85 results on '"M. Fatima Leite"'
Search Results
2. Burden of liver disease in Brazil, 1996–2022: a retrospective descriptive study of the epidemiology and impact on public healthcareResearch in context
- Author
-
João Sérgio Fonseca Guimarães, Jordana Almeida Mesquita, Thais Yuki Kimura, Ana Luíza Matos Oliveira, M. Fatima Leite, and André Gustavo Oliveira
- Subjects
Liver disease ,Liver transplant ,Public health ,Burden of disease ,Brazil ,Public aspects of medicine ,RA1-1270 - Abstract
Summary: Background: Liver disease is a major cause of mortality and morbidity worldwide and its epidemiology depends on the genetic background, exposure to risk factors, access to healthcare and other sociodemographic characteristics. Brazil is a large country with diverse multicultural and ethnic heritages and important socioeconomic inequalities. The burden of liver disease in Brazil, its regions and population is unknown. Methods: We retrieved data from the Unified Health System regarding liver diseases and analyzed the mortality and morbidity from 1996 to 2022 by gender, race/ethnicity, age, region and overall. We calculated the age-specific risk of deaths by liver disease, age-standardization of the data, mean hospitalization and liver transplant-associated costs. Findings: Malignant neoplasm of the liver and intrahepatic bile ducts, alcohol-associated liver disease, fibrosis, and cirrhosis of the liver, other diseases of the liver, hepatic failure, chronic viral hepatitis were identified as the major causes of death and morbidity in Brazil in the period analyzed. The epidemiology of these diseases was diverse, with variations according to geographic regions, gender and race/ethnicity. The major economic burden of liver disease is related to liver transplants, a common outcome of the progression of these diseases. Interpretation: Liver disease in Brazil is a serious issue for the public health system due to the high number of deaths and increasing mortality rate. Our study contributes as a necessary prerequisite for the development of tailored public health policies aimed at mitigating the increasing burden of liver diseases in specific populations and regions. Funding: CNPq, INCT, CAPES, FAPEMIG.
- Published
- 2024
- Full Text
- View/download PDF
3. Correlation Between Clinical and Pathological Findings of Liver Injury in 27 Patients With Lethal COVID‐19 Infections in Brazil
- Author
-
Monique Freire Santana, Mateus T. Guerra, Melanie A. Hundt, Maria M. Ciarleglio, Rebecca Augusta de Araújo Pinto, Bruna Guimarães Dutra, Mariana Simão Xavier, Marcus Vinicius Guimarães Lacerda, Anderson Jose Ferreira, David Campos Wanderley, Israel Júnior Borges do Nascimento, Roberto Ferreira de Almeida Araújo, Sérgio Veloso Brant Pinheiro, Stanley de Almeida Araújo, M. Fatima Leite, Luiz Carlos de Lima Ferreira, Michael H. Nathanson, and Paula Vieira Teixeira Vidigal
- Subjects
Diseases of the digestive system. Gastroenterology ,RC799-869 - Abstract
Liver test abnormalities are frequently observed in patients with coronavirus disease 2019 (COVID‐19) and are associated with worse prognosis. However, information is limited about pathological changes in the liver in this infection, so the mechanism of liver injury is unclear. Here we describe liver histopathology and clinical correlates of 27 patients who died of COVID‐19 in Manaus, Brazil. There was a high prevalence of liver injury (elevated alanine aminotransferase and aspartate aminotransferase in 44% and 48% of patients, respectively) in these patients. Histological analysis showed sinusoidal congestion and ischemic necrosis in more than 85% of the cases, but these appeared to be secondary to systemic rather than intrahepatic thrombotic events, as only 14% and 22% of samples were positive for CD61 (marker of platelet activation) and C4d (activated complement factor), respectively. Furthermore, the extent of these vascular findings did not correlate with the extent of transaminase elevations. Steatosis was present in 63% of patients, and portal inflammation was present in 52%. In most cases, hepatocytes expressed angiotensin‐converting enzyme 2 (ACE2), which is responsible for binding and entry of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), even though this ectoenzyme was minimally expressed on hepatocytes in normal controls. However, SARS‐CoV‐2 staining was not observed. Most hepatocytes also expressed inositol 1,4,5‐triphosphate receptor 3 (ITPR3), a calcium channel that becomes expressed in acute liver injury. Conclusion: The hepatocellular injury that commonly occurs in patients with severe COVID‐19 is not due to the vascular events that contribute to pulmonary or cardiac damage. However, new expression of ACE2 and ITPR3 with concomitant inflammation and steatosis suggests that liver injury may result from inflammation, metabolic abnormalities, and perhaps direct viral injury.
- Published
- 2022
- Full Text
- View/download PDF
4. Molecular Mechanism for Protection Against Liver Failure in Human Yellow Fever Infection
- Author
-
Fernanda de Oliveira Lemos, Andressa França, Antônio Carlos Melo Lima Filho, Rodrigo M. Florentino, Marcone Loiola Santos, Dabny G. Missiaggia, Gisele Olinto Libanio Rodrigues, Felipe Ferraz Dias, Ingredy Beatriz Souza Passos, Mauro M. Teixeira, Antônio Márcio de Faria Andrade, Cristiano Xavier Lima, Paula Vieira Teixeira Vidigal, Vivian Vasconcelos Costa, Matheus Castro Fonseca, Michael H. Nathanson, and M. Fatima Leite
- Subjects
Diseases of the digestive system. Gastroenterology ,RC799-869 - Abstract
Yellow fever (YF) is a viral hemorrhagic fever that typically involves the liver. Brazil recently experienced its largest recorded YF outbreak, and the disease was fatal in more than a third of affected individuals, mostly because of acute liver failure. Affected individuals are generally treated only supportively, but during the recent Brazilian outbreak, selected patients were treated with liver transplant. We took advantage of this clinical experience to better characterize the clinical and pathological features of YF‐induced liver failure and to examine the mechanism of hepatocellular injury in YF, to identify targets that would be amenable to therapeutic intervention in preventing progression to liver failure and death. Patients with YF liver failure rapidly developed massive transaminase elevations, with jaundice, coagulopathy, thrombocytopenia, and usually hepatic encephalopathy, along with pathological findings that included microvesicular steatosis and lytic necrosis. Hepatocytes began to express the type 3 isoform of the inositol trisphosphate receptor (ITPR3), an intracellular calcium (Ca2+) channel that is not normally expressed in hepatocytes. Experiments in an animal model, isolated hepatocytes, and liver‐derived cell lines showed that this new expression of ITPR3 was associated with increased nuclear Ca2+ signaling and hepatocyte proliferation, and reduced steatosis and cell death induced by the YF virus. Conclusion: Yellow fever often induces liver failure characterized by massive hepatocellular damage plus steatosis. New expression of ITPR3 also occurs in YF‐infected hepatocytes, which may represent an endogenous protective mechanism that could suggest approaches to treat affected individuals before they progress to liver failure, thereby decreasing the mortality of this disease in a way that does not rely on the costly and limited resource of liver transplantation.
- Published
- 2020
- Full Text
- View/download PDF
5. Polymorphism in the Promoter Region of NFE2L2 Gene Is a Genetic Marker of Susceptibility to Cirrhosis Associated with Alcohol Abuse
- Author
-
Kemper Nunes dos Santos, Rodrigo M. Florentino, Andressa França, Antônio Carlos Melo Lima Filho, Marcone Loiola dos Santos, Dabny Missiaggia, Matheus de Castro Fonseca, Igor Brasil Costa, Paula Vieira Teixeira Vidigal, Michael H. Nathanson, Fernanda de Oliveira Lemos, and M. Fatima Leite
- Subjects
Alcoholic liver disease ,Nrf2 ,Polymorphism ,NFE2L2 gene ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
Alcoholic liver disease (ALD) is a highly prevalent spectrum of pathologies caused by alcohol overconsumption. Morbidity and mortality related to ALD are increasing worldwide, thereby demanding strategies for early diagnosis and detection of ALD predisposition. A potential candidate as a marker for ALD susceptibility is the transcription factor nuclear factor erythroid-related factor 2 (Nrf2), codified by the nuclear factor erythroid 2-related factor 2 gene (NFE2L2). Nrf2 regulates expression of proteins that protect against oxidative stress and inflammation caused by alcohol overconsumption. Here, we assessed genetic variants of NFE2L2 for association with ALD. Specimens from patients diagnosed with cirrhosis caused by ALD were genotyped for three NFE2L2 single nucleotide polymorphisms (SNP) (SNPs: rs35652124, rs4893819, and rs6721961). Hematoxylin & eosin and immunohistochemistry were performed to determine the inflammatory score and Nrf2 expression, respectively. SNPs rs4893819 and rs6721961 were not specifically associated with ALD, but analysis of SNP rs35652124 suggested that this polymorphism predisposes to ALD. Furthermore, SNP rs35652124 was associated with a lower level of Nrf2 expression. Moreover, liver samples from ALD patients with this polymorphism displayed more severe inflammatory activity. Together, these findings provide evidence that the SNP rs35652124 variation in the Nrf2-encoding gene NFE2L2 is a potential genetic marker for susceptibility to ALD.
- Published
- 2019
- Full Text
- View/download PDF
6. The des-Arg9-bradykinin/B1R axis: Hepatic damage in COVID-19
- Author
-
Gabriel Moreira de M Mendes, Israel Júnior Borges Do Nascimento, Paulo HS. Marazzi-Diniz, Izabela B. Da Silveira, Matheus F. Itaborahy, Luiz E. Viana, Filipe A. Silva, Monique F Santana, Rebecca AA. Pinto, Bruna G. Dutra, Marcus Vinicius G. Lacerda, Stanley A. Araujo, David Wanderley, Paula VT. Vidigal, Paulo HC Diniz, Thiago Verano-Braga, Robson AS. Santos, and M Fatima Leite
- Subjects
Physiology ,Physiology (medical) - Abstract
Patients infected by the SARS-CoV-2 virus are commonly diagnosed with threatening liver conditions associated with drug-induced therapies and systemic viral action. RNA-Seq data from cells in bronchoalveolar lavage fluid from COVID-19 patients have pointed out dysregulation of kallikrein-kinin and renin-angiotensin systems as a possible mechanism that triggers multi-organ damage away from the leading site of virus infection. Therefore, we measured the plasma concentration of biologically active peptides from the kallikrein-kinin system, bradykinin and des-Arg9-bradykinin, and liver expression of its proinflammatory axis, bradykinin 1 receptor (B1R). We measured the plasma concentration of bradykinin and des-Arg9-bradykinin of 20 virologically confirmed COVID-19 patients using a liquid chromatography-tandem mass spectrometry-based methodology. The expression of B1R was evaluated by immunohistochemistry from post-mortem liver specimens of 27 COVID-19 individuals. We found a significantly higher blood level of des-Arg9-bradykinin and a lower bradykinin concentration in patients with COVID-19 compared to a healthy, uninfected control group. We also observed increased B1R expression levels in hepatic tissues of patients with COVID-19 under all hepatic injuries analyzed (liver congestion, portal vein dilation, steatosis, and ischemic necrosis). Our data indicate that des-Arg9-bradykinin/B1R is associated with the acute hepatic dysfunction induced by the SARS-CoV-2 virus infection in the pathogenesis of COVID-19.
- Published
- 2022
7. The des-Arg
- Author
-
Gabriel Moreira de M, Mendes, Israel Júnior Borges, Do Nascimento, Paulo Hs, Marazzi-Diniz, Izabela B, Da Silveira, Matheus F, Itaborahy, Luiz E, Viana, Filipe A, Silva, Monique F, Santana, Rebecca Aa, Pinto, Bruna G, Dutra, Marcus Vinicius G, Lacerda, Stanley A, Araujo, David, Wanderley, Paula Vt, Vidigal, Paulo Hc, Diniz, Thiago, Verano-Braga, Robson As, Santos, and M Fatima, Leite
- Abstract
Patients infected by the SARS-CoV-2 virus are commonly diagnosed with threatening liver conditions associated with drug-induced therapies and systemic viral action. RNA-Seq data from cells in bronchoalveolar lavage fluid from COVID-19 patients have pointed out dysregulation of kallikrein-kinin and renin-angiotensin systems as a possible mechanism that triggers multi-organ damage away from the leading site of virus infection. Therefore, we measured the plasma concentration of biologically active peptides from the kallikrein-kinin system, bradykinin and des-Arg
- Published
- 2022
8. Neutrophils regulate ITPR2 levels in epithelia by direct injection of elastase
- Author
-
N Ogino, M Fatima Leite, E Kruglov, H Asashima, DA Hafler, BE Ehrlich, and MH Nathanson
- Abstract
The destructive role of neutrophils in inflammation is well known1 but they also have less damaging effects such as tissue remodeling and modulation of metabolism2, 3. Usually, neutrophils in tissues release toxic or digestive compounds into the extracellular region4–8. Here we report that neutrophils can inject their granule contents directly into hepatocytes. Neutrophil elastase within the hepatocytes selectively degrades the inositol trisphosphate receptor (ITPR), especially the type 2 isoform which is the predominant intracellular calcium release channel in these cells9. This action reduces calcium signals and cell proliferation without cellular damage. In response, the hepatocytes increase expression of serpins E2 and A3, which block the effect of elastase. This phenomenon is also observed in liver biopsies from patients with alcoholic hepatitis, a condition characterized by infiltration of neutrophils10, 11. This non-destructive and reversible effect on hepatocytes defines a previously unappreciated role of neutrophils in transiently regulating signaling mechanisms in epithelia.
- Published
- 2022
9. Calcium signaling in chemotherapy-induced neuropathy
- Author
-
Matheus de Castro Fonseca, Paulo H S Marazzi-Diniz, M Fatima Leite, and Barbara E Ehrlich
- Subjects
Physiology ,Cell Biology ,Molecular Biology - Published
- 2023
10. Molecular Mechanism for Protection Against Liver Failure in Human Yellow Fever Infection
- Author
-
Dabny Goulart Missiaggia, Marcone Loiola dos Santos, Mauro M. Teixeira, Ingredy Passos, Antônio Carlos Melo Lima Filho, Andressa França, Matheus de Castro Fonseca, Vivian Vasconcelos Costa, Antonio Marcio De Faria Andrade, Michael H. Nathanson, Gisele Olinto Libanio Rodrigues, M. Fatima Leite, Rodrigo M. Florentino, Fernanda O. Lemos, Felipe F. Dias, Paula Vieira Teixeira Vidigal, and Cristiano Xavier Lima
- Subjects
Necrosis ,Hepatology ,business.industry ,medicine.medical_treatment ,Microvesicular Steatosis ,Original Articles ,Liver transplantation ,Jaundice ,medicine.disease ,Viral hemorrhagic fever ,medicine.anatomical_structure ,Hepatocyte ,Immunology ,medicine ,Original Article ,lcsh:Diseases of the digestive system. Gastroenterology ,medicine.symptom ,Steatosis ,lcsh:RC799-869 ,business ,Hepatic encephalopathy - Abstract
Yellow fever (YF) is a viral hemorrhagic fever that typically involves the liver. Brazil recently experienced its largest recorded YF outbreak, and the disease was fatal in more than a third of affected individuals, mostly because of acute liver failure. Affected individuals are generally treated only supportively, but during the recent Brazilian outbreak, selected patients were treated with liver transplant. We took advantage of this clinical experience to better characterize the clinical and pathological features of YF-induced liver failure and to examine the mechanism of hepatocellular injury in YF, to identify targets that would be amenable to therapeutic intervention in preventing progression to liver failure and death. Patients with YF liver failure rapidly developed massive transaminase elevations, with jaundice, coagulopathy, thrombocytopenia, and usually hepatic encephalopathy, along with pathological findings that included microvesicular steatosis and lytic necrosis. Hepatocytes began to express the type 3 isoform of the inositol trisphosphate receptor (ITPR3), an intracellular calcium (Ca2+) channel that is not normally expressed in hepatocytes. Experiments in an animal model, isolated hepatocytes, and liver-derived cell lines showed that this new expression of ITPR3 was associated with increased nuclear Ca2+ signaling and hepatocyte proliferation, and reduced steatosis and cell death induced by the YF virus. Conclusion: Yellow fever often induces liver failure characterized by massive hepatocellular damage plus steatosis. New expression of ITPR3 also occurs in YF-infected hepatocytes, which may represent an endogenous protective mechanism that could suggest approaches to treat affected individuals before they progress to liver failure, thereby decreasing the mortality of this disease in a way that does not rely on the costly and limited resource of liver transplantation.
- Published
- 2020
11. Ca2+Signaling in the Liver
- Author
-
M. Fatima Leite, Michael H. Nathanson, and Mateus T. Guerra
- Subjects
Bile flow ,Biochemistry ,Lipid metabolism ,Biology ,Cell cycle ,Inositol trisphosphate receptor ,Ca2 signaling - Published
- 2020
12. Inositol 1,4,5‑trisphosphate receptor type 3 is involved in resistance to apoptosis and maintenance of human hepatocellular carcinoma
- Author
-
Antônio Carlos Melo Lima Filho, Marcone Loiola dos Santos, Carlos Alberto Xavier Gonçalves, Rodrigo M. Florentino, Paula Vieira Teixeira Vidigal, Vitor Lima Coelho, Andressa França, M. Fatima Leite, Cristiano Xavier Lima, Paulo Henrique Costa Diniz, Giselle Foureaux, Michael H. Nathanson, and Fernanda O. Lemos
- Subjects
mitosis ,Cancer Research ,Oncogene ,Cell ,apoptosis ,Cancer ,Articles ,Biology ,Cell cycle ,calcium signaling ,medicine.disease ,Molecular medicine ,digestive system diseases ,chronic diseases ,liver cancer ,chemistry.chemical_compound ,medicine.anatomical_structure ,Oncology ,chemistry ,Apoptosis ,Hepatocellular carcinoma ,medicine ,Cancer research ,Inositol - Abstract
The expression of the inositol 1,4,5-trisphosphate receptor type 3 (ITRP3) in hepatocytes is a common event in the pathogenesis of hepatocellular carcinoma (HCC), regardless of the type of underlying liver disease. However, it is not known whether ITPR3 expression in hepatocytes is involved in tumor maintenance. The aim of the present study was to determine whether there is an association between ITPR3 expression and clinical and morphological parameters using HCC samples obtained from liver explants from patients (n=53) with different etiologies of underlying chronic liver disease (CLD). ITPR3 expression, mitosis and apoptosis were analyzed in human liver samples by immunohistochemistry. Clinical and event-free survival data were combined to assess the relationship between ITPR3 and liver cancer growth in patients. RNA sequencing analysis was performed to identify apoptotic genes altered by ITPR3 expression in a liver tumor cell line. ITPR3 was highly expressed in HCC tumor cells relative to adjacent CLD tissue and healthy livers. There was an inverse correlation between ITPR3 expression and mitotic and apoptotic indices in HCC, suggesting that ITPR3 contributed to the maintenance of HCC by promoting resistance to apoptosis. This was confirmed by the upregulation of CTSB, CHOP and GADD45, genes involved in the apoptotic pathway in HCC. The expression of ITPR3 in the liver may be a promising prognostic marker of HCC.
- Published
- 2021
13. Correlation Between Clinical and Pathological Findings of Liver Injury in 27 Patients With Lethal COVID‐19 Infections in Brazil
- Author
-
Anderson José Ferreira, Luiz Carlos de Lima Ferreira, Israel Júnior Borges do Nascimento, Paula Vieira Teixeira Vidigal, Mateus T. Guerra, Marcus V. G. Lacerda, Rebecca Augusta de Araújo Pinto, Stanley de Almeida Araújo, Melanie A. Hundt, David Campos Wanderley, Roberto Ferreira de Almeida Araújo, Monique Freire Santana, Sérgio Veloso Brant Pinheiro, Mariana Simão Xavier, Maria M. Ciarleglio, Michael H. Nathanson, Bruna Guimarães Dutra, and M. Fatima Leite
- Subjects
Adult ,Male ,medicine.medical_specialty ,Inflammation ,Complement factor I ,RC799-869 ,Gastroenterology ,Transaminase ,Liver Function Tests ,Internal medicine ,medicine ,Humans ,Platelet activation ,Pathological ,Aged ,Liver injury ,Aged, 80 and over ,Hepatology ,business.industry ,Liver Diseases ,COVID-19 ,Original Articles ,Diseases of the digestive system. Gastroenterology ,Middle Aged ,medicine.disease ,Liver ,Original Article ,Female ,Steatosis ,medicine.symptom ,business ,CD61 ,Brazil - Abstract
Liver test abnormalities are frequently observed in patients with coronavirus disease 2019 (COVID‐19) and are associated with worse prognosis. However, information is limited about pathological changes in the liver in this infection, so the mechanism of liver injury is unclear. Here we describe liver histopathology and clinical correlates of 27 patients who died of COVID‐19 in Manaus, Brazil. There was a high prevalence of liver injury (elevated alanine aminotransferase and aspartate aminotransferase in 44% and 48% of patients, respectively) in these patients. Histological analysis showed sinusoidal congestion and ischemic necrosis in more than 85% of the cases, but these appeared to be secondary to systemic rather than intrahepatic thrombotic events, as only 14% and 22% of samples were positive for CD61 (marker of platelet activation) and C4d (activated complement factor), respectively. Furthermore, the extent of these vascular findings did not correlate with the extent of transaminase elevations. Steatosis was present in 63% of patients, and portal inflammation was present in 52%. In most cases, hepatocytes expressed angiotensin‐converting enzyme 2 (ACE2), which is responsible for binding and entry of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), even though this ectoenzyme was minimally expressed on hepatocytes in normal controls. However, SARS‐CoV‐2 staining was not observed. Most hepatocytes also expressed inositol 1,4,5‐triphosphate receptor 3 (ITPR3), a calcium channel that becomes expressed in acute liver injury. Conclusion: The hepatocellular injury that commonly occurs in patients with severe COVID‐19 is not due to the vascular events that contribute to pulmonary or cardiac damage. However, new expression of ACE2 and ITPR3 with concomitant inflammation and steatosis suggests that liver injury may result from inflammation, metabolic abnormalities, and perhaps direct viral injury., New expression of ACE2 and ITPR3 in hepatocytes with concomitant inflammation and steatosis suggests that the frequently observed liver injury in patients with COVID‐19 may result from inflammation, metabolic abnormalities, and perhaps direct viral entry.
- Published
- 2021
14. A role for mast cells and mast cell tryptase in driving neutrophil recruitment in LPS-induced lung inflammation via protease-activated receptor 2 in mice
- Author
-
M. Fatima Leite, Geovanni Dantas Cassali, Aline Dias de Almeida, Luciana O. Andrade, Antônio Carlos Melo LimaFilho, Rodrigo M. Florentino, Ayslan Barra, Irismara Sousa Silva, André Klein, and Weslley Fernandes-Braga
- Subjects
0301 basic medicine ,Lipopolysaccharides ,Proteases ,Chemokine ,Chemokine CXCL1 ,Immunology ,Tryptase ,Inflammation ,Piperazines ,03 medical and health sciences ,Mice ,0302 clinical medicine ,medicine ,Animals ,Receptor, PAR-2 ,Calcium Signaling ,Mast Cells ,Lung ,Protease-activated receptor 2 ,Pharmacology ,biology ,medicine.diagnostic_test ,Chemistry ,Pneumonia ,respiratory system ,Macrophage Activation ,Mast cell ,respiratory tract diseases ,CXCL1 ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Bronchoalveolar lavage ,RAW 264.7 Cells ,Neutrophil Infiltration ,biology.protein ,Female ,Tryptases ,medicine.symptom ,Bronchoalveolar Lavage Fluid ,030215 immunology - Abstract
This study aims to investigate the role of protease-activated receptor (PAR) 2 and mast cell (MC) tryptase in LPS-induced lung inflammation and neutrophil recruitment in the lungs of C57BL/6 mice. C57BL/6 mice were pretreated with the PAR2 antagonist ENMD-1068, compound 48/80 or aprotinin prior to intranasal instillation of MC tryptase or LPS. Blood leukocytes, C-X-C motif chemokine ligand (CXCL) 1 production leukocytes recovered from bronchoalveolar lavage fluid (BALF), and histopathological analysis of the lung were evaluated 4 h later. Furthermore, we performed experiments to determine intracellular calcium signaling in RAW 264.7 cells stimulated with LPS in the presence or absence of a protease inhibitor cocktail or ENMD-1068 and evaluated PAR2 expression in the lungs of LPS-treated mice. Pharmacological blockade of PAR2 or inhibition of proteases reduced neutrophils recovered in BALF and LPS-induced calcium signaling. PAR2 blockade impaired LPS-induced lung inflammation, PAR2 expression in the lung and CXCL1 release in BALF, and increased circulating blood neutrophils. Intranasal instillation of MC tryptase increased the number of neutrophils recovered in BALF, and MC depletion with compound 48/80 impaired LPS-induced neutrophil migration. Our study provides, for the first time, evidence of a pivotal role for MCs and MC tryptase in neutrophil migration, lung inflammation and macrophage activation triggered by LPS, by a mechanism dependent on PAR2 activation.
- Published
- 2020
15. CPP-Ts: a new intracellular calcium channel modulator and a promising tool for drug delivery in cancer cells
- Author
-
Carlos Chávez-Olórtegui, Carla J. Aguiar, André Luís Branco de Barros, Gabriela Lago Biscoto, Bárbara Bruna Ribeiro de Oliveira-Mendes, Sued Eustáquio Mendes Miranda, Valbert Nascimento Cardoso, Anderson Oliveira do Carmo, Douglas Ferreira Sales-Medina, Hortênsia Gomes Leal, Carolina Campolina Rebello Horta, M. Fatima Leite, Evanguedes Kalapothakis, and Pedro Ferreira Pinto Brandão-Dias
- Subjects
0301 basic medicine ,Cytoplasm ,Tityus serrulatus ,Scorpion Venoms ,lcsh:Medicine ,Channel modulator ,Cell-Penetrating Peptides ,Calcium in biology ,Article ,Scorpions ,03 medical and health sciences ,Drug Delivery Systems ,Cell Line, Tumor ,Neoplasms ,Animals ,Humans ,Amino Acid Sequence ,lcsh:Science ,Multidisciplinary ,030102 biochemistry & molecular biology ,biology ,Voltage-dependent calcium channel ,Chemistry ,lcsh:R ,biology.organism_classification ,Cell biology ,030104 developmental biology ,Drug delivery ,Cancer cell ,Cell-penetrating peptide ,Calcium ,lcsh:Q ,Calcium Channels ,Intracellular - Abstract
Scorpion sting envenoming impacts millions of people worldwide, with cardiac effects being one of the main causes of death on victims. Here we describe the first Ca2+ channel toxin present in Tityus serrulatus (Ts) venom, a cell penetrating peptide (CPP) named CPP-Ts. We show that CPP-Ts increases intracellular Ca2+ release through the activation of nuclear InsP3R of cardiomyocytes, thereby causing an increase in the contraction frequency of these cells. Besides proposing a novel subfamily of Ca2+ active toxins, we investigated its potential use as a drug delivery system targeting cancer cell nucleus using CPP-Ts’s nuclear-targeting property. To this end, we prepared a synthetic CPP-Ts sub peptide14–39 lacking pharmacological activity which was directed to the nucleus of specific cancer cell lines. This research identifies a novel subfamily of Ca2+ active toxins and provides new insights into biotechnological applications of animal venoms.
- Published
- 2018
16. Inositol 1,4,5-trisphosphate receptor in the liver: Expression and function
- Author
-
Antônio Carlos Melo Lima Filho, M. Fatima Leite, Fernanda O. Lemos, Rodrigo M. Florentino, and Marcone Loiola dos Santos
- Subjects
Gene isoform ,Hepatocytes and cholangiocytes ,Calcium in biology ,Inositol 1,4,5-trisphosphate receptor ,chemistry.chemical_compound ,Animals ,Humans ,Inositol 1,4,5-Trisphosphate Receptors ,Protein Isoforms ,Inositol ,Receptor ,Calcium signaling ,Chemistry ,Liver Diseases ,Gastroenterology ,Minireviews ,General Medicine ,Cell biology ,Disease Models, Animal ,Bile Ducts, Intrahepatic ,Liver ,Second messenger system ,Hepatocytes ,Calcium ,Homeostasis ,Intracellular - Abstract
The liver is a complex organ that performs several functions to maintain homeostasis. These functions are modulated by calcium, a second messenger that regulates several intracellular events. In hepatocytes and cholangiocytes, which are the epithelial cell types in the liver, inositol 1,4,5-trisphosphate (InsP3) receptors (ITPR) are the only intracellular calcium release channels. Three isoforms of the ITPR have been described, named type 1, type 2 and type 3. These ITPR isoforms are differentially expressed in liver cells where they regulate distinct physiological functions. Changes in the expression level of these receptors correlate with several liver diseases and hepatic dysfunctions. In this review, we highlight how the expression level, modulation, and localization of ITPR isoforms in hepatocytes and cholangiocytes play a role in hepatic homeostasis and liver pathology.
- Published
- 2019
17. Polymorphism in the Promoter Region of NFE2L2 Gene Is a Genetic Marker of Susceptibility to Cirrhosis Associated with Alcohol Abuse
- Author
-
Rodrigo M. Florentino, Antônio Carlos Melo Lima Filho, Kemper Nunes dos Santos, Igor Brasil Costa, Matheus de Castro Fonseca, Paula Vieira Teixeira Vidigal, Marcone Loiola dos Santos, Andressa França, Michael H. Nathanson, Dabny Goulart Missiaggia, Fernanda O. Lemos, and M. Fatima Leite
- Subjects
Male ,Alcoholic liver disease ,Cirrhosis ,endocrine system diseases ,NFE2L2 gene ,Gene Expression ,Cirrose Hep?tica / patologia ,Hepacivirus ,lcsh:Chemistry ,0302 clinical medicine ,Liver Cirrhosis, Alcoholic ,Medicine ,Promoter Regions, Genetic ,lcsh:QH301-705.5 ,Spectroscopy ,General Medicine ,Middle Aged ,Hepatitis C ,Computer Science Applications ,Liver ,030220 oncology & carcinogenesis ,030211 gastroenterology & hepatology ,Female ,Adult ,endocrine system ,congenital, hereditary, and neonatal diseases and abnormalities ,NF-E2-Related Factor 2 ,Single-nucleotide polymorphism ,Polymorphism, Single Nucleotide ,Catalysis ,Article ,Nrf2 ,Fator 2 Relacionado a NF-E2 / gen?tica ,Inorganic Chemistry ,03 medical and health sciences ,Marcadores Gen?ticos ,SNP ,Humans ,Genetic Predisposition to Disease ,Physical and Theoretical Chemistry ,Polymorphism ,Molecular Biology ,Gene ,F?gado / anatomia & histologia ,Ethanol ,Polimorfismo Gen?tico / imunologia ,business.industry ,Organic Chemistry ,nutritional and metabolic diseases ,Promoter ,medicine.disease ,NFE2L2 ,Liver Transplantation ,Oxidative Stress ,Alcoolismo / diagn?stico ,lcsh:Biology (General) ,lcsh:QD1-999 ,Genes / imunologia ,Genetic marker ,Case-Control Studies ,Cancer research ,business - Abstract
Alcoholic liver disease (ALD) is a highly prevalent spectrum of pathologies caused by alcohol overconsumption. Morbidity and mortality related to ALD are increasing worldwide, thereby demanding strategies for early diagnosis and detection of ALD predisposition. A potential candidate as a marker for ALD susceptibility is the transcription factor nuclear factor erythroid-related factor 2 (Nrf2), codified by the nuclear factor erythroid 2-related factor 2 gene (NFE2L2). Nrf2 regulates expression of proteins that protect against oxidative stress and inflammation caused by alcohol overconsumption. Here, we assessed genetic variants of NFE2L2 for association with ALD. Specimens from patients diagnosed with cirrhosis caused by ALD were genotyped for three NFE2L2 single nucleotide polymorphisms (SNP) (SNPs: rs35652124, rs4893819, and rs6721961). Hematoxylin &, eosin and immunohistochemistry were performed to determine the inflammatory score and Nrf2 expression, respectively. SNPs rs4893819 and rs6721961 were not specifically associated with ALD, but analysis of SNP rs35652124 suggested that this polymorphism predisposes to ALD. Furthermore, SNP rs35652124 was associated with a lower level of Nrf2 expression. Moreover, liver samples from ALD patients with this polymorphism displayed more severe inflammatory activity. Together, these findings provide evidence that the SNP rs35652124 variation in the Nrf2-encoding gene NFE2L2 is a potential genetic marker for susceptibility to ALD.
- Published
- 2019
18. Effects of endotoxin on type 3 inositol 1,4,5-trisphosphate receptor in human cholangiocytes
- Author
-
Basile Njei, Meenakshisundaram Ananthanarayanam, Jittima Weerachayaphorn, Mateus T. Guerra, Antônio Carlos Melo Lima Filho, M. Fatima Leite, Paula Vieira Teixeira Vidigal, Jesus M. Banales, Michael H. Nathanson, Cristiano Xavier Lima, Andressa França, Marcone Loiola dos Santos, and Marie E. Robert
- Subjects
0301 basic medicine ,Agonist ,Adult ,Male ,Lipopolysaccharide ,medicine.drug_class ,Stimulation ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Cholestasis ,medicine ,Humans ,Inositol 1,4,5-Trisphosphate Receptors ,Inositol ,Calcium Signaling ,Receptor ,Hepatology ,Hepatitis, Alcoholic ,NF-kappa B ,Epithelial Cells ,Inositol trisphosphate receptor ,Middle Aged ,medicine.disease ,Molecular biology ,Endotoxemia ,Endotoxins ,030104 developmental biology ,chemistry ,Gene Expression Regulation ,TLR4 ,030211 gastroenterology & hepatology ,Female ,Bile Ducts - Abstract
Clinical conditions that result in endotoxemia, such as sepsis and alcoholic hepatitis (AH), often are accompanied by cholestasis. Although hepatocellular changes in response to lipopolysaccharide (LPS) have been well characterized, less is known about whether and how cholangiocytes contribute to this form of cholestasis. We examined effects of endotoxin on expression and function of the type 3 inositol trisphosphate receptor (ITPR3), because this is the main intracellular Ca2+ release channel in cholangiocytes, and loss of it impairs ductular bicarbonate secretion. Bile duct cells expressed the LPS receptor, Toll-like receptor 4 (TLR4), which links to activation of nuclear factor-κB (NF-κB). Analysis of the human ITPR3 promoter revealed five putative response elements to NF-κB, and promoter activity was inhibited by p65/p50. Nested 0.5- and 1.0-kilobase (kb) deletion fragments of the ITPR3 promoter were inhibited by NF-κB subunits. Chromatin immunoprecipitation (ChIP) assay showed that NF-κB interacts with the ITPR3 promoter, with an associated increase in H3K9 methylation. LPS decreased ITPR3 mRNA and protein expression and also decreased sensitivity of bile duct cells to calcium agonist stimuli. This reduction was reversed by inhibition of TLR4. ITPR3 expression was decreased or absent in cholangiocytes from patients with cholestasis of sepsis and from those with severe AH. Conclusion: Stimulation of TLR4 by LPS activates NF-κB to down-regulate ITPR3 expression in human cholangiocytes. This may contribute to the cholestasis that can be observed in conditions such as sepsis or AH.
- Published
- 2018
19. Inositol 1,4,5-trisphosphate receptor type 3 plays a protective role in hepatocytes during hepatic ischemia-reperfusion injury
- Author
-
Antônio Carlos Melo Lima Filho, Dabny Goulart Missiaggia, Meenakshisundaram Ananthanarayanan, Mateus T. Guerra, Fernanda O. Lemos, Roberta Cristelli Fonseca, Cristiano Xavier Lima, André G. Oliveira, M. Fatima Leite, Matheus de Castro Fonseca, Michael H. Nathanson, Marcone Loiola dos Santos, Rodrigo M. Florentino, Andressa França, and Paula Vieira Teixeira Vidigal
- Subjects
Male ,0301 basic medicine ,medicine.medical_specialty ,Necrosis ,Physiology ,medicine.medical_treatment ,Liver transplantation ,Protective Agents ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Internal medicine ,medicine ,Animals ,Humans ,Inositol 1,4,5-Trisphosphate Receptors ,Inositol ,Calcium Signaling ,Promoter Regions, Genetic ,Molecular Biology ,Mice, Knockout ,NFATC Transcription Factors ,Chemistry ,NFAT ,Cell Biology ,Inositol trisphosphate receptor ,medicine.disease ,DNA Demethylation ,Mice, Inbred C57BL ,Transplantation ,Disease Models, Animal ,030104 developmental biology ,Endocrinology ,Liver ,Reperfusion Injury ,Hepatocytes ,medicine.symptom ,Reperfusion injury ,Hepatic Thrombosis ,030217 neurology & neurosurgery - Abstract
Hepatic ischemia-reperfusion injury is seen in a variety of clinical conditions, including hepatic thrombosis, systemic hypotension, and liver transplantation. Calcium (Ca(2+)) signaling mediates several pathophysiological processes in the liver, but it is not known whether and how intracellular Ca(2+) channels are involved in the hepatocellular events secondary to ischemia-reperfusion. Using an animal model of hepatic ischemia-reperfusion injury, we observed a progressive increase in expression of the type 3 isoform of the inositol trisphosphate receptor (ITPR3), an intracellular Ca(2+) channel that is not normally expressed in healthy hepatocytes. ITPR3 expression was upregulated, at least in part, by a combination of demethylation of the ITPR3 promoter region and the increased transcriptional activity of the nuclear factor of activated T-cells (NFAT). Additionally, expression of pro-inflammatory interleukins and necrotic surface area were less pronounced in livers of control animals compared to liver-specific ITPR3 KO mice subjected to hepatic damage. Corroborating these findings, ITPR3 expression and activation of NFAT were observed in hepatocytes of liver biopsies from patients who underwent liver ischemia caused by thrombosis after organ transplant. Together, these results are consistent with the idea that ITPR3 expression in hepatocytes plays a protective role during hepatic injury induced by ischemia-reperfusion.
- Published
- 2020
20. Efficacy of Meglumine Antimoniate in a Low Polymerization State Orally Administered in a Murine Model of Visceral Leishmaniasis
- Author
-
Cynthia Demicheli, Kelly Cristina Kato, Ana Rabello, Eliane de Morais-Teixeira, M. Fatima Leite, Frédéric Frézard, José Dias Corrêa-Junior, Arshad Islam, and Whocely Victor de Castro
- Subjects
0301 basic medicine ,Drug ,media_common.quotation_subject ,Meglumine antimoniate ,Administration, Oral ,02 engineering and technology ,Pharmacology ,Intestinal absorption ,Polymerization ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,Oral administration ,medicine ,Animals ,Humans ,Pharmacology (medical) ,Experimental Therapeutics ,media_common ,Mice, Inbred BALB C ,Meglumine Antimoniate ,biology ,Antimony pentachloride ,Leishmaniasis ,021001 nanoscience & nanotechnology ,biology.organism_classification ,medicine.disease ,Disease Models, Animal ,030104 developmental biology ,Infectious Diseases ,Visceral leishmaniasis ,chemistry ,Leishmaniasis, Visceral ,Female ,Leishmania infantum ,Caco-2 Cells ,0210 nano-technology ,medicine.drug - Abstract
Progress toward the improvement of meglumine antimoniate (MA), commercially known as Glucantime, a highly effective but also toxic antileishmanial drug, has been hindered by the lack of knowledge and control of its chemical composition. Here, MA was manipulated chemically with the aim of achieving an orally effective drug. MA compounds were synthesized from either antimony pentachloride (MA-SbCl5) or potassium hexahydroxyantimonate [MA-KSb(OH)6] and prepared under a low polymerization state. These compounds were compared to Glucantime regarding chemical composition, permeation properties across a cellulose membrane and Caco-2 cell monolayer, and uptake by peritoneal macrophages. MA-SbCl5 and MA-KSb(OH)6 were characterized as less polymerized and more permeative 2:2 Sb-meglumine complexes than Glucantime, which consisted of a mixture of 2:3 and 3:3 Sb-meglumine complexes. The antileishmanial activities and hepatic uptake of all compounds were evaluated after oral administration in BALB/c mice infected with Leishmania infantum chagasi, as a model of visceral leishmaniasis (VL). The synthetic MA compounds given at 300 mg Sb/kg of body weight/12 h for 30 days significantly reduced spleen and liver parasite burdens, in contrast to those for Glucantime at the same dose. The greater activity of synthetic compounds could be attributed to their higher intestinal absorption and accumulation efficiency in the liver. MA-SbCl5 given orally was as efficacious as Glucantime by the parenteral route (80 mg Sb/kg/24 h intraperitoneally). These data taken together suggest that treatment with a less-polymerized form of MA by the oral route may be effective for the treatment of VL.
- Published
- 2018
21. Cholesterol-enriched membrane microdomains are needed for insulin signaling and proliferation in hepatic cells
- Author
-
André G. Oliveira, M. Fatima Leite, Andressa França, Laurent Dubuquoy, Rodrigo M. Florentino, Antônio Carlos Melo Lima Filho, Roberta Cristelli Fonseca, P.T. Vidigal, Matheus de Castro Fonseca, and Michael H. Nathanson
- Subjects
0301 basic medicine ,Physiology ,medicine.medical_treatment ,Cell membrane ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Membrane Microdomains ,Physiology (medical) ,medicine ,Animals ,Insulin ,Lipid raft ,Calcium signaling ,Cell Proliferation ,Hepatology ,biology ,Cholesterol ,Gastroenterology ,Liver regeneration ,Receptor, Insulin ,Cell biology ,Liver Regeneration ,Rats ,Insulin receptor ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,030220 oncology & carcinogenesis ,Hepatic stellate cell ,biology.protein ,Hepatocytes ,lipids (amino acids, peptides, and proteins) ,Calcium ,Signal Transduction ,Research Article - Abstract
Hepatocyte proliferation during liver regeneration is a well-coordinated process regulated by the activation of several growth factor receptors, including the insulin receptor (IR). The IR can be localized in part to cholesterol-enriched membrane microdomains, but the role of such domains in insulin-mediated events in hepatocytes is not known. We investigated whether partitioning of IRs into cholesterol-enriched membrane rafts is important for the mitogenic effects of insulin in the hepatic cells. IR and lipid rafts were labeled in HepG2 cells and primary rat hepatocytes. Membrane cholesterol was depleted in vitro with metyl-β-cyclodextrin (MβCD) and in vivo with lovastatin. Insulin-induced calcium (Ca2+) signals studies were examined in HepG2 cells and in freshly isolated rat hepatocytes as well as in whole liver in vivo by intravital confocal imaging. Liver regeneration was studied by 70% partial hepatectomy (PH), and hepatocyte proliferation was assessed by PCNA staining. A subpopulation of IR was found in membrane microdomains enriched in cholesterol. Depletion of cholesterol from plasma membrane resulted in redistribution of the IR along the cells, which was associated with impaired insulin-induced nuclear Ca2+signals, a signaling event that regulates hepatocyte proliferation. Cholesterol depletion also led to ERK1/2 hyper-phosphorylation. Lovastatin administration to rats decreased hepatic cholesterol content, disrupted lipid rafts and decreased insulin-induced Ca2+signaling in hepatocytes, and delayed liver regeneration after PH. Therefore, membrane cholesterol content and lipid rafts integrity showed to be important for the proliferative effects of insulin in hepatic cells.NEW & NOTEWORTHY One of insulin’s actions is to stimulate liver regeneration. Here we show that a subpopulation of insulin receptors is in a specialized cholesterol-enriched region of the cell membrane and this subfraction is important for insulin’s proliferative effects.
- Published
- 2018
22. Inositol 1, 4, 5-trisphosphate-dependent nuclear calcium signals regulate angiogenesis and cell motility in triple negative breast cancer
- Author
-
Clarissa Coelho de Carvalho, Márcio Flávio Dutra Moraes, Gabriel Fernandes, M. Fatima Leite, Carlos Alberto Gonçalves, Puebla Cassini, Ana Cândida Araújo e Silva, Barbara Hissa, Matheus de Castro Fonseca, Brígida Gomes Almeida, José Miguel Ortega, André G. Oliveira, Lucíola S. Barcelos, Andressa França, Luciana E. Drumond, Erika S. Guimarães, Rodrigo Matta Machado, and Marina De Brot
- Subjects
0301 basic medicine ,Adenoviruses ,Angiogenesis ,Physiology ,Tumor Physiology ,Cancer Treatment ,lcsh:Medicine ,Gene Expression ,Triple Negative Breast Neoplasms ,Inositol 1,4,5-Trisphosphate ,Cardiovascular Physiology ,Pathology and Laboratory Medicine ,Mice ,Cell Movement ,Breast Tumors ,Basic Cancer Research ,Medicine and Health Sciences ,lcsh:Science ,Triple-negative breast cancer ,Mice, Inbred BALB C ,Multidisciplinary ,Neovascularization, Pathologic ,Chemistry ,Cell migration ,Primary tumor ,Gene Expression Regulation, Neoplastic ,Cell Motility ,Oncology ,Medical Microbiology ,Tumor Angiogenesis ,Viral Pathogens ,Viruses ,Heterografts ,Female ,Pathogens ,Research Article ,medicine.medical_specialty ,Motility ,Microbiology ,Focal adhesion ,03 medical and health sciences ,Downregulation and upregulation ,Internal medicine ,Cell Line, Tumor ,Breast Cancer ,medicine ,Genetics ,Animals ,Humans ,Calcium Signaling ,Microbial Pathogens ,Cell Proliferation ,Cell Nucleus ,lcsh:R ,Organisms ,Cancers and Neoplasms ,Biology and Life Sciences ,Cell Biology ,medicine.disease ,Chemokine CXCL10 ,030104 developmental biology ,Endocrinology ,Tumor progression ,Cancer research ,lcsh:Q ,DNA viruses ,Developmental Biology - Abstract
Increases in nuclear calcium concentration generate specific biological outcomes that differ from those resulting from increased cytoplasmic calcium. Nuclear calcium effects on tumor cell proliferation are widely appreciated; nevertheless, its involvement in other steps of tumor progression is not well understood. Therefore, we evaluated whether nuclear calcium is essential in other additional stages of tumor progression, including key steps associated with the formation of the primary tumor or with the metastatic cascade. We found that nuclear calcium buffering impaired 4T1 triple negative breast cancer growth not just by decreasing tumor cell proliferation, but also by enhancing tumor necrosis. Moreover, nuclear calcium regulates tumor angiogenesis through a mechanism that involves the upregulation of the anti-angiogenic C-X-C motif chemokine 10 (CXCL10-IP10). In addition, nuclear calcium buffering regulates breast tumor cell motility, culminating in less cell invasion, likely due to enhanced vinculin expression, a focal adhesion structural protein. Together, our results show that nuclear calcium is essential for triple breast cancer angiogenesis and cell migration and can be considered as a promising strategic target for triple negative breast cancer therapy.
- Published
- 2017
23. Decoding Calcium Signaling Across the Nucleus
- Author
-
Lídia M. Andrade, Gustavo B. Menezes, M. Fatima Leite, André G. Oliveira, and Erika S. Guimarães
- Subjects
Cell Nucleus ,Calcium metabolism ,Physiology ,Reviews ,chemistry.chemical_element ,Biology ,Calcium ,Cell biology ,Repressor Proteins ,Cell nucleus ,Cytosol ,chemistry.chemical_compound ,Adenosine Triphosphate ,medicine.anatomical_structure ,chemistry ,TheoryofComputation_ANALYSISOFALGORITHMSANDPROBLEMCOMPLEXITY ,Second messenger system ,medicine ,Animals ,Humans ,Calcium Signaling ,Nucleus ,Adenosine triphosphate ,Calcium signaling - Abstract
Calcium (Ca2+) is an important multifaceted second messenger that regulates a wide range of cellular events. A Ca2+-signaling toolkit has been shown to exist in the nucleus and to be capable of generating and modulating nucleoplasmic Ca2+transients. Within the nucleus, Ca2+controls cellular events that are different from those modulated by cytosolic Ca2+. This review focuses on nuclear Ca2+signals and their role in regulating physiological and pathological processes.
- Published
- 2014
24. Calcium signalling from the type I inositol 1,4,5-trisphosphate receptor is required at early phase of liver regeneration
- Author
-
Flávia M. Melo, Gustavo B. Menezes, Miguel J. Ortega, M. Fatima Leite, Pedro Sousa, Erika S. Guimarães, Pedro Marques, Viviane A. Andrade, Rodrigo M. Florentino, and André G. Oliveira
- Subjects
Male ,Time Factors ,CHO Cells ,Biology ,Transfection ,Rats, Sprague-Dawley ,chemistry.chemical_compound ,Cricetulus ,Proliferating Cell Nuclear Antigen ,medicine ,Animals ,Hepatectomy ,Humans ,Inositol 1,4,5-Trisphosphate Receptors ,Lobules of liver ,Inositol ,Calcium Signaling ,Receptor ,Cell Proliferation ,Calcium signaling ,Gene knockdown ,Hepatology ,Regeneration (biology) ,Organ Size ,Liver regeneration ,Liver Regeneration ,Cell biology ,HEK293 Cells ,medicine.anatomical_structure ,Liver ,Biochemistry ,chemistry ,Hepatocyte ,RNA Interference ,Biomarkers - Abstract
Background & Aims Liver regeneration is a multistage process that unfolds gradually, with different mediators acting at different stages of regeneration. Calcium (Ca2+) signalling is essential for liver regeneration. In hepatocytes, Ca2+ signalling results from the activation of inositol 1,4,5-trisphosphate receptors (InsP3R) of which two of the three known isoforms are expressed (InsP3R-I and InsP3R-II). Here, we investigated the role of the InsP3R-I-dependent Ca2+ signals in hepatic proliferation during liver regeneration. Methods Partial hepatectomy (HX) in combination with knockdown of InsP3R-I (AdsiRNA-I) was used to evaluate the role of InsP3R-I on liver regeneration and hepatocyte proliferation, as assessed by liver to body mass ratio, PCNA expression, immunoblots and measurements of intracellular Ca2+ signalling. Results AdsiRNA-I efficiently infected the liver as demonstrated by the expression of β-galactosidase throughout the liver lobules. Moreover, this construct selectively and efficiently reduced the expression of InsP3R-I, as evaluated by immunoblots. Expression of AdsiRNA-I in liver decreased peak Ca2+ amplitude induced by vasopressin in isolated hepatocytes 2 days after HX. Reduced InsP3R-I expression prior to HX also delayed liver regeneration, as measured by liver to body weight ratio, and reduced hepatocyte proliferation, as evaluated by PCNA staining, at the same time point. At later stages of regeneration, control hepatocytes showed a decreased expression of InsP3R, as well as reduced InsP3R-mediated Ca2+ signalling, events that did not affect liver growth. Conclusion Together, these results show that InsP3R-I-dependent Ca2+ signalling is an early triggering pathway required for liver regeneration.
- Published
- 2014
25. The insulin receptor translocates to the nucleus to regulate cell proliferation in liver
- Author
-
André G. Oliveira, Albert Mennone, Sandhra M. Carvalho, Gustavo B. Menezes, M. Fatima Leite, Erika S. Guimarães, Rodrigo R. Resende, Mauro Cunha Xavier Pinto, Cleida A. Oliveira, Michael H. Nathanson, Maria Jimena Amaya, Lídia M. Andrade, and Marisa C. F. Casteluber
- Subjects
medicine.medical_specialty ,Hepatology ,biology ,Insulin ,medicine.medical_treatment ,GRB10 ,Liver regeneration ,IRS2 ,Insulin receptor ,Endocrinology ,medicine.anatomical_structure ,Insulin receptor substrate ,Internal medicine ,biology.protein ,medicine ,Nucleus ,Insulin-like growth factor 1 receptor - Abstract
Insulin’s metabolic effects in the liver are widely appreciated, but insulin’s ability to act as a hepatic mitogen is less well understood. Because the Insulin Receptor (IR) can traffic to the nucleus, and calcium (Ca2+) signals within the nucleus regulate cell proliferation, we investigated whether insulin’s mitogenic effects result from activation of Ca2+ signaling pathways by IRs within the nucleus. Insulin-induced increases in Ca2+ and cell proliferation depended upon clathrin- and caveolin-dependent translocation of the IR to the nucleus, as well as upon formation of inositol 1,4,5,-trisphosphate (InsP3) in the nucleus, whereas insulin’s metabolic effects did not depend on either of these events. Moreover, liver regeneration after partial hepatectomy also depended upon formation of InsP3 in the nucleus but not the cytosol, whereas hepatic glucose metabolism was not affected by buffering InsP3 in the nucleus. Conclusion: These findings provide evidence that insulin’s mitogenic effects are mediated by a subpopulation of IRs that traffic to the nucleus to locally activate InsP3-dependent Ca2+ signaling pathways. The steps along this signaling pathway reveal a number of potential targets for therapeutic modulation of liver growth in health and disease.
- Published
- 2013
26. Nuclear inositol 1,4,5-trisphosphate is a necessary and conserved signal for the induction of both pathological and physiological cardiomyocyte hypertrophy
- Author
-
Cibele Rocha-Resende, Silvia Guatimosim, Maria Jimena Amaya, Rodrigo R. Resende, M. Fatima Leite, Núbia C.G. Figueiró, Lídia M. Andrade, Carla J. Aguiar, K.G. Franchini, and Lilian Anee Muniz Arantes
- Subjects
medicine.medical_specialty ,Cardiomegaly ,Inositol 1,4,5-Trisphosphate ,Biology ,Histone Deacetylases ,Muscle hypertrophy ,chemistry.chemical_compound ,Internal medicine ,medicine ,Animals ,Inositol 1,4,5-Trisphosphate Receptors ,Myocyte ,Myocytes, Cardiac ,Inositol ,Insulin-Like Growth Factor I ,Rats, Wistar ,Molecular Biology ,Cell Proliferation ,Cell Nucleus ,Histone deacetylase 5 ,Endothelin-1 ,NFATC Transcription Factors ,Cell growth ,Calcineurin ,NFAT ,Rats ,Cell biology ,Endocrinology ,chemistry ,Calcium ,Signal transduction ,Cardiology and Cardiovascular Medicine ,Signal Transduction - Abstract
It is well established that inositol 1,4,5-trisphosphate (IP3) dependent Ca(2+) signaling plays a crucial role in cardiomyocyte hypertrophy. However, it is not yet known whether nuclear IP3 represents a Ca(2+) mobilizing pathway involved in this process. The goal of the current work was to investigate the specific role of nuclear IP3 in cardiomyocyte hypertrophic response. In this work, we used an adenovirus construct that selectively buffers IP3 in the nuclear region of neonatal cardiomyocytes. We showed for the first time that nuclear IP3 mediates endothelin-1 (ET-1) induced hypertrophy. We also found that both calcineurin (Cn)/nuclear factor of activated T Cells (NFAT) and histone deacetylase-5 (HDAC5) pathways require nuclear IP3 to mediate pathological cardiomyocyte growth. Additionally, we found that nuclear IP3 buffering inhibited insulin-like growth factor-1 (IGF-1) induced hypertrophy and prevented reexpression of fetal gene program. Together, these results demonstrated that nuclear IP3 is an essential and a conserved signal for both pathological and physiological forms of cardiomyocyte hypertrophy.
- Published
- 2012
27. Succinate modulates Ca2+ transient and cardiomyocyte viability through PKA-dependent pathway
- Author
-
M. Fatima Leite, Alvair P. Almeida, Dawidson Assis Gomes, Carla J. Aguiar, Vanessa L. Andrade, Rodrigo R. Resende, Márcia N.M. Alves, Ana Cristina do Nascimento Pinheiro, Enéas Ricardo de Morais Gomes, Silvia Guatimosim, Alfredo M. Goes, and Marina Ladeira
- Subjects
Male ,Cell Survival ,Physiology ,Succinic Acid ,Biology ,Receptors, G-Protein-Coupled ,Adenylyl cyclase ,chemistry.chemical_compound ,Animals ,Myocytes, Cardiac ,Calcium Signaling ,Viability assay ,RNA, Small Interfering ,Rats, Wistar ,Protein kinase A ,Molecular Biology ,Cells, Cultured ,G protein-coupled receptor ,Microscopy, Confocal ,Ryanodine receptor ,Calcium-Binding Proteins ,Ryanodine Receptor Calcium Release Channel ,Cell Biology ,Cyclic AMP-Dependent Protein Kinases ,Rats ,Phospholamban ,Cell biology ,Citric acid cycle ,chemistry ,Biochemistry ,Phosphorylation - Abstract
GPR91 is an orphan G-protein-coupled receptor (GPCR) that has been characterized as a receptor for succinate, a citric acid cycle intermediate, in several tissues. In the heart, the role of succinate is unknown. We now report that rat ventricular cardiomyocytes express GPR91. We found that succinate, through GPR91, increases the amplitude and the rate of decline of global Ca(2+) transient, by increasing the phosphorylation levels of ryanodine receptor and phospholamban, two well known Ca(2+) handling proteins. The effects of succinate on Ca(2+) transient were abolished by pre-treatment with adenylyl cyclase and cAMP-dependent protein kinase (PKA) inhibitors. Direct PKA activation by succinate was further confirmed using a FRET-based A-kinase activity reporter. Additionally, succinate decreases cardiomyocyte viability through a caspase-3 activation pathway, effect also prevented by PKA inhibition. Taken together, these observations show that succinate acts as a signaling molecule in cardiomyocytes, modulating global Ca(2+) transient and cell viability through a PKA-dependent pathway.
- Published
- 2010
28. RETRACTED: c-Jun Inhibits Thapsigargin-Induced ER Stress Through Up-Regulation of DSCR1/Adapt78
- Author
-
Agnes S Kim, Jun Ren, Xiaoyan Xiao, M. Fatima Leite, Xinglei Zhu, Peng Zhao, Jinxia Xu, and Ji Li
- Subjects
Programmed cell death ,chemistry.chemical_compound ,Thapsigargin ,Downregulation and upregulation ,Chemistry ,Kinase ,Endoplasmic reticulum ,Gene expression ,c-jun ,Unfolded protein response ,General Biochemistry, Genetics and Molecular Biology ,Cell biology - Abstract
The endoplasmic reticulum (ER) is exquisitely sensitive to changes in its internal environment. Various conditions, collectively termed “ER stress”, can perturb ER function, leading to the activation of a complex response known as the unfolded protein response (UPR). Although c-Jun N-terminal kinase (JNK) activation is nearly always associated with cell death by various stimuli, the functional role of JNK in ER stress-induced cell death remains unclear. JNK regulates gene expression through the phosphorylation and activation of transcription factors, such as c-Jun. Here, we investigated the role of c-Jun in the regulation of ER stress-related genes. c-Jun expression levels determined the response of mouse fibroblasts to ER stress induced by thapsigargin (TG, an inhibitor of sarco/endoplasmic reticulum Ca2+ ATPase). c-jun−/− mouse fibroblast cells were more sensitive to TG-induced cell death compared to wild-type mouse fibroblasts, while reconstitution of c-Jun expression in c-jun−/− cells (c-Jun Re) enhan...
- Published
- 2008
29. Nuclear Ca2+ regulates cardiomyocyte function
- Author
-
Mateus T. Guerra, Dawidson Assis Gomes, Silvia Guatimosim, Alfredo M. Goes, Carla J. Aguiar, Norma L. Gómez-Viquez, W. J. Lederer, Maria Jimena Amaya, Juliana Martins-Cruz, Michele Angela Rodrigues, and M. Fatima Leite
- Subjects
Cytoplasm ,medicine.medical_specialty ,Nuclear Envelope ,Physiology ,Blotting, Western ,Nuclear Localization Signals ,Nucleoplasmic reticulum ,Fluorescent Antibody Technique ,Inositol 1,4,5-Trisphosphate ,Biology ,Fluorescence ,Adenoviridae ,Internal medicine ,medicine ,Animals ,Inositol 1,4,5-Trisphosphate Receptors ,Myocytes, Cardiac ,Calcium Signaling ,Rats, Wistar ,Receptor ,Molecular Biology ,Calcium signaling ,Cell Nucleus ,Ryanodine receptor ,Ryanodine Receptor Calcium Release Channel ,NFAT ,Cell Biology ,Rats ,Cell biology ,Cell nucleus ,Parvalbumins ,medicine.anatomical_structure ,Endocrinology ,Animals, Newborn ,Calcium ,Nuclear localization sequence - Abstract
In the heart, cytosolic Ca(2+) signals are well-characterized events that participate in the activation of cell contraction. In contrast, nuclear Ca(2+) contribution to cardiomyocyte function remains elusive. Here, we examined functional consequences of buffering nuclear Ca(2+) in neonatal cardiomyocytes. We report that cardiomyocytes contain a nucleoplasmic reticulum, which expresses both ryanodine receptor (RyR) and inositol 1,4,5-trisphosphate receptor (InsP(3)R), providing a possible way for active regulation of nuclear Ca(2+). Adenovirus constructs encoding the Ca(2+) buffer protein parvalbumin were targeted to the nucleus with a nuclear localization signal (Ad-PV-NLS) or to the cytoplasm with a nuclear exclusion signal (Ad-PV-NES). A decrease in the amplitude of global Ca(2+) transients and RyR-II expression, as well as an increase in cell beating rate were observed in Ad-PV-NES and Ad-PV-NLS cells. When nuclear Ca(2+) buffering was imposed nuclear enlargement, increased calcineurin expression, NFAT translocation to the nucleus and subcellular redistribution of atrial natriuretic peptide were observed. Furthermore, prolongation of action potential duration occurred in adult ventricular myocytes. These results suggest that nuclear Ca(2+) levels underlie the regulation of specific protein targets and thereby modulate cardiomyocyte function. The local nuclear Ca(2+) signaling and the structures that control it constitute a novel regulatory motif in the heart.
- Published
- 2008
30. Insulin induces calcium signals in the nucleus of rat hepatocytes
- Author
-
M. Fatima Leite, Michele Angela Rodrigues, Viviane A. Andrade, Dawidson Assis Gomes, and Michael H. Nathanson
- Subjects
Male ,Phosphatidylinositol 4,5-Diphosphate ,medicine.medical_specialty ,medicine.medical_treatment ,Inositol 1,4,5-Trisphosphate ,Rats, Sprague-Dawley ,Internal medicine ,Insulin receptor substrate ,medicine ,Animals ,Insulin ,TRPM3 ,Phosphorylation ,Insulin-like growth factor 1 receptor ,Cell Nucleus ,Microscopy, Confocal ,Dose-Response Relationship, Drug ,Hepatology ,biology ,GRB10 ,Receptor, Insulin ,IRS2 ,Rats ,Insulin receptor ,Endocrinology ,Hepatocytes ,biology.protein ,Calcium ,Signal transduction ,Signal Transduction - Abstract
Insulin is an hepatic mitogen that promotes liver regeneration. Actions of insulin are mediated by the insulin receptor, which is a receptor tyrosine kinase. It is currently thought that signaling via the insulin receptor occurs at the plasma membrane, where it binds to insulin. Here we report that insulin induces calcium oscillations in isolated rat hepatocytes, and that these calcium signals depend upon activation of phospholipase C and the inositol 1,4,5-trisphosphate receptor, but not upon extracellular calcium. Furthermore, insulin-induced calcium signals occur in the nucleus, and are temporally associated with selective depletion of nuclear phosphatidylinositol bisphosphate and translocation of the insulin receptor to the nucleus. These findings suggest that the insulin receptor translocates to the nucleus to initiate nuclear, inositol 1,4,5-trisphosphate-mediated calcium signals in rat hepatocytes. This novel signaling mechanism may be responsible for insulin's effects on liver growth and regeneration.
- Published
- 2008
31. c-Met Must Translocate to the Nucleus to Initiate Calcium Signals
- Author
-
Michele Angela Rodrigues, Anton M. Bennett, M. Fatima Leite, Michael H. Nathanson, Marcus Vinicius Gomez, Tamas Balla, Péter Várnai, and Dawidson Assis Gomes
- Subjects
Fluorescent Antibody Technique ,Importin ,Biology ,Biochemistry ,Article ,Cell Line ,chemistry.chemical_compound ,medicine ,Humans ,Calcium Signaling ,Phosphatidylinositol ,RNA, Small Interfering ,Molecular Biology ,Calcium signaling ,Cell Nucleus ,Signal transducing adaptor protein ,Cell Biology ,Proto-Oncogene Proteins c-met ,Cell biology ,Protein Transport ,medicine.anatomical_structure ,chemistry ,Cytoplasm ,Hepatocyte growth factor ,Signal transduction ,Nucleus ,medicine.drug - Abstract
Hepatocyte growth factor (HGF) is important for cell proliferation, differentiation, and related activities. HGF acts through its receptor c-Met, which activates downstream signaling pathways. HGF binds to c-Met at the plasma membrane, where it is generally believed that c-Met signaling is initiated. Here we report that c-Met rapidly translocates to the nucleus upon stimulation with HGF. Ca(2+) signals that are induced by HGF result from phosphatidylinositol 4,5-bisphosphate hydrolysis and inositol 1,4,5-trisphosphate formation within the nucleus rather than within the cytoplasm. Translocation of c-Met to the nucleus depends upon the adaptor protein Gab1 and importin beta1, and formation of Ca(2+) signals in turn depends upon this translocation. HGF may exert its particular effects on cells because it bypasses signaling pathways in the cytoplasm to directly activate signaling pathways in the nucleus.
- Published
- 2008
32. Nucleoplasmic Calcium Is Required for Cell Proliferation
- Author
-
Wayne M. Grant, Dawidson Assis Gomes, Lei Zhang, Michele Angela Rodrigues, Anton M. Bennett, Yung-Chi Cheng, M. Fatima Leite, Wing Lam, and Michael H. Nathanson
- Subjects
Male ,Cytoplasm ,Fluorescent Antibody Technique ,Mice, Nude ,Mitosis ,Biology ,Biochemistry ,Article ,Cell Line ,Mice ,Prophase ,medicine ,Animals ,Humans ,Calcium Signaling ,Molecular Biology ,Cell Proliferation ,Calcium signaling ,Cell Nucleus ,Cell growth ,Cell Cycle ,Cell Biology ,Cell cycle ,Cell biology ,Cell nucleus ,medicine.anatomical_structure ,Cell culture ,Calcium - Abstract
Ca(2+) signals regulate cell proliferation, but the spatial and temporal specificity of these signals is unknown. Here we use selective buffers of nucleoplasmic or cytoplasmic Ca(2+) to determine that cell proliferation depends upon Ca(2+) signals within the nucleus rather than in the cytoplasm. Nuclear Ca(2+) signals stimulate cell growth rather than inhibit apoptosis and specifically permit cells to advance through early prophase. Selective buffering of nuclear but not cytoplasmic Ca(2+) signals also impairs growth of tumors in vivo. These findings reveal a major physiological and potential pathophysiological role for nucleoplasmic Ca(2+) signals and suggest that this information can be used to design novel therapeutic strategies to regulate conditions of abnormal cell growth.
- Published
- 2007
33. The Spatial Distribution of Inositol 1,4,5-Trisphosphate Receptor Isoforms Shapes Ca2+ Waves
- Author
-
Oscar Bruna-Romero, Emma A. Kruglov, Frank J. Giordano, Dawidson Assis Gomes, Michele Angela Rodrigues, Erick Hernandez, M. Fatima Leite, Michael H. Nathanson, Jonathan A. Dranoff, and Mateus T. Guerra
- Subjects
Gene isoform ,Vasopressins ,Molecular Sequence Data ,Receptors, Cytoplasmic and Nuclear ,Biology ,Biochemistry ,Article ,chemistry.chemical_compound ,Animals ,Inositol 1,4,5-Trisphosphate Receptors ,Protein Isoforms ,Inositol ,Calcium Signaling ,Receptor ,Molecular Biology ,Cells, Cultured ,Calcium signaling ,Membrane Glycoproteins ,Base Sequence ,Voltage-dependent calcium channel ,Cell Biology ,Inositol trisphosphate receptor ,Molecular biology ,Rats ,Cell biology ,chemistry ,Cell culture ,Second messenger system ,Hepatocytes ,Calcium Channels - Abstract
Cytosolic Ca(2+) is a versatile second messenger that can regulate multiple cellular processes simultaneously. This is accomplished in part through Ca(2+) waves and other spatial patterns of Ca(2+) signals. To investigate the mechanism responsible for the formation of Ca(2+) waves, we examined the role of inositol 1,4,5-trisphosphate receptor (InsP3R) isoforms in Ca(2+) wave formation. Ca(2+) signals were examined in hepatocytes, which express the type I and II InsP3R in a polarized fashion, and in AR4-2J cells, a nonpolarized cell line that expresses type I and II InsP3R in a ratio similar to what is found in hepatocytes but homogeneously throughout the cell. Expression of type I or II InsP3R was selectively suppressed by isoform-specific DNA antisense in an adenoviral delivery system, which was delivered to AR4-2J cells in culture and to hepatocytes in vivo. Loss of either isoform inhibited Ca(2+) signals to a similar extent in AR4-2J cells. In contrast, loss of the basolateral type I InsP3R decreased the sensitivity of hepatocytes to vasopressin but had little effect on the initiation or spread of Ca(2+) waves across hepatocytes. Loss of the apical type II isoform caused an even greater decrease in the sensitivity of hepatocytes to vasopressin and resulted in Ca(2+) waves that were much slower and delayed in onset. These findings provide evidence that the apical concentration of type II InsP3Rs is essential for the formation of Ca(2+) waves in hepatocytes. The subcellular distribution of InsP3R isoforms may critically determine the repertoire of spatial patterns of Ca(2+) signals.
- Published
- 2007
34. GPR91: expanding the frontiers of Krebs cycle intermediates
- Author
-
Carla J. Aguiar, M. Fatima Leite, Joao Antônio da Rocha Franco, Rafael N. Gingold, and Matheus de Castro Fonseca
- Subjects
0301 basic medicine ,Models, Molecular ,Cell signaling ,Succinate ,GPR91 ,Citric Acid Cycle ,Succinic Acid ,Gene Expression ,Blood Pressure ,White adipose tissue ,Review ,Biology ,Biochemistry ,Retina ,Receptors, G-Protein-Coupled ,Cell functions ,03 medical and health sciences ,0302 clinical medicine ,Extracellular ,Animals ,Humans ,Receptor ,Molecular Biology ,Myocardium ,Cell Biology ,Cell biology ,Citric acid cycle ,030104 developmental biology ,Liver ,Mitochondrial matrix ,030220 oncology & carcinogenesis ,Hepatic stellate cell ,Signal transduction ,Signal Transduction - Abstract
Since it was discovered, the citric acid cycle has been known to be central to cell metabolism and energy homeostasis. Mainly found in the mitochondrial matrix, some of the intermediates of the Krebs cycle are also present in the blood stream. Currently, there are several reports that indicate functional roles for Krebs intermediates out of its cycle. Succinate, for instance, acts as an extracellular ligand by binding to a G-protein coupled receptor, known as GPR91, expressed in kidney, liver, heart, retinal cells and possibly many other tissues, leading to a wide array of physiological and pathological effects. Through GPR91, succinate is involved in functions such as regulation of blood pressure, inhibition of lipolysis in white adipose tissue, development of retinal vascularization, cardiac hypertrophy and activation of stellate hepatic cells by ischemic hepatocytes. Along the current review, these new effects of succinate through GPR91 will be explored and discussed.
- Published
- 2015
35. Signaling pathways in biliary epithelial cells
- Author
-
M. Fatima Leite, Mateus T. Guerra, Viviane A. Andrade, and Michael H. Nathanson
- Published
- 2015
36. Hepatic DNA deposition drives drug-induced liver injury and inflammation in mice
- Author
-
Rafaela Vaz Sousa Pereira, Daniel O. Patricio, Daniele Araújo Pires, Sarah Chapman, Rafael Elias Marques, Zélia Menezes-Garcia, Daniel Cisalpino, M. Fatima Leite, Adriana Machado Saraiva, Gustavo B. Menezes, Bruna Araújo David, Rodrigo Guabiraba, Lindisley Ferreira Gomides, Pedro Marques, Júlia Tosta Novaes, W. Matthew Leevy, Mauro Martins Teixeira, André G. Oliveira, GermánArturo Mahecha, Danielle G. Souza, Vicente de Paulo Martins, Daniel S. Mansur, Laboratório de Imunobiofotônica, Departamento de Morfologia, Universidade Federal de Minas Gerais (UFMG), Departamento de Fisiologia e Biofísica, Departamento de Microbiologia, Imunologia e Parasitologia, Departamento de Microbiologia, Biological Imaging Core, Department of Biological Sciences, University of Notre Dame [Indiana] (UND), Departamento de Morfologia, Laboratório de Imunofarmacologia, Departamento de Bioquímica e Imunologia, Infectiologie et Santé Publique (UMR ISP), Institut National de la Recherche Agronomique (INRA)-Université de Tours, Departamento de Biologia Celular (CEL), Instituto de Ciências Biológicas (IB), Universidade de Brasilia [Brasília] (UnB), UR Infectiologie animale et Santé publique (UR IASP), Institut National de la Recherche Agronomique (INRA), Pôle Santé Animale de Tours PSAT, UMR 1282, Infectiologie et Santé Publique, ISP 213, and Institut National de la Recherche Agronomique (INRA)-Université de Tours (UT)
- Subjects
Programmed cell death ,acetaminophen overdose ,Necrosis ,Neutrophils ,Inflammation ,Biology ,Systemic inflammation ,Neutrophil Activation ,03 medical and health sciences ,TLR9 ,0302 clinical medicine ,medicine ,Animals ,Acetaminophen ,030304 developmental biology ,Liver injury ,0303 health sciences ,Innate immune system ,Hepatology ,[SDV.BA]Life Sciences [q-bio]/Animal biology ,DNA ,Analgesics, Non-Narcotic ,medicine.disease ,3. Good health ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Liver ,Toll-Like Receptor 9 ,030220 oncology & carcinogenesis ,Hepatocyte ,Immunology ,Hepatocytes ,Cancer research ,Chemical and Drug Induced Liver Injury ,medicine.symptom - Abstract
Drug-induced liver injury (DILI) is an important cause of acute liver failure, with limited therapeutic options. During DILI, oncotic necrosis with concomitant release and recognition of intracellular content amplifies liver inflammation and injury. Among these molecules, self-DNA has been widely shown to trigger inflammatory and autoimmune diseases; however, whether DNA released from damaged hepatocytes accumulates into necrotic liver and the impact of its recognition by the immune system remains elusive. Here we show that treatment with two different hepatotoxic compounds (acetaminophen and thioacetamide) caused DNA release into the hepatocyte cytoplasm, which occurred in parallel with cell death in vitro. Administration of these compounds in vivo caused massive DNA deposition within liver necrotic areas, together with an intravascular DNA coating. Using confocal intravital microscopy, we revealed that liver injury due to acetaminophen overdose led to a directional migration of neutrophils to DNA-rich areas, where they exhibit an active patrolling behavior. DNA removal by intravenous DNASE1 injection or ablation of Toll-like receptor 9 (TLR9)-mediated sensing significantly reduced systemic inflammation, liver neutrophil recruitment, and hepatotoxicity. Analysis of liver leukocytes by flow cytometry revealed that emigrated neutrophils up-regulated TLR9 expression during acetaminophen-mediated necrosis, and these cells sensed and reacted to extracellular DNA by activating the TLR9/NF-κB pathway. Likewise, adoptive transfer of wild-type neutrophils to TLR9−/− mice reversed the hepatoprotective phenotype otherwise observed in TLR9 absence. Conclusion: Hepatic DNA accumulation is a novel feature of DILI pathogenesis. Blockage of DNA recognition by the innate immune system may constitute a promising therapeutic venue. (Hepatology 2015;61:348–360)
- Published
- 2015
37. Biocompatibility Evaluation of Lithium-Hydroxyapatite Composites
- Author
-
M. Fatima Leite, Gultekin Goller, A.P.M. Shainberg, Faik N. Oktar, Patricia Valério, L.S. Ozyegin, and Alfredo M. Goes
- Subjects
Primary culture ,Materials science ,Biocompatibility ,Mechanical Engineering ,chemistry.chemical_element ,Sintering ,Osteoblast ,medicine.anatomical_structure ,chemistry ,Mechanics of Materials ,medicine ,Alkaline phosphatase ,General Materials Science ,Lithium ,Composite material - Abstract
Fifteen different lithium-hydroxyapatite composites were tested in this work. LiHA were prepared using 0.25 %, 1 % and 2 % of lithium and five different sintering temperature 900, 1000, 1100, 1200 and 1300oC. Primary culture of osteoblasts were used to evaluate the biocompatibility of the samples, concerning to cell viability and alkaline phosphatase production. The 1% LiHA samples sintered at 1100, 1200 and 13000C showed the best results.
- Published
- 2006
38. Biocompatibility of the Outer Prismatic and the Inner Nacreous Layers of Four Different Molluscs
- Author
-
Alfredo M. Goes, Simeon Agathopoulos, Patricia Valério, M. Fatima Leite, Faik N. Oktar, and Gultekin Goller
- Subjects
Materials science ,Biocompatibility ,biology ,Pecten jacobaeus ,Mechanical Engineering ,Aragonite ,Statistical difference ,Osteoblast ,engineering.material ,biology.organism_classification ,In vitro biocompatibility ,medicine.anatomical_structure ,Chemical engineering ,Mechanics of Materials ,engineering ,medicine ,Alkaline phosphatase ,General Materials Science ,Layer (electronics) - Abstract
The in vitro biocompatibility of aragonite material obtained from inner and out layers of four different molluscs was tested. After grinding and sieving, the obtained fine powders were put in contact with primary osteoblasts derived from rat calvariae. The viability of the cells increased at about 10% in the presence of powders derived from Vennus Gallina outer layer and from Pecten Jacobaeus inner layer. In the case of the presence of the other 6 tested powders, there was no statistical difference in cells’ viability. With regard to alkaline phosphatase production, all the tested powders induced a decrease of the production of this enzyme by osteoblasts. There was no evidence of any alterations in collagen production.
- Published
- 2006
39. Calcium signaling in the nucleusThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell
- Author
-
M. Fatima Leite, Dawidson Assis Gomes, Anton M. Bennett, and Michael H. Nathanson
- Subjects
Pharmacology ,Physiology ,Nucleoplasmic reticulum ,General Medicine ,Inositol trisphosphate receptor ,Biology ,Cell biology ,Cytosol ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Biochemistry ,Physiology (medical) ,Second messenger system ,medicine ,Inositol ,Nuclear pore ,Nucleus ,Calcium signaling - Abstract
Cytosolic Ca2+is a versatile secondary messenger that regulates a wide range of cellular activities. In the past decade, evidence has accumulated that free Ca2+within the nucleus also plays an important messenger function. Here we review the mechanisms and effects of Ca2+signals within the nucleus. In particular, evidence is reviewed that the nucleus contains the machinery necessary for production of inositol 1,4,5-trisphosphate and for inositol 1,4,5-trisphosphate receptor-mediated Ca2+release. The role of Ca2+signals within the nucleus is discussed including regulation of such critical cell functions as gene expression, activation of kinases, and permeability of nuclear pores.
- Published
- 2006
40. Calcium release from ryanodine receptors in the nucleoplasmic reticulum
- Author
-
M. Fatima Leite, Barbara E. Ehrlich, Michael H. Nathanson, Mateus T. Guerra, and Phedra Marius
- Subjects
Cytoplasm ,Nuclear Envelope ,Physiology ,Nucleoplasmic reticulum ,Receptors, Cytoplasmic and Nuclear ,Inositol 1,4,5-Trisphosphate ,Endoplasmic Reticulum ,Dantrolene ,Cell Line ,Mice ,chemistry.chemical_compound ,medicine ,Animals ,Inositol 1,4,5-Trisphosphate Receptors ,Inositol ,Calcium Signaling ,Muscle, Skeletal ,Receptor ,Molecular Biology ,Cell Nucleus ,Ryanodine receptor ,Chemistry ,Endoplasmic reticulum ,Skeletal muscle ,Ryanodine Receptor Calcium Release Channel ,Cell Biology ,musculoskeletal system ,Cell biology ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Biochemistry ,cardiovascular system ,Calcium ,Calcium Channels ,Nucleus ,medicine.drug - Abstract
Ca(2+) signals control DNA synthesis and repair, gene transcription, and other cell functions that occur within the nucleus. The nuclear envelope can store Ca(2+) and release it into the nucleus via either the inositol 1,4,5-trisphosphate receptor (InsP3R) or the ryanodine receptor (RyR). Furthermore, many cell types have a reticular network within their nuclei and InsP3Rs on this nucleoplasmic reticulum permit local subnuclear control of Ca(2+) signals and Ca(2+)-dependent intranuclear events. However, it is unknown whether RyR similarly is expressed on the nucleoplasmic reticulum and can control subnuclear Ca(2+) signals. Here we report that the type 1 RyR is expressed on intranuclear extensions of the sarcoplasmic reticulum of C2C12 cells, a skeletal muscle derived cell line. In addition, two-photon photorelease of caged Ca(2+) in the region of the nucleoplasmic reticulum evoked Ca(2+)-induced Ca(2+) release (CICR) within the nucleus, which could be suppressed by the RyR inhibitor dantrolene. These results show that intranuclear extensions of the nuclear envelope have functional RyR and provide a possible mechanism whereby cells expressing RyR can regulate Ca(2+) signals in discrete regions within the nucleus.
- Published
- 2006
41. Succinate causes pathological cardiomyocyte hypertrophy through GPR91 activation
- Author
-
Fernando Antônio Botoni, Thiago M. Cunha, Silvia Guatimosim, Marina Ladeira, Carla J. Aguiar, João A Rocha-Franco, Rodrigo R. Resende, Luiz Orlando Ladeira, Pedro Sousa, Gustavo B. Menezes, José M. Carballido, Anderson K. Santos, M. Fatima Leite, Cristiano Xavier Lima, Cibele Rocha-Resende, and Marcos B. Melo
- Subjects
Adult ,Liver Cirrhosis ,medicine.medical_specialty ,Succinate ,Heart Diseases ,Metabolite ,Succinic Acid ,Ischemia ,Blood Pressure ,Cardiomyocyte ,Biology ,Biochemistry ,Histone Deacetylases ,Receptors, G-Protein-Coupled ,Muscle hypertrophy ,chemistry.chemical_compound ,Internal medicine ,medicine ,Extracellular ,Animals ,Humans ,Myocyte ,Myocytes, Cardiac ,Rats, Wistar ,Receptor ,Molecular Biology ,Mice, Knockout ,Mitogen-Activated Protein Kinase Kinases ,Research ,Hypertrophy ,Cell Biology ,Middle Aged ,medicine.disease ,Citric acid cycle ,Endocrinology ,Animals, Newborn ,chemistry ,Succinic acid ,Calcium ,Calcium-Calmodulin-Dependent Protein Kinase Type 2 - Abstract
Background Succinate is an intermediate of the citric acid cycle as well as an extracellular circulating molecule, whose receptor, G protein-coupled receptor-91 (GPR91), was recently identified and characterized in several tissues, including heart. Because some pathological conditions such as ischemia increase succinate blood levels, we investigated the role of this metabolite during a heart ischemic event, using human and rodent models. Results We found that succinate causes cardiac hypertrophy in a GPR91 dependent manner. GPR91 activation triggers the phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), the expression of calcium/calmodulin dependent protein kinase IIδ (CaMKIIδ) and the translocation of histone deacetylase 5 (HDAC5) into the cytoplasm, which are hypertrophic-signaling events. Furthermore, we found that serum levels of succinate are increased in patients with cardiac hypertrophy associated with acute and chronic ischemic diseases. Conclusions These results show for the first time that succinate plays an important role in cardiomyocyte hypertrophy through GPR91 activation, and extend our understanding of how ischemia can induce hypertrophic cardiomyopathy. Electronic supplementary material The online version of this article (doi:10.1186/s12964-014-0078-2) contains supplementary material, which is available to authorized users.
- Published
- 2014
42. The Type III Inositol 1,4,5-Trisphosphate Receptor Preferentially Transmits Apoptotic Ca2+ Signals into Mitochondria
- Author
-
Dawidson Assis Gomes, Alfredo M. Goes, Mayerson Thompson, M. Fatima Leite, Natalia C. Souto, Carolina C.P. Mendes, Marcus Vinicius Gomez, T.S. Goes, Michele Angela Rodrigues, and Michael H. Nathanson
- Subjects
Gene isoform ,Small interfering RNA ,Fluorescent Antibody Technique ,Gene Expression ,Receptors, Cytoplasmic and Nuclear ,Apoptosis ,CHO Cells ,Biology ,Transfection ,Biochemistry ,Cell Line ,chemistry.chemical_compound ,Adenosine Triphosphate ,Cricetulus ,Cytosol ,Cricetinae ,Chlorocebus aethiops ,Animals ,Humans ,Inositol 1,4,5-Trisphosphate Receptors ,Protein Isoforms ,Gene silencing ,Inositol ,RNA, Small Interfering ,Receptor ,Molecular Biology ,Microscopy, Confocal ,Chinese hamster ovary cell ,Cell Biology ,Molecular biology ,Mitochondria ,Cell biology ,chemistry ,Calcium ,Calcium Channels ,Signal transduction ,Signal Transduction - Abstract
There are three isoforms of the inositol 1,4,5- trisphosphate receptor (InsP(3)R), each of which has a distinct effect on Ca(2+) signaling. However, it is not known whether each isoform similarly plays a distinct role in the activation of Ca(2+)-mediated events. To investigate this question, we examined the effects of each InsP(3)R isoform on transmission of Ca(2+) signals to mitochondria and induction of apoptosis. Each isoform was selectively silenced using isoform-specific small interfering RNA in Chinese hamster ovary cells, which express all three InsP(3)R isoforms. ATP-induced cytosolic Ca(2+) signaling patterns were altered, regardless of which isoform was silenced, but in a different fashion depending on the isoform. ATP also induced Ca(2+) signals in mitochondria, which were inhibited more effectively by silencing the type III InsP(3)R than by silencing either the type I or type II isoform. The type III isoform also co-localized most strongly with mitochondria. When apoptosis was induced by activation of either the extrinsic or intrinsic apoptotic pathway, induction was reduced most effectively by silencing the type III InsP(3)R. These findings provide evidence that the type III isoform of the InsP(3)R plays a special role in induction of apoptosis by preferentially transmitting Ca(2+) signals into mitochondria.
- Published
- 2005
43. Glutamate Release by Osteoblasts in the Presence of Ionic Products from Bioactive Glass 60S
- Author
-
Marivalda M. Pereira, Patricia Valério, C.C.P. Mendes, M. Fatima Leite, and Alfredo M. Goes
- Subjects
Materials science ,Mechanical Engineering ,Calcium channel ,Glutamate receptor ,Biomaterial ,Osteoblast ,Calcium in biology ,law.invention ,Cytosol ,medicine.anatomical_structure ,Biochemistry ,Mechanics of Materials ,law ,Bioactive glass ,Biophysics ,medicine ,General Materials Science ,Receptor - Abstract
Osteoblasts constitutively release glutamate and this release appears to be regulated by calcium entry. In this work we investigated if the bioactive glass with 60% of silicon (BG60S) could alter glutamate release by osteoblasts. We demonstrated that osteoblasts incubated with medium containing ionic products from the dissolution of BG60S showed lower release of glutamate when compared to control. Since intracellular calcium (Cai 2+) increase is required for glutamate release we investigated the subcellular distribution of the calcium channel inositol triphosphate receptors (InsP3Rs) in the presence of BG60S compared to control. We found that the type-III InsP3R was not expressed in osteoblast, while the type-II InsP3R was expressed mainly in the cytosol. We also found that the expression of type-II InsP3R decreased in BG60S treated osteoblasts compared to control. On the other hand, we found that the type-I InsP3R was expressed mainly in the nucleus and its expression increased in the presence of the biomaterial.
- Published
- 2005
44. Evaluation of Osteoblast Viability, Alkaline Phosphatase Production and Collagen Secretion in the Presence of Hydroxyapatite Reinforced with Oxide Glasses
- Author
-
Alfredo M. Goes, Gultekin Goller, M. Fatima Leite, B.V. Sampaio, Patricia Valério, and Faik N. Oktar
- Subjects
Materials science ,Mechanical Engineering ,Collagen secretion ,Metallurgy ,Oxide ,Osteoblast ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Mechanics of Materials ,medicine ,Alkaline phosphatase ,General Materials Science ,Viability assay ,Nuclear chemistry - Abstract
Two (45.5 wt % P2O5, 54.5 wt% CaO) and three (45 wt % P2O5, 28 wt% CaO, 27wt% Na2O) oxide glasses, sinterizing at 1200 and 1300oC[2] showed an increase on cell viability and proliferation.
- Published
- 2005
45. Attachment of Blood Cells onto ZrO2 and SiO2-Containing Glass
- Author
-
António J. Calado, M. Fatima Leite, Simeon Agathopoulos, Patricia Valério, and Alfredo M. Goes
- Subjects
Materials science ,Settling ,Mechanics of Materials ,Scanning electron microscope ,Settling time ,Mechanical Engineering ,Surface roughness ,Analytical chemistry ,General Materials Science ,Cubic zirconia ,Adhesion ,Composite material ,Whole blood - Abstract
Samples of zirconia and a bioinert SiO2-containing glass with different surface roughness were immersed into human whole blood for different settling times to investigate the adhesion and attachment of blood cells onto these materials. The cell/material interface was directly observed by scanning electron microscopy (SEM). The results indicate that the blood cells preserved their physiology and attaching capability regardless the type of material, surface roughness, and settling time. The SEM images strongly indicate the normal function of adhesion proteins.
- Published
- 2005
46. Biocompatibility Evaluation of Zeolite Compared to Bone HA, Calcium Phosphate (Ca2PO4) and Eugenol Paste
- Author
-
Patricia Valério, A.P.M. Schainberg, M. Fatima Leite, Alfredo M. Goes, P. Kursuoğlu, and L.S. Ozyegin
- Subjects
Clinoptilolite ,Materials science ,Biocompatibility ,Mechanical Engineering ,chemistry.chemical_element ,Osteoblast ,Vacuole ,Calcium ,Eugenol ,chemistry.chemical_compound ,medicine.anatomical_structure ,chemistry ,Biochemistry ,Mechanics of Materials ,medicine ,Alkaline phosphatase ,General Materials Science ,Zeolite ,Nuclear chemistry - Abstract
The biocompatibility of Zeolite was evaluated, in vitro, compared to a control and to three different biomaterials: hydroxyapatite from bovine bone, calcium phosphate and a commercial eugenol paste. The Zeolite did not affect cellular proliferation neither the alkaline phosphatase and collagen production. The apoptosis index of the zeolite groups were similar to control and optical microscopy observations did not show any morphological cell change, except the some cytoplasmatic vacuole formation.
- Published
- 2005
47. Biocompatibility Evaluation of Three Different Titanium-Hydroxyapatite Composites
- Author
-
M. Fatima Leite, Alfredo M. Goes, Faik N. Oktar, Patricia Valério, Gultekin Goller, and B.V. Sampaio
- Subjects
Materials science ,medicine.anatomical_structure ,Biocompatibility ,chemistry ,Mechanics of Materials ,Mechanical Engineering ,medicine ,chemistry.chemical_element ,Alkaline phosphatase ,General Materials Science ,Osteoblast ,Composite material ,Titanium - Abstract
Titanium reinforced with hydroxyapatite (TiHA) prepared using 15% of titanium and 3 different sinterizing temperatures 1100, 1200 and 1300 oC showed a significant increase in cell proliferation, when compared to the control.
- Published
- 2005
48. Endotoxin unmasks the role of gap junctions in the liver
- Author
-
Mateus T. Guerra, Paulo Renato A.V. Correa, David C. Spray, M. Fatima Leite, and Michael H. Nathanson
- Subjects
Lipopolysaccharides ,Male ,Receptors, Vasopressin ,Vasopressin ,medicine.medical_specialty ,Biophysics ,Biology ,Polymerase Chain Reaction ,Biochemistry ,Connexins ,Mice ,Internal medicine ,medicine ,Animals ,Lobules of liver ,Calcium Signaling ,education ,Receptor ,Molecular Biology ,DNA Primers ,Vasopressin receptor ,Mice, Knockout ,education.field_of_study ,Base Sequence ,urogenital system ,Wild type ,Gap junction ,Gap Junctions ,Cell Biology ,Endotoxins ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Endocrinology ,Hepatocyte ,Hepatocytes ,Connexin 32 - Abstract
Gap junctions are thought to be necessary for proper tissue function. However, no clear hepatic phenotype has been described in patients lacking connexin 32 (Cx32), the principal gap junction in liver. To determine the physiological role of Cx32 in liver, we compared the response of wild type and Cx32-deficient mice to endotoxin, since this stress increases serum levels of hormones that bind to receptors that are asymmetrically distributed across the hepatic lobule. In hepatocyte couplets isolated from wild type mice, most hepatocytes could transfer microinjected dye to their neighbor even after treatment with endotoxin. Dye transfer was not observed in Cx32-deficient couplets. Treatment of hepatocyte couplets from wild type mice with vasopressin induced calcium (Ca2+) waves that crossed the couplets in a concentration-dependent fashion, but the delay in transmission was markedly prolonged at all concentrations in Cx32-deficient couplets. Expression of the vasopressin receptor and the inositol 1,4,5-trisphosphate receptor was not decreased by endotoxin or in Cx32-deficient couplets. Finally, endotoxin caused transient hypoglycemia and cholestasis in wild type animals, but hypoglycemia was slightly prolonged and cholestasis was much worse in Cx32-deficient mice treated with endotoxin. The hepatic response to endotoxin is markedly impaired in the absence of Cx32. Thus, an important role of gap junctions in the liver is to assure integrated and uniform tissue response in times of stress.
- Published
- 2004
49. The effect of ionic products from bioactive glass dissolution on osteoblast proliferation and collagen production
- Author
-
Alfredo M. Goes, M. Fatima Leite, Patricia Valério, and Marivalda M. Pereira
- Subjects
Calcium Phosphates ,Materials science ,Cell Survival ,Biophysics ,Bioengineering ,Bioceramic ,Vacuole ,law.invention ,Nitric oxide ,Biomaterials ,chemistry.chemical_compound ,law ,medicine ,Animals ,Viability assay ,Rats, Wistar ,Dissolution ,Cells, Cultured ,Cell Size ,Ions ,Osteoblasts ,Osteoblast ,Silicon Dioxide ,Rats ,medicine.anatomical_structure ,Animals, Newborn ,Biochemistry ,chemistry ,Mechanics of Materials ,Bioactive glass ,Ceramics and Composites ,Alkaline phosphatase ,Collagen ,Glass ,Cell Division - Abstract
Bioactive ceramics developed during the past few decades have interesting properties from the biological standpoint, but their effects on cellular events remain partially unknown. In the current work, we investigated cellular viability, proliferation, morphology changes and metabolic activity of rat primary culture osteoblasts in contact with the ionic products from the dissolution of a bioactive glass with 60% of silica (BG60S) and a biphasic calcium phosphate (BCP). We observed that although osteoblasts cultured with BG60S showed vacuole formation, cell viability was increased when compared to BCP and control. The vacuole formation was not due to the presence of high calcium concentration in the ionic products from the dissolution of BG60S and was not related to nitric oxide production from the osteoblasts. We did find that high silicon concentration could induce cellular vacuole formation. Additionally, energy dispersive spectroscopy analysis indicated that vacuole contained 75% more silicon than other regions in the cell outside the vacuole. We further found that collagen production was higher in osteoblast cultured in the presence of BG60S compared to BCP and control, while alkaline phosphatase production was similar among cells incubated with BG60S, BCP and control. Together, our results indicate that osteoblast vacuole formation was due to high silicon contents in the dissolution of BG60S and we can suggest that despite the vacuole formation, there is no significant alteration in the bioceramic cell interaction.
- Published
- 2004
50. Evaluation of Osteoblasts Viability, Alkaline Phosphatase Production and Collagen Secretion in the Presence of TiHA
- Author
-
Alfredo M. Goes, M. Fatima Leite, Patricia Valério, Gultekin Goller, and Faik N. Oktar
- Subjects
medicine.anatomical_structure ,Materials science ,Mechanics of Materials ,Mechanical Engineering ,Collagen secretion ,medicine ,Alkaline phosphatase ,General Materials Science ,Osteoblast ,Cell biology - Published
- 2003
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.