Relação in vitro do citomegalovírus com receptor do fator de crescimento epidérmico (EGFR) em células de glândula salivar humana

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Programa de Pós-Graduação em Virologia. Ananindeua, PA, Brasil.O citomegalovírus humano (HHCMV) é um herpesvírus humano que infecta uma variedade de tipos celulares sugerindo que o mesmo utilize múltiplos receptores para a entrada na célula hospedeira, a exemplo do receptor do fator de crescimento epidérmico (EGFR). Esse receptor quando sobreexpresso desempenha um importante papel no desenvolvimento de cânceres. A função desregulada do EGFR pode ser resultado também de uma mutação, a mais comum é o EGFRvIIII, ou ativação autócrina a partir da superprodução de ligantes, os quais podem induzir a secreção de metaloproteinases da matriz (MMPs), que são enzimas proteolíticas facilitadoras dos mecanismos de invasão tumoral e metástases. Estudos mostram que a ativação da via de sinalização do EGFR pelo HCMV tem sido associada a etiologia e patogênese de tumor nas glândulas salivares. Dessa forma, o objetivo do presente estudo foi avaliar a possível relação entre o HCMV e o EGFR em células de glândula salivar humana, in vitro, a fim de melhor compreender a patogenia de neoplasias possivelmente induzidas pelo HCMV. Para isso, foi utilizada uma linhagem celular derivada de glândula salivar humana (células HSG) e antes da inoculação do HCMV, as células HSG foram submetidas à técnica de imunofluorescência para detectar a expressão das proteínas: EGF, EGFR, EGFRvIII, MMP-2 e MMP-9. Meios condicionados com diferentes concentrações de EGF foram submetidos a técnica de zimografia para avaliar a secreção de proteases. Posteriormente, as células HSG foram inoculadas com a cepa AD169 e a observação do efeito citopático foi realizada em diferentes momentos pós-infecção. Os meios condicionados das células infectadas foram examinados por zimografia. E pela técnica de western blot avaliou-se a relação do HCMV com o EGFR nas células HSG. Antes da inoculação do HCMV, as células HSG expressaram EGF, EGFR, EGFRvIII, MMP-2 e MMP-9. Diferentes concentrações de EGF induziram um aumento na secreção de MMP-2 e MMP-9. As células HSG inoculadas com a cepa AD169 apresentaram efeito citopático com sete dias de inoculação e os meios condicionadas dessas células infectadas mostraram um aumento significativo na expressão de MMP-2 e MMP-9. Por western blot, verificou-se a fosforilação do receptor EGFR (pEGFR) e um aumento na expressão do EGFRvIII. Esses dados sugerem que o EGFR pode ser um receptor para o HCMV nas células HSG e que o aumento na expressão do EGFRvIII, bem como das MMPs, nas células infectadas poderia ser um dos mecanismos pelo qual o HCMV promoveria o desenvolvimento de tumores nas glândulas salivares

A protocol of hepatic volume measurement using magnetic resonance imaging in individuals from the Eastern Brazilian Amazon population.

Determination of hepatic volume is an important preoperative procedure and is done through imaging exams or standard liver volume (SLV) formulas developed based on the biotype of each population. In the absence of a specific SLV formula for the Brazilian Eastern Amazon population, the measurement of liver volume is made with reference values from other populations. The aim of study was to compare the hepatic volume in healthy residents from the Brazilian Eastern Amazon population obtained with magnetic resonance imaging (MRI) and recommended SLV formulas validated to other populations. This was a Observational, cross-sectional study. Anthropometric data of 42 healthy individuals aged 18-60 years of both sexes was collected to measure the liver volume through SLV formulas calculations and MRI. MRI shows similarity with the Western European SLV liver volume values and significant differences with the Japan SLV formula, mainly for women, with a moderate-to-weak correlation with the MRI measurements. There was a strong correlation between weight and body surface area in male patients analysed with measurements of the liver volume by the MRI and SLV formulas. The SLV formula based on the Western European population could be used in the absence of a specific formula for individuals living in the Amazon region. The results suggest that liver measurement formulas should take into consideration the sex of individuals, as well as the development of a specific SLV formula for the Eastern Amazon population and the conduction of similar studies in other Brazilian regions

Doppler ultrasonography: A non‐invasive method used to diagnose and follow up patients with chronic hepatitis C

National Health and Medical Research Council (61856616.1.3001.5172)Universidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brazil / Santa Casa de Misericórdia. Belém, PA, Brazil.Universidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brazil / Universidade do Estado do Pará. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Santa Casa de Misericórdia. Belém, PA, BrazilUniversidade Federal do Pará. Núcleo de Medicina Tropical. Belém, PA, Brazil / Universidade do Estado do Pará. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Background and Aim: This study aimed to investigate the association between the findings of Doppler ultrasonography and transient elastography using FibroScan and to determine the cut-off points, sensitivity, and specificity of resistance indices, and pulsatility of the hepatic vessels to predict significant hepatic fibrosis. Methods: This is a transversal, observational, and analytical study that includes 30 patients with chronic hepatitis C who were admitted at a public referral hospital. Transient elastography and ultrasonographic data were collected, and the linear association between these methods was evaluated using the Pearson test. Various Doppler velocimetric indices were compared according to the presence/absence of significant (≥ F2) fibrosis. Results: There was a moderate-strong linear association between the FibroScan data and the Doppler velocimetric indices and splenic index in the hepatic vessels; the mean values of the indices differed between groups with absent/mild (F0/F1) and significant (≥ F2) hepatic fibrosis. There was an association between the monophasic and biphasic wave pattern of the suprahepatic veins and the stratification of hepatic fibrosis estimated by the values of kilopascal in FibroScan. Conclusion: Doppler ultrasonography is a non-invasive method used to evaluate liver fibrosis, and it presents acceptable sensitivity/specificity for the prediction of fibrosis ≥ F2 in patients with chronic hepatitis C

Mechanisms of human cytomegalovirus infection with a focus on epidermal growth factor receptor interactions

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Divisão de Doenças Infecciosas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Virologia e Biologia Molecular. Ananindeua, PA, Brasil.Federal University of Pará. School of Dentistry. Department of Oral and Maxillofacial Pathology. Belém, PA, Brazil / University of São Paulo. Institute of Biomedical Sciences. Department of Cell and Developmental Biology. São Paulo, SP, Brazil.Pará State University. Center for Biological Sciences and Health. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Divisão de Doenças Infecciosas. Ananindeua, PA, Brasil / Pará State University. Center for Biological Sciences and Health. Belém, PA, Brazil / Federal University of Pará. Tropical Medicine Center. Division of Infectious Diseases. Belém, PA, Brazil.Human cytomegalovirus (HCMV) is a widespread opportunistic herpesvirus that causes severe diseases in immunocompromised individuals. It has a high prevalence worldwide that is linked with socioeconomic factors. Similar to other herpesviruses, HCMV has the ability to establish lifelong persistence and latent infection following primary exposure. HCMV infects a broad range of cell types. This broad tropism suggests that it may use multiple receptors for host cell entry. The identification of receptors used by HCMV is essential for understanding viral pathogenesis, because these receptors mediate the early events necessary for infection. Many cell surface components have been identified as virus receptors, such as epidermal growth factor receptor (EGFR), which is characterized by tyrosine kinase activity and plays a crucial role in the control of key cellular transduction pathways. EGFR is essential for HCMV binding, signaling, and host cell entry. This review focuses on HCMV infection via EGFR on different cell types and its implications for the cellular environment, viral persistence, and infection

Nerve Growth Factor and Pathogenesis of Leprosy: Review and Update

Neurotrophins are a family of proteins that regulate different aspects of biological development and neural function and are of great importance in neuroplasticity. This group of proteins has multiple functions in neuronal cells, as well as in other cellular populations. Nerve growth factor (NGF) is a neurotrophin that is endogenously produced during development and maturation by multiple cell types, including neurons, Schwann cells, oligodendrocytes, lymphocytes, mast cells, macrophages, keratinocytes, and fibroblasts. These cells produce proNGF, which is transformed by proteolytic cleavage into the biologically active NGF in the endoplasmic reticulum. The present review describes the role of NGF in the pathogenesis of leprosy and its correlations with different clinical forms of the disease and with the phenomena of regeneration and neural injury observed during infection. We discuss the involvement of NGF in the induction of neural damage and the pathophysiology of pain associated with peripheral neuropathy in leprosy. We also discuss the roles of immune factors in the evolution of this pathological process. Finally, we highlight avenues of investigation for future research to broaden our understanding of the role of NGF in the pathogenesis of leprosy. Our analysis of the literature indicates that NGF plays an important role in the evolution and outcome of Mycobacterium leprae infection. The findings described here highlight an important area of investigation, as leprosy is one of the main causes of infection in the peripheral nervous system

Detection and activity of MMP-2 and MMP-9 in Leishmania amazonensis and Leishmania braziliensis promastigotes

Institute Evandro Chagas (IEC), Department of Health Surveillance, Ministry of Health: Laboratory of Electron Microscopy and Laboratory of Superficial and Systemic Mycoses, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—finance code 001.Ministério da Saúde. Secretaria de Vigilância em Saúde e Ambiente. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde e Ambiente. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde e Ambiente. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Federal University of Para. School of Dentistry. Department of Oral Pathology. Belém, PA, Brazil.Federal University of Para. School of Dentistry. Department of Oral Pathology. Belém, PA, Brazil.Federal University of Para. School of Dentistry. Department of Oral Pathology. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde e Ambiente. Instituto Evandro Chagas. Laboratório de Microscopia Eletrônica. Ananindeua, PA, Brasil.Metalloproteinases (MMPs) are remarkable zinc-dependent endopeptidases, critical for degrading components of the extracellular matrix, thus actively influencing cell migration. Their impact on intracellular parasites, such as the enigmatic protozoan Leishmania, elicits intriguing queries. This study explores into the untapped territory of MMP-2 and MMP-9 within Leishmania spp. promastigotes. Notably, we successfully detected and quantified these MMPs, while also evaluating their activity in two distinct Leishmania species—L. amazonensis (La) and L. braziliensis (Lb)—at various growth stages and isolated from distinct clinical tegumentar disease forms. The results unveiled the presence of MMP-2 and MMP-9 in both species, albeit with distinct localization patterns. Specifically, MMP-9 exhibited significantly higher gelatinolytic activity in La when compared to Lb. Moreover, our data cleverly illustrated the presence and release of MMP-2 and MMP-9 by La and Lb promastigotes, exposing their ability to invade and migrate within a collagen matrix. This pioneering study establishes a compelling correlation between MMP-2 and MMP-9 and their potential role in the dynamics of La and Lb infection. Suggesting their potential as prognostic markers for severe leishmaniasis and promising target molecules for therapeutic interventions, this research opens new avenues for combatting this debilitating parasitic disease

Detection and activity of MMP-2 and MMP-9 in Leishmania amazonensis and Leishmania braziliensis promastigotes

Abstract Metalloproteinases (MMPs) are remarkable zinc-dependent endopeptidases, critical for degrading components of the extracellular matrix, thus actively influencing cell migration. Their impact on intracellular parasites, such as the enigmatic protozoan Leishmania, elicits intriguing queries. This study explores into the untapped territory of MMP-2 and MMP-9 within Leishmania spp. promastigotes. Notably, we successfully detected and quantified these MMPs, while also evaluating their activity in two distinct Leishmania species—L. amazonensis (La) and L. braziliensis (Lb)—at various growth stages and isolated from distinct clinical tegumentar disease forms. The results unveiled the presence of MMP-2 and MMP-9 in both species, albeit with distinct localization patterns. Specifically, MMP-9 exhibited significantly higher gelatinolytic activity in La when compared to Lb. Moreover, our data cleverly illustrated the presence and release of MMP-2 and MMP-9 by La and Lb promastigotes, exposing their ability to invade and migrate within a collagen matrix. This pioneering study establishes a compelling correlation between MMP-2 and MMP-9 and their potential role in the dynamics of La and Lb infection. Suggesting their potential as prognostic markers for severe leishmaniasis and promising target molecules for therapeutic interventions, this research opens new avenues for combatting this debilitating parasitic disease

Correction: CT Chest and pulmonary functional changes in patients with HTLV-associated myelopathy in the Eastern Brazilian Amazon.

[This corrects the article DOI: 10.1371/journal.pone.0186055.]

A novel cell line derived from pleomorphic adenoma expresses MMP2, MMP9, TIMP1, TIMP2, and shows numeric chromosomal anomalies.

Pleomorphic adenoma is the most common salivary gland neoplasm, and it can be locally invasive, despite its slow growth. This study aimed to establish a novel cell line (AP-1) derived from a human pleomorphic adenoma sample to better understand local invasiveness of this tumor. AP-1 cell line was characterized by cell growth analysis, expression of epithelial and myoepithelial markers by immunofluorescence, electron microscopy, 3D cell culture assays, cytogenetic features and transcriptomic study. Expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) was also analyzed by immunofluorescence and zymography. Furthermore, epithelial and myoepithelial markers, MMPs and TIMPs were studied in the tumor that originated the cell line. AP-1 cells showed neoplastic epithelial and myoepithelial markers, such as cytokeratins, vimentin, S100 protein and smooth-muscle actin. These molecules were also found in vivo, in the tumor that originated the cell line. MMPs and TIMPs were observed in vivo and in AP-1 cells. Growth curve showed that AP-1 exhibited a doubling time of 3.342 days. AP-1 cells grown inside Matrigel recapitulated tumor architecture. Different numerical and structural chromosomal anomalies were visualized in cytogenetic analysis. Transcriptomic analysis addressed expression of 7 target genes (VIM, TIMP2, MMP2, MMP9, TIMP1, ACTA2 e PLAG1). Results were compared to transcriptomic profile of non-neoplastic salivary gland cells (HSG). Only MMP9 was not expressed in both libraries, and VIM was expressed solely in AP-1 library. The major difference regarding gene expression level between AP-1 and HSG samples occurred for MMP2. This gene was 184 times more expressed in AP-1 cells. Our findings suggest that AP-1 cell line could be a useful model for further studies on pleomorphic adenoma biology