dias e o menor tempo foi de 1 dia. O custo médio por paciente por dia, em toda a amostra, foi de U$581,80. Conclusão: Faz-se necessário a formulação e aplicação de estratégias que promovam a redução dos acidentes motociclísticos na cidade de Campinas. Nível de evidência II; Estudo retrospectivo.