Coupling Kinesin Spindle Protein and Aurora B Inhibition with Apoptosis Induction Enhances Oral Cancer Cell Killing.

Many proteins regulating mitosis have emerged as targets for cancer therapy, including the kinesin spindle protein (KSP) and Aurora kinase B (AurB). KSP is crucial for proper spindle pole separation during mitosis, while AurB plays roles in chromosome segregation and cytokinesis. Agents targeting KSP and AurB selectively affect dividing cells and have shown significant activity in vitro. However, these drugs, despite advancing to clinical trials, often yield unsatisfactory outcomes as monotherapy, likely due to variable responses driven by cyclin B degradation and apoptosis signal accumulation networks. Accumulated data suggest that combining emerging antimitotics with various cytostatic drugs can enhance tumor-killing effects compared to monotherapy. Here, we investigated the impact of inhibiting anti-apoptotic signals with the BH3-mimetic Navitoclax in oral cancer cells treated with the selective KSP inhibitor, Ispinesib, or AurB inhibitor, Barasertib, aiming to potentiate cell death. The combination of BH3-mimetics with both KSP and AurB inhibitors synergistically induced substantial cell death, primarily through apoptosis. A mechanistic analysis underlying this synergistic activity, undertaken by live-cell imaging, is presented. Our data underscore the importance of combining BH3-mimetics with antimitotics in clinical trials to maximize their effectiveness.

Cannabidiol Attenuates In Vivo Leukocyte Recruitment to the Spinal Cord Microvasculature at Peak Disease of Experimental Autoimmune Encephalomyelitis.

Introduction: Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system characterized by neuroinflammation leading to demyelination. The associated symptoms lead to a devastating decrease in quality of life. The cannabinoids and their derivatives have emerged as an encouraging alternative due to their management of symptom in MS. Objective: The aim of the study was to investigate the mechanism of action of cannabidiol (CBD), a nonpsychoactive cannabinoid, on molecular and cellular events associated with leukocyte recruitment induced by experimental autoimmune encephalomyelitis (EAE). Materials and Methods: C57BL/6 female mice were randomly assigned to the four experimental groups: C (control group), CBD (cannabidiol-treated group, 5 mg/kg i.p.; 14 days), EAE (experimental autoimmune encephalomyelitis-induced group), and EAE+CBD (experimental autoimmune encephalomyelitis-induced plus cannabidiol-treated group). Results: The results indicated that 5 mg/kg of CBD injected intraperitoneally between the 1st and 14th days of EAE could reduce the leukocyte rolling and adhesion into the spinal cord microvasculature as well cellular tissue infiltration. These results were supported by a decreased mRNA expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in the spinal cord. Conclusion: Purified CBD reduces in vivo VCAM and ICAM-mediated leukocyte recruitment to the spinal cord microvasculature at EAE peak disease.

Combination Therapy as a Promising Way to Fight Oral Cancer.

Oral cancer is a highly aggressive tumor with invasive properties that can lead to metastasis and high mortality rates. Conventional treatment strategies, such as surgery, chemotherapy, and radiation therapy, alone or in combination, are associated with significant side effects. Currently, combination therapy has become the standard practice for the treatment of locally advanced oral cancer, emerging as an effective approach in improving outcomes. In this review, we present an in-depth analysis of the current advancements in combination therapies for oral cancer. The review explores the current therapeutic options and highlights the limitations of monotherapy approaches. It then focuses on combinatorial approaches that target microtubules, as well as various signaling pathway components implicated in oral cancer progression, namely, DNA repair players, the epidermal growth factor receptor, cyclin-dependent kinases, epigenetic readers, and immune checkpoint proteins. The review discusses the rationale behind combining different agents and examines the preclinical and clinical evidence supporting the effectiveness of these combinations, emphasizing their ability to enhance treatment response and overcome drug resistance. Challenges and limitations associated with combination therapy are discussed, including potential toxicity and the need for personalized treatment approaches. A future perspective is also provided to highlight the existing challenges and possible resolutions toward the clinical translation of current oral cancer therapies.

Increased neutrophil derived chemokines (CXCL10 and CCL2) in human trichinellosis as possible serological markers of the polarization of the immune response against the parasite.

Trichinella britovi is a widely distributed parasitic nematode, transmitted through ingestion of raw or poorly cooked meat containing muscle larvae. This helminth can regulate the host immune system during the early phase of infection. The immune mechanism mainly involves the interaction of Th1 and Th2 responses and related cytokines. Chemokines (C-X-C or C-C) and matrix metalloproteinases (MMPs) have also shown to be implicated in a number of parasitic infections, mainly malaria, neurocysticercosis, angiostronyloidosis, and schistosomiasis, but poor is known about their role in human Trichinella infection. We previously found that serum MMP-9 levels were significantly increased in T. britovi infected patients with relevant symptoms such as diarrhea, myalgia, and facial oedema, which makes these enzymes a potential reliable indicator of inflammation in trichinellosis patients. These changes were also observed in T. spiralis/T. pseudospiralis experimentally infected mice. No data are available about circulating levels of two pro-inflammatory chemokines, CXCL10 and CCL2, in trichinellosis patients with or w/o clinical signs of the infection. In this study, the association of serum level of CXCL10 and CCL2 with clinical outcome of T. britovi infection and their relation to MMP-9 were investigated. Patients (median age 49 ± 0.33 years) acquired infection by consuming raw sausages prepared with wild boar and pork meat. Sera were collected during the acute and the convalescent phases of the infection. A positive significant association (r = 0.61, p = 0.0004) was observed between MMP-9 and CXCL10 levels. The CXCL10 level significantly correlated with the severity of symptoms in patients being particularly higher in patients suffering diarrhea, myalgia, and facial oedema, thus suggesting a positive association of this chemokine with symptomatologic traits, especially myalgia (and increased LDH and CPK levels) (p < 0.005). No correlation was found between levels of CCL2 and the clinical symptoms.

Comparison Between Surgical and Conservative Treatment for AOSpine Type A3 and A4 Thoracolumbar Fractures without Neurological Deficit: Prospective Observational Cohort Study.

Objective To compare the clinical results between conservative (CS) and surgical treatment (CXS) of A3 and A4 fractures without neurological deficit. Methods Prospective observational study of patients with thoracolumbar fractures type A3 and A4. These patients were separated between the surgical and conservative groups, and evaluated sequentially through the numeric rating scale (NRS), Roland-Morris disability questionnaire (RMDQ), EuroQol-5D (EQ-5D) quality of life questionnaire, and Denis work scale (DWS) up to 2.5 years of follow-up. Results Both groups showed significant improvement, with no statistical difference in pain questionnaires (NRS: CXS 2.4 ± 2.6; CS 3.5 ± 2.6; p > 0.05), functionality (RMDQ: CS 7 ± 6.4; CXS 5.5 ± 5.2; p > 0.05), quality of life (EQ-5D), and return to work (DWS). Conclusion Both treatments are viable options with equivalent clinical results. There is a tendency toward better results in the surgical treatment of A4 fractures.

Comparison Between Surgical and Conservative Treatment for AOSpine Type A3 and A4 Thoracolumbar Fractures without Neurological Deficit: Prospective Observational Cohort Study / Comparação entre tratamento cirúrgico e conservador para fraturas toracolombares AOSpine tipo A3 e A4 sem déficit neurológico: Estudo de coorte prospectivo observacional

Abstract Objective To compare the clinical results between conservative (CS) and surgical treatment (CXS) of A3 and A4 fractures without neurological deficit. Methods Prospective observational study of patients with thoracolumbar fractures type A3 and A4. These patients were separated between the surgical and conservative groups, and evaluated sequentially through the numeric rating scale (NRS), Roland-Morris disability questionnaire (RMDQ), EuroQol-5D (EQ-5D) quality of life questionnaire, and Denis work scale (DWS) up to 2.5 years of follow-up. Results Both groups showed significant improvement, with no statistical difference in pain questionnaires (NRS: CXS 2.4 ± 2.6; CS 3.5 ± 2.6; p> 0.05), functionality (RMDQ: CS 7 ± 6.4; CXS 5.5 ± 5.2; p> 0.05), quality of life (EQ-5D), and return to work (DWS). Conclusion Both treatments are viable options with equivalent clinical results. There is a tendency toward better results in the surgical treatment of A4 fractures.

Resumo Objetivo Comparar os resultados clínicos entre os tratamentos conservador (CS) e cirúrgico (CXS) das fraturas A3 e A4 sem déficit neurológico. Métodos Estudo prospectivo observacional de paciente com fraturas toracolombares tipo A3 e A4. Esses pacientes foram separados entre os grupos cirúrgico e conservador e avaliados sequencialmente através da escala numérica de dor (NRS), do questionário de incapacidade de Roland-Morris (RMDQ), do EuroQol-5D (EQ-5D) e da escala de trabalho de Denis (DWS) até 2,5 anos de acompanhamento. Resultados Ambos os grupos apresentaram melhora significante, sem diferença estatística nos questionários de dor (NRS: CXS 2,4 ± 2,6; CS 3,5 ± 2,6; p> 0,05), funcionalidade (RMDQ: CS 7 ± 6,4; CXS 5,5 ± 5,2; p> 0,05), qualidade de vida (EQ-5D) e retorno ao trabalho (DWS). Conclusão Ambos os tratamentos são opções viáveis e com resultados clínicos equivalentes. Há uma tendência a melhores resultados no tratamento cirúrgico das fraturas A4.

Maximizing Anticancer Response with MPS1 and CENPE Inhibition Alongside Apoptosis Induction.

Antimitotic compounds, targeting key spindle assembly checkpoint (SAC) components (e.g., MPS1, Aurora kinase B, PLK1, KLP1, CENPE), are potential alternatives to microtubule-targeting antimitotic agents (e.g., paclitaxel) to circumvent resistance and side effects associated with their use. They can be classified into mitotic blockers, causing SAC-induced mitotic arrest, or mitotic drivers, pushing cells through aberrant mitosis by overriding SAC. These drugs, although advancing to clinical trials, exhibit unsatisfactory cancer treatment outcomes as monotherapy, probably due to variable cell fate responses driven by cyclin B degradation and apoptosis signal accumulation networks. We investigated the impact of inhibiting anti-apoptotic signals with the BH3-mimetic navitoclax in lung cancer cells treated with the selective CENPE inhibitor GSK923295 (mitotic blocker) or the MPS1 inhibitor BAY1217389 (mitotic driver). Our aim was to steer treated cancer cells towards cell death. BH3-mimetics, in combination with both mitotic blockers and drivers, induced substantial cell death, mainly through apoptosis, in 2D and 3D cultures. Crucially, these synergistic concentrations were less toxic to non-tumor cells. This highlights the significance of combining BH3-mimetics with antimitotics, either blockers or drivers, which have reached the clinical trial phase, to enhance their effectiveness.

Epidemiological patterns of adolescent idiopathic scoliosis in a spinal center in curitiba / Epidemiologia da escoliose idiopática do adolescente de um centro especializado em curitiba / Epidemiología de la escoliosis idiopática en adolescentes en un centro especializado en curitiba

ABSTRACT: Objective: Evaluation of the profile of the population with adolescent idiopathic scoliosis (AIS) treated at a center specializing in spine surgery in Curitiba-PR. Methods: Prospective multicenter study, being analyzed, at the moment, only data from the center in question. All patients referred from the basic health service for evaluation of deformity underwent panoramic radiography of the total spine to evaluate the curvature using the Cobb method, evaluation of skeletal maturity using the Risser classification, and the classification of scoliosis using the Lenke classification. Clinical photographs of all patients and an assessment of the quality of life using the SRS-30 questionnaire were also taken. Results: Thirty patients with scoliosis and a mean age of 14.63 ± 3 years were evaluated. Prevalence of female:male 1.5:1. Patients had a mean Cobb of 45.96°, and most were close to Risser 4 skeletal maturity (48.3%). 60% had a thoracic curve (Lenke 1), and 13.3% had a double curve (Lenke 3 or 6). 63.3% of cases had a delay in medical care, taking an average of 18 months between referral and consultation with a specialist. 60% of patients were referred for surgical treatment after the first appointment. Conclusion: The population sample of this center follows the literature. A Cobb >40° in the first consultation with a specialist and a high referral rate to surgery suggest the failure of early diagnosis and the need for public policies for better knowledge and assistance for adolescent idiopathic scoliosis. Level of Evidence IV; Descriptive Epidemiological Study.

RESUMO: Objetivo: Avaliação do perfil da população com escoliose idiopática do adolescente (EIA) atendida no centro especializado em cirurgia de coluna em Curitiba-PR. Métodos: Estudo multicêntrico prospectivo, sendo analisados, no momento, somente os dados do centro em questão. Todos os pacientes encaminhados do serviço de saúde básica para avaliação de deformidade foram submetidos a radiografia panorâmica de coluna total para avaliação da curvatura através do método de Cobb, avaliação da maturidade esquelética pela classificação de Risser e classificação da escoliose por Lenke. Foram, também, realizadas fotos clínicas de todos os pacientes e avaliação da qualidade de vida pelo questionário SRS-30. Resultados: Foram avaliados 30 pacientes com escoliose e idade média de 14,63 ± 3 anos. Prevalência de sexo feminino:masculino 1,5:1. Os pacientes apresentavam Cobb médio 45,96° e a maioria estava próxima da maturidade esquelética Risser 4 (48,3%). 60% apresentavam curva torácica (Lenke 1) e 13,3% dupla curva (Lenke 3 ou 6). 63,3% dos casos apresentaram atraso na assistência médica, demorando em média 18 meses entre o encaminhamento e a consulta com especialista. 60% dos pacientes foram encaminhados para tratamento cirúrgico após a primeira consulta com um especialista. Conclusão: A amostra populacional deste centro encontra-se consoante a literatura. Pelo Cobb >40° na primeira consulta com o especialista e a alta taxa de encaminhamento para cirurgia, fica demonstrado a falha do diagnóstico precoce e necessidade de políticas públicas para melhor conhecimento e assistência da escoliose idiopática do adolescente. Nível de evidência: IV; Estudo Epidemiológico Descritivo.

RESUMEN: Objetivo: Evaluación del perfil de la población con escoliosis idiopática del adolescente (EIA) tratada en un centro especializado en Curitiba-PR. Métodos: Estudio multicéntrico prospectivo, analizándose, por el momento, únicamente los datos del centro en cuestión. A todos los pacientes derivados del servicio básico de salud para evaluación de deformidad se les realizó radiografía panorámica de columna total para evaluación de la curvatura mediante el método de Cobb, evaluación de la madurez esquelética mediante la clasificación de Risser y clasificación de escoliosis mediante la clasificación de Lenke. También se tomaron fotografías clínicas de todos los pacientes y evaluación de la calidad de vida mediante el cuestionario SRS-30. Resultados: Se evaluaron 30 pacientes con escoliosis y edad media de 14,63 ± 3 años. Prevalencia femenino:masculino 1,5:1. Los pacientes tenían un Cobb medio de 45,96° y la mayoría estaba cerca de la madurez esquelética Risser 4 (48,3%). El 60% tenía curva torácica (Lenke 1) y el 13,3% tenía doble curva (Lenke 3 o 6). El 63,3% de los casos tuvo retraso en la atención médica, transcurriendo en promedio 18 meses entre la derivación y la consulta con un especialista. El 60% de los pacientes fueron derivados para tratamiento quirúrgico después de la primera cita. Conclusión: La muestra poblacional de este centro está de acuerdo con la literatura. Por Cobb > 40° en la primera consulta con un especialista y una alta tasa de derivación a cirugía, se sugiere el fracaso del diagnóstico precoz y la necesidad de políticas públicas para mejorar el conocimiento y la asistencia de la escoliosis idiopática del adolescente. Nivel de Evidencia IV; Estudio Epidemiológico Descriptivo.

Navitoclax Enhances the Therapeutic Effects of PLK1 Targeting on Lung Cancer Cells in 2D and 3D Culture Systems.

The efficacy of antimitotics is limited by slippage, whereby treated cells arrested in mitosis exit mitosis without cell division and, eventually, escape apoptosis, constituting a serious resistance mechanism to antimitotics. Strategies to overcome slippage should potentiate the cancer cell killing activity of these antimitotics. Such strategies should accelerate cell death in mitosis before slippage. Here, we undertook a mechanistic analysis to test whether the apoptosis activator Navitoclax potentiates apoptosis triggered by the antimitotic BI2536, a potent inhibitor of Polo-like kinase 1 (PLK1) with the goal of overcoming slippage. We found that cancer cells in 2D cultures treated with BI2536 alone accumulate in mitosis, but a significant fraction of arrested cells undergo slippage and survive. Remarkably, combining BI2536 with Navitoclax dramatically reduces slippage, shifting the cell fate to accelerated death in mitosis. The results are confirmed in 3D spheroids, a preclinical system that mimics in vivo tumor drug responses. Importantly, in 3D spheroids, the effect of the BI2536/Navitoclax combination requires a lower therapeutic dosage of each drug, underlying its potential to improve the therapeutic index. Our results highlight the relevance of apoptosis potentiators to circumvent slippage associated with antimitotics. The combination of BI2536 with Navitoclax shows in vitro synergy/additive effect, which warrants further clinical research.

Inhalation of dimethyl fumarate-encapsulated solid lipid nanoparticles attenuate clinical signs of experimental autoimmune encephalomyelitis and pulmonary inflammatory dysfunction in mice.

RATIONALE: The FDA-approved Dimethyl Fumarate (DMF) as an oral drug for Multiple Sclerosis (MS) treatment based on its immunomodulatory activities. However, it also caused severe adverse effects mainly related to the gastrointestinal system. OBJECTIVE: Investigated the potential effects of solid lipid nanoparticles (SLNs) containing DMF, administered by inhalation on the clinical signs, central nervous system (CNS) inflammatory response, and lung function changes in mice with experimental autoimmune encephalomyelitis (EAE). MATERIALS AND METHODS: EAE was induced using MOG35-55 peptide in female C57BL/6J mice and the mice were treated via inhalation with DMF-encapsulated SLN (CTRL/SLN/DMF and EAE/SLN/DMF), empty SLN (CTRL/SLN and EAE/SLN), or saline solution (CTRL/saline and EAE/saline), every 72 h during 21 days. RESULTS: After 21 days post-induction, EAE mice treated with DMF-loaded SLN, when compared with EAE/saline and EAE/SLN, showed decreased clinical score and weight loss, reduction in brain and spinal cord injury and inflammation, also related to the increased influx of Foxp3+ cells into the spinal cord and lung tissues. Moreover, our data revealed that EAE mice showed signs of respiratory disease, marked by increased vascular permeability, leukocyte influx, production of TNF-α and IL-17, perivascular and peribronchial inflammation, with pulmonary mechanical dysfunction associated with loss of respiratory volumes and elasticity, which DMF-encapsulated reverted in SLN nebulization. CONCLUSION: Our study suggests that inhalation of DMF-encapsulated SLN is an effective therapeutic protocol that reduces not only the CNS inflammatory process and disability progression, characteristic of EAE disease, but also protects mice from lung inflammation and pulmonary dysfunction.

Adhesive systems applied to dentin substrate under electric current: systematic review.

OBJECTIVES: The purpose of this systematic review was to collect and discuss the technique of adhesive systems application on dentin substrate under electric current. MATERIALS AND METHODS: The first search strategy was based on data available at PubMed, LILACS, Scielo, Scopus, and Cochrane Library, using a combination of descriptors such as "dentin bond agents OR adhesive system AND electric current OR electrobond" or "dentin bonding agents OR dentin bonding agent application OR adhesive system AND electric current OR electrobond", with no limit regarding the publication year. The second search strategy was based on the articles' references found previously. An additional search strategy was applied that concerned the proposed theme in the SBU-UNICAMP (Unicamp's Library System Institutional Repository). RESULTS: Twelve studies published between 2006 and 2020 were found. The analyses of the selected studies showed that the use of electric current during adhesive systems application on dentin, whether conventional or self-conditioning, increases resinous monomer infiltration in the dentin substrate, which improves the hybridization processes and the bond strength of the restorative material to dentin. CONCLUSIONS: Despite the favorable results related to the use of this technique, there is still no specific protocol for the application of adhesive systems under electric current.

LyeTx I-b Peptide Attenuates Tumor Burden and Metastasis in a Mouse 4T1 Breast Cancer Model.

Cationic anticancer peptides have exhibited potent anti-proliferative and anti-inflammatory effects in neoplastic illness conditions. LyeTx I-b is a synthetic peptide derived from Lycosa erythrognatha spider venom that previously showed antibiotic activity in vitro and in vivo. This study focused on the effects of LyeTxI-b on a 4T1 mouse mammary carcinoma model. Mice with a palpable tumor in the left flank were subcutaneously or intratumorally injected with LyeTx I-b (5 mg/kg), which significantly decreased the tumor volume and metastatic nodules. Histological analyses showed a large necrotic area in treated primary tumors compared to the control. LyeTxI-b reduced tumor growth and lung metastasis in the 4T1 mouse mammary carcinoma model with no signs of toxicity in healthy or cancerous mice. The mechanism of action of LyeTx I-b on the 4T1 mouse mammary carcinoma model was evaluated in vitro and is associated with induction of apoptosis and cell proliferation inhibition. Furthermore, LyeTx I-b seems to be an efficient regulator of the 4T1 tumor microenvironment by modulating several cytokines, such as TGF-ß, TNF-α, IL-1ß, IL-6, and IL-10, in primary tumor and lung, spleen, and brain. LyeTx I-b also plays a role in leukocytes rolling and adhesion into spinal cord microcirculation and in the number of circulating leukocytes. These data suggest a potent antineoplastic efficacy ofLyeTx I-b.

Trichinella spiralis excretory-secretory products downregulate MMP-9 in Dark Agouti rats affected by experimental autoimmune encephalomyelitis.

Matrix metalloproteinases (MMPs), are implicated in the pathogenesis of multiple sclerosis (MS) and in its animal model, experimental autoimmune encephalomyelitis (EAE). Our aim was to investigate whether amelioration of EAE in Dark Agouti (DA) rats, induced by Trichinella spiralis muscle larvae excretory-secretory products (ES L1), could be related to the level and activity of gelatinases, MMP-9 and MMP-2. Serum levels of MMP-9, MMP-2, NGAL/MMP-9, TIMP-1, and cytokines, evaluated by gel-zymography or ELISA, as well as gelatinases and TIMP-1 expression in the spinal cord (SC), were determined in: i) EAE induced, ii) ES L1-treated EAE induced animals. Milder clinical signs in ES L1-treated EAE induced DA rats were accompanied with lower serum levels of MMP-9 and NGAL/MMP-9 complex. However, the correlation between the severity of EAE and the level of serum MMP-9 was found only in the peak of the disease, with MMP-9/TIMP-1 ratio higher in EAE animals without ES L1 treatment. Lower expression of MMP-9 in SC of ES L1-treated, EAE induced rats, correlated with the reduced number of SC infiltrating cells. In SC infiltrates, in the effector and the recovery phase, production of anti-inflammatory cytokines IL-4 and IL-10 was higher in animals treated with ES L1 prior to EAE induction, compared to untreated EAE animals. Reduced expression of MMP-9 in SC tissue, which correlated with the reduced number of infiltrating cells, might be ascribed to regulatory mechanisms, among which is IL-10.

Glutathione-S-transferase omega 1 and nurse cell formation during experimental Trichinella infection.

The glutathione-S-transferases omega (GSTO) are multifunctional enzymes involved in cellular defense. During the nurse cell (NC) formation in Trichinella spiralis infection, the structural and regulatory genes of the skeletal muscle cell are downregulated and a new phenotype is acquired which advances parasite growth and survival. Previous studies showed that the GSTO1 is overexpressed in the NC during T. spiralis infection. To clarify the role of GSTO1 during NC formation, we evaluated the production of this enzyme by immunohistochemistry (IHC) in the diaphragms of mice experimentally infected with T. spiralis at 15, 28 and 60 days post infection (dpi); phosphorylation of Akt (p-Akt) and JNK1 (p-JNK1) were also evaluated. Furthermore, we evaluated the in vitro effects of T. spiralis excretory/secretory (ES) products from muscle larvae on specific functions (viability, proliferative response, apoptosis) in two cell lines (HeLa and U937), as well as its ability to induce GSTO1, p-AkT, p-ERK1/2 and p-JNK1. Results showed that GSTO1 was elevated in NC present in the diaphragms of T. spiralis experimentally infected mice at 15 dpi and progressively increased up to 60 dpi. The activation pattern of Akt in NC was similar to that of GSTO1, whereas JNK1 was never phosphorylated. ES induced a dose-dependent proliferative response in U937 cells, at 24 h and 48 h of treatment, but not in HeLa cells. However, after 72 h following treatment, significant cell death was observed in both cell lines at all doses. The apoptotic index (a.i.) was significantly higher than in untreated cells in both cell lines but only at the highest concentration of ES tested. Furthermore, Western Blots revealed that cells treated with ES for 24, 48 and 72 h, exhibited time-dependent overexpression of GSTO1, whereas p-Akt appeared only after 24 h of treatment. The p-ERK-1/2 peaked at 24 h then declined at 48 h and 72 h after treatment; however, it remained significantly higher than in untreated cells. No changes were observed in p-JNK1 at 24 and 48 h after treatment but a sharp increase in p-JNK1 was observed at 72 h. Also in HeLa cells, ES induced a small but significant increase in GSTO1 expression after 24 and 48 h of treatment where p-JNK1 was present only after 72 h of treatment. In conclusion, T. spiralis ES can reproduce in vitro the modifications observed inside the NC during experimental infection in mice.

Carcinoma ductal in situ associado à fibroadenoma: relato de caso / Carcinoma ductal in situ arising in a fibroadenoma: report of a rare case

Apesar de fibroadenoma ser uma das lesões mais comuns da mama, sua correlação a alteração maligna é rara.1,2,3,5,7 Quando ocorre tem apresentação clínica similar a dos fibroadenomas típicos, diagnóstico anatomopatológico em geral pós-cirúrgico e tratamento conforme do carcinoma do mesmo tipo histológico isolado.1,2,3,4,6 Apresentamos caso de mulher de 52 anos, apresentando em propedêutica investigativa nódulo em mama direita. Core-biopsy sugeriu carcinoma ductal in situ desenvolvido dentro de fibroadenoma, confirmado em análise anatomopatológica pósoperatória. Tratamento e seguimento se deu conforme já estabelecido pela literatura para carcinoma in situ. Devido baixa incidência, há poucas evidências científicas quanto ao diagnóstico, tratamento e prognóstico desse tipo de lesão. Portanto, apresentar à comunidade científica casos de carcinoma inclusos a fibroadenoma se faz relevante.

Although fibroadenoma is one of the most common lesions of the breast, its correlation with malignant changes is rare. When it occurs, the clinical presentation is similar to typical fibroadenomas, anatomopathological diagnosis is after surgery and treatment goes according to the carcinoma histological type. A case of a 52-year-old woman is presented, with an impalpable nodule in the right breast, diagnosed from screening exams. Corebiopsy suggested carcinoma ductal in situ developed within fibroadenoma, confirmed in the postoperative anatomopathological analysis. Treatment and follow-up followed as established in the literature for carcinoma in situ. Due the low incidence, there are poor scientific evidence regarding the diagnosis, treatment and prognosis of this type of injury. Therefore, presenting cases of Carcinoma ductal in situ arising in a fibroadenoma to the scientific community is so relevant.

Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research.

Most cancer biologists still rely on conventional two-dimensional (2D) monolayer culture techniques to test in vitro anti-tumor drugs prior to in vivo testing. However, the vast majority of promising preclinical drugs have no or weak efficacy in real patients with tumors, thereby delaying the discovery of successful therapeutics. This is because 2D culture lacks cell-cell contacts and natural tumor microenvironment, important in tumor signaling and drug response, thereby resulting in a reduced malignant phenotype compared to the real tumor. In this sense, three-dimensional (3D) cultures of cancer cells that better recapitulate in vivo cell environments emerged as scientifically accurate and low cost cancer models for preclinical screening and testing of new drug candidates before moving to expensive and time-consuming animal models. Here, we provide a comprehensive overview of 3D tumor systems and highlight the strategies for spheroid construction and evaluation tools of targeted therapies, focusing on their applicability in cancer research. Examples of the applicability of 3D culture for the evaluation of the therapeutic efficacy of nanomedicines are discussed.

Toxoplasma gondii Monitoring in Liver Transplantation Patients: A Single Center Cross-Sectional Study in an Italian Hospital.

Toxoplasma gondii (TG) is one of the most widespread intracellular parasites in the world, despite the slight declining trend in industrialized countries. Whilst the infection is often asymptomatic in immunocompetent hosts, in immunocompromised patients such as organ transplant recipients it can have important clinical sequels with even fatal consequences. We retrospectively reviewed 568 primary liver transplants (LT) from deceased donors from 2012 to 2017. Data were analyzed adjusting for year, gender, and age. The study objective was to assess the incidence of post-transplant TG infection and adherence to international guidelines for primary chemoprophylaxis. Prior to transplantation, 42.4% of recipients tested seronegative and 56.5% seropositive, while 36.6% of donors were seropositive and 40.4% showed undetermined serology. Anti-TG antibody titer was higher in patients born abroad (71.4%) versus Italy (54.8%). Among recipients at high risk of post-transplant TG infection, 82.7% of them received chemoprophylaxis, while in 17.3% of cases no prophylaxis was administered. At a mean (SD) follow-up of 21.2 (12.4) months no case of TG infection has been observed. Despite the low rate of adherence to recommendations, prophylaxis of high-risk LT recipients provides control of post-transplant TG infection risk. Review of current guidelines is warranted for low-risk populations.

Assessing the cytotoxic/genotoxic activity and estrogenic/antiestrogenic potential of essential oils from seven aromatic plants.

Alternative therapies with new drugs are needed because the clinical efficacy of conventional chemotherapy is often reduced due to collateral effects. Many natural products of plant origin, including essential oils (EOs) have proved to be effective in prevention and therapy of several diseases such as bacterial infections, chronic diseases and cancer. In the present study, we investigated some biological activities of EOs extracted from seven plants: Rosmarinus officinalis, Salvia somalensis, Thymus vulgaris, Achillea millefolium, Helichrysum italicum, Pistacia lentiscus, Myrtus communis. In particular, we evaluated the cytotoxic and genotoxic activity using the cytochalasin B-blocked micronucleus assay (CBMN) in human peripheral lymphocytes, cytotoxicity in a human ovarian carcinoma cell line (A2780), and the estrogenic/antiestrogenic activity using a yeast strain expressing the human estrogen receptor alpha (ERα). Our results show that most EOs can have a strong cytotoxic and a slight/moderate genotoxic effect on human peripheral lymphocytes, and also a pronounced cytotoxic effect in A2780 cells. In addition, some EOs seem to have a marked antiestrogenic activity that could potentially perturb the estrogen-dependent tissues.

Environmental heterogeneity influences plant resource use - A case study in a rural community of NE Brazil.

In regions of high environmental heterogeneity, it is important to observe if different landscape units are used differently by human populations, which can have consequences for the conservation of these units. Thus, the present study seeks to answer the following questions: (1) are different native vegetation units used for different purposes by the local population? (2) is there a difference between the conservation status of native vegetation units according to the local perception? The study was implemented in the rural community of Morrão de Cima, in the municipality of São Desidério (Northeast Brazil). Data were collected through semi-structured interviews and participatory workshops. The main native vegetation units recognized by the interviewees were Brejo, Mata Seca, Serra and Chapada. The community uses the different units of native vegetation for different purposes. The extraction of medicinal resources was important in the Chapada, while the exploitation of food resources was important in the Brejo, and the collection of wood resources was more substantial in Mata Seca and in Serra. We also found differences in the perceived conservation status of the native vegetation units. Our findings indicate the need of caution when proposing conservation strategies in heterogeneous environments, since each landscape unit may be undergoing specific processes, requiring different strategies.

Effects of Azadirachta indica seed kernel extracts on early erythrocytic schizogony of Plasmodium berghei and pro-inflammatory response in inbred mice.

BACKGROUND: Medicinal plant research may contribute to develop new pharmacological control tools for vector borne diseases, such as malaria. METHODS: The effects of methanol extracts (ME) obtained from seed kernel of ripe and unripe Azadirachta indica fruits were studied on erythrocytic proliferation of the rodent malaria parasite Plasmodium berghei strain ANKA and on mice pro-inflammatory response, as evaluated by measuring the matrix-metalloproteinase-9 (MMP-9) and tumour necrosis factor (TNF) plasma levels, in two mouse strains (C57BL/6 and BALB/c) which are considered as prototypical of Th1 and Th2 immune response, respectively. RESULTS: ME obtained from seed kernel of unripe Azadirachta indica fruits decreased by about 30% the proportion of erythrocytes infected with the malaria parasite in C57BL/6 mice in the 4 days suppressive test. In this treatment group, MMP-9 and TNF levels were notably higher than those measured in the same mouse strain treated with the anti-malarial drug artesunate, Azadirachta indica kernel extracts from ripe fruits or solvent. In BALB/c mice, treatment with kernel extracts did not influence parasitaemia. MMP-9 and TNF levels measured in this mouse strain were notably lower than those recorded in C57BL/6 mice and did not vary among treatment groups. CONCLUSIONS: The effects of the ME on the parasite-host interactions appeared to be mouse strain-dependent, but also related to the ripening stage of the neem fruits, as only the unripe fruit seed kernel extracts displayed appreciable bioactivity.