http://repositorio.ufc.br/handle/riufc/390 2026-06-21T14:30:00Z http://repositorio.ufc.br/handle/riufc/86843 Título: Implementação e padronização da cultura de fibroblastos derivados de pele para obtenção de DNA genômico para aplicações em técnicas de sequenciamento e análise de cariótipo Autor(es): Laurindo, Lucas Oliveira Abstract: Cancer represents a major global public health challenge, with high incidence, morbidity, and mortality, characterized by the accumulation of genetic and epigenetic alterations that compromise genomic integrity and promote malignant transformation. Among the various types of cancer, hematological neoplasms stand out, including leukemias, lymphomas, and myelodysplastic syndromes, associated with mutations in genes involved in the cell cycle and DNA repair. The distinction between germline and somatic variants is essential for diagnosis, prognosis, and clinical management, with Next Generation Sequencing (NGS) being an efficient tool for detecting these alterations. This study aimed to implement and standardize a protocol for fibroblast culture from skin biopsies to obtain high-quality genomic DNA for NGS. This is an experimental and descriptive study using samples from individuals with and without a history of hematological neoplasms, recruited at the Walter Cantídio University Hospital (HUWC), older than 17 years, processed at the Cancer Cytogenomics (LCC), at the Center for Research and Development of Drugs (NPDM). A cell culture protocol adapted to local conditions was standardized. Under a laminar flow hood, tissue fragments were processed in supplemented AmnioMax-II medium (30% SFB and 1% antibiotic), with dermis isolation and culture in T25 flasks at 37 °C and 5% CO2 until adhesion and growth. The medium was renewed until 80% confluence, followed by passaging. Culture conditions were standardized regarding medium, temperature, CO2, and number of passages. Cell morphology, growth curve, doubling time, cytogenetic stability by G-banding, and DNA integrity by gel electrophoresis were evaluated. The sample included 20 participants, 65% female, and eight samples were excluded due to contamination or lack of growth. The median age was 31 years (17–75) and the mean BMI was 24.96 ± 3.66 kg/m2. The mean growth time was 11.50 ± 3.80 days and processing time was 42.92 ± 23.75 days. DNA showed a median of 83.10 ng/μL (spectrophotometry) and 56.60 ng/μL (Qubit), with purity (A260/A280) of 1.91 ± 0.09. Cultures were successfully established with adequate and predictable growth, presenting typical phases of adaptation, exponential growth, and plateau, with maintenance of chromosomal stability even in late passages. Dermal fibroblasts proved suitable for germline variant analysis due to lower influence of somatic and environmental mutations. A t(9;22) translocation was observed in bone marrow from one patient, absent in fibroblasts, reinforcing its somatic nature and the genetic stability of the model. Despite the limited number of samples, the results demonstrate the feasibility of the model as a reliable source of DNA for NGS with potential applications in research and clinical practice, contributing to the standardization of laboratory workflows in cancer genetics studies, as well as supporting investigations into genetic predisposition. Tipo: Dissertação 2026-01-01T00:00:00Z http://repositorio.ufc.br/handle/riufc/86785 Título: Esclerose sistêmica em população de baixa renda: perfil clínico, epidemiológico e funcional em região de baixa latitude Autor(es): Alcantara, Vitória Myria Moura Arruda Abstract: Systemic sclerosis (SSc) is a rare autoimmune disease characterized by vasculopathy, inflammation, and progressive fibrosis of the skin and internal organs, with heterogeneous clinical presentations and outcomes influenced by socioeconomic factors. This study aimed to describe the clinical, sociodemographic, and epidemiological profile of patients with SSc, as well as to analyze associations with clinical outcomes and functional capacity. This is an observational, descriptive, cross-sectional study with retrospective components, conducted at the Rheumatology Outpatient Clinic of the Walter Cantídio University Hospital in Fortaleza, Brazil. A total of 90 patients diagnosed with SSc were included after providing informed consent. Data were organized in electronic spreadsheets and analyzed using descriptive and inferential statistics according to variable distribution, with categorical variables expressed as absolute and relative frequencies. The study was approved by the Research Ethics Committee (CAAE: 77968023.7.0000.5054). The sample was predominantly female (92%) and self-reported as mixed-race (91.1%), with a mean age of 51.7±12.7 years. A higher frequency of the diffuse cutaneous subtype (55.5%) was observed, in contrast to the literature, which typically reports predominance of the limited form. Regarding social determinants, 30.1% had income below one minimum wage and 26.7% had low educational level. In terms of functional capacity, 60.7% of patients had mild impairment, approximately one-third had moderate impairment, and a small proportion had severe impairment, reflecting disease heterogeneity. Clinical manifestations included cutaneous involvement (skin thickening 90%), gastrointestinal involvement (gastroesophageal reflux 77.8%), and interstitial lung disease (65.4%). Raynaud’s phenomenon was observed in 75.6% of patients, a lower frequency than reported in other cohorts. Lower educational level was associated with higher inflammatory activity and higher modified Rodnan skin score, while lower income showed a tendency toward longer time to diagnosis. Systemic sclerosis in this cohort presented a more severe profile, with predominance of the diffuse subtype and high frequency of pulmonary involvement. The lower prevalence of Raynaud’s phenomenon and the inversion in subtype distribution compared to the literature suggest possible regional clinical particularities. Socioeconomic factors showed a potential impact on disease progression, particularly regarding inflammatory profile and diagnostic delay, highlighting the need for a multidisciplinary approach and strategies that integrate clinical and social aspects to promote early diagnosis and greater equity in care. Tipo: Dissertação 2026-01-01T00:00:00Z http://repositorio.ufc.br/handle/riufc/86780 Título: Características clínicas e socioeconômicas de pacientes com psoríase e artrite psoriásica em uma população de baixa renda Autor(es): Montenegro, Mariana Lima Abstract: Psoriasis (PsO) is a chronic inflammatory disease affecting the integumentary system and may also involve the joints, characterizing psoriatic arthritis (PsA). Inflammatory mediators associated with obesity and metabolic syndrome, such as cytokines and adipokines, have been implicated in the pathogenesis of PsO and PsA. Environmental and socioeconomic factors also influence both conditions; however, there are few studies involving low-income, non-predominantly White populations outside the Northern Hemisphere. We described the clinical profile of PsO and PsA in a low-income population, evaluating the impact of socioeconomic and environmental factors, including low latitude. Patients with PsO and PsA (CASPAR criteria), seen between January 2023 and January 2025, were examined, and medical records were reviewed to exclude other causes of arthritis and comorbidities. We recorded body mass index (BMI), serum C-reactive protein (CRP), Psoriasis Area and Severity Index (PASI), Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Ankylosing Spondylitis Disease Activity Score (ASDAS-CRP), Disease Activity in Psoriatic Arthritis (DAPSA), Health Assessment Questionnaire (HAQ-DI), use of disease-modifying antirheumatic drugs (DMARDs), and Short Form-36 (SF-36) scores, as well as monthly family income and educational level. A total of 303 patients were included (149 with PsO and 154 with PsA), with a median age of 53 years (IQR: 38–61), of whom 188 (62.05%) were female. Plaque psoriasis (133; 89.26%) and polyarticular involvement (91; 59.09%) predominated. Most participants (192; 63.37%) reported a monthly income ≤ 1 minimum wage. The median BMI was 28.20 kg/m2 (IQR: 24.91–32.42), similar between PsO and PsA groups. Median CRP levels were 0.30 mg/dL (IQR: 0.20–0.70) in PsO and 0.84 mg/dL (IQR: 0.3–2.0) in PsA. The median PASI was 2.20 (IQR: 0.58–4.25) in PsO and 1.00 (IQR: 0–2.80) in PsA. Overall, disease activity was low, with ASDAS-CRP 2.20 (IQR: 1.00–2.90), BASDAI 3.50 (IQR: 0.75–5.30), and DAPSA 12.75 (IQR: 2.92–23.88). Quality of life, assessed by SF-36, was worse in PsA compared to PsO. Women with PsO had worse HAQ-DI scores than men (p = 0.005). Patients with lower income showed worse performance in the domains of physical functioning (p = 0.020), role physical (p = 0.028), and social functioning (p = 0.045). Most patients were receiving biologic therapies, with 111 (72.08%) in the PsA group and 93 (62.42%) in the PsO group. Despite low income and educational levels, with 116 (38.28%) having fewer than 8 years of schooling and only 70 (23.1%) having higher education, we observed low disease activity in both PsO and PsA. Women, despite more frequent use of DMARDs, had worse quality of life scores and greater symptom burden. This is the first description of a low-income, low-education population living at low latitude with PsO and PsA. These findings are noteworthy, as they suggest a better prognosis than that observed in populations with higher income and education levels living at higher latitudes. Tipo: Dissertação 2026-01-01T00:00:00Z http://repositorio.ufc.br/handle/riufc/86546 Título: O receptor P2X7 microglial orquestra neuroinflamação, disfunção autofágica e sinucleinopatia em ratos parkinsonianos Autor(es): Nascimento, Tyciane de Souza Abstract: Parkinson’s disease (PD) is a multisystem neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons in the substantia nigra and the presence of protein inclusions known as Lewy bodies, which are primarily composed of aggregated α-synuclein. Under physiological conditions, these proteins are targeted for autophagic degradation. Hyperactivity of purinergic P2X7 receptors has been extensively implicated in the pathophysiology of PD, with growing evidence supporting their role in autophagic dysfunction and protein aggregation. Thus, the present study aimed to investigate the neuroprotective effect of the P2X7 receptor antagonist Brilliant Blue G (BBG), administered during a prodromal phase of PD, focusing on its impact on α-synuclein accumulation and dysfunction of autophagic pathways in an experimental model of rotenone-induced parkinsonism. For this purpose, male Wistar rats were divided into four groups: control, control treated, parkinsonian (rotenone 2.75 mg/kg for 21 days, intraperitoneally), and parkinsonian treated with BBG (50 mg/kg for 15 days, intraperitoneally). Animals were subjected to behavioral tests and, at the end of the protocol, euthanized for brain collection for immunohistochemical and molecular analyses. P2X7 receptor blockade significantly protected the animals against deficits in volatile odor detection, locomotor activity (open field test), motor coordination (rotarod test), depressive-like behavior (sucrose preference test), and working memory (Y-maze test) induced by rotenone exposure. BBG prevented the loss of dopaminergic neurons in the substantia nigra and striatum, reduced neuroinflammation, and decreased the accumulation of total and phosphorylated α- synuclein. Additionally, BBG modulated autophagic and mitophagic pathways, as evidenced by reduced expression of proteins such as LC3 and PINK1. The effect of BBG occurred by decreasing the number of microglial P2X7 receptors. The results indicate that rotenone-induced microglial purinergic hyperactivity acts as a multifaceted pathogenic integration center, amplifying neuroinflammation, disrupting autophagic processes, and promoting the accumulation of pathogenic α-synuclein, thereby creating a vicious cycle that culminates in dopaminergic neuronal death. Thus, P2X7 receptor blockade emerges as a promising therapeutic approach for targeting multiple pathological pathways in PD. Tipo: Tese 2025-01-01T00:00:00Z