Chagas Disease (CD) is caused by the flagellated protozoan Trypanosoma cruzi, which is transmitted between hosts by triatomines (Hemiptera: Reduviidae). CD is a neglected and endemic disease in the Americas. Triatomine salivary molecules counteract hemostasis and modulate the host's immune system to facilitate hematophagy. Among them, apyrases catabolize nucleotides and nucleosides and interfere with ADP-mediated platelet aggregation. Rhodnius prolixus is an important vector of CD. The aims of this study were to perform an in silico analysis of R. prolixus apyrases and produce recombinant proteins in a heterologous prokaryotic expression system. Two R. prolixus apyrase sequences (T1H8D6 and T1H852) were characterized in silico, cloned into the expression vector pET17b (Novagen) fused with a 6xHis tail for expression in Escherichia coli BL21(DE3) strain. After transformation, selection and cultivation of bacterial clones, SDS-PAGE and Western blotting were performed to confirm the expression of recombinant R. prolixus apyrases. The apyrase activity was evaluated in solution and by zymography. Of the two R. prolixus apyrases, only the T1H8D6 protein of approximately 40 kDa was expressed. The recombinant protein was observed only in the insoluble fraction in the form of cellular aggregates, forming inclusion bodies. The inclusion bodies were prepared and purified by affinity chromatography. The amino acid sequences of R. prolixus apyrases have sites for N-glycosylation, O-glycosylation, acetylation, phosphorylation and the presence of a signal peptide. Furthermore, the comparative analysis between the R. prolixus apyrase models with members of the CD39, 5'-nucleotidase and Cimex families showed that they are similar to the Cimex family, which are dependent on Ca2+ for their activity in the catabolism of nucleotides. Apyrases facilitate hematophagy through the inhibition of ADP-mediated platelet aggregation in the vertebrate host and their study may assist in the development of approaches to prevent CD transmission.

: Capitulo 1: The SMYD family, which contains the conserved SET and MYND domains, comprises five members that have a lysine methyltransferase function. A growing number of evidences indicate the involvement of SMYD family genes in different types of cancer. Here, the aim was to investigate the most common genetic alterations of the SMYD family among the most frequent solid tumors and determine their potential use as biomarkers. A multiplatform integrative analysis was performed on the most common mutations, copy number alterations (CNA) and mRNA expression in SMYD family genes. Data from cohorts of invasive breast carcinoma (BRCA), prostate adenocarcinoma (PRAD), colorectal carcinoma (CRC), stomach adenocarcinoma (STAD), lung adenocarcinoma (LUAD), lung squamous cell carcinoma (LUSC), uterine corpus endometrial carcinoma (UCEC) and uterine carcinosarcoma (UCS) available on The Cancer Genome Atlas (TCGA) and cBioPortal were analyzed. SMYD genes were found to have a lower overall mutation frequency among the tumors examined. Most are missense mutations scattered between their domains. Different mutations were identified in SMYD1 (78), SMYD3 (44), SMYD5 (44), SMYD4 (39) and SMYD2 (33). It is interesting to note that mutations in all SMYD genes were found more frequently in UCECs. Therefore, no mutation was detected frequently in the same cohort and no specific mutation had sufficient stratification power to be used as a biomarker in the cancers examined. With regard to CNA, the low-level gain alteration was the most common among the genes, with the exception of SMYD4, which had heterozygous loss as the most frequent alteration in all the tumors analyzed. With regard to mRNA expression, each member of the SMYD family had a distinct expression pattern. SMYD1 was divided between downregulated expression and basal level, while SMYD2 and SMYD3 had alterations depending on the tumor analyzed, but most tended to upregulation; SMYD4 showed a great tendency to downregulation, suggesting tumor suppressor action; SMYD5 was both upregulated and downregulated, depending on the tumor evaluated. With regard to survival analysis, SMYD3 and SMYD4 in STAD, SMYD3 in BC and SMYD2 and SMYD4 in LUAD showed differences in survival time, depending on the level of expression. Therefore, the results show that SMYD family members have different functions depending on the tumor, and can act as oncogene and tumor suppressor, which makes it even more evident the importance of a better understanding of the involvement of these genes during carcinogenesis. Capitulo 2: Leukemias are a group of malignant diseases that affect blood cells, leading to a high proliferation rate of non-functional white blood cells and inhibiting normal hematopoietic development. They are characterized by the type of cell affected and its stage of maturation and are classified into four main subtypes: chronic myeloid leukaemia (CML), acute myeloid leukaemia (AML), chronic lymphocytic leukaemia (CLL) and acute lymphocytic leukaemia (ALL), the last of which is the subject of this study. ALL is generally characterized by the high proliferation of blasts, with the capacity to invade other regions of the body. It mostly affects children between the ages of 1 and 4. The SMYD family of methyltransferases has so far had few studies correlating them with ALL, but there are studies in the literature showing SMYD2 altered in samples from children with ALL, which is a possible prognostic indicator. SMYD5, on the other hand, was found to be upregulated in these same samples and, as a result of this finding, SMYD5 was chosen as the subject of this study. It was proposed to evaluate the number of altered copies (CNA) and mRNA expression levels of the SMYD family from cohorts obtained through cBioPortal and BloodSpot. In relation to CNA, there were differences in alterations between the SMYD family, with SMYD1 and SMYD5 showing heterozygous loss as the most frequent alteration, while the others showed a low-level gain as the most frequent alteration in ALL. In terms of mRNA expression, SMYD1 and SMYD2 were downregulated while SMYD5 was upregulated in ALL when compared to healthy bone marrow samples. The other members showed no statistically significant difference in expression. Therefore, despite the limited information, the SMYD family has been correlated not only with ALL but also with other blood malignancies. Previous studies have shown that SMYD5 has a function in hematopoiesis and that it was altered in ALL. It has also been reported that SMYD5 forms a repressor complex with NCoR and is responsible for H4K20me3, thus maintaining the repression of pro-inflammatory genes. In the literature, it has been described that its depletion can lead to chromosomal aberrations. Therefore, there is evidence that SMYD5 plays a role in leukemogenesis due to its involvement in the above-mentioned functions. It was then proposed to evaluate the effects caused by SMYD5 more precisely by starting stable models of ALL-B cell lines expressing Cas9, in order to perform a gene knockout using the CRISPR-Cas9 technique.

Introduction: Stroke is one of the main causes of morbidity and mortality in the world. With the advancement of the SARS-CoV-2 pandemic, cases of stroke in patients hospitalized with COVID-19 have accumulated in the scientific literature. Objectives: To investigate the association between exposure to SARS-CoV-2 and the presence of pro thrombotic factors in stroke patients treated in public neurology services of the SUS. Methodology: This is a prospective, hospital-based case-control study. ,. The study is nested within the multicenter case-control titled NeuroCOVID: Association of SARS CoV-2 with the occurrence, prognosis and pathogenesis of cerebrovascular diseases and other neurological manifestations, funded by the Inova Fiocruz Program (Fundação Oswaldo Cruz - Fundo Emergencial de Combate to COVID-19: VPPCB-005-FIO-20-2 22). Results: 148 patients were enrolled for this study at a single center. Among the participants, there was a predominance of female individuals (51.4%, n=76), white ethnicity (29.1%, n=43) and specifically the economically active population (43.9%, n=65). The majority of admitted patients had acute/subacute exposure to SARS-CoV-2 (72.3%, n=107), most of whom were previously vaccinated (80.4%, n=86). Stroke constituted the majority of stroke cases in this study (77%, n=114). The average age was 55.3 years, with a median of 55.5. The average length of stay was 12 days, with a median of 10 days. Conclusion: This study is one of the pioneers in demonstrating clinical epidemiological and laboratory characteristics of patients investigated with stroke. The studied sample presents characteristics similar to those described in other clinical studies. Through the division into acute/subacute exposed, warning exposed and non-exposed, it was possible to draw an overview of severity and outcomes.

Background Recently, next-generation sequencing (NGS) technology is revolutionizing

studies on circulating miRNAs in cancer, as it enables global analysis of these small non-
coding RNAs in body fluids, allowing both the detection of those with low expression

and new ones. In this study, we identified circulating miRNAs associated with oral cavity
and oropharyngeal SCC, and further investigated their expression levels in silico,
predicted their target genes, and identified the biological pathways they regulate.
Results The RNA-seq technique was applied to plasma samples for total miRNAs
identification. The bioinformatic analyses enabled the identification of six prominent
miRNAs significantly involved with oral cavity and oropharyngeal squamous cell
carcinoma (SCC): miR-30d-5p, miR-122-5p, miR-126-3p, miR-150-5p, miR-191-5p and
miR-223-3. A total of 194 target genes, predicted for these miRNA's, common to the

TargetScan and miRDB prediction platforms, were identified. Afterwards, a functional
analysis of these genes, performed using the DAVID tool, highlighted pathways and
annotations with significant enrichment. Among them, the pathway microRNAs in
cancer, from the Kyoto Encyclopedia of Genes and Genomes (KEGG) was enriched.
From the Gene Ontology (GO), the nucleus (Cell Compartment), protein binding
(Molecular Function) and negative regulation of transcription DNA-templated
(Biological Process) annotations were enriched. In addition, the Protein-Protein
Interaction Network analysis with STRING and Cytoscape allowed the identification of
genes relevant to the studied disease: PRKCA, RRAS2, RASA1, CALML4, PAX5 and
FOXO1.
Conclusion Among the complex network of miRNAs associated with oral cavity and
oropharyngeal SCC,.our miRNoma experiment and in silico prediction of target genes,
functional analyses, and construction of the PPI network of specific miRNAs highlighted
the miR-150-5p and miR-223-3p, providing important insights into how they can
modulate the expression and function of genes in oral and oropharyngeal cancer. These
results lay the foundation for further investigations, including studies to validate the
potential of these miRNAs as diagnosis or prognosis biomarkers, or targets for the
development of novel therapeutic approaches.

Background: At the beginning of 2020, during the COVID-19 pandemic, workers working in health services were under great pressure, exposing themselves to the risk of physical and mental illness. Objectives: To evaluate absenteeism in hospital health service workers, for all causes, not directly attributable to suspected or confirmed SARS-CoV-2 infection, comparing with data from the same population in a period prior to the COVID19 pandemic event. Methods: A longitudinal observational study was carried out with repeated measures of absenteeism in the same population group, in the pre-pandemic period (March to July 2019) and during the pandemic (March to July 2020), of healthcare workers from 40 hospitals public universities in Brazil. All causes of absenteeism, not directly attributed to COVID-19, were analyzed. Absenteeism data from health workers who were involved in both periods of interest were included and those who were in only one of the periods were excluded. Results: 32.691 workers were included in the study, with healthcare professionals making up 82,5% of the sample. Comparison of periods before and during the pandemic showed a 26,6% reduction in absence from work for all causes, with the exception of COVID-19 and absence related to mental health. Regarding absence from work related to mental health, the odds ratio was 39,0% higher during the pandemic. At the beginning of thepandemic, there was anincrease in absenteeism (all causes), followed by a progressive reduction until the end of the observation period. Conclusion: Absenteeism related to mental illness among healthcare workers increased during the COVID-19 pandemic, highlighting the need for healthcare managers to prioritize and implement support strategies to minimize absenteeism.

Objective: To evaluate the impact of head and neck cancer, as well as treatment outcomes, on health-related quality of life (HRQoL) in patients with head and neck cancer who underwent different combinations of surgery, chemotherapy, and radiotherapy. Methods: Two prospective, multicentric studies were conducted in 18 Brazilian medical institutions. Included were patients diagnosed with squamous cell carcinoma of the head and neck in stages I to IVB, with primary tumors located in various regions, excluding patients with a history of prior radiotherapy and those with distant metastasis. Treatment protocols followed the guidelines of each participating institution, including surgeries, systemic therapies, and radiotherapy. Data collection occurred from June 2014 to December 2019, and patients provided written informed consent. HRQoL was assessed using the cancer-specific questionnaire from the European Organization for Research and Treatment of Cancer (EORTC QLQ-C30) and a specialized module for head and neck cancer patients (EORTC QLQ-H&N43). Results: The findings of this thesis emphasize the need for a holistic approach in treating head and neck cancer, which considers the need to adapt and evolve treatment strategies to not only extend survival but also to improve the quality of that survival, taking into account the socioeconomic context of patients. The choice of radiotherapy technique has direct implications on the HRQoL and, possibly, on the OS. Equitable access to advanced treatments is essential for improving health outcomes. Furthermore, recovery of HRQoL after treatment requires ongoing support and long-term monitoring. The studies presented provide evidence to support clinical and health policy decisions and reinforce the need to improve access to and the quality of radiotherapy treatments and care for patients with head and neck cancer in Brazil.

Background: SARS-CoV-2 infection and perinatal neurological outcomes are still not completely understood. However, there is recent evidence of white matter disease and impaired neurodevelopment in newborns after maternal SARS-CoV-2 infection. These appear to occur as consequence of both direct viral effects and systemic inflammatory response, with glial cells / myelin involvement and regional hypoxia / microvasculature disruption. We sought to characterize consequences of the maternal and fetal inflammatory condition in the central nervous system of newborns post SARS-CoV-2 maternal infection. Methods: We conducted a longitudinal prospective cohort study, from June 2020 to December 2021, with follow-up of newborns from mothers exposed or not exposed to SARS-CoV-2 infection during pregnancy. Brain analysis included data from cranial ultrasound scan (CUS) with grayscale, Doppler studies (color and spectral), and ultrasound-based brain elastography (shear-wave mode), in specific regions of interest (ROIs): deep white matter, superficial white matter, corpus callosum, basal ganglia, and cortical grey matter. Brain elastography was used for estimation of brain parenchymal stiffness, which is an indirect quantifier for myelin cerebral tissue content. Results: A total of 219 single-pregnancy children were enrolled, including 201 from mothers exposed to SARS-CoV-2 infection and 18 from unexposed controls. Neuroimaging evaluation was performed at 6 months of adjusted chronological age, and it demonstrated 18 grayscale and 21 Doppler abnormal exams. Predominant findings were hyper echogenicity of deep brain white matter and basal ganglia (caudate nuclei / thalamus), and reduction in resistivity and pulsatility index of intracranial arterial flow. Anterior brain circulation (middle cerebral and pericallosal arteries) displayed wider range of flow variation than posterior circulation (basilar artery). Shear-wave US elastography analysis demonstrated reduction of stiffness values in SARS-CoV-2 exposed group at all analyzed regions of interest, especially in the deep white matter elasticity coefficients (3.98 ± 0.62) compared to control group (7.76 ± 0.77); pvalue<0.001. Conclusion: This study further characterizes pediatric structural encephalic changes associated with SARS-CoV-2 infection during pregnancy. Maternal infection has shown to be related to cerebral deep white matter predominant involvement, with regional hyper echogenicity and reduction of elasticity coefficients, suggesting zonal impairment of myelin content. Morphological findings may be subtle, and functional studies such as Doppler and elastography may be a valuable tool for identifying more accurately which infants are at risk of neurological damage.

Melatonin is a promising antioxidant substance that can minimize the effects of ototoxicity-induced hearing loss. Experimental studies have been performed to test the otoprotective efficacy of melatonin. Objective: To analyze the methodology presented by primary studies on determining the otoprotective efficacy of melatonin. The study is a systematic review with meta-analysis to clarify if the hormone is promising in preventing/minimizing human ototoxicity. Methods: Altogether, 154 articles were found in four databases. The PICOS strategy (Population, Intervention, Comparison, and Outcome) was used to define the eligibility criteria. Studies that met the inclusion criteria for the second step were included in a qualitative synthesis. Each study type was analyzed with the Critical Appraisal Checklist for Quasi-experimental Studies and the Critical Appraisal Checklist for Randomized Controlled Trials of the Joanna Briggs Institute. Results: Seven articles were selected, and four were included in the meta-analysis. It was possible to obtain seven outcomes according to the standard auditory frequencies presented among the studies, considering a minimum of three standard frequencies. The outcomes analyzed were for the frequencies of 1500, 2000, 3000, 4000, 5000, 6000, and 8000 Hz. Conclusion: Melatonin can provide protection against the ototoxic effects of cisplatin and aminoglycosides at 5000 Hz, 6000 Hz, and 8000 Hz, thereby minimizing the reduction in Otoacustic Emissions (OAE) amplitude. The same effect was not observed in the lower frequencies. Despite the limited number of studies that were evaluated, the results appeared consistent in higher frequencies. However, the methodology of the available studies did not meet the necessary methodological rigor that promotes the safe replicability of these studies.

With an increase in infertility, there is a growing demand for assisted reproduction techniques and in vitro embryo production. However, specific treatments are required for the underlying causes of infertility. Mesenchymal stem cells (MSCs) have been used to treat ovarian dysfunction. Because part of the mechanism of action of MSCs is due to their paracrine action, in this study, we investigated the therapeutic potential of extracellular defeats derived from MSCs (MSC-EVs) resulting from ultrafiltration (UF) and ultracentrifugation (UC), evaluating their effects on bovine ovarian stromal cell migration and their use in an animal model of acute ovarian injury. MSCs were acquired from bovine adipose tissue and characterized according to the International Society for Cell Therapy guidelines. MSC-VEs were instructed on the secretome of these cells using UF and UC and evaluated for size, concentration, induction potential, and migration of bovine ovarian stromal cells using experiments and cell migration assays. After comparing the isolation methods, we evaluated the rate of viable oocyte retrieval and the number of embryos produced in cows treated with three different therapeutic regimens of MSCEVs in an acute injury model. Four follicular puncture sessions were performed for each group. The first group received only the vehicle after the first follicular puncture. In the second group, proteins produced by 7.5 × 106 MSCs were injected after the first follicular puncture. In the third group, fruit trees produced by 2.5 × 106 MSCs were injected after the first session, and in the fourth group, the same amount of vengeful was injected after sessions 1, 2, and 3. The characterization of the experimental experiments showed that there was no difference in particle size between the isolation methods, but UF showed a higher yield. Regardless of the method used, these possibilities were able to induce cell migration and anticipation. Considering that there was no difference between the methodologies in functional assays, the yield obtained through UF proved to be a determining factor for large-scale use in vivo. We evaluated three therapeutic regimens for holiday treatment, observed follicular punctures, and observed that the group treated with the least amount of insufficiencies in the three doses maintained oocyte production and increased in vitro embryo production. Therefore, MSC-EVs demonstrated beneficial effects on both cell and external migration in vitro and on vacation fertiertility with follicular puncture injury in vivo.

Considering the relationship between Central Auditory Processing (PAC) and disfluencies, this study aims to analyze auditory processing in individuals with stuttering disfluencies. Methods: 30 individuals, with a mean age off 22.4 years and minimum age of 12 years, 26 males (86.67%) and four females (13.33%), were submitted to anamnesis, meatoscopy, basic audiological evaluation (tonal audiometry, vocal audiometry and impedance metry), specific assessment of stuttering (SSI-3) and behavioral tests of the PAC. For the evaluation of PAC, the following tests were applied: Binaural Interaction Processing (MLD), Temporal Processing (RGDT, PPS and DPS), Integration and binaural separation in dichotic listening (TDD) and monotic listening of low redundancy (SSI). Results: Regarding the degree of classification of stuttering, nests and study, the mild degree prevailed. In the overall result of all the skills evaluated, it was observed that 76.67% (23 individuals) presented pca alteration. In the analysis of all tests of behavioral evaluation of auditory processing performed, no correlation was found between the degree of stuttering and the alteration of the PAC. Conclusion: It was found that most of the gaga sample presentes PAC disorder, especially in auditory abilities of auditory closure and temporal ordering. In the comparison of the degrees of severity of stuttering,a change in the abilities of auditory closure, binaural integration and figure-background was -identified.

Background: Colorectal cancer is the second most common and deadliest tumor around the world. In recent decades, the incidence of colorectal cancer has progressively increased among young patients. Previous analyzes have shown that colorectal cancer in this population has a worse prognosis due to more aggressive characteristics and more advanced stages at the time of diagnosis. However, these findings are not uniform across different parts of the globe. Objective: To evaluate the effect of age on the clinical presentation, pathological staging, therapy and oncological outcomes of colorectal cancer. Methods: The medical records of 434 patients with colorectal adenocarcinoma were reviewed. Patients were divided into two groups according to age. The young group (age < 50 years) consisted of 78 patients, and the older group (age ≥ 50 years) consisted of 356 patients. Clinical variables, time interval between onset of symptoms and diagnosis, pathological stage, operative mortality and oncological stages were compared between groups. Results: The average follow-up in months was 35.59 ± 16.27. Age was an independent factor for longer hospital stays in patients > 50 years old [PR = 0.11 (95% CI: 0.01; 0.21), p = 0.0336]. Patients < 50 years old had more undifferentiated histological types > 50 years old [PR= 2.38 (95% CI: 1.18; 4.78; p = 0.0147)]. There were no other differences on surgical and clinical presentation and oncological outcome. Conclusion: Young patients have a higher frequency of undifferentiated tumors. Age has not been shown to impact oncological outcomes.

Rhodnius neglectus, an insect from the Reduviidae family (Hemiptera), is a potential vector of Trypanosoma cruzi, the causative protozoan of Chagas disease. Triatomine saliva comprises a rich mixture of molecules synthesized by the salivary glands to counter host responses, aid the uptake of blood and, indirectly, T. cruzi transmission. Therefore, the saliva of this hematophagous insect has been a relevant subject of studies due to the presence of these molecules with high biotechnological potential. The goal of this work was to study an Odorant Binding Protein from R. neglectus saliva (RnOBP) using computational and experimental approaches aiming, to contribute to the understanding of its structural and functional characteristics. RnOBP molecular modeling, computational molecular dynamics and clustering tests were performed to obtain information about the protein's flexibility. Next, molecular docking assays were carried out to verify the affinity of RnOBP with agonists of vertebrate coagulation pathways. Finally, putative RnOBP oligomers were observed. To obtain the recombinant RnOBP, its nucleotide sequence was obtained from the R. neglectus transcriptome, cloned into the pET100/D-TOPO expression vector and produced by Escherichia coli Rosetta(DE3)pLysS bacteria. The recombinant RnOBP was purified by affinity chromatography, refolded, and used in biophysical characterization assays. This protein is modeled to be entirely formed by α-helices and have a significant internal cavity, potentially a ligand-binding site. Compared with insect OBPs, regardless of the composition or classification of the OBP, the binding site region is well conserved. The recombinant RnOBP purified from inclusion bodies revealed a secondary structure composition different than expected. Initial functional indicated rRnOBP could act to inhibit platelet aggregation, but not coagulation. This study showed RnOBP structural properties and may contribute to the understanding of its functions and biotechnological potential. 

Introduction. Breast cancer globally represents one of the leading causes of mortality among women, which underscores the need for effective pathways for the treatment of individuals with this disease. The collection, storage, and processing of survival data with the interest of understanding the correlation between treatment and survival demands time from researchers and institutions. Objective. The present study was designed to create an innovative real-time survival data network, with the aim of correlating survival time with used treatments, with the purpose of generating hypotheses for future prospective studies regarding the survival benefits of treatment in different clinical, social, and geographic contexts. Methods: After defining the data conventionally considered as crucial in the treatment of breast cancer, a data network was designed that returns to the physician the overall survival, progression-free survival, and event-free survival in real time. The criteria for each survival metric were defined and a real-time chart was generated where all survivals are represented and, when combined, result in the overall survival. Results: With free and simple registration, the network was created and allows access to any Brazilian oncologist. Patient entry is fully parameterized, allowing the quick and safe input of information about the disease and treatment. The physician can access patient data in real time and thus can compare it with results from the literature, especially survival data, using advanced filters. Consent for the submission of information is obtained electronically from the patient, also in real time, from an email address or phone. Conclusion: This data network is feasible and can be a tool of interest in research institutions, particularly as a screening for larger studies.

The progress of molecular techniques was essential for the previous diagnosis of leprosy. However, current studies in the area have not been able to clarify the performance of these techniques in clinical practice yet. The main objective of this project was to test in clinical practice the accuracy, sensitivity and specificity of a new pair of primers developed for the repetitive element target (RLEP) of Mycobacterium leprae. Furthermore, as a secondary objective, was sought to identify the transmission potential of patients with multibacillary and paucibacillary leprosy. Thus, a crosssectional cohort study of diagnostic accuracy was carried out with a total of one hundred patients with a clinical picture compatible with leprosy. In this study, real-time Polymerase Chain Reaction (qPCR) was used to quantify M. leprae in different layers of the skin and in nasal swabs. Skin samples were divided into four layers: epidermis, upper dermis, lower dermis and hypodermis. The results of quantification of the bacillus in nasal swab samples were related to the number of household contacts. As a positivity criterion for the qPCR reaction, three cutoff points were established: 1 - any quantifiable bacillus DNA, 2 - quantification greater than or equal to 0.1 bacillus, 3 - quantification greater than or equal to 1 bacillus. Validation of the in vitro quantification of the reaction resulted in a quantification limit of 0.03 bacilli. The best sensitivity was observed in the upper dermis according to the first established cut-off point [sensitivity = 59.26% (95% CI = 45.97 – 71.32)]. In the epidermis, the third cutoff point resulted in 100% specificity (95% CI = 92.29 - 100). The best accuracy was found in the hypodermis at the first cutoff point [accuracy = 68.09% (95% CI = 58.11 – 76.64)]. For swab samples, the best sensitivity was 20.83% (95% CI = 11.73 – 34.26), and was achieved at the first cutoff point. At the three tested cutoff points, 100% specificity was obtained in the swab samples. The number of bacilli found in nasal swabs was not significantly related to the number of household contacts also diagnosed with leprosy. Paucibacillary patients tested positive only for bacillus fragments in nasal swabs, but not for the entire bacillus. This fact demonstrates that paucibacillary patients may not be a relevant source of disease transmission. It was concluded that different types of skin samples have little influence on the accuracy of the qPCR test. However, in contrast to this result, different cutoff points may influence the accuracy, sensitivity and specificity of the qPCR test. xviii Keywords: L

Introduction: Hematopoietic stem cells (HSC) have numerous therapeutic applications in
the treatment of hematologic malignancies, autoimmune diseases and in regenerative
medicine. The clinical, biochemical and immunological characteristics of individuals are
relevant factors in the quality of HSC collected and cryopreserved, thus interfering with the
results of hematopoietic stem cell transplants. Culture media and culture substrates are in
vitro models necessary to enable experimental research and therapeutic use. This study
aimed to characterize and evaluate the viability and immune markers of human HSC, as
well as to evaluate the role of platelet-rich fibrin (PRF) as a substrate for their cultivation.
Methods: This is a retrospective, descriptive and experimental study conducted with 14
HSC samples obtained by apheresis from 14 healthy adult individuals. The samples were
submitted to typing for HLA antigens by the SSO technique and ABO/Rh systems by
immunohematology. Viability and types of regulated killing were assessed by flow
cytometry. Reactive species were evaluated by flow cytometry (ROS/DCFDA;
ERNs/DAFFM Diacetate). HSC differentiation potential was evaluated using a PRF as
substrate; the results were tabulated and analyzed using the Prism 5.0 program. Results:
HSC donors were young adults with hematological and biochemical parameters within
normal reference values. The cryopreservation time of 34 ± 15 months reduced in 2% the
total HSC-CD34+. There was greater expression of the A*02, DRB1*04, DRB1*07 loci for
HLA antigens. As for the type of regulated cell death, there was no difference between the
percentages of initial apoptosis, late apoptosis and by unknown mechanism. As for the ABO
system, there was a predominance of type O (6 individuals), followed by type A (5
individuals). For the Rh factor there was a predominance of the positive type (10
individuals). After cryopreservation, there was great individual variability in the production
of reactive species and greater production of ERNs than ROS by the group of individuals.
The fibrin mesh favored the anticipation of cell grouping (homing / 2 days of incubation),
while in the absence of fibrin, grouping was observed after 3 days. The fibrin mesh favored
the appearance of signs of cell differentiation (pseudopods and evident nucleolus) after 3
days of incubation. Conclusion: The set of results showed that HSC cryopreservation
parameters should be considered to ensure their quality, as there was a reduction in cell
viability when thawed and some samples had mitochondrial metabolism close to 0 (zero).
Considering that, the PRF mesh provided adhesion, homing and cell differentiation signals,
it is believed that the parameters related to the freezing/thawing method and the
cryopreservation time were appropriate to guarantee the quality of the samples. However,
it is suggested the continuity of studies to determine the effect of the curve of time x cell
quality to help the decision making of national banks of stem cell cryopreservation.

Early detection of subclinical atherosclerosis on patients with Framingham intermediate risk is a major unmet clinical need and additional tests are recommended to personalize the risk estimates. A prognostic revelation of CT-adapted Leaman score coupled with the analysis of potentials candidates for biomarkers by comprehensive metabolomic profile can deepen the current knowledge of the phenotype of patients and be used as a complementary technique for re-stratification of the risk. To test the hypothesis that CT-LeSC is able to discriminate different metabolomic profiles in intermediate risk subjects and aid physicians in the decision-making in order to better stratify these patients. We studied 40 subjects with an intermediate CV risk profile, subdividing them according to the presence or absence of CAD, assessed by computed tomography angiography. They were recruited into three groups according to CT-LeSc score (Low: < 5 and High > 5), and Control, and another analysis in four groups according to CT-LeSc tertiles (T1: 0.3-3.7; T2: 3.8-8.2; T3: 8.3-24.1 and Control). Metabolomic analysis of serum from patients in each group was performed with a global approach based on undirected performance liquid chromatography coupled with time-of-flight mass spectrometry (LC-QTOF/MS) and gas chromatography (GC MSMS). Data were analyzed between January 2018 and July 2020. Among the patients, 28 (70%) had identifiable plaques in the coronary tree; of these, 16 patients (40%) were high-risk, 12 (30%) were low-risk, and 12 (30%) were controls. In the GC MSMS approach, high concentrations of pipecolic acid, 2-hydroxy-3-methylbutyric acid, palmitic acid, oleic acid, asparagine, linoleic acid, L-phenylalanine, L-glutamic acid were found in the control group, while L-5- oxoproline was expressed more intensely in the High CT-LeSc group when compared to the Low CT-LeSc and Control groups. The sPLS-DA model had a low misclassification error rate and the value of the Area Under Curve (AUC) from ROC curves was 0.789 (95% CI: 0.559-0.952), 0.734 (95% CI: 0.424-0.955) and 0.654 (95% CI: 0.389-0.812) in the plaque group vs control, High-CT LeSc group vs control and High-CT LeSc group vs Low-CT LeSc, respectively. In the LC MSMS approach, the data show a discriminating metabolite profile between the extreme phenotypes of the intermediate group (High CT-LeSc versus Control and T3 versus Control), with AUC of 0.926 and 0.967, respectively. In this pilot, prospective, translational study, metabolomics analysis revealed discriminatory metabolic fingerprints of high CT-LeSc and T3 phenotypes, despite comparable intermediate CV risk profiles with controls, which seems to indicate, even in this global analysis, a hint of there may be a discriminating molecular signature for these patients. Ultimately, a specific grouping of metabolites, rather than a single marker, may improve the biological understanding of CVD risk.

Real-world, primary data on the treatment of psoriasis are scarce,
especially concerning the role of soluble biomarkers as outcome predictors. Objective:
We evaluated the utility of Th1/Th17 serum cytokines along with clinical characteristics
as predictors of drug survival in the treatment of psoriasis. Methods: We consecutively
included participants with moderate to severe psoriasis who were followed up for 6
years. Baseline interferon-gama (IFN-γ), tumor necrosis factor-alpha and interleukin
(IL)-2, IL-4, IL-6, IL-10 and IL-17A were measured using a cytometric bead array
(CBA); clinical data were assessed. We calculated hazard ratios (HRs) for drug survival
using a Cox proportional hazards model. Results: We included 262 patients, most of
whom used systemic immunosuppressants or biologics. In the multivariate model, poor
quality of life measured by the Dermatology Life Quality Index (HR = 1.04; 95% CI =
1.01-1.07; p = 0.012) and elevated baseline IL-6 (HR = 1.99; 95% CI = 1.29-3.08; p =
0.002) were associated with treatment interruption. Conclusions: Poor quality of life
and elevated baseline serum IL-6 level predicted treatment interruption in patients with
moderate to severe psoriasis. Although IL-6 is not the most important mediator of the
inflammatory pathway in the skin environment, it is an interesting candidate biomarker
for predicting psoriasis treatment response.

Otologic surgery plays a crucial role in the treatment of hearing loss, infections, and lateral skull base tumors. Accurate segmentation of otologic structures from computed tomography (CT) scans can significantly enhance surgical planning and intraoperative guidance. This manuscript presents two experiments that leverage deep learning algorithms for automated segmentation of key temporal bone structures in CT scans.

In the first experiment, three convolutional neural network (CNN) models (AH-Net, U-Net, ResNet) were compared using 150 manually segmented CT scans. The performance of the models was evaluated based on Dice coefficient, Hausdorff distance, and segmentation speed for the inner ear, ossicles, facial nerve, and sigmoid sinus. AH-Net achieved the best results, with Dice coefficients of 0.91, 0.85, 0.75, and 0.86 for the respective structures. The automated segmentation pipeline demonstrated good correlation with manual segmentation performed by a specialist, opening up possibilities for augmented reality, simulation, and automation applications in otologic procedures.

In the second experiment, a state-of-the-art deep learning algorithm (Swin UNETR) was used to build a prediction model for rapid segmentation of nine key temporal bone structures in 325 clinical CT scans. The model achieved high Dice coefficients, Balanced Accuracy, Volume Similarity, Average Symmetric Surface Distance, and 95th Hausdorff Distance, with a mean processing time of 9.1 seconds per study. This robust model offers valuable new datasets for otologic surgical planning and navigation.

In conclusion, the application of deep learning algorithms in this project demonstrates the potential for improved preoperative evaluation and intraoperative guidance in otologic surgery through automated segmentation of otologic structures from temporal bone CT scans.

- In most patients who undergo phacoemulsification, the cataracts are immature, there are no anatomical changes and, therefore, the surgeries don’t have a high risk of complications. In these patients, is the addition of the femtosecond laser beneficial based on the parameters of Cumulative Dissipated Energy (CDE) and endothelial loss? Purpose: To analyze whether femtosecond laser-assisted surgery leads to less CDE and decreased endothelial cell loss compared to conventional surgery. Patients and Methods: This non-blinded, non-randomized, quasi-experimental clinical trial was conducted at one center and involved one surgeon. Patients with cataracts and 50–80 years old were included, and the exclusion criteria were radial keratotomy, trabeculectomy, drain tube implant, corneal transplant, posterior vitrectomy, and reimplantation of intraocular lens. In total, 298 patients were recruited between October 2020 and April 2021, and the data collected included sex, laterality, age, ocular comorbidities, systemic comorbidities and CDE. An endothelial cell count was performed before and after surgery. Patients were divided according to femtosecond laser-assisted phacoemulsification or conventional phacoemulsification. The femtolaser patients were submitted to the equipment, and then, immediately after treatment, phacoemulsification surgery was performed. In the conventional method, the “divide and conquer” technique was used. The statistical analysis was made using an analysis of covariance linear model, using SAS version 9.4 (SAS Institute, Inc., 1999). Values with p < 0.05 were considered significant. Results: A total of 132 patients were analyzed. The only statistically relevant predictors of CDE were the severity of the cataract (p < 0.0001) and age of ≥75 years (p = 0.0003). The following factors were not significant: technique with or without laser (p = 0.6862), sex (p = 0.8897), systemic arterial hypertension (p = 0.1658), and diabetes (p = 0.9017). Grade 4 cataracts were associated with higher CDE than grade 3 cataracts, which in turn were associated with higher CDE than grade 2 cataracts. A comparison of pre- and post-operative specular microscopy with and without laser revealed no significant discrepancy (p = 0.5017). Conclusion: FLACS didn’t reduce CDE or endothelial cell loss compared to conventional surgery regardless of severity. This result is important for hospitals to be able to make precise indications for the use of FLACS, optimizing costs without reducing the quality of results.

Background: The description of the maternal, fetal and pediatric consequences of
SARS-CoV-2 infection occurring during pregnancy and investigating the underlying
mechanisms is crucial for a better understanding of the disease and providing evidence
for better care and treatments for the mother dyad -son, based on scientific evidence.
Objectives: To characterize the physiological changes that occur in the profile of
immunological mediators during pregnancy, to study changes in the immunological
response in pregnant women infected by SARS-CoV-2 in different trimesters of
pregnancy and to correlate them with clinical and anthropometric outcomes in their
newborns. born in the perinatal period and in the first year of the infants&#39; lives.
Methods: Study carried out in two phases. In the first phase, a prospective
observational study designed to characterize the immunological status of pregnant
women with convalescent COVID-19 and panoramic analysis of the interaction
between soluble serum inflammatory mediators took place. A sample of 141 pregnant
women from all gestational periods comprised patients with convalescent SARS-CoV-2
infection between 3 and 20 weeks after the onset of symptoms (COVID, n=89) and a
control group of uninfected pregnant women whose material for analysis of serum 

inflammatory mediators was collected in the pre-pandemic period (CG, n=52). In the
second phase, a total of 262 newborns born to pregnant women who had SARS-CoV-2
infection with the onset of symptoms during the gestational period were divided into
three groups according to the trimester in which the maternal infection occurred: 1st
trimester (n=58), 2nd trimester (n=80) and 3rd trimester (n=124). In this group,
correlations were studied between the immunological profile of pregnant women and
the severity of maternal COVID with clinical and anthropometric outcomes of their
children at birth and during the first year of life.
Results: In the first phase of the study, levels of chemokines, pro-inflammatory
cytokines, regulatory cytokines and growth factors were quantified using a high-
throughput microsphere array. In the GC group, the majority of serum soluble
inflammatory mediators showed a progressive physiological decrease in the 2nd and
3rd trimester of pregnancy, while higher levels of chemokines, cytokines and growth
factors were observed in the COVID-19 patient group. Soluble serum inflammatory
mediator signatures and heatmap analysis indicated that the greatest increase
observed in the COVID group was related to pro-inflammatory cytokines (IL-6, TNF-,
IL-12, IFN- and IL-17 ) . Analysis of a larger set of biomarkers showed an increase in
the ratio of inflammatory mediators in the COVID/GC groups in the 2nd (three-fold
increase) and 3rd (three- to 15-fold increase) trimesters. Integrative network analysis
demonstrated that pregnancy in GC evolves with decreasing connectivity between
pairs of serum soluble inflammatory mediators towards the 3rd trimester. Although the
COVID group had a similar profile, the number of connections was notably lower
throughout the pregnancy. In the second phase of the study, a higher incidence of
pregnant women who developed more severe conditions when infected in the 3rd
trimester (p&lt;0.001) and a higher incidence of prematurity (p=0.0246) was found. There
were no differences in anthropometric measurements at birth when neonates were
compared by the gestational trimester in which the maternal infection occurred or by
the severity of maternal COVID-19. Regarding longitudinal anthropometry follow-up in
the first year of life of this group of children, less weight gain was found in the first
month of life among infants whose mothers had COVID-19 in the 2nd and 3rd
trimesters of pregnancy compared to the 1st trimester group. (p=0.0152) and less
weight gain in the group whose mothers had severe COVID (p=0.0148). In relation to
length, there was no difference in gains over the first year of life between the groups,
but in the intra-group assessment, lower gains were observed in the first month of life
among children of mothers with severe COVID and with infection in the 3rd trimester of
pregnancy. . In relation to head circumference, the intragroup assessment showed a
smaller gain in the first month in all groups, which persisted in the 6th month of life

among infants, except in the group whose mothers had COVID in the 1st gestational
trimester. The improvement in head circumference gain occurred at 12 months, except
in the group of children of mothers with non-severe COVID.
Conclusion: In the group of pregnant women with COVID-19, a pronounced increase
in serum levels of soluble inflammatory mediators was found in the 3rd trimester,
associated with a decrease in the network interaction between them. Greater severity
of COVID in pregnant women was linked to a higher incidence of prematurity and
referral of newborns to the Neonatal Intensive Care Unit. There were no differences in
the anthropometric profile of the newborns at birth, regardless of the group. However,
the anthropometric profile of infants during the first year of life revealed distinct patterns
in terms of weight gain, length, and head circumference, when comparing the groups
based on the severity of maternal COVID and the gestational period during which the
SARS-CoV-2 infection occurred.

Background. Fatigue, cognitive complaints, and olfactory disfunction are the most frequent

persistent symptoms in patients after severe acute respiratory syndrome coronavirus 2 (SARS-
CoV-2) infection. This study aimed to assess fatigue and neuropsychological performance and

investigate changes in the thickness and volume of gray matter and microstructural
abnormalities in the white matter in a group of patients with mild-to-moderate coronavirus
disease 2019 (COVID-19). Additionally, we aimed to investigate changes in olfactory bulb
volume and brain network in the white matter in patients with persistent hyposmia following
COVID-19.
Methods. We studied 56 COVID-19 patients and 37 matched controls using magnetic
resonance imaging (MRI). Cognition was assessed using Montreal Cognitive Assessment and
Cambridge Neuropsychological Test Automated Battery, and fatigue was assessed using
Chalder Fatigue Scale (CFQ-11). The Sniffin' Sticks smell identification test (SS-16) was used
to evaluate participants' ability to identify odors. T1-weighted MRI was used to assess gray
matter thickness and volume. The olfactory bulbs were manually segmented from T2-weighted
MRI. Fiber-specific apparent fiber density (FD), free water index, and diffusion tensor imaging
data were extracted using diffusion-weighted MRI (d-MRI). Network-Based Statistics (NBS)
and graph theoretical analysis were used to explore the WM. d-MRI data were correlated with
clinical, cognitive measures, and SS-16 score using partial correlations and general linear
modeling.
Results. COVID-19 patients had mild-to-moderate acute illness (95% non-hospitalized). The
average period between real-time quantitative reverse transcription polymerase chain

reaction-based diagnosis and clinical/MRI assessments was 93.3 (±26.4) days. The COVID-
19 group had higher total CFQ-11 scores than the control group (p < 0.001). There were no

differences in neuropsychological performance between groups. The COVID-19 group had
lower FD in the association, projection, and commissural tracts, but no change in gray matter.
The corona radiata, corticospinal tract, corpus callosum, arcuate fasciculus, cingulate, fornix,
inferior fronto-occipital fasciculus, inferior longitudinal fasciculus, superior longitudinal
fasciculus, and uncinate fasciculus were involved. CFQ-11 scores, performance in reaction

time, and visual memory tests correlated with microstructural changes in patients with COVID-
19.

The COVID-19 persistent hyposmia group had reduced olfactory bulb volume compared to
controls. In NBS, COVID-19 patients showed increased structural connectivity in a subnetwork
comprising parietal brain regions. Regarding global network topological properties, patients
exhibited lower global and local efficiency and higher assortativity than controls. Concerning
local network topological properties, patients had reduced local efficiency (left lateral orbital
gyrus and pallidum), increased clustering (left lateral orbital gyrus), increased nodal strength
(right anterior orbital gyrus), and reduced nodal strength (left amygdala). SS-16 test score was
negatively correlated with clustering of whole-brain white matter in the COVID-19 group.
Conclusions. Quantitative d-MRI detected changes in the white matter microstructure of
patients recovering from COVID-19. This study suggests a possible brain substrate underlying
the symptoms caused by SARS-CoV-2 during medium- to long-term recovery. Additionally,
patients with olfactory disfunction after COVID-19 had relevant white matter network
dysfunction with increased connectivity in the parietal sensory cortex. Reduced integration and
increased segregation are observed within olfactory-related brain areas might be due to
compensatory plasticity mechanisms devoted to recovering olfactory function.

The cholinergic vasodilation plays an important role in the blood pressure (BP) and systemic vascular resistance (SVR) responses at the onset of isometric handgrip exercise. Herein, we tested the hypothesis that women present an attenuated fall in SVR and increased pressor response compared to men at the onset of isometric handgrip exercise due to a blunted sympathetic cholinergic vasodilation. Twenty (10 women) young and healthy volunteers (average age: 22 ± 1 years old) were recruited. Participants performed 20s bout of isometric handgrip exercise at 40% of maximal voluntary contraction. Heart rate (HR), mean BP, stroke volume (SV), cardiac output (CO), and SVR responses to exercise were expressed as a change from 10s immediately before the voluntary contraction. Men demonstrated a drop in SVR at 3s (Δ-0.9 ± 0.3 mmHg.L-1 .min), 10s (Δ -1.6 ± 0.4 mmHg.L-1 .min) and 20s (Δ-1.9 ± 0.7 mmHg.L-1 .min) of the onset of isometric exercise, and these responses tend to be attenuated in women (3s: Δ-0.3 ± 0.3 mmHg.L-1 .min; 10s: Δ-1.2 ± 0.4 mmHg.L-1 .min; 20s: Δ-0.6 ± 0.5 mmHg.L-1 .min; P=0.131). While men present a typical exercise-evoked biphasic mean BP response, women show a progressive increase in BP at the onset of exercise. There are no sex differences in HR, SV and CO responses to exercise, which all increased progressively over the 20s of handgrip bout. These findings demonstrated a lack of sex differences in SVR and BP responses at the onset of isometric handgrip exercise, suggesting that the sympathetic cholinergic vasodilation contribution to circulatory responses at the beginning of exercise is irrespective of sex.

Introduction. Uroflowmetry is an often-performed urological test, providing relevant
diagnostic information for many voiding disorders. Home-based uroflowmetry can
potentially better estimate the patient’s voiding pattern and could be more widely
adopted if an accurate low-cost device was available.
Objective. Development and initial evaluation of an open-platform, open-source
ambulatory uroflowmeter.
Patients and methods. A portable uroflowmeter was designed and built with a load
cell and a Secure Digital (SD) memory card unit connected to a Arduino Uno
microcontroller board. The device was powered by a 9-volt battery and mounted upon
a 3D-printed frame. Its output consisted of comma-separated value (CSV) tables, which
were plotted and analyzed with the R programming language. Twenty healthy male
urologists volunteered to test and assess the device with regards to ease of use and
portability. Each participant answered the International Prostatic Symptom Score (I-
PSS) and Peeling diagram, completed a 24 hour bladder diary, and a questionnaire to
evaluate user experience. Two experienced urodynamicists rated the quality of
uroflowmetric tracings with a 3-degree ordinal scale (1. Interpretable without artifacts;
2. Interpretable with artifacts; 3. Uninterpretable).
Results. Participants’ median age was 36 years old. None had an I-PSS above five or a
Peeling score greater than two. Overall, 138 voidings were recorded (77 daily and 61
nighly episodes). Assessments regarding ease of use, robustness and portability
obtained overall maximum score in 80% of the evaluations. Most uroflowmetry
tracings had no artifacts (91.30% and 84.06% according to urodynamicist A and B,
respectively). About 98% were considered interpretable by both evaluators.
Discussion. A wider adoption of ambulatory uroflowmetry has so far been hampered
by cost and limited device availability, which we conjectured could be circumvented by
a 3D printed portable uroflowmeter using widely accessible programmable
microcontroller boards as processing unit.
Conclusion. The construction of an accurate, easy-to-use, low-cost uroflowmeter
suitable for ambulatory use is feasible. This device can potentially increase access to
home-based and ambulatory uroflowmetry and improve knowledge about the
patients’ voiding patterns.

The present study aimed to evaluate the general toxicity of the use of piplartine (Piper
tuberculatum), as well as to analyze its teratogenic potential in the embryonic development of mice (Mus muscullus). For the execution of the work, the mice were arranged in the proportion of three females for one male, for the accomplishment of the coitus. Following the confirmation of the female's pregnancy, through the presence of the vaginal plug, treatment with piplartine at concentrations of 25, 50 and 100 mg/kg was started. Treatment of pregnant females was performed via gavage, once a day, between the sixth and eighteenth day of gestation, and the animals were monitored. In this work, aspects such as maternal toxicity, embryo-fetal toxicity, fetal visceral analysis, fetal skeletal analysis, hematological and biochemical analysis and finally, DNA fragmentation and cell cycle were analyzed. Overall, maternal toxicity was assessed through visual observations regarding the weight of organs, such as liver, spleen, heart, intestine, lung and placenta, from which they were removed and weighed. These data demonstrate that even with no statistical difference between the groups of Organs analyzed organs, the heart showed the greatest weight gain in the experimental group of 100mg/kg and the other organs in the other concentrations tested. In addition, embryo-fetal toxicity showed no statistical difference between the mice studied. Regarding fetal anomalies, and fetal skeletal analyses, there were also no changes related to piplartine. Hematological and biochemical analyzes were also performed,
where we can observe that there was no significant change in the erythograms, leukograms and plateletograms evaluated. In the evaluation of DNA fragmentation in bone marrow cells, the results expressed fragmentation rates of less than 22% in the experimental groups, similar to fragmentation of approximately 19% in the control. The distribution of subpopulation of cells in the cell cycle shows a statistically significant difference in the S phase of the experimental group treated at the concentration of 50 mg/kg of piplartine. The results corroborate the safety of piplartine exposure to the concentrations and animal model evaluated, and it is possible to observe that piplartine did not demonstrate significant toxic effects and/or correlated with the exposure of the experimental groups. The study demonstrated innovative potential regarding the molecule, providing promising results for further studies and corroborating biomedical, nanotechnological and pharmaceutical applications.

Introduction: Vascular cognitive decline is one of the varieties of dementia syndromes resulting from cerebrovascular impairment and its ischemic manifestations with a high prevalence in the elderly. Calcifications of cardiac structures such as the mitral annulus, aortic valve, papillary muscles and aortic root, in addition to being prevalent in the elderly population, are an expression of systemic atherosclerosis, also associated with cardiovascular risk factors common to those for cerebrovascular atherogenesis. Thus, patients with a higher degree of cardiac calcifications are expected to have lower cognitive performance and a higher prevalence of cognitive decline.

Objective: To verify the association between cardiac calcium index and cognitive decline in octogenarians free of overt cerebrovascular disease. Methods: Cross-sectional analysis of an observational cohort study evaluating the prevalence of risk factors and the development of atherosclerotic disease in patients aged 80 years and older free of overt cerebrovascular disease at baseline. For cognitive assessment, the Mini Mental State Examination (MMSE) was performed. The cardiac calcium index was obtained through a two-dimensional transthoracic echocardiogram according to the semi-quantitative score proposed by Gaibazzi et al. with analysis of calcifications in the mitral annulus, aortic valve, papillary muscles and aortic root. The final score was the sum of all calcium deposits identified in a range from 0 (no visible calcium) to 8 (extensive cardiac and aortic root calcium deposits). Non-contrast-enhanced cranial magnetic resonance imaging was performed at T1, T2 and FLAIR views to assess neuroimaging alterations compatible with vascular dementia in a sample of the group. The collected data were analyzed using Student's T test, Mann-Whitney test, chi-square test, multivariate analysis by logistic regression and linear regression.

Results: In the study, 255 elderly people were evaluated. The median age of the enrolled individuals was 83 years, composed of 61% female and presence of cognitive decline in 52 individuals (20.4% of the participants). Mitral annulus calcification was statistically significantly correlated with cognitive decline in both the unadjusted model (p=0.024), the linear regression model adjusted for age, sex and time of study (p=0.033) and the adjusted model for age, sex, time of study, aortic valve sclerosis and total calcification score (p=0.033). The independent variables total calcium score and aortic valve sclerosis were analyzed in multivariate models, which did not correlate with the MMSE score characteristic of cognitive decline. No statistically significant differences on cranial magnetic resonance variables were found between groups. Conclusion: The presence of mitral annulus calcification was independently associated with cognitive decline in this sample of octogenarians free of overt cerebrovascular disease. Total calcium score, aortic valve sclerosis, papillary muscle calcification, and aortic root calcification did not correlate with the MMSE score characteristic of cognitive decline after adjustments.

Introduction: Leprosy is a chronic infectious disease caused by
Mycobacterium leprae and Mycobacterium lepromatosis that mainly affects the
skin and peripheral nervous system. It constitutes a public health problem, and
its manifestations are a result of the interaction between pathogen-host. Its
diagnosis is basically clinical, but complementary tests are great tools to
increase the accuracy of the diagnosis and impact on earlier treatment and
contributes to stop the transmission chain. The molecular tests are the most
promising.
Methodology: This is an analytical cross-sectional study aimed at evaluating
the diagnostic accuracy of real-time polymerase chain reaction (qPCR) in the
diagnosis of leprosy. It consists of identifying the presence of a genetic
fragment of M. leprae in the peripheral serum of immunocompetent patients
treated at the Hansenology outpatient clinic of the HUB from 06/01/2020 to
06/01/2021. The chosen gene in the study was RLEP, and the reactions were
carried out in a Quantstudio 1 thermocycler.
Results: 56 patients were included in the study, being 20 cases and 36
controls. Ages had a normal distribution in both groups, with a mean of 39.9
years (SD 17.3) in the case group and 45.19 years in the controls (SD 14.75).
Most patients were diagnosed with the Virchowian form (multibacillary - 50%),
followed by borderline (30%), tuberculoid (15%) and pure neural forms (5%).
Previous work used 200µl of serum, while the analysis of the present study
used the content of up to 1500µl, the so-called concentrated serum. Only 2
patients had the expression of the RLEP gene in the concentrated serum
analysis, generating a sensitivity of 10%. The evaluation of the concentrated
serum showed better accuracy, since the specificity was 100%, while in the
normal serum the specificity was 94.44%. Comparing the two analysis
techniques in serum, the Mcnemar test showed that the techniques are
equivalent (p = 0.633), despite the better specificity in concentrated serum.
However, the agreement evaluation by the Kappa method resulted in 0.3
(0.05465 - 0.5453) showing that the theses do not have good agreement.
Conclusion: Blood is not the best tissue to be used in the laboratory diagnosis
of leprosy. Even with the improvement of techniques and sample concentration,
the qPCR of RLPE in the serum of affected patients maintains low sensitivity,
limiting its indication on a large scale. Even so, its high specificity can contribute
to the evaluation of difficult-to-manage cases.

Resumo em inglês: Cervical cancer is the third most common cancer in incidence and mortality in women. The natural history of cervical cancer initially involves HPV infection with progression to pre-cancer and invasive cancer. Vaccination against HPV was implemented in the Federal District in 2013, with the quadrivalent vaccine, which targets only high-risk types 16 and 18, unlike the ninth-valent vaccine, which also protects against high-risk types 31, 33, 45, 52 and 58. In Brazil, after the implementation of the vaccine, there was a reduction in the incidence only of HPV types associated with the quadrivalent vaccine and there are no studies showing the impact of HPV vaccination on the development of precancer and cervical cancer. Cervical cancer screening in Brazil is still performed by cytopathological examination, thus, the objective of this study is to compare the frequency of altered cervical cytopathological exams in vaccinated and unvaccinated patients against HPV in the Federal District. Results of tests performed in the years 2018 to 2021 of patients eligible for HPV vaccination in the Federal District since the beginning of the campaigns in 2013 were included, that is, patients born after the year 2000. The vaccination status of the patients was obtained in list made available by the Health Department of the Federal District. Data analysis was performed by calculating the percentage of the difference for less or for more, using the Fisher test, to compare frequencies, and using the t test, to compare the mean age of the patients, using the Graphpad Prism program for Windows. A total of 9629 cervical Pap smears from the population eligible for HPV vaccination were analyzed from 2018 to 2021. Only 48.17% of the exams were from vaccinated patients and most (93.05%) of them received 3 or 2 doses of the vaccine. Thus, 44.83% of the cervical cytopathological exams were from patients who received at least 2 doses of the vaccine.The frequency of abnormal cervical cytopathological examinations in the population eligible for HPV vaccination was 6.93%. The most frequent diagnostic categories of abnormal exams in both groups, vaccinated and unvaccinated, were atypical squamous cells of undetermined significance (ASC-US) and low-grade squamous intraepithelial lesion (LSIL), corresponding to 53.67% and 38.41% of the abnormal exams in unvaccinated and 60.50% and 30.89% in vaccinated. The frequency of abnormal exams was 7.09% and 6.76%, respectively, in unvaccinated and vaccinated patients, being 4.65% lower in exams of vaccinated patients, but this difference was not significant. The frequency of LSIL was 23.16% lower in vaccinated. The number of exams with LSIL was significantly lower in vaccinated, but there was no significant difference between vaccinated and nonvaccinated regarding the number of exams with high-grade squamous intraepithelial lesion (HSIL) and other diagnostic categories. The total frequency of examinations in the diagnostic categories for which colposcopy is recommended (ASC-H, HSIL and AGC) was not different between unvaccinated and vaccinated (0.56% vs. 0.56%). The age of patients in the population eligible for HPV vaccination when undergoing cervical cytopathological examination in the years 2018 to 2021 ranged from 9 to 22 years, with a mean (± SD) of 18.21 (±0.01) years . The mean age of vaccinated patients when undergoing cervical cytopathological examination was significantly higher when compared to that of unvaccinated patients (18.45± 0.02 x 17.98 ± 0.02 x, t test, p<0 .0001), regardless of the test result (negative or positive). The types and respective frequency of microorganisms were similar in vaccinated and unvaccinated and in accordance with previous studies carried out before the introduction of vaccination. Lactobacillus sp was the most frequent organism regardless of vaccination status. Regarding the abnormal findings of organisms considered endogenous, supracytoplasmic bacilli and Candida sp were the most frequent and there was no significant difference in frequency between vaccinated and unvaccinated. In terms of abnormal findings in the microbiology suggestive of STIs, presence of Trichomonas vaginalis and cytopathic effect by the Herpes virus, no significant difference was observed between vaccinated and unvaccinated When comparing abnormal and negative tests for malignancy within the same group (vaccinated or unvaccinated), in vaccinated, Lactobacillus sp were significantly more frequent in negative while cocci and other bacilli were more frequent in abnormal ones; and, in unvaccinated, supracytoplasmic bacilli were more frequent in abnormal tests than in negative ones. Abnormal microbiology findings analyzed together (excluding the finding of Lactobacillus sp without other microorganisms) were more frequent in abnormal tests than in negative tests in vaccinated and unvaccinated, but the difference was significant in vaccinated. When analyzed by diagnostic category, a significant difference in the frequency of abnormal findings in microbiology was observed only when comparing the ASCUS category with negative for malignancy. Abnormal findings were significantly more frequent in ASCUS category than in negative test in vaccinated . It can therefore be concluded that: i) the frequency of vaccinated and unvaccinated patients in the population eligible for vaccination against HPV and who underwent cervical cytology examination in the last 4 years is similar to vaccination coverage, which suggests that the vaccination status did not change the search for care; ii) the greatest impact after 9 years of initiation of HPV vaccination in the cytopathological examination for cervical cancer screening is in the development of low-grade squamous intraepithelial lesion (LSIL) which was less frequent in vaccinated, indicating a vaccine protection against HPV in the development of this type of lesion; iii) the diagnosis of intraepithelial lesion in the cytopathological examination of the cervix of vaccinated patients, mainly in the diagnostic category of high squamous intraepithelial lesion (HSIL), emphasizes the importance of defining which types of HPV not associated with the quadrivalent vaccine are the most commonresponsible for the development of these lesions and, consequently, what would be the cost/benefit of implementing the nonavalent vaccine in Brazil; iv, HPV vaccination did not change the frequency of microbiological findings, including those associated with with sexually transmitted infections and, consequently, it can be concluded that vaccination did not result in an increase in risky sexual behavior in vaccinated women; v, the association of abnormal findings in microbiology with abnormal result, in both vaccinated and unvaccinated groups, suggests a possible effect of other microorganisms, in addition to HPV, on the appearance of pre-cancer lesions; considering that the difference in abnormal microbiology findings between abnormal and negative is significant only in the ASCUS category, it can be said that the reactive alterations caused by certain microorganisms resulted in diagnostic difficulty; vi) the higher mean age of vaccinated patients compared to unvaccinated patients, which was observed regardless of the test result (negative or altered) and the most frequent findings in microbiology, suggests a later search for care at the UBS, which may be associated to a feeling of protection from the vaccine. Palavras chaves em português: HPV, câncer, v

Introduction: Neuromuscular diseases represent a heterogeneous group of diseases with diverse phenotypes that share common pathophysiological pathways and generate various respiratory manifestations, such as reduced lung volumes, greater collapse in extrathoracic airways, impaired ability to cough, reduced clearance of airway secretions and recurrent infections. In order to reduce morbidity and mortality, the use of cough assistants can optimize the clearing of secretions and reduce the incidence of recurrent infections in these patients. Recently, optimized assistant maneuvers have been described which, in an experimental setting, were able to displace more secretion through a rigid tube. However, the influence of the complex dynamics of the extrathoracic airway on the effectiveness of the maneuvers has not been considered. In individuals affected by these diseases, the upper airways may be pathologically compromised by a combination of hypotonia of the pharyngeal dilator muscles and reduced longitudinal tissue tension resulting from the caudal traction exerted by the thoracic structures. Both changes culminate in reduced airway diameter and a tendency to collapse, which severely limits the application of this technique. Objectives: To analyze the influence of chamber pressure and longitudinal strain in determining flows and pressures during traditional and optimized mechanical insufflationexsufflation maneuvers in an experimental analog Starling resistor model. Methodology: An experimental study was conducted in the Respiratory Physiology Laboratory of the University of Brasilia with a Starling resistor coupled to a cough assistant and fixed compliance test lung. Combinations of chamber pressure (Pchamber), 0 cmH2O, -3 cmH2O and -5 cmH2O, and longitudinal strain (strain), 0%, 12.5%, 25%, 37.5% and 50%, were applied to standard and optimized maneuvers with pressures of +30/-30 and +40/-40 cmH2O configured in the cough assistant. Pressure and flow curves generated by the maneuvers were recorded, from which we extracted the data of interest: peak inspiratory flow (PFI), proximal pressure at peak inspiratory flow (Pupstream Insp), chamber pressure at peak inspiratory flow (Pcamber Insp), distal pressure at peak inspiratory flow (Pdownstream Insp), proximal pressure at zero flow (Pupstream V0); chamber pressure at zero flow (Pchamber V0); distal pressure at zero flow (Pdownstream V0), peak expiratory flow (PEF); proximal pressure at peak expiratory flow (Pupstream Exp); distal pressure at peak expiratory flow (Pdownstream Exp). Subsequently, bias flow was calculated, through the PEF/PFI difference PEF-PFI ratio. For statistical analysis, Split-plot design was used to evaluate the effect of Pcamber, strain and Pcamber*strain interaction in determining the flows and pressures studied. Results: During insufflation, there was a statistical difference between the peak flows obtained and between the peak pressures recorded, and this difference was due in most maneuvers and combinations to the interaction between Pcamber and strain. For all maneuvers performed, the mean peak flow was higher around the 12.5% and 25% strain and Pcamber between 0 and -3 cmH2O, representing better coupling for adequate insufflation with these combinations. In the zero flow phase, between inspiration and expiration, the interaction between Pcamber and strain was also a determinant of the equilibrium pressures achieved. However, the effective values reached were close to the pressures configured in the cough assistant, exerting little influence on airway patency in this phase. Finally, in the exsufflation phase there was no statistical difference between the peak flow obtained and between most of the peak pressures recorded. Conclusions: During the insufflation phase, the competence of the airway dilator mechanisms is able to significantly influence the effectiveness of the cough assistant in promoting adequate inspiration. The combinations between Pcamber and strain that generate higher PFI are in agreement with previously described literature data, which found an ideal strain around 25%, where there was no airflow limitation. In the exhaled phase, the absence of statistical significance in relation to the manipulations of Pcamber and strain suggest that regardless of the adjustment made, the system is more susceptible to flow limitation and collapse due to the high negative pressures usually applied by the cough assistant in this phase. This inability of the compensatory adjustments of the airway to withstand high negative pressures may represent a greater limitation of the application of the cough assistant in the expiratory phase. Our study therefore suggests the need to evaluate in further studies strategies to optimize the expiratory phase of the cough assistant maneuvers, such as the effect of lower exsufflation pressures on flow limitation and airway collapse, and the effectiveness of clearing secretions.

Introduction: Tungiasis is an ectoparasitis caused by Tunga penetrans. It inhabits rural soils, communities with low social development or indigenous communities. Despite being a selflimited infection, the disease may be the gateway to secondary complications. The control of infestation at the community level is complex and there is no accepted intervention. The most widely used treatment in tungiasis is the manual extraction of fleas causing consequences such as bacterial infections. Objectives: We aimed to evaluate the scientific knowledge through a systematic review of the literature and to evaluate the knowledge of health professionals and students regarding the control of tungiasis at both individual and environmental epidemiological and therapeutic levels. Methods: The first part of the research consisted of a systematic review of the literature with international scope. The second consisted of a qualiquantitative research, using the methodology of exploratory factor analysis with factor Q. Results: The systematic review resulted in 3376 references. with the final inclusion of 19 articles. In the research using the Q method, there were 119 participants. Most of them were female, and the mean age was 33.82 years. The median classification of each statement was not related to the academic degree of the participants in most cases. Only the third statement showed consensus among the participants. Conclusion: Although tungiasis is a neglected disease and mechanical extraction is the most widely used type of treatment, there is sufficient evidence to support the use of occlusive agents, especially products manufactured based on dimeticone. There is a lack of knowledge of endemic areas and factors that contribute to the environmental control of tungiasis through research using the Q method.