Induction of SK-MEL-28 Invasion by Brain Cortical Cell-Conditioned Medium Through CXCL10 Signaling.

Melanoma, an infrequent yet significant variant of skin cancer, emerges as a primary cause of brain metastasis among various malignancies. Despite recognizing the involvement of inflammatory molecules, particularly chemokines, in shaping the metastatic microenvironment, the intricate cellular signaling mechanisms underlying cerebral metastasis remain elusive. In our pursuit to unravel the role of cytokines in melanoma metastasis, we devised a protocol utilizing mixed cerebral cortical cells and SK-MEL-28 melanoma cell lines. Contrary to expectations, we observed no discernible morphological change in melanoma cells exposed to a cerebral conditioned medium (CM). However, a substantial increase in both migration and proliferation was quantitatively noted. Profiling the chemokine secretion by melanoma in response to the cerebral CM unveiled the pivotal role of interferon gamma-induced protein 10 (CXCL10), inhibiting the secretion of interleukin 8 (CXCL8). Furthermore, through a transwell assay, we demonstrated that knockdown CXCL10 led to a significant decrease in the migration of the SK-MEL-28 cell line. In conclusion, our findings suggest that a cerebral CM induces melanoma cell migration, while modulating the secretion of CXCL10 and CXCL8 in the context of brain metastases. These insights advance our understanding of the underlying mechanisms in melanoma cerebral metastasis, paving the way for further exploration and targeted therapeutic interventions.

Perspectivas de estudantes da área de saúde sobre participação em programa intergeracional: potencialidades e desafios / Health students' perspectives on participation in an intergenerational program: potentialities and challenges

Resumo: Introdução: As habilidades de comunicação (HC) são competências indispensáveis para qualquer profissional da área de saúde, pois permitem maior adesão, compreensão e satisfação geral do paciente. O constante envelhecimento da população brasileira resulta cada vez mais na presença do público idoso no sistema de saúde. Esse cenário tem levado as escolas médicas a pensar em uma reformulação dos seus componentes curriculares no âmbito das HC, visto que esse assunto ainda apresenta lacunas. Nesse contexto, os programas intergeracionais possuem um valioso papel no desenvolvimento das HC por permitirem a interação entre diferentes gerações. Objetivo: Este estudo teve como objetivo investigar as perspectivas de estudantes da área de saúde sobre a participação em um programa intergeracional no município de Paulo Afonso, na Bahia, intitulado "[Tec-Idoso]: utilização de tecnologia como ferramenta de inclusão digital e de apoio psicossocial ao idoso", vinculado à Universidade Federal do Vale do São Francisco (Univasf). Método: Trata-se de um estudo qualitativo e exploratório, realizado a partir de entrevistas semiestruturadas, utilizando o método de análise de conteúdo. Resultado: As seguintes categorias temáticas foram definidas: contato intergeracional, percepção e estigma com pessoas idosas, HC, impacto no desempenho acadêmico e impacto nas práticas profissionais futuras. Conclusão: Os resultados da presente pesquisa incluem melhora do desempenho acadêmico e da interação social por meio do desenvolvimento das HC, mudança de perspectivas perante o envelhecimento e preparação para o enfrentamento com mais confiança de situações da vida pessoal e profissional.

Abstract: Introduction: Communication skills (CS) are indispensable competencies for any healthcare professional as such skills lead to greater patient compliance, understanding, and overall satisfaction. The constant aging of the Brazilian population increasingly results in the presence of the older people in the healthcare system. This has led medical schools to think about a reformulation of their curricular components within the scope of CS, since this subject still presents gaps. In this context, intergenerational programs play a valuable role in the development of CS by allowing interaction between different generations. Objective: To investigate the perspectives of health students about their participation in an intergenerational program in the city of Paulo Afonso/BA entitled "[Tec-Idoso]: utilização de tecnologia como ferramenta de inclusão digital e de apoio psicossocial ao idoso" [Tec-Idoso: use of technology as a tool for digital including and psychosocial support for the elderly], linked to the Federal University of Vale of São Francisco (UNIVASF). Method: This is an exploratory qualitative study based on semi-structured interviews, using the data analysis method. Results: The following thematic categories were defined: intergenerational contact, perception and stigma about old age, CS, impact on academic performance and impact on future professional practices. Conclusion: Given the above, the results obtained from this research include improved academic performance and social interaction through the development of CS, change of paradigms facing aging, and preparation for handling personal and professional life situation with more confidence.

Overexpression of UDP-Glucose 4-Epimerase Is Associated with Differentiation Grade of Gastric Cancer.

The UDP-glucose 4-epimerase (GALE) is a glycosyltransferase, which acts on protein and lipid glycosylation in normal and neoplastic cells. This study is aimed at investigating the differential tissue expression of GALE and its possible association with clinical-pathological parameters and the outcome of gastric adenocarcinoma patients. Seventy-one patients were evaluated in relation to GALE expression by immunohistochemistry. Our results showed that 48 (67.6%) patients were GALE positive and 23 (32.4%) negative. Positive staining was present on well-differentiated and moderate-differentiated histological grade of gastric adenocarcinomas (p < 0.0001). There was no significant association with outcome parameters (p > 0.05). Besides that, our results corroborated with the validation cohort analysis, where the expression of GALE mRNA was also associated with the histological grade (p < 0.001). These results suggest a possible use of this enzyme as a biomarker for well and moderately differentiated tumors.

Dexamethasone inhibits cytokine production in PBMC from systemic sclerosis patients.

Glucocorticoids (GC) are widely used in the treatment of SSc, although there is not much evidence to prove the benefits offered by these drugs in this disease. In this study, we evaluated the effects of a GC on cytokine production in peripheral blood mononuclear cells (PBMC) of SSc patients. The effect of dexamethasone (DEX) was evaluated in PBMC of 21 SSc patients and 10 healthy volunteers after stimulation of cells with anti-CD3 and anti-CD28. Cytokines IL-2, IL-4, IL-6, IL-10, IL-17A, IL-17F, IFN-γ, TNF, and IL-1ß were quantified in the culture supernatant by CBA or ELISA. Of the patients evaluated in this study, 8 (38%) were taking corticosteroids, and esophageal dysfunction was more frequent in these patients when compared to those who did not take corticosteroids. DEX (1.000 nM) treatment in PBMC of SSc patients stimulated with anti-CD3 and anti-CD28 promoted a significant reduction in IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, TNF, IL-1ß (p < 0.001 for all), and IL-17F (p = 0.023) cytokines levels. We did not observe differences in response to in vitro treatment with DEX between groups of patients taking or not taking corticosteroids. In PBMC from healthy volunteers, we observed that DEX treatment significantly reduced IL-4, IFN-γ (p = 0.003 for both), IL-6, IL-10, IL-17A, and TNF (p = 0.002 for all) cytokines. These results show that DEX treatment in PBMC of SSc patients reduced the production of important cytokines involved in the pathogenesis of the disease, suggesting a possible mechanism of action of the CG in the treatment of SSc.

Organic Extract of Justicia pectoralis Jacq. Leaf Inhibits Interferon-γ Secretion and Has Bacteriostatic Activity against Acinetobacter baumannii and Klebsiella pneumoniae.

Justicia pectoralis Jacq. (Acanthaceae) leaves currently found in the Brazilian north-east are widely used to treat diabetes, menstrual pains, asthma, and other disorders. This work aimed to identify the phytochemical characterization and biological activities of J. pectoralis leaf extracts. The plant material was ground and the crude extracts were obtained with water or acetone: water (7:3 v/v), yielding aqueous (JPA), and organic (JPO) extracts. Phytochemical characterization was performed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). Cytotoxicity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay and trypan blue (TB) exclusion assay in peripheral blood mononuclear cells (PBMCs), BALB/c splenocytes, and neoplastic cells (TOLEDO, K562, DU-145, and PANC-1) at 1, 10, and 100 µg/mL. Antibacterial activity was evaluated using the microdilution test to obtain the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Cytokines, IFN-γ, and IL-17A from culture supernatants of BALB/c mice splenocytes were measured by sandwich ELISA. In the TLC analysis, both JPA and JPO extracts presented coumarin and flavonoids. In addition, HPLC was able to identify coumarin, apigenin, and ellagic acid in both extracts. JPO IC50 was 57.59 ± 1.03 µg/mL (MTT) and 69.44 ± 8.08 µg/mL (TB) in TOLEDO. MIC value of JPO against Acinetobacter baumannii and Klebsiella pneumoniae was 500 µg/mL. JPO (100 µg/mL) significantly inhibited IFN-γ levels (p=0.03). J. pectoralis is a potential candidate to be further investigated as an IFN-γ inhibitory agent and against Acinetobacter baumannii and Klebsiella pneumoniae.

Different profile of cytokine production in patients with systemic sclerosis and association with clinical manifestations.

Immune dysregulation is a central process in the pathogenesis of systemic sclerosis (SSc). Cytokines produced by lymphocytes and monocytes are important mediators and induce tissue damage, recruit additional inflammatory cells, and promote extracellular matrix production and fibrosis. In the present research, we aimed to study the associations between levels of cytokines in serum and culture supernatants from peripheral blood mononuclear cells (PBMCs) and clinical manifestations in SSc patients. Serum samples were obtained from 56 SSc patients and 56 unrelated age- and gender-matched healthy individuals. Resting and anti-CD3/CD28-stimulated PBMC cultures were obtained from 19 SSc patients and 8 healthy controls. IL-2, IL-4, IL-6, IL-10, IL-17A, TNF, and IFN-γ levels were measured by ELISA or CBA. Serum cytokines, except IL-17A, were below the kit detection limit in most of the patients and controls. In unstimulated PBMC, the production of TNF(p = 0.004), IL-10(p = .048), IL-2(p < 0.001), and IL-6 (p = 0.01) was higher in SSc patients than in healthy controls. After anti-CD3/CD28 stimulation, scleroderma PBMCs had lower concentrations of TNF(p = 0.009), IL-10(p = .018), and IL-2(p = .002) than HC. In unstimulated PBMC, IL-2 concentration was higher in patients with esophageal dysmotility (p = 0.04), and IL-10 levels had a positive correlation with modified Rodnan score (p = 0.03). After anti-CD3/CD28 stimulation, higher levels of IL-2 and IL-4 were observed in SSc patients with lung fibrosis (p = 0.01 and 0.006, respectively), and higher levels of IL-10 (p = 0.04) and IL-4 (p = 0.04) in patients with digital ulcers. In conclusion, SSc patients have a different profile of cytokine production and this was associated with clinical manifestations.

Corticosteroid inhibits chemokines production in systemic sclerosis patients.

In this study, we evaluated glucocorticoids (GC) effects on cytokine/chemokine levels in serum samples and peripheral blood mononuclear cell (PBMC) production from systemic sclerosis (SSc) patients. We evaluated cytokine and chemokine levels in serum samples from SSc patients taking or not taking systemic glucocorticoids. PBMCs response to methylprednisolone (MP) was examined from 15 SSc patients and 8 healthy control subjects following PBMC stimulation with anti-CD3/CD28. Cytokine (IFN-γ, TNF, IL-2, IL-4, IL-6, IL-10, and IL-17A) and chemokine (CXCL8/IL-8, CCL5/RANTES, CXCL9/MIG, CCL2/MCP-1, and CXCL10/IP-10) levels were quantified in serum and in PBMC culture supernatants by CBA or ELISA. Compared with patients not taking corticosteroids, we did not observe any significant differences in cytokines/chemokines serum levels in patients using systemic corticosteroids. After stimulation with anti-CD3/CD28, PBMCs treated with MP (100µM), showed a significant reduction of CCL2/MCP-1 (p=0.001), CCL5/RANTES (p=0.04), and CXCL8/IL-8 (p=0.003) levels in SSc patients. In PBMC from healthy controls, we observed decreased IFN-γ, TNF, IL-2, and IL-10 levels after MP treatment, compared with stimulated condition (p<0.01 for all). However in SSc patients, we did not find any significant reduction in these cytokine levels after MP treatment. In conclusion, CCL2/MCP-1, CCL5/RANTES, and CXCL8/IL-8 are chemokines that are potentially modulated by corticosteroids in vitro in SSc patients, but no effect was observed on IL-2, IL-4, IL-6, IL-10, IL-17A, TFN, and IFN-γ secretion. These results suggest a potential effect of GCs on SSc treatment and may reflect the benefit of their use in some patients.

Evaluation of Antibacterial, Antineoplastic, and Immunomodulatory Activity of Paullinia cupana Seeds Crude Extract and Ethyl-Acetate Fraction.

Paullinia cupana (Guarana) is a native plant of Amazon region that has very traditional importance. Its seeds are rich in bioactive compounds, including tannins, which exhibit relevant properties. Objective. This study aimed to evaluate antibacterial, antineoplastic, and immunomodulatory activity of P. cupana seeds crude extract (CE) and ethyl-acetate fraction (EAF). Methods. Antibacterial activity was evaluated by determination of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Antineoplastic activity was evaluated by MTT assays in hepatocellular carcinoma (HepG2), breast adenocarcinoma (MCF-7), ductal carcinoma (T47-D), non-Hodgkin's B cell lymphoma (Toledo), T cell leukemia (Jukart), and Acute Leukemia (HL-60) cell lines. BALB/c mice splenocytes were treated to assess IFN-γ, IL-6, IL-17, and IL-10 levels by sandwich ELISA. Results. CE and EAF were not toxic to peripheral blood cells and splenocytes. CE and EAF fractions showed a bacteriostatic activity (MIC = 250 µg/mL) and presented IC50 values of 70.25 µg/mL and 61.18 µg/mL in HL-60 leukemia cell line. All cytokines evaluated had their levels reduced after treatment, following dose-response model. Discussion and Conclusion. Different biological activities were observed for both CE and EAF, suggesting P. cupana as a source of bioactive substances, especially tannins that may be used for several diseases treatments.

The glycomic profile of invasive ductal carcinoma of the breast is altered in patients with hypoxic regions: implications for tumor behavior.

Hypoxic areas in solid tumors are often associated with poor prognosis and resistance to chemotherapy. The aim of the study was to analyze the expression of galectin-1 (Gal-1), galectin-3 (Gal-3), sialic acid and b1-6 branched glycan structures in hypoxic environment of invasive ductal carcinoma (IDC) of the breast. We performed lectin histochemistry with phytohemag glutinin-L (L-PHA) and Sambucus nigra lectin (SNA); and immunohistochemistry for Gal-1, Gal-3, carbonic anhydrase IX, hypoxia-inducible factor, estrogen receptor (ER), progesterone receptor and human epidermal growth factor receptor type-2 for 86 IDC samples. Patients with markers positive for hypoxia were mostly ER-negative (p = 0.003) and presented with more nodal invasion than the non-hypoxic group (p = 0.0439). Concerning the glycobiological aspects, the hypoxic group expressed more of Gal-3 (p = 0.0021) and SNA ligands (p = 0.0498), however, there was no association between lectin- and galectin-staining and clinical and histopathological data. Our results suggest a change in the glycomic profile of patients within hypoxic regions of IDC. However, further studies are needed to evaluate the role of lectin- and galectin-ligands in tumor's hypoxic environment, as well as their potential to be used as therapeutic targets.

Immunohistochemiluminescence detection: a quantitative tool in breast cancer HER-2 status evaluation.

Her-2 status evaluation in breast cancer has prognostic and treatment response value but its interobserver variation among pathologists is a problem since it is not quantitatively assayed. This study presents an immunohistochemiluminescence method to quantify Her-2 in breast cancer. Anti-Her-2 antibody was conjugated to acridinium ester (AE) and used to evaluate/quantify Her-2 status in breast Invasive Ductal Carcinoma (IDC, n=50) comparing with traditional immunohistochemistry. Anti-HER-2-AE results were expressed in Relative Lights Units (RLU) and showed to be able to distinguish and quantify the differences between the three groups of Her-2 status. 3+ Her-2 status presented the highest RLU (246,982 × 10(3) ± 2.061 × 10(3)) compared to 2+ (76,146 × 10(3) ± 0.290 × 10(3)), negative (27,415 × 10(3) ± 1.445 × 10(3)) and normal tissues (27,064 × 10(3) ± 2.060). Status differences were significant between 3+ and 2+ (p=0.0025); 2+ and negative (p=0.0003), and +3 and +1 (p=0.0001) beside this, normal breast control RLU was 27,064 × 10(3) ± 2,060 × 10(3), similar to negative cases. Results showed that anti-HER-2-AE conjugate was effective in breast tumors Her-2 status evaluation, allowing its quantitative establishment to consequently decrease the subjectivity in prognostic and predictive information intrinsic to this test.

Análise da expressão de carboidratos no estroma do carcinoma ductal invasivo da mama e sua correlação com dados clínicos e histopatológicos / Carbohydrates expression analysis in stroma of breast invasive ductal carcinoma and its correlation with clinical and histopathological datas

Objetivos: Analisar o perfil de carboidratos no estroma do carcinoma ductal invasivo, usando histoquímica com lectinas, e correlacionar os achados com dados clínicos e histopatológicos. Métodos: Estudo retrospectivo baseado na análise dos casos de 30 pacientes diagnosticados com carcinoma ductal invasivo durante o período de Maio a Outubro de 2008, As mostras tumorais foram submetidas a histoquímica com lectinas, na qual foram utilizadas as lectinas conjugadas à peroxidase: PNA-Per, 25 mg/mL, UEA-I-Per, 40 mg/mL e Con A-Per, 40 mg/mL específicas para D-galactose, L-fucose e glicose/manose, respectivamente. Posteriormente, os achados histoquímicos foram relacionados com os dados clínicos e histopatológicos (idade, invasão linfonodal, status para p53, tamanho tumoral e variante histológica) das pacientes. Resultados: Na histoquímica com lectinas, a Con A reconheceu a matriz extracelular em 53,33% dos casos, enquanto a PNA e a UEA-I reconheceram 30 e 40%. Já o endotélio de vasos sanguíneos foi mais reconhecido pela UEA-I (40% dos casos) que a Con A (10% dos casos). Biópsias de pacientes cuja matriz extracelular foi reconhecida pela Con A foram positivas também para p53 (p = 0,029), e todos os casos positivos para p53 (p = 0,041) tiveram os respectivos endotélios vasculares reconhecidos por essa lectina. O reconhecimento do estroma pela UEA-I em relação à variante histológica pouco diferenciada mostrou-se significativo (p = 0,034) em relação às demais variáveis analisadas. Conclusões: Os resultados indicaram o potencial das lectinas como sondas eficientes e os glicoconjugados de estroma como biomarcadores ou fonte de informações da biologia tumoral do carcinoma ductal invasivo.

Objectives: To analyze the carbohydrates profile in the stroma of invasive ductal carcinoma, using lectin histochemistry, and correlate the findings with clinical and histopathological data. Methods: Retrospective study based on analysis of 30 cases from patients previously diagnosed with invasive ductal carcinoma during the period from May to October 2008. Tumour samples were subjected to the lectin histochemistry technique, in which the peroxidase-conjugated lectins were used: PNA-Per, 25 mg/mL, UEA-I-Per, 40 mg/mL e Con A-Per, 40 mg/mL specifics for D-galactose, L-fucose and glucose/mannose respectively. Subsequently the histochemical findings were correlated with clinical and histopathological data (age, lymph node invasion, status to p53, tumor size and histological variant) of the patients. Results: In lectin histochemistry, the Con A stained the extracellular matriz of 53.33% of cases, while the PNA and UEA-I recognized 30 and 40% of the ECM, respectively. The blood vessels endothelium was most recognized by the lectin UEA-I (40%) than Con A (10%). Biopsies from patients with ECM stained by Con A were also positive for p53 (p = 0.029), and all positive cases to p53 (p = 0.041) had the respective endothelium recognized by the same lectin. The ECM staining by the UEA-I in relation to poorly differentiated histological variant showed a significant difference (p = 0.034) between the variants analyzed. Conclusions: The results indicated the potential of lectins as efficient probres and stromal glycoconjugates as biomarker or source of information in the diagnosis of invasive ductal carcinoma.