EVALUATION OF THE IMMUNOLOCALIZATION OF THE PHOSPHATIDYL INOSITOL-3 RECEPTOR (IP3R) IN BASAL CELL AND SQUAMOUS CELL SKIN CARCINOMAS / AVALIAÇÃO DA IMUNOLOCALIZAÇÃO DO RECEPTOR DO FOSFATIDIL INOSITOL-3 (IP3R) EM CARCINOMAS BASOCELULAR E DE CÉLULAS ESCAMOSAS DE PELE

Introduction: Non-melanoma skin neoplasms (basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) form the most common group of all types of neoplasms. Although cancer is not yet being classified as a channelopathy, it has been suggested that pumps and ionic channels contribute to its progression by affecting autophagy, which could become an important therapeutic target in this context. Objective: To evaluate the immunohistochemical expression of IP3R in both skin tumors. Method: Immunohistochemistry was performed on 60 slides of non-melanoma cancer using primary anti-IP3R antibodies, verifying their presence and quantifying Results: For the first time, IP3R immunolocalization was identified in non-melanoma skin neoplasms, being evident above 90% of neoplastic cells was observed in all slides studied, regardless of the histological pattern, invasion and other tumor characteristics. Unlike what was seen in the internal control of normal skin, in which there was immunolocalization of IP3R in basal cell, in tumors, immunohistochemical expression occurred throughout the entire body of the neoplasm. Conclusion: There was immunolocalization of IP3R in tumor cells in both BCC and SCC. It was not possible to establish a correlation between tumor characteristics and IP3R expression, as immunostaining was similar in all analyzed samples. Despite this, IP3R to be associated with the pathophysiology of non-melanoma skin cancer, but its expression does not seem to be associated with tumor aggressiveness.

Introdução: As neoplasias de pele não melanoma (carcinoma basocelular (CBC) e carcinoma espinocelular (CEC) formam o grupo mais comum de todos os tipos de neoplasias. Embora o câncer ainda não esteja sendo pautado como canalopatia, tem sido sugerido que bombas e canais iônicos contribuem para a sua progressão por afetar a autofagia. Proteínas iônicas, em especial as de canais de cálcio, como o receptor da fosfatidil inositol 3 (IP3R) participam de maneira ativa na autofagia por estarem envolvidas na etapa inicial de formação do autofagossomo, podendo se tornar em alvo terapêutico importante neste contexto. Objetivo: Avaliar a expressão imuno histoquímica do IP3R em ambos os tumores de pele. Método: Foi realizada imuno histoquímica em 60 lâminas de câncer não melanoma utilizando anticorpos primários anti-IP3R verificando a sua expressão. Resultados: Pela primeira vez foi identificada a imunolocalização do IP3R em câncer de pele não melanoma, sendo evidente em mais de 90% das células neoplásicas em todas as lâminas estudadas, independentemente do padrão histológico, invasão e demais características tumorais. Diferentemente do visualizado no controle interno de pele normal, no qual houve imunolocalização de IP3R em células basais, nos tumores, a expressão imuno histoquímica ocorreu em todo o corpo da neoplasia. Conclusão: Houve imunolocalização de IP3R em células tumorais tanto em CBC quanto em CEC. Não foi possível estabelecer correlação entre as características tumorais e a expressão de IP3R, pois a imunomarcação apresentou-se de forma similar em todas as amostras analisadas. Apesar disso, IP3R está associado à fisiopatologia do câncer de pele não melanoma, mas a sua expressão não parece estar associada à agressividade tumoral.

Platelet-rich fibrin improves the osteoneogenesis in non-critical defects in calvaria: a histological and histometric study.

PURPOSE: The aim of this study was to evaluate the effect of platelet-rich fibrin (PRF) and autograft on non-critical bone repair. METHODS: Four bone defects (8.3 × 2 mm) were produced on the calvarium of 15 rabbits. The surgical defects were treated with either autograft, autograft associated to PRF, PRF alone, and sham. Animals were euthanized on the second, fourth or sixth posteoperative week. Histological analyses for presence of bone development on deffect was evaluated comparing the groups treated with autograft and without the autograft separately within the same period. Mann-Whitney's tests were used to compare the percentage of bone repair in each post-operative period for autograft × autograft + PRF groups and also for control × PRF groups (α = 5%). RESULTS: No differences were observed between the groups that received autograft and autograft associated to PRF on the second and fourth postoperative week, but areas treated with PRF demonstrated significant osteogenesis when compared to sham group on the fourth and sixth weeks. The groups that received PRF (with autograft or alone) demonstrated an enlarged bone deposition when compared to their control group. CONCLUSIONS: The use of PRF may influence bone repair and improve the bone deposition in late period of repair demonstrating osteoconductive and osteogenic properties.

The Nervous System Development Regulator Neuropilin-1 as a Potential Prognostic Marker and Therapeutic Target in Brain Cancer.

Neuropilins are transmembrane glycoproteins that regulate developmental processes in the nervous system and other tissues. Overexpression of neuropilin-1 (NRP1) occurs in many solid tumor types and, in several instances, may predict patient outcome in terms of overall survival. Experimental inhibition of NRP1 activity can display antitumor effects in different cancer models. Here, we review NRP1 expression and function in adult and pediatric brain cancers, particularly glioblastomas (GBMs) and medulloblastomas, and present analyses of NRP1 transcript levels and their association with patient survival in GBMs. The case of NRP1 highlights the potential of regulators of neurodevelopment as biomarkers and therapeutic targets in brain cancer.

Prolong intermittent injection of human parathyroid hormone in rats modulates the expression of several proteins in the neuropil of the cerebellar cortex.

INTRODUCTION: The intermittent use of recombinant human parathyroid hormone (iPTH) alters calcium metabolism and induces osteogenesis in experimental models. However, the real effects of iPTH in excitable cells and neurons that require membrane receptors to undergo membrane depolarization/repolarization (Na+K+ATPase) to generate ATP, voltage-gated calcium channel (calcium-IP3R-calponin) as well as GABAergic (GABAA) signaling remains unclear. OBJECTIVES: In this study, the expression of IP3R, Na+K+-ATPase, GABAA and calmodulin proteins were evaluated in histological sections of the cerebellum of rats following prolonged injection of iPTH. METHODS: Twenty Wistar rats were used in this study and randomly assigned as either or control group. The test group were subcutaneously injected with 20 µg/kg of iPTH, 3×/week for 8 weeks, while the control group received 1 ml/kg of 0.9% saline solution. The rats were euthanized on the 60th day after the first administration, and their cerebellar vermis was removed and submitted to histological and immunohistochemical evaluation for detection of IP3R, Na+K+-ATPase, GABAA and calmodulin proteins. The expression of proteins was evaluated in the areas corresponding to the Purkinje cells as well as in neuropil of molecular layer of cerebellum. All results were transformed into a percentage for each area analyzed to verify significance between groups. RESULTS: Rats that received iPTH demonstrated significant reduction of IP3R, calmodulin and GABAA in Purkinje cells and neuropil of molecular layer while the expression of Na+K+-ATPase was similar. CONCLUSION: It was concluded that iPTH decreased the expression of IP3R and calmodulin while it did not alter the expression of Na+K+-ATPase. These changes insinuate the ionic activity of calcium and sodium/potassium. Yet, the iPTH alters GABAergic signaling in Purkinje cells, suggesting neurotransmission activity changes in the cerebellum.

Câncer de pele não melanoma: revisão integrativa / Non-melanoma skin cancer: an integrative review

Introdução: O câncer de pele não melanoma compeende grupo de neoplasias com alta incidência na população mundial. É dividido em carcinoma basocelular e de células escamosas. Por ser de grande prevalência, entender o processo de oncogênese e a relação com íons, proteínas e receptores celulares no câncer de pele não melanoma pode contribuir para que novas terapêuticas sejam avaliadas. Objetivo: Entender o processo da oncogênese dos tumores de pele não melanomas e sua relação com a imunolocalização do IP3R. Métodos: Revisão integrativa da literatura com síntese de evidências. A base de dados foi o PubMed; a estratégia de busca: "carcinoma espinocelular, AND/ OR carcinoma basocelular, AND/OR IP3R, AND/OR imunoistoquímica". Foram considerados para revisão os trabalhos publicados entre 2018 e 2023. Foram incluídos 40 trabalhos, integralmente lidos e resumidos. Resultados: Câncer de pele não melanoma são os tumores malignos mais comuns em todo o mundo, sendo 75-80% o carcinoma basocelular, e até 25% o de células escamosas. As interações moleculares de forma geral, envolvem grande participação de moléculas supressoras tumorais, assim como de procto-oncogenes. Além disso, canais iônicos voltagem dependente controlam o fluxo citosólico de íons, dentre eles o cálcio. O IP3R (receptor do fosfatidil inositol-3) permite a saída de cálcio do retículo endoplasmático para que seja utilizado pela célula para atividades fisiológias como proliferação, angiogênese, motilidade e capacidade de invasão. Conclusão: O IP3R, pelas características de expressão imunoistoquímica, parece estar relacionado também, à fisiopatologia do câncer de pele não melanoma.

Introduction: Non-melanoma skin cancer comprises a group of neoplasms with a high incidence in the world population. It is divided into basal cell carcinoma and squamous cell carcinoma. As it is highly prevalent, understanding the process of oncogenesis and the relationship with ions, proteins and cellular receptors in nonmelanoma skin cancer can contribute to the evaluation of new therapies. Objective: To understand the oncogenesis process of non-melanoma skin tumors and its relationship with the immunolocalization of IP3R. Methods: Integrative literature review with evidence synthesis. The database was PubMed; the search strategy: "squamous cell carcinoma, AND/OR basal cell carcinoma, AND/OR IP3R, AND/OR immunohistochemistry". Works published between 2018 and 2023 were considered for review; 40 works were included, fully read and summarized. Results: Non-melanoma skin cancer is the most common malignant tumor worldwide, with 75-80% being basal cell carcinoma, and up to 25% being cell carcinoma. Molecular interactions in general involve a large participation of tumor suppressor molecules, as well as procto-oncogenes. Furthermore, voltage-dependent ion channels control the cytosolic flow of ions, including calcium. The IP3R (phosphatidyl inositol-3 receptor) allows the exit of calcium from the endoplasmic reticulum so that it can be used by the cell for physiological activities such as proliferation, angiogenesis, motility and invasion capacity. Conclusion: The IP3R, due to its immunohistochemical expression characteristics, appears may also be related to the pathophysiology of nonmelanoma skin cancer.

Periodontitis in rheumatoid arthritis: a case-control study in a brazilian sample

To investigate the presence of periodontitis in RA patients comparing it with controls in a Brazilian sample. Methods: This is a case control study conducted in a public health rheumatologic center. One hundred and sixteen RA patients and 68 paired controls were compared for epidemiological data and presence of periodontal disease evaluated by number of remaining teeth, presence of bacterial plaque, bleeding on probing, probing depth and clinical level of gingival insertion. In addition, data on comorbidities was collected. Results: RA patients and controls have the same amount of teeth loss (P = 0.84). RA patients had more calculus (P = 0.02); dental plaques (P = 0.04); gingival recession (P = 0.02) and bleeding (P = 0.01). Although the number of individuals with periodontitis was higher in RA patients, the severity of periodontitis was similar in both groups (P = ns). Presence of diabetes and hypothyroidism also associated with periodontitis (P = 0.01 and 0.02 respectively). In a model of logistic regression built to assess the independence of association of RA and its comorbidities with periodontitis, only diabetes and RA remained independent. Conclusion: This case control study shows higher frequency of periodontitis in RA patients than controls

Platelet-rich fibrin improves the osteoneogenesis in non-critical defects in calvaria: a histological and histometric study

ABSTRACT Purpose: The aim of this study was to evaluate the effect of platelet-rich fibrin (PRF) and autograft on non-critical bone repair. Methods: Four bone defects (8.3 × 2 mm) were produced on the calvarium of 15 rabbits. The surgical defects were treated with either autograft, autograft associated to PRF, PRF alone, and sham. Animals were euthanized on the second, fourth or sixth posteoperative week. Histological analyses for presence of bone development on deffect was evaluated comparing the groups treated with autograft and without the autograft separately within the same period. Mann-Whitney's tests were used to compare the percentage of bone repair in each post-operative period for autograft × autograft + PRF groups and also for control × PRF groups (α = 5%). Results: No differences were observed between the groups that received autograft and autograft associated to PRF on the second and fourth postoperative week, but areas treated with PRF demonstrated significant osteogenesis when compared to sham group on the fourth and sixth weeks. The groups that received PRF (with autograft or alone) demonstrated an enlarged bone deposition when compared to their control group. Conclusions: The use of PRF may influence bone repair and improve the bone deposition in late period of repair demonstrating osteoconductive and osteogenic properties.

EVALUATION OF THE OMENTUM AS A STEM CELL DONOR ISCHEMIC MYOCARDIAL THROUGH CD34 IMMUNOHISTOCHEMICAL ANALYSIS / AVALIAÇÃO DO OMENTO COMO DOADOR DE CÉLULAS TRONCO NO MIOCÁRDIO ISQUÊMICO ATRAVÉS DA ANÁLISE IMUNOHISTOQUÍMICA DO CD34

Background: Cardiovascular diseases are the leading cause of death in the world. The use of hematopoietic precursor cells and recent advances made in heart graft bioengineering offer a new therapeutic modality for post-myocardial infarction (MI)  and cardiac tissue regeneration. CD34 is a marker expressed on all hematopoietic and endothelial precursor cells, and functions as a cell adhesion factor. The antibody corresponding to this marker is used in immunohistochemistry to assess the formation of new vessels and the presence of stem cells. Aim: To evaluate the efficacy of omentopexy as stem cell donor, on previously infarcted myocardium, using immunohistochemically analysis of CD34. Method: Myocardial infarction was generated in four pigs, by ligature of the 1st and 2nd marginal branches of the circumflex artery. In three animals, abrasion of the infarcted epicardium was performed followed by multiple myocardial perforations and the mobilization of the omentum from the abdominal cavity to the mediastinum, sutured on the infarcted area. In the fourth animal, omentopexy was not performed and only the abrasion and perforation of the infarcted area were performed. All hearts were removed for CD34 immunohistochemically evaluation. Results:  In the samples from the group submitted to omentopexy, there was a 60% increase in angiogenesis, and in the samples from the control animal there was minimal staining.  Four samples from different sites of each animal, totaling 16 histopathological samples were evaluated. All samples were immunolabelled for CD34. Conclusions: Omentopexy proved to be effective in seeding previously infarcted myocardium with stem angiogenic cells, seen through immunohistochemistry, using CD34 marker.

Racional: As doenças cardiovasculares são a principal causa de morte no mundo. O uso de células precursoras hematopoiéticas e os recentes progressos feitos na bioengenharia de enxertos cardíacos oferecem uma nova modalidade terapêutica para a regeneração do tecido cardíaco pós-infarto do miocárdio (IM). O CD34 é um marcador expresso em todas as células precursoras hematopoiéticas e endoteliais, e funciona como fator de adesão celular. O anticorpo que correspondente a este marcador é utilizado na imunohistoquímica para avaliar a formação de novos vasos e a presença de células-tronco. Objetivo: O estudo teve por objetivo avaliar a eficácia da omentopexia na neovascularização e na doação de células tronco de corações suínos previamente infartados, a partir da análise imunohistoquímica do CD34. Método: O infarto do miocárdio foi gerado em 4 suínos, por ligadura do 1°e 2° ramos marginais da artéria circunflexa. Em 3 animais realizou-se abrasão cuidadosa do epicárdio infartado seguido de múltiplas perfurações miocárdicas e a mobilização do omento da cavidade abdominal para o mediastino, envolvendo a área infartada e as perfurações. No quarto animal não foi realizado a omentopexia sendo realizado apenas a abrasão e perfuração da área infartada. Todos os animais foram eutanasiados ao 30º dia pós operatório e os corações retirados para avaliação macroscópica, microscópica e Imunohistoquímica do CD34. Resultados: Nas amostras do grupo submetido a omentopexia, ocorreu um aumento de 60% da angiogênese, sendo que nas amostras do animal controle houve marcação mínima. Foram avaliadas quatro amostras de diferentes sítios de cada coração dos animais, totalizando 16 amostras histopatológicas. Todas as amostras foram imunomarcadas para CD-34. Conclusões: O omento mostrou-se eficiente na indução de neovascularização pela presença de células tronco, vista através da marcação do CD34, demonstrando grande potencial como futura terapêutica para restaurar áreas de miocárdio isquêmico.

TISSUE RESPONSES AND HEALING PATTERN OF PERIOSTAL DEFECTS IN CALVARY BETWEEN TWO TYPES OF MEMBRANES PRODUCED THROUGH BIOENGINEERING: STUDY IN RABBITS / RESPOSTAS TECIDUAIS E PADRÃO DE CICATRIZAÇÃO DE DEFEITOS PERIOSTAIS NA CALVÁRIA ENTRE DOIS TIPOS DE MEMBRANAS PRODUZIDAS POR MEIO DE BIOENGENHARIA: ESTUDO EM COELHOS

Introduction: Simultaneously with the development of new biomaterials, numerous surgical modalities have been proposed, with the aim of promoting adequate regeneration of bone defects, without providing fenestrations or dehiscence. Among the regenerative techniques currently accepted for this purpose, guided tissue regeneration (GTR) stands out. Objective: To evaluate the healing pattern and the histological response of the organism to the insertion of a type A gelatin-based membrane, and to compare it with a commercial membrane. Methods: Fifteen adult rabbits were selected. In the parietal region of the calvaria, flaps were performed to create a supraperiosteal pouch and implantation of 2 types of resorbable membranes: Marquette membrane and Bio-Gide® membrane. After the surgical procedure, the animals were allocated into 3 randomly selected groups (n=5) for the period of euthanasia, which occurred at 2, 4 and 6 postoperative weeks. Subsequently, the calvaria areas were removed and stained by H&E and analyzed for periosteum formation, inflammatory infiltrate in the area of membrane insertion, membrane degradation/resorption, and perimembrane fibrosis. Results: A slight inflammatory infiltration occurred around the Bio-Gide® membrane and it underwent resorption during the postoperative period with periosteum reconstruction, predominantly of a fibrous nature. The Marquette membrane also did not trigger an important inflammatory reaction. However, there was no significant resorption during the period; there was also significant formation of fibrous capsule surrounding the entire membrane area. Conclusion: The Marquette membrane produces tissue isolation reaction, minimizing periosteal formation while not providing its degradation

Introdução: Simultaneamente ao desenvolvimento de novos biomateriais, abundantes modalidades cirúrgicas têm sido propostas, com a finalidade de promover adequada regeneração de defeitos ósseos, sem proporcionar 2 fenestrações ou deiscências. Entre as técnicas regenerativas atualmente aceitas para este propósito, destaca-se a regeneração tecidual guiada (RTG). Objetivo: Avaliar o padrão de cicatrização e a resposta histológica do organismo frente à inserção de uma membrana a base de gelatina tipo A, e compará-la com membrana comercial. Métodos: Foram selecionadas 15 coelhas adultas. Na região parietal da calvária foram realizados retalhos para criação de uma bolsa supraperiostal e implantação de 2 tipos de membranas reabsorvíveis: membrana Marquette e membrana Bio-Gide®. Após o procedimento cirúrgico os animais foram alocados em 3 grupos randomicamente selecionados (n=5) para o período de eutanásia, que ocorreu em 2, 4 e 6 semanas pós-operatórias. Posteriormente, as áreas da calvária foram removidas e coradas por H&E e analisadas para averiguar formação de periósteo, infiltrado inflamatório na área de inserção de membrana, degradação/reabsorção da membrana, e fibrose perimembranar. Resultados: Ao redor da membrana Bio-Gide® ocorreu discreta infiltração inflamatória e a membrana sofreu reabsorção ao decorrer do período pósoperatório com reconstrução de periósteo prevalentemente de caráter fibroso. A membrana Marquette também não desencadeou importante reação inflamatória. Contudo, não foi verificada reabsorção significativa durante o período, também ocorreu significante formação de cápsula fibrosa cerceando toda área membranar. Conclusão: A membrana Marquette produz reação de isolamento tecidual, minimizando a formação periostal ao mesmo tempo que não proporciona sua degradação.

Effects of a local single dose administration of growth hormone on the osseointegration of titanium implants

Background: The aim of the present study was to evaluate the effect of different concentrations of growth hormone (GH) on endosteal implant’s surface at the early stages of osseointegration. Material and Methods: Sixty tapered acid-etched titanium implants were divided into four groups: i) Collagen, used as a control group; and three experimental groups, where after collagen coating, GH was administered directly to the surface in varying concentrations: ii) 0.265 mg, iii) 0.53 mg, and iv) 1 mg. Implants were placed in an interpolated fashion in the anterior flange of C3, C4 or C5 of 15 sheep with minimum distance of 6 mm between implants. After 3-, 6- and 12-weeks of healing samples were harvested, histologically processed, qualitatively and quantitatively assessed for bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO). Results: Statistical analysis as a function of time in vivo and coating resulted in no significant differences for BIC and BAFO at any evaluation time point. Histological evaluation demonstrated similar osseointegration features for all groups with woven bone formation at 3 weeks and progressive replacement of woven for lamellar bone in close contact with the implant surface and within the implant’s threads. Conclusions: A single local application of growth hormone to the surface of titanium implants did not yield improved implant osseointegration independent of healing time.(AU)

Effects of local single dose administration of parathormone on the early stages of osseointegration: A pre-clinical study.

The present study aimed to evaluate the effect of parathormone (PTH) administered directly to the implant's surface prior to insertion, using a large translational animal model. Sixty titanium implants were divided into four groups: (i) Collagen, control group, where implants were coated with Type-I Bovine-collagen, and three experimental groups, where implants received varying doses of PTH: (ii) 12.5, (iii) 25, and (iv) 50 µg, prior to placement. Fifteen female sheep (~2 years old, weighing ~65 kg) received four implants in an interpolated fashion in C3, C4 or C5 vertebral bodies. After 3-, 6- and 12-weeks, samples were harvested, histologically processed, qualitatively and quantitatively assessed for bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO). BIC yielded lower values at 6-weeks for 50 µg relative to the control group, with no significant differences, when compared to the 12.5- and 25-µg. No significant differences were detected at 6-weeks between collagen, 12.5- and 25-µg groups. At 3- and 12-weeks, no differences were detected for BIC among PTH groups. With respect to BAFO, no significant differences were observed between the control and experimental groups independent of PTH concentration and time in vivo. Qualitative observations at 3-weeks indicated the presence of a more mature bone near the implant's surface with the application of PTH, however, no significant differences in new bone formation or healing patterns were observed at 6- and 12-weeks. Single local application of different concentrations of PTH on titanium implant's surface did not influence the osseointegration at any time-point evaluation in low-density bone.

Congestive Hepatopathy Secondary to Right Ventricular Hypertrophy Related to Monocrotaline-Induced Pulmonary Arterial Hypertension.

Heart dysfunction and liver disease often coexist. Among the types of cardiohepatic syndrome, Type 2 is characterized by the chronic impairment of cardiac function, leading to chronic liver injury, referred to as congestive hepatopathy (CH). In this study, we aimed to establish a rat model of CH secondary to right ventricular hypertrophy (RVH) related to monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH). Fifty male Wistar rats were divided into four groups and randomly assigned to control and experimental groups. Three experimental groups were submitted to intraperitoneal MCT inoculation (60 mg/kg) and were under its effect for 15, 30 and 37 days. The animals were then sacrificed, obtaining cardiac and hepatic tissues for anatomopathological and morphometric analysis. At macroscopic examination, the livers in the MCT groups presented a nutmeg-like appearance. PAH produced marked RVH and dilatation in the MCT groups, characterized by a significant increase in right ventricular free wall thickness (RVFWT) and chamber area. At histological evaluation, centrilobular congestion was the earliest manifestation, with preservation of the hepatocytes. Centrilobular hemorrhagic necrosis was observed in the groups exposed to prolonged MCT. Sinusoidal dilatation was markedly increased in the MCT groups, quantified by the Sinusoidal Lumen Ratio (SLR). The Congestive Hepatic Fibrosis Score and the Centrilobular Fibrosis Ratio (CFR) were also significantly increased in the MCT30 group. Hepatic atrophy, steatosis, apoptotic bodies and, rarely, hydropic swelling were also observed. SLR correlated strongly with CFR and RVFWT, and CFR correlated moderately with RVFWT. Our rat model was able to cause CH, related to monocrotaline-induced PAH and RVH; it was feasible, reproducible, and safe.

SOFTWARE-ASSISTED IMAGE ANALYSIS FOR IDENTIFICATION AND QUANTIFICATION OF HEPATIC SINUSOIDAL DILATATION AND CENTRILOBULAR FIBROSIS.

BACKGROUND: Heart dysfunction and liver disease often coexist because of systemic disorders. Any cause of right ventricular failure may precipitate hepatic congestion and fibrosis. Digital image technologies have been introduced to pathology diagnosis, allowing an objective quantitative assessment. The quantification of fibrous tissue in liver biopsy sections is extremely important in the classification, diagnosis and grading of chronic liver disease. AIM: To create a semi-automatic computerized protocol to quantify any amount of centrilobular fibrosis and sinusoidal dilatation in liver Masson's Trichrome-stained specimen. METHOD: Once fibrosis had been established, liver samples were collected, histologically processed, stained with Masson's trichrome, and whole-slide images were captured with an appropriated digital pathology slide scanner. After, a random selection of the regions of interest (ROI's) was conducted. The data were subjected to software-assisted image analysis (ImageJ®). RESULTS: The analysis of 250 ROI's allowed to empirically obtain the best application settings to identify the centrilobular fibrosis (CF) and sinusoidal lumen (SL). After the establishment of the colour threshold application settings, an in-house Macro was recorded to set the measurements (fraction area and total area) and calculate the CF and SL ratios by an automatic batch processing. CONCLUSION: Was possible to create a more detailed method that identifies and quantifies the area occupied by fibrous tissue and sinusoidal lumen in Masson's trichrome-stained livers specimens.

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-ß1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin.

PURPOSE: Herein we evaluated the effects of platelet concentrate (PC) and platelet-poor plasma (PPP) on bone repair using noncritical defects in the calvaria of rabbits and compared them to the presence of TGF-ß1 and osteocalcin on reparative sites. METHODS: Five noncritical defects of 8.7 mm in diameter were created on the calvaria of 15 animals. Each defect was treated differently, using autograft (ABG), ABG associated with PC (ABG + PC), ABG with PPP (ABG + PPP), isolated PPP, and blood clot (control). The animals were submitted to euthanasia on the second, fourth and sixth week post-surgery. RESULTS: The defects that received ABG+PC or PPP demonstrated lower bone formation when compared to specimens that received ABG in the same period. These results coincided to significant higher immunopositivity for TGF-ß1 for specimens that received PC, and lower presence of cytokine in the group PPP. However, either higher or lower presence of TGF-ß1 were also correlated to lower presence of osteocalcin. Likewise, these results were similar to findings in specimens treated only with PPP when compared to control. CONCLUSIONS: PC and PPP were not effective when applied in association with ABG. Similarly, isolated use of PPP was not beneficial in optimizing the bone repair.

Co-Occurrence of Pineal-Region and Pituitary-Stalk Hemangioblastomas in a Patient Presenting with Von Hippel-Lindau Disease ­ A Case Report

Introduction Hemangioblastomas of the pineal region or pituitary stalk are extremely rare. Only two cases of hemangioblastomas involving the pineal region have been reported, and four involving the pituitary stalk. The purpose of the present manuscript is to describe an unusual case of supposed hemangioblastoma found concomitantly in the pineal region and pituitary stalk of a patient diagnosed with Von Hippel-Lindau (VHL) disease. Case Report A 35-year-old female patient with a previous diagnosis of VHL complaining of occipital headaches and balance disturbances for three weeks, who previously had a cerebellar hemangioblastoma resected. The visual characteristics of the tumor suggested a friable vascular lesion with a reddish-brown surface, and an incisional biopsy was performed. The tumor consisted of a dense vascular network surrounded by fibrous stroma abundant in reticulin and composed by both fusiform and dispersed xanthomatous cells; the immunohistochemistry was immunopositive for neuronspecific enolase and immunonegative for epithelial membranous antigen. The patient has been monitored closely for 2 years, and the supratentorial masses have not presented any volume alteration. Conclusion This rare association must be taken into account in patients with VHL disease, or at least be suspected in patients who present a thickening of the pituitary stalk and a pineal-region mass. We believe a biopsy of our asymptomatic patient could have been dangerous due to inherent complications like intraoperative bleeding. We recommend close observation of asymptomatic lesions with MRIs every six months or until the lesions become symptomatic. If the pineal-region tumor does become symptomatic, gross resection via a transcallosal approach would be ideal.

Are healthcare workers immunized after receiving hepatitis B vaccination according to recommended guidelines? A systematic review and meta-analysis.

OBJECTIVES: Nowadays, it is believed that more than 240 million people worldwide are infected with the hepatitis B virus (HBV), associated with irreversible health-related consequences, represented by hepatic failure, cirrhosis, and hepatocellular carcinoma, and already are ranked as 15th cause of human death around the world. METHODS: A systematic review and a meta-analysis were performed to evaluate the effectiveness of vaccination and immunization on health professionals against HBV. The review was registered at the PROSPERO database (CRD42017075643). A search for cross-sectional studies was performed in PubMed, Scopus, Web of Science, LILACS, BBO, Cochrane Library, and in the gray literature. The Effective Public Health Practice Project Modified Scale was used to evaluate the internal quality of the studies included. RESULTS: A total of 1865 articles were identified after the removal of duplicates. Of these, 790 studies remained after screening the titles and abstracts. Finally, ten studies remained after full-text reading for qualitative analysis, all of which were used in the meta-analysis. A significant difference was found in the vaccine protocol for health professionals immunized against hepatitis B, compared to those not immunized. The risk ratio was 7.37 (95% confidence interval = 3.92-13.83; P < 0.00001). CONCLUSION: This study showed that the vaccine protocol is effective in immunizing health professionals against hepatitis B.

Software-assisted image analysis for identification and quantification of hepatic sinusoidal dilatation and centrilobular fibrosis / Análise da imagem histológica assistida por software para identificação e quantificação de dilatação sinusoidal e fibrose hepática centrolubular

ABSTRACT Background: Heart dysfunction and liver disease often coexist because of systemic disorders. Any cause of right ventricular failure may precipitate hepatic congestion and fibrosis. Digital image technologies have been introduced to pathology diagnosis, allowing an objective quantitative assessment. The quantification of fibrous tissue in liver biopsy sections is extremely important in the classification, diagnosis and grading of chronic liver disease. Aim: To create a semi-automatic computerized protocol to quantify any amount of centrilobular fibrosis and sinusoidal dilatation in liver Masson's Trichrome-stained specimen. Method: Once fibrosis had been established, liver samples were collected, histologically processed, stained with Masson's trichrome, and whole-slide images were captured with an appropriated digital pathology slide scanner. After, a random selection of the regions of interest (ROI's) was conducted. The data were subjected to software-assisted image analysis (ImageJ®). Results: The analysis of 250 ROI's allowed to empirically obtain the best application settings to identify the centrilobular fibrosis (CF) and sinusoidal lumen (SL). After the establishment of the colour threshold application settings, an in-house Macro was recorded to set the measurements (fraction area and total area) and calculate the CF and SL ratios by an automatic batch processing. Conclusion: Was possible to create a more detailed method that identifies and quantifies the area occupied by fibrous tissue and sinusoidal lumen in Masson's trichrome-stained livers specimens.

Resumo Racional: Tecnologias de imagem digital têm sido introduzidas ao diagnóstico patológico, permitindo avaliações quantitativas objetivas. A quantificação de tecido fibroso em biópsias de fígado é extremamente importante para a classificação, diagnóstico e graduação de doenças crônicas hepáticas. Objetivo: Criar um protocolo computadorizado semi-automático para quantificação de fibrose centrolobular e dilatação sinusoidal em amostras de fígado coradas por Tricrômico de Masson. Método: Uma vez instaurada a fibrose, amostras de fígado foram coletadas, processadas histologicamente, coradas por Tricrômico de Masson e WSI (Whole Slide Images) foram capturadas por scanner digital patológico apropriado. Uma seleção aleatória das regiões de interesse (ROI) foi realizada. Os dados foram submetidos a uma análise de imagem assistida por software (ImageJ®). Resultados: A análise de 250 ROIs permitiu obter-se empiricamente as melhores configurações capazes de identificar fibrose centrolobular (FC) e lúmen sinusoidal (LS). Após o estabelecimento das configurações de padrão de cor, uma Macro de autoria própria foi gravada para definir as medidas (área da fração e área total) e calcular as razões de FC e LS por processamento em grupo/lote (batch mode). Conclusão: Foi possível criar um método detalhado capaz de identificar e quantificar a área ocupada por tecido fibroso e lúmen sinusoidal em espécimes de fígado coradas por Tricrômico de Masson.

Dano histológico renal em modelo experimental de hipertrofia ventricular direita induzida por monocrotalina / Renal histological damage in an experimental model of monocrotalin-induced right ventricular hypertrophy

Monocrotalina, alcalóide tóxico obtido de plantas do gênero crotalaria, pode ter potencial efeito tóxico em órgãos do corpo humano, como rins, pulmões, coração, fígado e outros efeitos. O objetivo deste estudo foi avaliar o dano renal causado pela exposição à monocrotalina. Trata-se de estudo experimental em ratos divididos em 4 grupos, um dos quais recebeu injeção de soro fisiológico e os outros três inoculação de monocrotalina, com tempos diferentes para sacrifício; subsequentemente, estudo histológico foi feito a fim de evidenciar as lesões renais. Em conclusão, constatou-se que houve lesão renal. Contudo, não foi possível afirmar o mecanismo exato responsável por elas, ou seja, se foram decorrentes da ação tóxica direta da monocrotalina, ou se, também, esteve relacionado a outros fatores sistêmicos.

Monocrotaline, a toxic alkaloid obtained from plants of the Crotalaria genus, may have a potential toxic effect on human body organs, such as kidneys, lungs, heart, liver and other effects. The aim of this study was to evaluate the kidney damage caused by exposure to monocrotaline. This is an experimental study in rats divided into 4 groups, one of which received saline injection and the other three received monocrotaline inoculation, with different times for sacrifice; subsequently, a histological study was performed in order to evidence renal lesions. In conclusion, it was found that there was kidney damage. However, it was not possible to state the exact mechanism responsible for them, that is, if they were due to the direct toxic action of monocrotaline, or if it was also related to other systemic factors.

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin

ABSTRACT Purpose Herein we evaluated the effects of platelet concentrate (PC) and platelet-poor plasma (PPP) on bone repair using noncritical defects in the calvaria of rabbits and compared them to the presence of TGF-β1 and osteocalcin on reparative sites. Methods Five noncritical defects of 8.7 mm in diameter were created on the calvaria of 15 animals. Each defect was treated differently, using autograft (ABG), ABG associated with PC (ABG + PC), ABG with PPP (ABG + PPP), isolated PPP, and blood clot (control). The animals were submitted to euthanasia on the second, fourth and sixth week post-surgery. Results The defects that received ABG+PC or PPP demonstrated lower bone formation when compared to specimens that received ABG in the same period. These results coincided to significant higher immunopositivity for TGF-β1 for specimens that received PC, and lower presence of cytokine in the group PPP. However, either higher or lower presence of TGF-β1 were also correlated to lower presence of osteocalcin. Likewise, these results were similar to findings in specimens treated only with PPP when compared to control. Conclusions PC and PPP were not effective when applied in association with ABG. Similarly, isolated use of PPP was not beneficial in optimizing the bone repair.

Growth hormone effects on healing efficacy, bone resorption and renal morphology of rats: histological and histometric study in rat calvaria.

Previous reports demonstrated the utility of systemic application of growth hormone (GH) in the treatment of bone defects. Very few studies correlated bone repair efficacy with hepatic and renal side effects promoted by locally-delivered GH. The objectives of this study were to assess the bone repair properties along with hepatic and renal adverse effects promoted by local application of GH in a rat model. Thirty-two rats were randomly divided (4 groups; n = 8/group), as follows: (i) AB (autogenous bone + local application of saline solution [SS]), (ii) AB+ (autogenous bone + SS local application + SS irrigation), (iii) AB/GH+ (autogenous bone + SS local application + GH irrigation) and (iv) AB/GHL+ (autogenous bone + GH local application + GH irrigation). Critical-sized defects (diameter = 5.0 mm) were surgically created by a single operator in the calvaria of rats. Defects were filled with ground autogenous bone. Defects pertaining to AB+ and AB/GH+ received a mixture of autogenous bone and a SS-saturated (0.02 mL) collagen sponge covered with bovine cortical membrane. Defects in group AB/GHL+, were filled with the same biomaterials saturated with GH (0.02 mL). SS (0.1 mL) or GH (0.1 mL, equivalent to 0.4 IU) were applied locally on alternate days (8 weeks) in animals in groups AB, AB+ and AB/GH+ or AB/GHL+, respectively. Bone repair properties was determined in hematoxylin/eosin-stained slices using traditional histologic and histomorphometric techniques along with optical microscopy and digital image analysis. Statistical differences among groups was determined using Kruskal-Wallis and Tukey post hoc tests (α = 0.05). Histology results indicated that AB and AB+ displayed greater presence of autogenous bone as compared to AB/GH+ and AB/GHL+. Histomorphometric results indicated significantly higher osteoid matrix formation in AB and AB+ when compared to AB/GHL+ (p = 0.009). Kidneys and livers were found to have their glomeruli preserved in AB and AB+. Strong glomeruli necrosis and large areas of protein deposition were found in AB/GH+. Abnormal small-sized glomeruli were found in AB/GHL+. The utilization of autogenous bone graft associated with local application and irrigation with GH was shown to not improve the bone repair in calvarial critical-sized defects in a rat model.