Immunoexpression of PD-L1, CD4+ and CD8+ cell infiltrates and tumor-infiltrating lymphocytes (TILs) in the microenvironment of actinic cheilitis and lower lip squamous cell carcinoma

Abstract Lower lip squamous cell carcinomas (LLSCC) could be associated with a previous history of potentially malignant oral diseases (PMOD), especially actinic cheilitis (AC), with high sun exposure being a well-described risk factor. Immune evasion mechanisms, such as the PD-1/PD-L1 (programmed cell death protein 1/programmed death-ligand 1) pathway has been gaining prominence since immunotherapy with immune checkpoint inhibitors showed a positive effect on the survival of patients with different types of neoplasms. Concomitant with the characterization of the tumor microenvironment, the expression of either or both PD-1 and PD-L1 molecules may estimate mutual relations of progression or regression of the carcinoma and prognostic values of the patient. Objective: Considering the importance of tumor microenvironment characterization, this study aims to determine the immunoexpression of PD-L1 and correlate with the frequency of CD4+ and CD8+ cells in AC and LLSCC lesions and with tumor-infiltrating lymphocytes (TILs) in LLSCC and its relationship with histopathological characteristics. Methodology: This sample includes 33 cases of AC and 17 cases of LLSCC. The cases were submitted to histopathological analysis and to CD4+, CD8+, and PD-L1+ cell determination by immunohistochemistry. Results: There was a significant difference among the frequencies of CD4+, CD8+, and PD-L1+ cells between AC and LSCC cases, higher in the last group. Moreover, histopathological and atypical changes in AC and LLSCC were correlated with the frequencies of PD-L1+, CD4+, and CD8+ cells. In AC, PD-L1+ cases had a low frequency of CD4+ cells, but on the other hand, PD-L1+ cases of LLSCC had a higher frequency of CD4+ and CD8+ cells. Conclusion: Therefore, the PD-L1 molecule may be a potential escape route for the immune response in oral lesions, but the mechanisms differ between AC and LLSCC. Future studies related to immune evasion and immunotherapy in oral lesions should consider the analysis of inflammatory infiltrate and TILs.

Effects of Stryphnodendron adstringens extracts on murine 4T1 tumor line

Breast cancer appears as the main gynecological cancer and presents high morbidity and mortality. Because most diagnoses are made belatedly, it is necessary to seek therapeutic options that aim for advanced stages of the disease. This study aims to evaluate the antitumoral action of Stryphnodendron adstringens fruit extracts on 4T1 murine mammary carcinoma cell culture. The inhibitory potential of S. adstringens fruit extract on the metalloproteinases (MMPs) 2 and 9 was evaluated through zymography. From these results, MTT assays were performed to evaluate the extracts' effects on the murine mammary carcinoma 4T1 line cell viability. From the crude extract, the following extracts were obtained: hydroalcoholic (SAFCEA), hexane (SAFCEB), chloroform (SAFCEC), and ethyl acetate (SAFCED). Lastly, the migration of the cells treated with extracts SAFCEA and SAFCED was verified. The hydroalcoholic extract (SAFCEA) was the most efficient in inhibiting gelatinases. During the phytochemical study, it was noted that alkaloids were present in all partitions. The 50 % growth inhibition (IC50) concentrations found were: 40.1 µg/mL (SAFCEA) and 70.14 µg/mL (SAFCED). After the cellular cytotoxicity assay, cell morphology was altered by treatment with the selected partitions (SAFCEA and SAFCED), obtaining morphology consistent with apoptosis. The results demonstrate that S. adstringens extracts exhibit the inhibitory activity of MMP-2 and MMP-9 as well as cytotoxicity toward 4T1 tumor cells. These findings indicate that follow-up studies of the partitions from S. adstringens may lead to the development of novel chemotherapeutics for oncological treatments.

Assessment of the Toxicological Effects of Pesticides and Detergent Mixtures on Zebrafish Gills: a Histological Study

Abstract Pesticides, as Temephos and Roundup, and surfactants, like Sodium Dodecyl Sulfate (SDS), when discharged into the environment, are harmful to non-target wildlife, especially fish. To evaluate the effects of those chemicals alone and their binary combinations, we assessed the histological alterations in Danio rerio gills after 15 days of exposure to Temephos (1ppm), Roundup (10ppm), SDS (2ppm), SDS plus Temephos (2ppm+1ppm), SDS plus Roundup (2ppm+10ppm) and Temephos plus Roundup (1ppm+10ppm). Hyperplasia, a regressive change, was significant in all exposed groups. The progressive change, complete fusion of lamellae, was significantly intensive in Roundup plus Temephos group. Regarding circulatory disturbances, vascular congestion was significant intensive in SDS group, while severe aneurysm was observed in Roundup group. Although xenobiotics in the blend are biologically complex, mainly for long periods, combinatorial exposure incremented only complete fusion of lamellae. Our results reinforce the rationalization of indiscriminate use of those compounds alone or in combination.

Immunohistochemical Analysis of P-gp, LC3-II, and Cathepsin-D Associated with Histological Changes in Fish Liver: from the Impacted Environment to Clean Water

Abstract Herein we evaluated the histopathological alterations and expression patterns of multixenobiotic resistence (MXR) and autophagic proteins in liver samples of fish chronically exposed to anthropogenic contaminants in a highly polluted river, and then again after they had been transferred to good quality water. Two groups were established: euthanized on the day of capture (0 h), and maintained for 30 days in a tank (30 d). The fish of 0 h presented liver with vacuolated and hypertrophic hepatocytes. Also, it was observed strong immunostaining of cathepsin-D, LC3-II and P-gp. Necrosis and apoptosis were also observed throughout the liver. Conversely, the second group (30 d) showed recovery of the liver normal histology and weak immunoreaction of the studied proteins. So, our results indicated that there was a hepatic recovery in the fish kept in good quality water, as showed by the decreased expression of cathepsin-D, LC3-II, and the MXR (P-gp). Therefore, the alterations here observed could be proposed as potential biomarkers to be tested for following the impacts of remediation or mitigation measures to environmental impacts.

Effects of high fat diet on kidney lipid content and the Na, K-ATPase activity

ABSTRACT It is widely known that high fat diet (HFD) can contribute to the advent of health problems. Recent studies have indicated that obesity imposes a hemodynamic overload to the kidneys. In order to further investigate such injuries, two groups of six Swiss mice each were fed with a controlled AIN93G diet or a high fat (AIN93G modified) diet for eight weeks. Blood samples were collected to determine the hormonal, lipid profile, glucose, urea, and creatinine levels. Histopathological and immunohistochemical analysis were carried out to analysis the kidney damage. Fractions of renal membranes were prepared to assess the Na,K-ATPase activity, lipid peroxidation, total cholesterol, and phospholipid content. The results indicated that the blood lipid profile, urea and creatinine was not altered by the HFD. On the other hand, it was observed in HFD diet mice elevated glucose blood levels along with an augment on insulin and a decrease on corticosterone release. HFD provoked a reduction in the diameter of the convoluted tubules and cell volume in Bowman's capsule and an increased number of positive cells with Na,K-ATPase, but reduced the Na,K-ATPase activity and the cholesterol content in the kidney cell membrane but favored the lipid peroxidation.

Inhibition of gelatinase activity of MMP-2 and MMP-9 by extracts of Bauhinia ungulata L / Inibição da atividade gelatinolítica de MMP-2 e MMP-9 a partir de extratos de Bauhinia ungulata l

Metastasis is responsible for the majority of cancer-related deaths. Tumour invasion and metastasis result from processes that include the proteolytic degradation of the extracellular matrix adjacent to the tumour. The matrix metalloproteinases (MMPs), particularly MMP-2 and MMP-9, have prognostic influence in human cancers after they cleave the main structural components of the basal membrane. These actions make MMPs an attractive target for cancer and metastasis studies. This study evaluated the inhibitory potency of extracts of Bauhinia ungulata L. (BU) on the gelatinolytic activity of MMP-2 and -9 and recognized the group of secondary compounds responsible for this property. The zymographic analysis of the BU stem revealed that the ethyl acetate partition (D) caused a higher inhibition of MMP-2 and MMP-9. The phytochemical study of D showed the presence of steroids, tannins, and coumarins and the significant presence of alkaloids and flavonoids. The phytochemical study of the fractions obtained through the column chromatography of partition D revealed a significant presence of flavonoids and alkaloids in the fractions that showed better inhibition of the gelatinolytic activity of MMPs. In conclusion, these results suggest that the stem fraction of BU has the potential to inhibit MMP-2 and MMP-9 and should be used in studies on the recognition of active biomolecules.

A metástase é a responsável pela maioria das mortes relacionadas ao câncer. Tanto a invasão tumoral quanto a metástase resultam de processos que incluem a degradação proteolítica da matriz extracelular adjacente ao tumor. As metaloproteinases de matriz (MMPs), especialmente as MMP-2 e MMP-9, têm influência direta no prognóstico dos diversos tipos de câncer humano, pois clivam os principais componentes estruturais da membrana basal. Estas ações fazem das MMPs alvos atraentes para estudos envolvendo câncer e metástase. Este trabalho teve como objetivo avaliar o potencial inibidor dos extratos de Bauhinia ungulata L. (BU) sobre a atividade gelatinolítica das MMP-2 e 9 e reconhecer o grupo de compostos secundários responsáveis por esta propriedade. Análises por zimograma do ramo de BU revelaram que a partição de acetato de etila (D) causou maior inibição de MMP-2 e MMP-9. O estudo fitoquímico de D mostrou a presença de esteróides, taninos e cumarinas e a presença significativa de alcaloides e flavonoides. As frações da coluna cromatográfica da partição D e seus estudos fitoquímicos revelaram uma grande presença de flavonoides e alcaloides nas frações que apresentaram maior inibição da atividade gelatinolítica de MMPs. Em conclusão, estes resultados sugerem que frações do ramo de BU têm o potencial inibidor de MMP-2 e MMP-9 e devem ser utilizadas em estudos envolvendo o reconhecimento de biomoléculas ativas.

Osteoradionecrosis of the jaw infected with Actinomyces sp: a challenge for clinical practice / Osteonecrose de mandíbula infectada com Actinomyces sp: um desafio para a prática clínica

The Actinomyces sp has been linked to osteoradionecrosis of the jaw. The identification of these bacteria on histopathological findings confers a determining factor of poor prognosis and is related to difficulties of local control of bone necrosis and risk of recurrent infections. The treatment is complicated due to insidious growth pattern of Actinomyces sp. The methods for diagnosis and therapeutic approaches constitute challenges that underscore the need for understanding the risk factors for infection and knowledge about clinical conditions associated with this pathology. The objective of this study is to report two cases of cancer patients suffering from an osteoradionecrosis infection by Actinomyces sp, to describe approaches and discuss therapeutic options in the light of the current literature.

O Actinomyces sp tem sido associado a osteorradionecrose. A identificação desta bactéria confere um fator determinante de mau prognóstico e está relacionado a dificuldades do controle da necrose óssea local e a risco de infecções recorrentes. O tratamento é complicado devido ao padrão de crescimento insidioso do Actinomyces sp. Os métodos de diagnóstico e as abordagens terapêuticas constituem desafios que reforçam a necessidade de compreender os fatores de risco para a infecção e o conhecimento sobre as condições clínicas associadas a esta patologia. O objetivo deste trabalho é relatar dois casos clínicos de pacientes com câncer que sofrem infecção osteorradionecrose por Actinomyces sp, para descrever as abordagens e discutir as opções terapêuticas à luz da literatura atual.

Efeito da natacao associada a diferentes tratamentos sobre o musculo soleo de ratos: estudo histologico e morfometrico / Effect of swimming associated with different treatments on rat soleus muscle: histological and morphometric study / Efecto de la natacion vinculada a diversos tratamientos sobre el musculo soleo de ratones: estudio histologico y morfometrico

INTRODUÇÃO: O uso de suplementos ergogênicos farmacológicos e nutricionais cresce a cada dia no ambiente esportivo. A creatina e os esteroides anabólicos são os recursos ergogênicos mais procurados ultimamente. OBJETIVO: Avaliar as alterações histológicas e morfométricas do músculo sóleo de ratos que receberam suplementos e foram submetidos a treinamento físico (TF) de natação. MÉTODOS: Foram utilizados 50 ratos, distribuidos em cinco grupos: três não submetidos à atividade física (controles) e dois que receberam treinamento de natação (treinados). Um grupo controle recebeu apenas ração (grupo controle ração), dois grupos (um controle e outro treinado) receberam diariamente 3,3 mg de creatina por grama de dieta dissolvida em água aplicada oralmente via gavagem, e os outros dois grupos (um controle e outro treinado) receberam injeção intramuscular do esteroide anabolizante decanoato de nandrolona (Deca-durabolin(r) - Organon) 5 mg/kg duas vezes por semana. Os grupos treinados foram submetidos ao TF com duração de 60 min/sessão de segunda a sexta-feira por nove semanas, e foram sacrificados no final deste período. Após o sacrifício, o músculo sóleo foi retirado, fixado em formalina (10%) tamponada e incluído em parafina. Os cortes histológicos foram corados pela técnica de hematoxilina-eosina para avaliação de hiperplasia e hipertrofia muscular. Para análise dos dados aplicou-se a análise de variância ANOVA One-Way com post hoc de Tukey-Kramer com nível de significância de 5%. RESULTADOS: Foi demonstrado que a creatina associada ao exercício provavelmente aumente o espaço intersticial no músculo esquelético sem alterar o tamanho ou número de fibras musculares, enquanto ...

INTRODUCTION: The use of pharmacologic and nutritional ergogenic supplements is growing every day in sports environment. Creatine and anabolic steroids are the most popular ergogenic resources used lately. OBJECTIVE: To evaluate the histological and morphometric changes of the soleus muscle of rats receiving supplements and submitted to physical training (swimming). METHODS: 50 rats were allocated into five groups: three did not undergo physical activity (controls) and two that received swimming training (trained). A control group received only rat chow (control chow group), two groups (one control and one trained) received 3.3mg of creatine per gram of diet dissolved in water daily applied orally via gavage, and the other two groups (control and other trained) received intramuscular injections of the anabolic steroid nandrolone decanoate (Deca-durabolin(r) - Organon) 5mg/kg twice weekly. The trained groups were submitted to the physical training 60min/day lasting from Monday to Friday for 9 weeks and were sacrificed at the end of the period. After sacrifice, the soleus muscle was removed, fixed in buffered formalin (10%) and embedded in paraffin. The histological sections were stained by hematoxylin-eosin for evaluation of muscle hyperplasia and hypertrophy. For data analysis we applied the One-Way ANOVA with post hoc Tukey-Kramer significance level of 5%. RESULTS: It was shown that creatine during exercise probably increase the interstitial space in skeletal muscle without changing the size or number of muscle fibers, whereas the anabolic agent increased the number of muscle fibers per muscle area examined, though no changes have been observed in the area of the fiber. CONCLUSION: The muscle of rats submitted to swimming training responds differently opposite type of ergogenic aid used. .

INTRODUCCIÓN: El uso de suplementos ergogénicos farmacológicos y nutricionales crece, cada día, en el ambiente deportivo. La creatina y los esteroides anabólicos son los recursos ergogénicos más procurados en los últimos tiempos. OBJETIVO: Evaluar las alteraciones histológicas y morfométricas del músculo sóleo de ratones que recibieron suplementos y fueron sometidos a entrenamiento físico (EF) de natación. MÉTODOS: Se utilizaron 50 ratones, distribuidos en cinco grupos: tres de estos no sometidos a la actividad física (controles) y dos que recibieron entrenamiento de natación (entrenados). Un grupo de control recibió solamente ración (grupo de controle por ración), dos grupos (uno de control y otro entrenado) recibieron diariamente 3,3 mg de creatina, por gramo de dieta, disuelta en agua aplicada oralmente vía gavaje, y los otros dos grupos (uno de control y otro entrenado) recibieron inyección intramuscular del esteroide anabolizante decanoato de nandrolona (Deca-durabolin(r) - Organon), 5 mg/kg dos veces por semana. Los grupos entrenados fueron sometidos al EF, con duración de 60 min/sesión, de lunes a viernes durante nueve semanas, y fueron sacrificados al final de este período. Después del sacrificio, el músculo sóleo fue retirado, fijado en formalina (10%) tamponada e insertado en parafina. Los cortes histológicos fueron coloridos mediante la técnica de hematoxilina-eosina para evaluación de hiperplasia e hipertrofia muscular. Para el análisis de los datos se aplicó el análisis de variancia ANOVA One-Way con post hoc de Tukey-Kramer, con nivel de significancia de 5%. RESULTADOS: Se demostró que la creatina, vinculada al ejercicio, aumenta probablemente el espacio intersticial en el músculo esquelético, sin alterar el tamaño ni el ...

Carcinoma neuroendócrino de pequenas células de laringe: relato de caso erevisão da literatura / Small cell neuroendocrine carcinoma of the larynx: case report and literature revie

O presente relato de caso refere-se ao carcinomaneuroendócrino de pequenas células de laringe acometendouma mulher de 52 anos. Este carcinoma é umaneoplasia altamente maligna com metástases frequentese taxa de sobrevida baixa. Depois dos carcinomasespinocelulares, os tumores de laringe mais frequentessão os neuroendócrinos, que podem ser derivados decélulas neurais ou epiteliais. Os tumores neuroendócrinosderivados de epitélio podem ser classificados emquatro grupos patológicos bem definidos: tumor carcinoide,tumor carcinoide atípico, carcinoma neuroendócrinode pequenas células e carcinoma neuroendócrinode células grandes. Os carcinomas neuroendócrinos têmmorfologia distinta, entretanto apresentam perfil imunohistoquímicomarcado por algumas combinações depositividade para citoqueratina, cromogranina, sinaptofisina,CD56, CD57, enolase, serotonina, somatostatinae bombesina. Esses tumores da laringe apresentam umgrande espectro de evolução clínica, dependente dosubtipo histológico, que influencia significativamente oplanejamento de todo o tratamento.

This case report refers to small cell neuroendocrinecarcinoma of the larynx affecting a 52-year-old female.This is a highly malignant neoplasm with frequentmetastases and a lower survival rate. After squamouscarcinomas, the most frequent tumors of the larynx arethe neuroendocrine, these tumors can be derived fromneural or epithelial cells. Neuroendocrine tumors derivedfrom epithelium can be classified into four pathologicalcategories defined: carcinoid tumor, atypical carcinoidtumor, small cell neuroendocrine carcinoma andlarge cell neuroendocrine carcinoma. It is known thatdespite of having distinct morphology, neuroendocrinecarcinomas have immunohistochemical profile markedby some combination of positive for cytokeratin, chromogranin,synaptophysin, CD56, CD57, enolase, serotonin,somatostatin and bombesin. These laryngeal tumorsshow a broad spectrum of clinical, dependent onthe histological subtype, which influences the planningof the whole treatment.

Expressão de metaloproteinases de matriz e de seus inibidores teciduais em carcinomas basocelulares / Expression of matrix metalloproteinasis and their tissue inhibitors in basal cell carcinoma

INTRODUÇÃO: Aproximadamente 80 por cento das neoplasias malignas de pele não-melanomas são carcinomas basocelulares (CBC). Apesar das raras metástases, esses tumores são localmente agressivos. As metaloproteinases de matriz (MMPs), especialmente as MMP-2 e 9, são importantes no processo de invasão. Em contrapartida, os inibidores teciduais das MMPs (TIMPs) têm como principal função a inibição dessas enzimas. OBJETIVO: Investigar a associação de variáveis clinicopatológicas de pacientes portadores de CBC com a expressão de MMP-2, MMP-9, TIMP-1 e TIMP-2. MATERIAL E MÉTODOS: Foram selecionados 31 casos de CBC, sendo então obtidos, retrospectivamente, os dados referentes a idade, sexo e tamanho da lesão. Cortes histológicos das lesões foram submetidos a reação imuno-histoquímica pela técnica estreptavidina-biotina-peroxidase para detecção dos antígenos de interesse. Índices de imunomarcação foram construídos e comparados com os dados previamente obtidos. RESULTADOS: Observou-se correlação significativa entre idade e tamanho da lesão (R = 0,532; p = 0,008). Não foram observadas correlações significativas entre as outras variáveis e a expressão imuno-histoquímica dos antígenos de interesse. CONCLUSÃO: A expressão das metaloproteinases e de seus inibidores teciduais não parece ser influenciada pelos parâmetros investigados. Estudos adicionais são necessários para melhor compreensão de sua associação com o comportamento biológico do CBC.

INTRODUCTION: Approximately 80 percent of non-melanoma skin neoplasias are basal cell carcinomas (BCC). Although metastasis is rare, BBC carcinomas are locally aggressive tumors. Matrix metalloproteinases (MMPs), mainly MMP-2 and MMP-9, play an important role on the invasion process. On the other hand, tissue inhibitors of MMPs (TIMPs) have the main function of inhibiting these enzymes. OBJECTIVE: To investigate the association of clinical-pathological variables of BCC patients with the expression of MMP-2 and MMP-9, TIMP-1 and TIMP-2. Methods: Thirty-one BCC cases were selected. Gender, age of the patients and size of the lesions were obtained retrospectively. Histological cuts of the lesions were exposed to immunohistochemistry reaction by use of the streptavidine-biotin peroxidase technique in order to detect antigens. Immunomarking parameters were established and compared with previous data. RESULTS: A significant correlation between age and size of the lesion was observed (R = 0.532; p = 0.008). No significant correlations between other variables and immunohistochemical expression of antigens were observed. CONCLUSION: The expression of MMPs and TIMPs does not seem to be influenced by the parameters investigated in this work. Additional studies should be made to better understand its association with the biological behavior of basal cell carcinomas.