Ribeirao Preto Research Articles

P093 PLASMA PROTEINS PROFILE AND THERAPEUTIC RESPONSIVENESS IN PATIENTS WITH PREECLAMPSIA

Preeclampsia (PE) is a hypertensive syndrome of pregnancy, characterized by target organ damage, related to a high incidence of cardiovascular sequelae. PE requires antihypertensive drug therapy, however, about 40% of patients do not respond to this treatment, leading to worse clinical outcomes. Through the proteomic strategy, we 1) compared the profile of circulating proteins expressed in 10 responsive (R-PE) and 10 non-responsive to antihypertensive therapy (NR-PE), besides 10 healthy pregnant (HP); 2) investigated which proteins are differentially expressed between the groups and 3) found out which signaling pathways are altered between them. The study followed the Declaration of Helsinki and was approved by the Research Ethics Committee of the Ribeirao Preto Medical School, University of Sao Paulo (FMRP-USP, protocol code CAAE-37738620.0.0000.5440). We performed plasma protein quantification using mass spectrometry, the obtained data underwent bioinformatics analyses based on Uniprot, PatternLab for Proteomics, String and MetaboAnalyst softwares. Considering a fold change of 1.5, three proteins showed as significantly differentially expressed between HP and R-PE, one downregulated (transtyretin) and two upregulated (apolipoprotein C1 and hemoglobin subunit beta). Between HP and NR-PE, there were six, two downregulated (clusterin and plasmin heavy chain A) and four upregulated (apolipoprotein A-IV; heparin cofactor II; complement C4B and haptoglobin related protein). Between R-PE and NR-PE there were four, one downregulated (fibronectin) and three upregulated (pregnancy-specific beta-1-glycoprotein 1 (PSG1); complement C4B (C4B) and complement C4A (C4A). Regarding PE, C4A and C4B intensity peaks were correlated with blood pressure and creatinine levels. Both, together with PSG1, showed significant correlations with the newborn's weight and gestational week (all p< 0.05). These findings help to understand the pathophysiological mechanisms between the two subgroups of the disease, since the under regulated circulating proteins act in the blood pressure modulation and the upregulated may occur as compensatory mechanisms due to an inflammatory scenario. These data give insights to explain the complexity and clinical consequences of this disorder.

Lung Ultrasound Score at Hospital Discharge as a Predictor of Emergency Department Visit and Hospital Readmission in Patients With Acute Heart Failure.

Purpose The number of B-lines on lung ultrasound at hospital discharge in patients admitted with acute heart failure (AHF) is associated with poor outcomes. Assessing B-lines can be challenging to execute and replicate, depending on the clinical context. This study aims to determine whether the lung ultrasound score (LUS) at discharge predicts hospital readmission or emergency department (ED) visits in the 30 days after an AHF hospital admission. Methods We conducted an observational study at the medical ward of the emergency unit of the Clinics Hospital of the Ribeirao Preto Medical School, University of Sao Paulo, a tertiary university hospital in Ribeirao Preto, Sao Paulo, Brazil, where consecutive adults admitted with AHF were included. On the day of hospital discharge, we measured the LUS and tracked these patients for up to 30 days to monitor emergency department visits, hospital readmission, and the number of days free from hospital stay. Results A total of 46 patients were included in the study. A composite outcome of ED visits or hospital readmission in the 30 days after hospital discharge was achieved for 22 (47.8%) patients. The LUS at hospital discharge had a receiver operating characteristic (ROC) area of 0.93 (95% CI, 0.82-0.99) to predict the composite outcome, against 0.67 (95% CI, 0.52-0.81) for the clinical congestion score (CCS). A LUS ≥ 7 at discharge had a sensitivity of 95.5% and a specificity of 87.5% to predict the composite outcome. The average exam duration was 176±65 (sd) seconds. Conclusions The LUS at hospital discharge following admission for AHF proves to be an accurate tool for predicting the likelihood of return to the ED and/or hospital readmission within 30 days post discharge.

Exploring the Chikungunya virus landscape in a dengue-endemic Brazilian area

We aimed to describe the landscape, including molecular, epidemiological, and clinical aspects of CHIKV infections in the Ribeirao Preto region, an area endemic to dengue. We randomly screened 3744 plasma samples that had undergone DENV diagnosis to evaluate CHIKV-RNA using an in-house RT-PCR assay. Positive samples were followed clinically, and RNA samples were submitted to whole genome sequencing. Seventeen cases (0.5 %) were positive for CHIKV-RNA despite being negative for DENV-RNA. Notably, half of the patients experienced prolonged arthralgia lasting more than 90 days. Compared with the healthy control group, leukopenia and thrombocytopenia were observed in all CHIKV-positive individuals with statistically significant P values (P < 0.0001 and P = 0.0003, respectively). The genomic analysis revealed that the CHIKV strains being studied are classified within the East-Central-South-African (ECSA) genotype. This analysis identified new mutations, E1: K211E and E2: V264A, while the previously known mutation E1: A226V was not detected among these strains. This study highlights the need for epidemiological surveillance and preparedness for potential CHIKV epidemics in Brazil, particularly where other arboviruses co-circulate.

30 Ethnobotany applied to ruminant feeding inserted in countryside property redesign

Abstract Adoption of multi-species herbal leys, planting trees and integrating livestock can increase agricultural productivity and carbon storage on ruminant livestock farms. Most of the forage produced in pastures during the year is available in hot and rainy seasons, so the cold and dry season becomes a critical point, where forage production is insufficient. Thus, there is a need to use other complementary feed sources. This study aims to characterize bibliographically the plants that are part of ethnobotanical science, according to their bio compounds, based on the perception of the livestock farmers interviewed, aimed at increasing productivity, and that are used in the feeding of ruminant animals by livestock farmers in the regions between the cities of Araras and Ribeirao Preto in the state of Sao Paulo, Brazil. The research took place in a qualitative way adopting the use of questionnaires and interviews with rural producers, and the answers used by them were characterized in an empirical way, according to occurrences based on the experience and experience of errors and successes in animal nutrition and health, in which it was later possible to perform a textual analysis of what was described by the producers, aligned with the literature. Semi-structured interviews were conducted with farmers, involving questions about: i) which plant species exist on the property; - ii) what is the importance, value or function associated with these species. The species to be surveyed will be those: plant species that are in some way related to the production system (present on and/or off pasture), and have been planted or maintained by farmers to: i) provide animal welfare; ii) positively influence forage or soil quality; iii) complement animal nutrition; iv) provide ecosystem services (related to ecological/environmental factors; v) generate complementary income; vi) bring scenic, ornamental or landscape benefit on the pastures. The wisdom of the farmers is in line with the information in the literature, since the purpose of the food has characteristics such as being drought-tolerant, having increased dry matter, a wealth of minerals and vitamins, flavonoids, tannins, antioxidant and anti-inflammatory substances, energy and protein, as well as good digestibility by ruminant animals. Therefore, the productive potential of rural properties in Brazil goes beyond grass and crop treatments. It is necessary to explore the species present on rural properties and show farmers the importance of identifying plants and for what purpose they can be used, such as animal feed subsidized by the farm's own ruminants. Especially when there is grazing of the animals and the presence of these foods in the paddock.

Emergency and Urgent Care Network: Analysis of the Stroke Care Line According to Care Times and Outcomes

Background: Given the importance of acute stroke care for public health in terms of its burden and consequences like disability and death, evidence-based practice has directed the development of clinical protocols and operational guidelines to care of the populations affected by this pathology.&#x0D; Aim: To evaluate the impact of implementing the stroke care line in the Urgency and Emergency Care Network of a large Brazilian municipality.&#x0D; Methods: This is a retrospective study, conducted between 2014 and 2019 in public health services in Ribeirao Preto, SP, a large municipality of Brazil. Data was collected through secondary sources. A total of 403 patients were included in the study, of which: 172 were treated between 2014 and 2016 and 231 between 2017 and 2019; 118 were served by the mobile pre-hospital care services and 285 by the fixed pre-hospital care services. Data comprise clinical, epidemiological characteristics and severity of cases, as well as stroke care times and outcomes and were analyzed using descriptive techniques, Mann-Whitney and Chi-squared tests.&#x0D; Results: First care provided by a mobile pre-hospital care unit increased from 16.9% to 38.5%. The service times for people undergoing thrombolysis were shorter in the period from 2017 to 2019 compared to 2014 to 2016 in the door-to-needle, stroke onset-to-call, stroke onset-to-ambulance dispatch and regulation-to-door times. Stroke victims treated first by a mobile (ambulance) unit had shorter care times when compared to those who received care in a fixed unit, except in ambulance dispatch-to-door time. No difference was identified in the percentage of outcomes studied in the period from 2017 to 2019 compared to the period from 2014 to 2016. There was a higher craniectomy occurrence in people treated at home than in those attended in health services. The percentage of unfavorable stroke outcomes increased depending on the case severity.&#x0D; Conclusion: The care flow organization with defined responsibilities for each care point through the creation of a care line and the use of pre-established protocols reduced pre- and in-hospital care times; however, they did not have a significative influence on the case outcomes.

An assessment of alterations to human sperm methylation patterns in coronavirus disease 2019 infected and healthy control males

To determine whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection affects male reproductive health, considering the many potential factors that contribute to declines in male fertility on a semiglobal scale. In total, 64 human semen samples-32 treatment and 32 control-were laboratory processed and bioinformatically analyzed to assess differences in DNA methylation patterns. Implementing multiple bioinformatic tools, the analyses conducted will elicit between-group differences with respect to epigenetic age, epigenetic instability, semiglobal, and regional methylation, in addition to methylation patterns as a function of time since infection. Universityhospital. The study cohort of 64 individuals was drawn from a larger population of 94 volunteer participants recruited at the Human Reproduction Center at the Clinical Hospital of the Ribeirao Preto Medical School-University of São Paulo between June 2021 and January 2022 as well as in accordance with the ethical guidelines established by the Declaration of Helsinki. Exposure to SARS-CoV-2. Effects on male reproductive health were reported as differences in DNA methylation measured using an array. Mean β values at key regulatory loci for human spermatocytes were analyzed and compared between groups. Further analysis of β values using epigenetic age, instability, semiglobal, and regional methylation tools provided an analysis with substantial breadth and depth. In all analyses, there were no differences between groups. Considering these results, it can be inferred that infection with SARS-CoV-2 does not alter the epigenome of human spermatocytes in significant and/or persistent ways. Tangentially, these data also suggest that human male reproductive health is minimally altered by the virus, or that it is altered in a way that is independent of epigenetic programming. Infection with SARS-CoV-2 has been reportedly associated with alterations in male fertility. This study asserts that such alterations do not have an epigenetic basis but are likely a result of concomitant symptomatology, i.e., fever and inflammation. Across the multiple bioinformatic analyses conducted, the results of this test did not detect any differences in DNA methylation patterns between coronavirus disease 2019 and noncoronavirus disease semen donor groups.

PHYSICAL ACTIVITY ASSOCIATED TO LOWER PREVALENCE OFDEMENTIA IN BRAZILIAN OLDER ADULTS

AbstractBackgroundEvidences indicate that physically active older individuals maintain healthy functioning longer than do sedentary peers. Some lifestyle could impact the risk of dementia including Alzheimer’s. In spite of this, the effect of physical activity on the relationship between age and the risk of dementia is unknown. This is a two‐phase study that aims to investigate the association of physical exercise and age with the prevalence of dementia in community older adults, from Ribeirao Preto city, state of Sao Paulo, Brazil.MethodCross‐sectional study of a randomized community‐based sample of 928 elderly subjects aged 60 years or older was performed. Standardized instruments for screening dementia and a socio‐economic questionnaire were applied to the subjects at home in phase I. The dementia diagnose was made at hospital in phase II, according to DSM‐V criteria.ResultOveral Dementia was 6.0%. Alzheimer’s disease was the most frequent dementia (2.8%), followed by Vascular dementia (1.3%), and Mixed dementia (1.1%). Initial multivariate analysis showed sociodemographic factors associated to dementia. After adjusting for all the variables, multivariate analysis showed an association with age group ≥80 (OR = 6.7; p&lt;0.001); household chores (OR = 2.7; p = 0.004); no employment (OR = 4.3; p&lt;0.001); and stroke (OR = 4.5; p = 0.001). There was no association with gender, socioeconomic class, educational level, and alcohol use. Secondary analysis showed an inverse relationship between physical activity and age (OR = 0.4; p&lt;0.001). After deliberately exclude age, the final model showed a significant association between higher rates of dementia and lack of physical activity (OR = 2.0; p = 0.024).ConclusionAccording to previous studies, in our sample, lack of exercise was associated with two fold higher risk of dementia, only after exclude age of the analysis. This result could be due to decreasing level of physical activity found in the oldest subjects. We suggest that relatively simple lifestyle changes such as increasing the practice of exercise could enhance cognitive performance in elderly people. Future investigations are needed to clarify the real impact of physical activity on the relationship between dementia and age.

Microcosting Analysis of Hematopoietic Stem Cell Transplantation and Chemotherapy with Intermediate Doses of Cytarabine in the Treatment of Acute Myeloid Leukemia

Adult patients with Acute Myeloid Leukemia (AML) are stratified according to clinical and laboratory features at diagnosis into favorable-, intermediate-, or unfavorable-risk groups and may be treated differently according to risk. The European LeukemiaNet panel of experts' recommendation for the post-induction treatment of the favorable subgroup is either chemotherapy with intermediate doses of cytarabine (IDAC, e.g. 1.5 g/m2 q12h for 3 days) or autologous hematopoietic stem-cell transplantation (auto-HSCT). Both alternatives are costly and few studies have compared health economics aspects, which are particularly relevant in source-restrained regions. Moreover, AML treatment in low- and middle-income countries was associated with a higher frequency of invasive fungal diseases (IFD) and infections by multi-resistant bacteria compared to high-income countries and this may directly affect the treatment cost. Studies comparing IFD incidence in patients with hematological malignancies showed lower rates in patients treated with auto-HSCT compared to intensive chemotherapy. Material &amp; Methods: we performed the micro-costing analysis patients with intermediate-risk AML who received auto-HSCT (n=5) or IDAC (n=4) as post-induction treatment at University of Sao Paulo Hospital, Ribeirao Preto, Brazil. For cost analysis, we collected information about all resources and services consumed by patients such as medicines, materials, laboratory analysis, units of blood transfusion components, apheresis, hospital admissions, and outpatient care, in the period that began on the date of diagnosis until the end of treatment. The two groups had similar clinical and laboratorial characteristics at diagnosis and were treated according to the ICAML2015 protocol, in which induction (ID-1) is based on daunorubicin + standard-dose cytarabine (‘3+7‘), followed by a second cycle (ID-2) with daunorubicin + IDAC and, for those patients that achieved CR, consolidation was done with two cycles of IDAC (CONS-1 and CONS-2) or one cycle of IDAC followed by auto-HSCT (CONS-1 and auto-HSCT). The choice between the two alternatives of post-induction regimens was pre-defined by the participating center of the ICAML2015 study. Results: the mean cost of auto-HSCT and IDAC was US$ 4,188.12 and 2,368.22, respectively. Patients who received auto-HSCT stayed longer in-hospital compared to those in the IDAC group (21.8 versus 12.5 days). When the whole treatment is analyzed, the mean cost of the treatment with four cycles of chemotherapy was US$ 121,980.93 and that with three cycles followed by auto-HSCT was US$ 84,619.16. The most costly phase of treatment was the ID-1 (US$ 9,690.10). Regardless of the type of treatment, we observed that the inputs with the greatest economic impact were hospital admissions, which represented 51.2% of the total costs. . The second input with the greatest economic impact was drugs, and in that regard, treatment of infections played an important role in increasing the use of supplies in general. All patients in the study had febrile neutropenia that required hospital admission. Three patients were treated for IFD episodes, all of them were considered to be probable according to the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group, and the most frequently used antifungals were amphotericin B (deoxycholate) and voriconazole. All patients received prophylaxis with fluconazole. The frequency of IFD considering the whole treatment was 19.4 % (7/36 episodes of febrile neutropenia) and, there was no significant difference between patients who received auto-HSCT or IDAC. In conclusion, our results show that the cost of the post-induction treatment using auto-HSCT was higher than with IDAC and that cost with hospital stay.

Intravitreal bevacizumab plus propranolol for neovascular age-related macular degeneration (the BEVALOL study): a phase I clinical trial

BackgroundGiven the persistently large public health impact of neovascular age-related macular degeneration (nARMD) despite many years of anti-VEGF therapy as the first-line treatment and the demonstrated ability of b-blockers to reduce neovascularization, a synergistic effect between an anti-VEGF agent and an intravitreal beta-blocker is important to investigate in the quest for therapeutic alternatives that maximize efficacy and/or reduce costs. The main purpose of this study is to investigate the safety of a 0.1 ml intravitreal injection of a combination of bevacizumab (1.25 mg/0.05 ml) and propranolol (50 g/0.05 ml) to treat nARMD.MethodsProspective phase I clinical trial that included patients with nARMD. Comprehensive ophthalmic evaluation was performed at baseline and included Early Treatment Diabetic Retinopathy Study (ETDRS) best-corrected visual acuity (BCVA), biomicroscopy of the anterior and posterior segments, binocular indirect ophthalmoscopy, color fundus photography, spectral domain optical coherence tomography (OCT), OCT angiography (OCT-A), fluorescein angiography (Spectralis, Heidelberg), and full-field electroretinography (ERG). All eyes were treated with a 0.1 ml intravitreal injection of a combination of bevacizumab (1.25 mg/0.05 ml) and propranolol (50 g/0.05 ml) within 1 week of baseline evaluation. The patients were reexamined at weeks 4, 8 and 12, and clinical evaluation and SD-OCT were performed at all follow-up visits. Additional injections of combination bevacizumab (1.25 mg/0.05 ml) and propranolol (50 g/0.05 ml) were administered at weeks 4 and 8. At the final study evaluation (week 12), color fundus photography, OCT-A, fluorescein angiography, and full-field ERG were repeated.ResultsEleven patients (11 eyes) completed all study visits of the 12 week study. Full field ERG b-waves did not show significant (p < 0.05) changes at week 12 compared to baseline. During the 12 week follow-up period, none of the study eyes developed intraocular inflammation, endophthalmitis or intraocular pressure elevation more than 4 mmHg over baseline. Mean ± SE BCVA (logMAR) was 0.79 ± 0.09 at baseline and was significantly (p < 0.05) improved to 0.61 ± 0.10 at week 4; 0.53 ± 0.10 at week 8; and 0.51 ± 0.09 at week 12. Mean ± SE central subfield thickness (CST) (μm) was 462 ± 45 at baseline and was significantly (p < 0.05) lower at 4, 8 and 12 weeks (385 ± 37; 356 ± 29 and 341 ± 24, respectively).ConclusionsIn this 12 week trial of a combination of intravitreal bevacizumab and propranolol for treatment of nARMD, no adverse events or signals of ocular toxicity were observed. Further studies using this combination therapy are warranted.Trial Registration Project registered in Plataforma Brasil with CAAE number 28108920.0.0000.5440 and approved in ethics committee of Clinics Hospital of Ribeirao Preto Medicine School of São Paulo University—Ribeirão Preto, São Paulo, Brazil (appreciation number 3.999.989 gave the approval).

Leveraging cellular immunotherapy at the cutting edge of autoimmune disease R&amp;D

David McCall, Commissioning Editor,&#160;BioInsights, talks to Silvio Manfredo-Vieira, Associate Director for Correlative and Translational Studies at the Department of Dermatology, University of PennsylvaniaSilvio Manfredo-Vieira is the Associate Director for Correlative and Translational Studies at the Department of Dermatology of the University of Pennsylvania, where he is responsible for direct translational research necessary to move novel cell therapies developed in the lab into early-phase clinical trials under an academic-industry alliance. He received his PhD at the School of Medicine of Ribeirao Preto, Sao Paulo, Brazil, followed by postdoctoral training at Harvard University and Yale University, where he discovered that gut bacteria translocate to distal tissues and induce autoantibodies typical of lupus and autoimmune hepatitis, indicating a critical role for intestinal microbes in initiating or perpetuating local or systemic autoimmunity.

Integrating Methylome and Transcriptome Signatures Expands the Molecular Classification of the Pituitary Tumors.

To explore pituitary tumors by methylome and transcriptome signatures in a heterogeneous ethnic population. In this retrospective cross-sectional study, clinicopathological features, methylome, and transcriptome were evaluated in pituitary tumors from 77 patients (61% women, age 12-72 years) followed due to functioning (FPT: GH-secreting n = 18, ACTH-secreting n = 14) and nonfunctioning pituitary tumors (NFPT, n = 45) at Ribeirao Preto Medical School, University of São Paulo. Unsupervised hierarchical clustering analysis (UHCA) of methylome (n = 77) and transcriptome (n = 65 out of 77) revealed 3 clusters each: one enriched by FPT, one by NFPT, and a third by ACTH-secreting and NFPT. Comparison between each omics-derived clusters identified 3568 and 5994 differentially methylated and expressed genes, respectively, which were associated with each other, with tumor clinical presentation, and with 2017 and 2022 WHO classifications. UHCA considering 11 transcripts related to pituitary development/differentiation also supported 3 clusters: POU1F1-driven somatotroph, TBX19-driven corticotroph, and NR5A1-driven gonadotroph adenomas, with rare exceptions (NR5A1 expressed in few GH-secreting and corticotroph silent adenomas; POU1F1 in few ACTH-secreting adenomas; and TBX19 in few NFPTs). This large heterogenic ethnic Brazilian cohort confirms that integrated methylome and transcriptome signatures classify FPT and NFPT, which are associated with clinical presentation and tumor invasiveness. Moreover, the cluster NFPT/ACTH-secreting adenomas raises interest regarding tumor heterogeneity, supporting the challenge raised by the 2017 and 2022 WHO definition regarding the discrepancy, in rare cases, between clinical presentation and pituitary lineage markers. Finally, making our data publicly available enables further studies to validate genes/pathways involved in pituitary tumor pathogenesis and prognosis.

Neutrophil Extracellular Traps Contribute to Perivascular Adipose Tissue Dysfunction in High‐Fat Diet Obese Mice

BackgroundObesity triggers functional changes in the perivascular adipose tissue (PVAT), favoring the release of vasoconstrictor factors and the activation of contractile mechanisms in vascular smooth muscle cells. In the PVAT of obese individuals, there is infiltration of immune cells, including neutrophils. A neutrophil defense mechanism is the formation of neutrophil extracellular traps (NETs), a cytotoxic framework that allows maintenance and expansion of inflammatory processes. We hypothesize that PVAT dysfunction in obese mice is due to increased neutrophil infiltration, NETs formation and inflammation.MethodsSix weeks‐old male Balb/C mice received control (CD) or high fat diet (HFD) for 18 weeks. Vascular function was assessed in endothelium‐intact mesenteric arteries in the presence or absence of PVAT. Mice were treated with DNAse type 1 (10 mg/kg for 14 days, every other day). Molecular mechanisms were investigated. Experiments were approved by the Ethics Committee on Animal Research of the Ribeirao Preto Medical School, University of Sao Paulo (protocol nº 209/2016)ResultsValues are expressed relatively to the contraction triggered by 120 mM KCl [Emax (% KCl)]. In Balb/C mice, the PVAT decreased the contractile response in control arteries [Control PVAT (‐): 123.2±8.1 n=6; Control PVAT (+): 64.4±4.0 n=6]. However, HFD promoted partial loss of PVAT anti‐contractile effect [HFD PVAT (‐): 127.9±6.9 n=6; HFD PVAT (+): 98.3±7.4 n=6]. In PVAT of obese mice there was a greater amount of neutrophils, compared to PVAT from control mice [Control: 8.0% n=5; HFD: 18.9% n=5]. In PVAT of obese mice there was greater formation of NETs, and production of interleukin 6 (IL‐6), compared to PVAT from control mice [NETs in pg/ml – Control: 30.5±2.7 n=6; HFD: 71.4±3.2 n=6; IL‐6 in pg/100 mg of PVAT – Control: 45.6±1.9 n=6; HFD: 98.4±3.1 n=6]. DNAse treatment reduced the formation of NETs and IL‐6 in the PVAT of obese mice [NETs in pg/ml – HFD+DNAse: 18.3±1.2 n=6; IL‐6 in pg/100 mg of PVAT – HFD: 48.1±3.4 n=6]. DNAse treatment restored PVAT anticontractile function in obese mice [HFD+DNAse PVAT (+): 71.3±7.4 n=6]. Adipose tissue of obese humans exhibited greater formation of NETs and production of IL‐6 compared to adipose tissue of eutrophic humans [NETs in pg/ml – Eutrophic: 20.5±3.7 n=7; Obese: 79.2±1.3 n=7; IL‐6 in pg/100 mg of adipose tissue – Eutrophic: 9.7±2.9 n=7; Obese: 27.7±2.6 n=7].ConclusionThese results indicate that, in obesity, there is a greater neutrophils infiltration in the PVAT and, consequently, greater formation of NETs and inflammation, events that contribute to PVAT dysfunction.

Supraphysiological Levels of Testosterone Induce Cardiac Dysfunction via NLRP3 Inflammasome Activation in Macrophages

IntroductionTestosterone (Testo) modulates vascular tone and cardiac performance. Athletes who use Testo at supraphysiological doses exhibit increased blood pressure, higher inflammatory marker levels, vascular dysfunction, and cardiac hypertrophy. NLRP3 inflammasome activation, as part of the innate immune system response, contributes to proinflammatory cytokines production, leading e.g to cardiac dysfunction.HypothesisWe hypothesized that supraphysiological levels of Testo promotes cardiac dysfunction via generation of mitochondrial reactive oxygen species (mROS) and activation of the NLRP3 inflammasome.MethodsMale, 12 week‐old wild type (WT) and NLRP3 knockout (NLRP3‐/‐) mice were used. Mice were treated with testosterone propionate [Testo‐P (10 mg/kg)] or vehicle for 30 days. Cardiac function was evaluated using echocardiography. After in vivo experiments, western blot and ELISA were performed to evaluate NLRP3 inflammasome components. ROS generation was evaluated by lucigenin. Bone marrow‐derived macrophages (BMDMs) were isolated, primed with lipopolysaccharide (LPS [500 ng/mL 4 h]) and stimulated with Testo [10‐7 M], for 4, 6, 12 and 24 h. ROS generation was evaluated by DHE fluorescence and MitoSOX.ResultsEchocardiography showed cardiac dysfunction in WT mice treated with Testo‐P, characterized by reduced ejection fraction, shortening fraction, cardiac output and systolic volume. Testo‐P also increased interventricular septum, left ventricle posterior wall and decreased left ventricle internal diameter. All these effects were observed in NLRP3‐/‐mice. Furthermore, WT mice treated with Testo‐P showed increased cardiac expression of NLRP3 receptor, active Caspase‐1 and IL‐1β levels. These effects were prevented in NLRP3‐/‐. In addition, increased ROS generation was observed in the left ventricle of WT mice, but not NLRP3‐/‐ mice, treated with TP. Testo‐P‐treated WT mice showed increased macrophage infiltrate in the left ventricle, and this effect was not seen in the WT vehicle mice. In in vitro experiments, BMDMs primed with LPS and stimulated with Testo showed increased expression of the NLRP3 receptor and IL‐1β levels after 12 and 24 h. In addition, Testo‐stimulated BMDMs exhibited increased mROS generation.ConclusionSupraphysiological levels of testosterone induce cardiac dysfunction via mROS generation and NLRP3 inflammasome activation. This study was approved by the Ethics Committee on Animal Experimentation of the Ribeirao Preto Medical School (020/2021).

Sagittal balance and intervertebral disc composition in patients with low back pain.

The aim of this study was to verify the relationship between quantitative T2 relaxation measurements of lumbar intervertebral discs (IVDs) and spinopelvic parameters in patients with chronic low back pain. The study was approved by the Clinical Hospital of the Ribeirao Preto Medical School (USP) Ethics Committee, and written consent was obtained from all patients. A total of 455 IVDs from 91 consecutive patients with chronic low back pain were included in this prospective study. All subjects were assessed using the Oswestry Disability Index and visual analogue scale questionnaires and were confirmed to have no other spine diseases except disc degeneration. Spinopelvic parameters including the pelvic incidence (PI), pelvic tilt (PT), sacral slope (SS), sagittal vertical axis (SVA), global tilt (GT), T1 pelvic angle (TPA), lumbar lordosis (LL), thoracic kyphosis (TK), pelvic incidence minus lumbar lordosis mismatch (PI-LL), and lack of lumbar lordosis (LLL) were measured. The study group was categorized according to the Roussouly classification. Sagittal T2 maps were acquired to extract the IVD relaxation times, and the complete manual segmentation of IVDs at all levels was performed using Display® software. Lumbar IVD T2 relaxation times showed significant correlation with PT (P<0.01), GT (P<0.01), TPA (P<0.01), PI-LL (P=0.01), and LLL (P=0.01). No difference was noted between Roussouly subtypes regarding T2 relaxation times at any disc level. Data from questionnaires showed no correlation with T2 relaxation times. Global tilt and T1 pelvic angle were correlated with IVD composition changes (T2 relaxometry). There was no correlation between clinical symptoms and IVD T2 relaxation times.

Cohort Profile: The 1978-79 Ribeirao Preto (Brazil) birth cohort study.

Cohort Profile: The 1978-79 Ribeirao Preto (Brazil) birth cohort study.

Molecular surveillance of the on-going SARS-COV-2 epidemic in Ribeirao Preto City, Brazil.

The Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of an unprecedented worldwide pandemic. Brazil demonstrates one of the highest numbers of confirmed SARS-CoV-2 cases, and São Paulo State is the epicenter of the pandemics in the country. Nevertheless, little is known about the SARS-CoV-2 circulation in other cities in the State than São Paulo city. The objective of this study was to analyze phylogenetically SARS-CoV-2 strains circulating in city of Ribeirão Preto at the beginning of the pandemic and during the actual second wave. Twenty-nine nasopharyngeal SARS-CoV-2 RNA positive samples were sequenced by nanopore technology (18 obtained at the initial period of the pandemic and 11 during the second wave) and analyzed them phylogenetically. The performed analysis demonstrated that the majority of the strains obtained in the initial period of the pandemic in Ribeirão Preto belonged mainly to the B1.1.33 lineage (61.1%), but B.1.1 (27.8%) and B.1.1.28 (11.1%) lineages were also identified. In contrast, the second wave strains were composed exclusively by the Brazilian variant of concern (VOC) P.1 (91%) and P.2 (9%) lineages. The obtained phylogenetic results were suggestive of successive SARS-CoV-2 lineage substitution in this Brazilian region by the P.1 VOC. The performed study examines the SARS-CoV-2 genotypes in Ribeirão Preto city via genomic surveillance data. The obtained findings can contribute for continuous long-term genomic surveillance of SARS-CoV-2 due to the accelerated dynamics of viral lineage substitution, predict further waves and examine lineage behavior during SARS-CoV-2 vaccination.

Prevalência de competições terapêuticas na farmacoterapia de usuários das Unidades de Saúde da Família de Ribeirão Preto, São Paulo

No atual cenário epidemiológico verifica-se elevada frequência de doenças crônicas não transmissíveis com o aumento no uso de medicamentos e competições terapêuticas. Assim, o objetivo deste estudo foi estimar a prevalência e identificar as competições terapêuticas em usuários que faziam seguimento em seis Unidades de Saúde da Família de Ribeirão Preto, São Paulo. Trata-se de um estudo retrospectivo, longitudinal, descritivo e exploratório com amostra constituída por 226 participantes. A maioria era idoso (69,5%), pertencia ao sexo feminino (63,7 %), era sedentário (48,2%) e apresentava quatro ou cinco doenças crônicas não transmissíveis (50,8%), sendo mais frequentes a hipertensão arterial sistêmica, o diabetes mellitus tipo 2 e dislipidemia. O risco potencial de ocorrência de competições terapêuticas foi estimado em 64,2%. Dentre elas, destacou-se a competição terapêutica que envolvia os betabloqueadores, utilizados para o tratamento da hipertensão arterial. Os dados evidenciam a necessidade de uma abordagem integral no cuidado às pessoas com multimorbidades na Atenção Primária à Saúde. Além disso, alerta os riscos das competições terapêuticas e a importância do monitoramento das complicações da doença índice na presença de uma competição terapêutica potencial.

Intravitreal bevacizumab (IVB) versus IVB in combination with pars plana vitrectomy for vitreous hemorrhage secondary to proliferative diabetic retinopathy: a randomized clinical trial

BackgroundThe main purpose of this study is to compare the vitreous hemorrhage (VH) score reduction and visual acuity outcomes in patients with VH secondary to proliferative diabetic retinopathy (PDR) treated with intravitreal injections of bevacizumab (IVB) versus IVB and pars plana vitrectomy (IVB and PPV).MethodsPatients with VH secondary to PDR were randomized into 2 groups: in Group A, patients were treated with a total of 3 IVB (1.5 mg/0.06 ml) at 8-week intervals; and in Group B, patients received a single IVB (1.5 mg/0.06 ml) and, 7 days later, underwent PPV. Patients received an ophthalmic evaluation that included best-corrected visual acuity (BCVA), indirect ophthalmoscopy, and mode B echography at weeks 8, 16 and 24. VH was classified according to the Diabetic Retinopathy Vitrectomy Study classification as grade 1, 2 or 3. Change in VH score was the primary outcome measure and change in BCVA was the secondary outcome.ResultsSeventy-three eyes of 66 patients were randomized and 70 eyes completed the 24-week follow-up visit. Mean VH score reduction (± SEM) of 0.4571 ± 0.0283 (p = 0.0014), 1.3429 ± 0.0393 (p < 0.0001) and 1.8286 ± 0.0438 (p < 0.001) was observed in Group A at 8, 16 and 24 weeks after treatment, respectively (Table 2; Fig. 2). In Group B, the reduction of VH score (± SEM) was 2.2571 ± 0.0720 (p = 0.0014), 2.2857 ± 0.0606 (p < 0.0001) and 2.2286 ± 0.0726 (p < 0.001) at 8, 16 and 24 weeks after treatment, respectively. Group comparison revealed a significantly greater reduction in mean VH score in Group B at 8 and 16 weeks after treatment (p < 0.0001). However, at 24 weeks this difference was no longer statistically significant (p = 0.1854). In Group A, mean (± SEM) BCVA showed an improvement of 0.00285 ± 0.0004 (p = 0.971), 0.5371 ± 0.0072 (p < 0.0001), 0.8143 ± 0.0001 (p < 0.0001) and 0.8543 ± 0.0008 (p < 0.0001) compared to baseline at 1, 8, 16 and 24 weeks after treatment, respectively. In Group B, mean (± SEM) BCVA showed an improvement of 0.3657 ± 0.0507 (p = 0.0002), 0.8857 ± 0.0385 (p < 0.0001), 0.9457 ± 0.0499 (p < 0.0001) and 0.9629 ± 0477 (p < 0.0001) compared to baseline at 1, 8, 16 and 24 weeks after treatment, respectively. No significant difference in BCVA improvement was observed between groups at 24 weeks after treatment.ConclusionPPV with preoperative IVB is associated with more rapid clearance of VH and improvement in BCVA than IVB injections alone. However, after 24 weeks of follow-up, the reduction in VH score and BCVA were similar between both treatment strategies.Trial RegistrationThe project is registered in Plataforma Brasil with CAAE number 927354.7.0000.5440 and was approved by the Ethics Committee of the Clinics Hospital of Ribeirao Preto Medicine School of São Paulo University—Ribeirão Preto, São Paulo, Brazil (appreciation number 3.053.397 gave the approval).

Virtual Microscopy as a Learning Tool in Brazilian Medical Education

Virtual microscopy (VM) is a widely used teaching method in Medical Education in many developed countries. In Brazil, however, this is not the case for most medical schools, considering Brazilian social inequality and uneven access to technology. Recently, the Covid-19 pandemic has also challenged Universities to seek and make a transition toward more effective methods of full-time online education. Thus, the main goal of this work was to verify student's perception and academic performance, assessed upon VM implementation in a Brazilian Medical School. Ribeirao Preto Medical School students answered a 26-question survey with regards to optical microscopy (OM) and VM. Academic performance was compared between participants that were (year of 2019) or were not (year of 2015) exposed to VM. Taken the results together, subjective impressions such as handling, suitability, learning effectiveness, and pleasure using the tools, have shown a higher score for virtual microscopy (median=29), when compared to optical microscopy (median=24) with a P-value<0.001 by Wilcoxon rank test, upon measurement using an ordinal scale. Regarding academic performance, no statistically significant differences were found between groups (P-value=0.38, Cohen's d=0.19). Therefore, VM proved to be adequate to the Brazilian medical education in light of Brazilian social contexts and Covid-19 pandemic.

ARTERIAL HYPERTENSION MAY INVOLVE SARCOPLASMIC RETICULUM CALCIUM ATPASE (SERCA) PROTEOLYSIS BY MATRIX METALLOPROTEINASE (MMP)-2

Objective: Aortic SERCA protein level and activity are reduced in hypertension. Increased MMP-2 activity may cleave SERCA in acute myocardial ischemia and reperfusion injury. Our objective was to verify whether increased vascular MMP-2 activity in hypertension proteolyzes SERCA to alter cytosolic calcium, thus causing enhanced vasoconstriction or maladaptive remodeling. Design and method: Male Sprague-Dawley rats were submitted to two kidney-one clip (2K-1C) or sham surgery and treated with doxycycline (Doxy, 30 mg/kg/day) by gavage from the third to seventh day post-surgery. Systolic blood pressure (SBP; tail-cuff plethysmography) was daily measured, and after seven days aortas were collected and processed for gelatin and in situ zymography, SERCA protein levels by Western blot and calcium immunofluorescence using Rhod-2AM. Aortas were isolated from normal rats were incubated at 37°C for 6 or 12 hours in DMEM with angiotensin (Ang) II (0.1 μM) +/- MMP inhibitors, ONO-4817 (10 μM) or Doxycycline (30 μM). Aorta segments were processed as above and used for co-immunoprecipitation of SERCA and MMP-2. Statistical analysis was done by two-way ANOVA followed by Tukey post-hoc test. The Ethics Committee for Animal Research of the Ribeirao Preto Medical School approved all protocols (122/2019). Results: MMP-2 activity in 2K-1C aorta was elevated 1.5-fold vs. controls and Doxy decreased it by 1.3-fold (n = 5–8; p &lt; 0.05). SERCA proteolysis was 3-fold increase in 2K-1C aorta (n = 4–8; p &lt; 0.05 vs. controls). Treatment with Doxy seemed to prevent this effect (0.49 ± 0.07 2K1C+Doxy vs 1.42 ± 0.42 2K1C). These effects were not associated with changes in SBP as Doxy did not alter it. Cytosolic calcium concentrations trended to increase in 2K1C aorta from 13 to 19 a.u. (n = 3; p = 0.08) and Doxy decreased it (n = 3; p &lt; 0.05). In AngII treated aorta, MMP-2 activity also increased by 1.5-fold (n = 4-5; p &lt; 0.05 vs. controls) and both ONO-4817 and Doxy reduced its activity by the half (n = 4-5; p &lt; 0.05). MMP-2 was co-immunoprecipitated with SERCA in the incubated aortas, and AngII seemed to increase it. Conclusions: SERCA levels may be decreased by MMP-2 activity in the aortas of hypertensive rats which may contribute to calcium-induced vascular alterations in hypertension. These effects are currently being investigated.