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1.
Nutrients ; 15(19)2023 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-37836476

RESUMO

There is evidence that the association of protein intake and frailty may depend on the source of dietary protein. The mechanism underlying this association is not clear. In this study, we explore circulating metabolites as mediators of the relationship between dietary protein and of frailty in participants of the Baltimore Longitudinal Study of Aging (BLSA). Cross-sectional analyses in 735 BLSA participants of associations between plant and animal protein intake and frailty. Usual protein intake from plant and animal sources were estimated with a Food Frequency Questionnaire (FFQ) and frailty was assessed with a 44-item Frailty Index (FI). Compared with the lowest quartile, higher quartiles of plant, but not animal, protein were associated with lower FI. Twenty-five plasma metabolites were associated with plant protein intake; of these, fifteen, including phosphatidylcholines, cholesterol esters, sphingomyelins, and indole metabolites, mediated the association between plant protein intake and FI. The protective association between plant protein consumption and FI is mediated by lower abundance of lipid metabolites and higher abundance of tryptophan-related metabolites.


Assuntos
Fragilidade , Humanos , Idoso , Estudos Longitudinais , Proteínas de Plantas , Estudos Transversais , Proteínas Alimentares , Idoso Fragilizado
2.
J Cancer Res Clin Oncol ; 148(10): 2881-2891, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35678885

RESUMO

Treatment options for brain metastatic breast cancer are limited because the molecular mechanism for how breast cancer cells infiltrate the brain is not fully understood. For breast tumors to metastasize to the brain first, cells need to detach from the primary tumor, enter in the blood circulation, survive within the microvascular niche, and then cross the blood-brain barrier (BBB) to colonize into the brain. It is critical to understand how breast cancer cells transmigrate through the BBB to prevent brain metastasis. Nuclear respiratory factor 1 (NRF1) transcription factor has been reported to be highly active in several human cancers and its aberrant expression facilitates in the acquisition of breast cancer stem cells (BCSCs). Inhibitor of differentiation protein 3 (ID3), a transcription regulating protein, induces pluripotent endothelial stem cells (ESCs). Herein, we investigated if NRF1-induced BCSCs could cross a BBB model and guiding of BCSCs by ID3-induced ESCs across the BBB. BCSCs and ESCs were subjected to functional gain/loss experiments to determine if NRF1/ID3 contributed to lineage-specific BCSCs organ entry. First, we tested whether NRF1 promoted migration of breast cancer using a BBB model consisting of BCSCs or MDA-MB231 cells, brain endothelial cell layer, and astrocytes. NRF1 overexpression increased the propensity for BCSCs and NRF1-induced MDA-MB231 cells to adhere to brain endothelial cells and migrate across a human BBB model. Increased adhesion of NRF1-induced BCSCs to ESCsID3 was detected. NRF1-induced BCSCs crossed through the BBB model and this was promoted by ESCsID3. We also showed that environmental relevant exposure to PCBs (PCB153 and PCB77) produced differential effects. Treatment with PCB153 showed increased growth of NRF1-induced BCSCs tumor spheroids and increased in vivo migration of ESCsID3. Exosomal ID3 released from endothelial cells also supported the growth of NRF1-induced BCSCs and provide the basis for paracrine effects by ESCsID3 associated with breast tumors. Xenograft experiments showed that ID3 overexpressing brain ESCs not only supported the growth of BCSC tumor spheroids but guided them to the neural crest in zebrafish. These findings show for the first time a novel role for ID3 and NRF1 by which ESCsID3 help guide BCSCsNRF1 to distant metastatic sites where they most likely facilitate the colonization, survival, and proliferation of BCSCs. This knowledge is important for pre-clinical testing of NRF1/ID3 modifying agents to prevent the spread of breast cancer to the brain.


Assuntos
Encéfalo , Neoplasias da Mama , Proteínas Inibidoras de Diferenciação , Proteínas de Neoplasias , Células-Tronco Neoplásicas , Fator 1 Nuclear Respiratório , Animais , Encéfalo/patologia , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Células Endoteliais/patologia , Feminino , Humanos , Proteínas Inibidoras de Diferenciação/genética , Proteínas de Neoplasias/genética , Células-Tronco Neoplásicas/metabolismo , Fator 1 Nuclear Respiratório/genética , Comunicação Parácrina , Peixe-Zebra/metabolismo
3.
J Cancer Res Clin Oncol ; 148(7): 1641-1682, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35441887

RESUMO

PURPOSE: The mechanisms contributing to recurrence of glioblastoma (GBM), an aggressive neuroepithelial brain tumor, remain unknown. We have recently shown that nuclear respiratory factor 1 (NRF1) is an oncogenic transcription factor and its transcriptional activity is associated with the progression and prognosis of GBM. Herein, we extend our efforts to (1) identify influential NRF1-driven gene and microRNA (miRNA) expression for the aggressiveness of mesenchymal GBM; and (2) understand the molecular basis for its poor response to therapy. METHODS: Clinical data and RNA-Seq from four independent GBM cohorts were analyzed by Bayesian Network Inference with Java Objects (BANJO) and Markov chain Monte Carlo (MCMC)-based gene order to identify molecular drivers of mesenchymal GBM as well as prognostic indicators of poor response to radiation and chemotherapy. RESULTS: We are the first to report sex-specific NRF1 motif enriched gene signatures showing increased susceptibility to GBM. Risk estimates for GBM were increased by greater than 100-fold with the joint effect of NRF1-driven gene signatures-CDK4, DUSP6, MSH2, NRF1, and PARK7 in female GBM patients and CDK4, CASP2, H6PD, and NRF1 in male GBM patients. NRF1-driven causal Bayesian network genes were predictive of poor survival and resistance to chemoradiation in IDH1 wild-type mesenchymal GBM patients. NRF1-regulatable miRNAs were also associated with poor response to chemoradiation therapy in female IDH1 wild-type mesenchymal GBM. Stable overexpression of NRF1 reprogramed human astrocytes into neural stem cell-like cells expressing SOX2 and nestin. These cells differentiated into neurons and form tumorospheroids. CONCLUSIONS: In summary, our novel discovery shows that NRF1-driven causal genes and miRNAs involved in cancer cell stemness and mesenchymal features contribute to cancer aggressiveness and recurrence of aggressive therapy-resistant glioblastoma.


Assuntos
Neoplasias Encefálicas , Glioblastoma , MicroRNAs , Fator 1 Nuclear Respiratório , Teorema de Bayes , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/terapia , Feminino , Glioblastoma/genética , Glioblastoma/metabolismo , Glioblastoma/patologia , Glioblastoma/terapia , Humanos , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Recidiva Local de Neoplasia/genética , Recidiva Local de Neoplasia/patologia , Células-Tronco Neoplásicas/patologia , Fator 1 Nuclear Respiratório/genética , Prognóstico , Transcriptoma
4.
Indian J Public Health ; 64(3): 277-284, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32985430

RESUMO

BACKGROUND: Improving quality of health services and providing safe care require well-trained and skilled workforce. The inclusion of components of patient safety in graduate training curricula, followed by adherence to curricula in teaching programs, can improve the quality of health-care services. OBJECTIVES: To review the existing training curricula for five subgroups of health workforce (Allopathic doctors, nurses, laboratory technicians, pharmacists, and nurse midwives) and to document the components and identified variables of patient safety covered. METHODS: A mixed-methods study was conducted during July 2017-March 2018. Data were collected through desk review, field visits, in-depth interviews, self-administered questionnaires, and focused group discussions (FGDs). A total of 24 variables were identified by the experts to review the training curricula. RESULTS: Seven states, 28 institutes, and 42 health-care facilities were visited. A total of 516 staff from different health cadres participated in the study through 54 interviews, 156 self-administered questionnaires, and 24 FGDs. Of 24 patient safety variables considered, 16 were covered in the medical and nursing, 9 in laboratory technician and pharmacist, and 5 in midwives' curricula. The teaching material on the patient safety, for most categories of staff, was not available in consolidated form, and there was no standardization. CONCLUSION: There is a need for the development of comprehensive training material cum operational modules on patient safety, suitably adopted as per the learning needs of different subgroups of health staff. The need for strengthening patient safety has been further underscored as the health workforce is fighting the coronavirus disease 19 (COVID-19) pandemic. The initiatives on patient safety will contribute to improved overall quality of health services, which in turn would advance universal health coverage.


Assuntos
Educação de Pós-Graduação/métodos , Pessoal de Saúde/educação , Segurança do Paciente , Betacoronavirus , COVID-19 , Infecções por Coronavirus/epidemiologia , Currículo , Coleta de Dados/métodos , Feminino , Humanos , Índia , Masculino , Pandemias , Pneumonia Viral/epidemiologia , Melhoria de Qualidade , SARS-CoV-2
5.
Indian J Ophthalmol ; 68(6): 974-980, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32461408

RESUMO

Oculoplastic surgeries encompass both emergency surgeries for traumatic conditions and infectious disorders as well as elective aesthetic procedures. The COVID-19 pandemic has brought about a drastic change in this practice. Given the highly infectious nature of the disease as well as the global scarcity of medical resources; it is only prudent to treat only emergent conditions during the pandemic as we incorporate evidence-based screening and protective measures into our practices. This manuscript is a compilation of evidence-based guidelines for surgical procedures that oculoplastic surgeons can employ during the COVID-19 pandemic. These guidelines also serve as the basic framework upon which further recommendations may be based on in the future, as elective surgeries start being performed on a regular basis.


Assuntos
Betacoronavirus , Blefaroplastia/métodos , Consenso , Infecções por Coronavirus/epidemiologia , Doenças do Aparelho Lacrimal/cirurgia , Oftalmologia/organização & administração , Pandemias , Pneumonia Viral/epidemiologia , Padrões de Prática Médica/normas , COVID-19 , Humanos , Índia , Medição de Risco , SARS-CoV-2 , Sociedades Médicas , Cirurgia Plástica/organização & administração
6.
Am J Physiol Gastrointest Liver Physiol ; 318(4): G705-G716, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32090604

RESUMO

Anemia is a frequent diagnosis in critically ill infants, but the clinical implications of severe anemia in these patients remain unclear. In this study, we examined preweaned mice to investigate the effects of severe anemia during early infancy on gut mucosal permeability. C57BL/6 mice were subjected to timed phlebotomy between postnatal days (P) 2-10 to induce severe anemia (hematocrits 20%-24%), and intestinal permeability was tracked longitudinally between P10 and P20 as intestine-to-plasma translocation of enteral macromolecules and bacterial translocation. Epithelial junctions were evaluated by electron microscopy, polymerase chain reactions, immunohistochemistry, and/or enzyme immunoassays on intestinal tissues, Caco-2 intestinal epithelial-like cells, and colonic organoids. Preweaned mouse pups showed an age-related susceptibility to severe anemia, with increased intestinal permeability to enteral macromolecules (dextran, ovalbumin, ß-lactoglobulin) and luminal bacteria. Electron micrographs showed increased paracellular permeability and ultrastructural abnormalities of the adherens junctions. These findings were explained by the loss of E-cadherin in epithelial cells, which was caused by destabilization of the E-cadherin (Cdh1) mRNA because of microRNA let-7e-5p binding to the 3'-untranslated region. Severe anemia resulted in a disproportionate and persistent increase in intestinal permeability in preweaned mice because of the disruption of epithelial adherens junctions. These changes are mediated via microRNA let-7e-mediated depletion of Cdh1 mRNA.NEW & NOTEWORTHY This research article shows that newborn infants with severe anemia show an age-related susceptibility to developing increased intestinal permeability to ingested macromolecules. This abnormal permeability develops because of abnormalities in intestinal epithelial junctions caused by a deficiency of the molecule E-cadherin in epithelial cells. The deficiency of E-cadherin is caused by destabilization of its mRNA precursor because of increased expression and binding of another molecule, the microRNA let-7e-5p, to the E-cadherin mRNA.


Assuntos
Junções Aderentes/patologia , Anemia Neonatal/patologia , Mucosa Intestinal/patologia , Intestinos/patologia , Junções Aderentes/ultraestrutura , Animais , Animais Recém-Nascidos , Células CACO-2 , Caderinas/genética , Caderinas/metabolismo , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Camundongos , MicroRNAs/antagonistas & inibidores , MicroRNAs/genética , MicroRNAs/metabolismo , Permeabilidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
8.
Cells ; 7(12)2018 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-30486409

RESUMO

We have previously shown nuclear respiratory factor 1 (NRF1)-mediated transcriptional programming of mitobiogenesis contributes to estrogen-induced breast cancer through modulating cell cycle progression. In this study, we report a new role of NRF1 that goes beyond that of programming mitobiogenesis. Specifically, we report a novel oncogenic function of NRF1 supporting its causative role in breast cancer development and progression. The gain of NRF1 and/or treatment with 17ß-estradiol (E2) produced heterogeneous breast cancer stem cell (BCSC)-like subsets composed of more than 10 distinct cell sub-populations. Flow sorting combined with confocal imaging of markers for pluripotency, epithelial mesenchymal transition (EMT), and BCSCs phenotypically confirmed that the BCSC-like subset arise from cell re-programming. Thus, we determined the molecular actions of NRF1 on its target gene CXCR4 because of its known role in the acquisition of the BCSC-like subset through EMT. CXCR4 was activated by NRF1 in a redox-dependent manner during malignant transformation. An NRF1-induced BCSC-like subset was able to form xenograft tumors in vivo, while inhibiting transcription of CXCR4 prevented xenograft tumor growth. Consistent with our observation of NRF1-driven breast tumorigenesis in the experimental model, higher protein levels of NRF1 were also found in human breast cancer tissue specimens. This highly novel role of NRF1 in the stochastic acquisition of BCSC-like subsets and their progression to a malignant phenotype may open an entirely new research direction targeting NRF1 signaling in invasive breast cancer. Our discovery of targeting transcriptional activation of CXCR4 to inhibit NRF1-induced oncogenic transformation provides a mechanistic explanation for estrogen-dependent breast carcinogenesis and opens new avenues in strategic therapeutics to fight breast cancer.

9.
Indian J Ophthalmol ; 66(10): 1508-1511, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30249857

RESUMO

Hypereosinophilic syndrome (HES) is a spectrum of myeloproliferative disorder, which is characterized by persistent and marked blood eosinophilia and damage to multiple organs due to eosinophilic infiltration. Idiopathic HES is identified after ruling out all other causes of eosinophilia. Poor prognosis is usually associated with cardiac involvement and malignant transformation of blood cells. We report a rare case of HES in an 8-year-old boy who presented with unilateral proptosis and torticollis. The patient responded well to corticosteroid therapy with reduction of proptosis and torticollis and normalization of serum eosinophil count.


Assuntos
Exoftalmia/etiologia , Síndrome Hipereosinofílica/complicações , Órbita/diagnóstico por imagem , Torcicolo/etiologia , Criança , Diagnóstico Diferencial , Exoftalmia/diagnóstico , Humanos , Síndrome Hipereosinofílica/diagnóstico , Imageamento por Ressonância Magnética , Masculino , Doenças Raras , Torcicolo/diagnóstico
10.
Bioorg Med Chem ; 25(5): 1608-1621, 2017 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-28159485

RESUMO

The total synthesis of a number of representative natural products isolated from Leucetta and Clathrina sponges containing a polysubstituted 2-aminoimidazole are described. These syntheses take advantage of the site specific metallation reactions of 4,5-diiodoimidazoles resulting in the syntheses of three different classes of Leucetta derived natural products. The cytotoxicities of these natural products, along with several precursors in MCF7 cells were determined through an MTT growth assay. For comparative purposes a series of naphthimidazole-containing family members are included.


Assuntos
Alcaloides/síntese química , Alcaloides/farmacologia , Alcaloides/química , Animais , Produtos Biológicos/síntese química , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Feminino , Humanos , Células MCF-7 , Poríferos , Análise Espectral/métodos , Relação Estrutura-Atividade
11.
Toxicology ; 378: 10-16, 2017 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-28069514

RESUMO

More than 26 million people in West Bengal, India, are exposed to arsenic through drinking water, leading to several deleterious endpoints including precancerous and cancerous skin lesions and other non-dermatological health effects. Here, our aim was to identify whether miR21 is associated with such dermatological and non-dermatological health outcomes in chronically exposed humans. A total of 123 subjects from West Bengal were recruited for this study (45 exposed individuals with skin lesions, 38 exposed individuals without skin lesions and 40 unexposed individuals). The miR21 expression patterns in the lymphocytes were studied by quantitative realtime PCR and the effects on downstream targets were validated by Western blotting. Associations between the miR21 expression patterns and non-dermatological health effects were determined from epidemiological survey data. In vitro studies were done with low dose (0.05ppm) of chronic arsenic exposure to HaCaT cells for 15 passages. Interestingly, within the exposed group, the skin lesion individuals showed almost 4.5 fold up-regulation of miR21 compared to the no skin lesion group. The expression of the downstream targets of miR21 (PTEN and PDCD4) varied inversely, while the expression of pAKT and PI3K varied proportionately with its expression levels. Results of in vitro studies showed similar trends. Again miR21 was 2.03 fold up-regulated in the exposed individuals with respiratory diseases compared to the individuals without the same. This study for the first time shows that miR21 plays an important role in contributing to arsenic induced dermatological and non-dermatological health outcomes in an exposed population.


Assuntos
Arsênio/toxicidade , MicroRNAs , Transtornos Respiratórios , Dermatopatias , Poluentes Químicos da Água/toxicidade , Adulto , Arsênio/análise , Arsênio/urina , Ciclo Celular/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Água Potável/análise , Exposição Ambiental , Feminino , Predisposição Genética para Doença , Humanos , Índia/epidemiologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Masculino , MicroRNAs/genética , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Transtornos Respiratórios/induzido quimicamente , Transtornos Respiratórios/epidemiologia , Transtornos Respiratórios/genética , Transtornos Respiratórios/urina , Dermatopatias/induzido quimicamente , Dermatopatias/epidemiologia , Dermatopatias/genética , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/urina
12.
Indian J Sex Transm Dis AIDS ; 38(2): 121-127, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-30148264

RESUMO

CONTEXT: In India, the HIV positivity among injecting drug users (IDUs) stands at a staggering 7.71%. Among the states, HIV positivity among IDUs is highest in Punjab and Delhi, 21.2% and 18.3%, respectively. Interestingly, these two states are near to the "Golden Crescent." AIMS: The aim of this study was to examine the similarities and differences between the IDUs in Punjab and Delhi, in the context of vulnerability to HIV. SETTINGS AND DESIGN: This study uses data from the HIV Sentinel Surveillance-2010-2011 (HSS). The HSS is a cross-sectional data collection process for HIV surveillance in India. HSS, apart from collecting the blood samples from the respondents, also collects basic sociodemographic as well as some information on the drug use patterns of the IDUs. DATA AND METHODS: The raw data from HSS 2010-2011 were used for this study. Bivariate and multivariate analyses performed to obtain the results. RESULTS: Descriptive analyses revealed that the IDUs of Punjab and Delhi are very different from each other. In Delhi, 62.4% of IDUs inject drugs for more than 5 years; whereas in Punjab, it was only 32.4%. Majority of the Delhi IDUs (86.5%) inject more than three times a week whereas the corresponding percentage in Punjab was only 29.5%. The profiles of the HIV positives also differ between these two states. CONCLUSIONS: It would be prudent to adopt state-specific strategies to prevent the spread of HIV among the IDUs.

13.
Microvasc Res ; 98: 126-38, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25665868

RESUMO

While significant progress has been made to advance our knowledge of microvascular lesion formation, yet the investigation of how stem-like cells may contribute to the pathogenesis of microvascular diseases is still in its infancy. We assessed whether the inhibitor of DNA binding and differentiation 3 (ID3) contributes to the acquisition of a molecular stem cell-like signature in microvascular endothelial cells. The effects of stable ID3 overexpression and SU5416 treatment - a chemical inducer of microvascular lesions, had on the stemness signature were determined by flow cytometry, immunoblot, and immunohistochemistry. Continuous ID3 expression produced a molecular stemness signature consisting of CD133(+) VEGFR3(+) CD34(+) cells. Cells exposed to SU5416 showed positive protein expression of ID3, VEGFR3, CD34 and increased expression of pluripotent transcription factors Oct-4 and Sox-2. ID3 overexpressing cells supported the formation of a 3-D microvascular lesion co-cultured with smooth muscle cells. In addition, in vivo microvascular lesions from SuHx rodent model showed an increased expression of ID3, VEGFR3, and Pyk2 similar to SU5416 treated human endothelial cells. Further investigations into how normal and stem-like cells utilize ID3 may open up new avenues for a better understanding of the molecular mechanisms which are underlying the pathological development of microvascular diseases.


Assuntos
Células Endoteliais/citologia , Proteínas Inibidoras de Diferenciação/metabolismo , Microcirculação , Proteínas de Neoplasias/metabolismo , Células-Tronco/citologia , Antígeno AC133 , Animais , Antígenos CD/metabolismo , Antígenos CD34/metabolismo , Ciclo Celular , Diferenciação Celular , Separação Celular , Citometria de Fluxo , Glicoproteínas/metabolismo , Humanos , Indóis/química , Proteínas Inibidoras de Diferenciação/genética , Proteínas de Neoplasias/genética , Peptídeos/metabolismo , Fenótipo , Pirróis/química , Ratos , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/metabolismo
14.
J Mol Neurosci ; 55(3): 618-31, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25129100

RESUMO

Severe symptoms of cerebral and cardiorenal vascular diseases can be triggered when cerebral, coronary, or glomerular arterioles grow inappropriately as a result of abnormal cell proliferation. The risk factor(s) and molecular mechanisms responsible for microvascular lesion formation are largely unknown. Although controversial, both animal and epidemiological studies have shown that estrogen increases the risk of stroke which may be due to microvascular lesions. Since microvascular diseases are characterized by excessive vessel growth, it is plausible that estrogen-induced neovascularization contributes to the growth of microvascular lesions. We present evidence for how ID3 overexpression in endothelial cells contributes to the development of an estrogen-induced neovascular phenotype with an additional focus on Pyk2 kinase. Our data showed that ID3 overexpression increased neovascularization, cell migration, and spheroid growth of human cerebral microvascular endothelial cells, hCMEC/D3. ID3-overexpressing cells showed significant estrogen-induced G2/M phase transition. Estrogen treatment increased both ID3 phosphorylation; total protein that was inhibited by tamoxifen, and Pyk2-mediated estrogen-induced ID3 mRNA expression. These findings suggest that Pyk2 signals ID3 expression and ID3 is necessary for estrogen-induced neovascularization in hCMEC/D3 cells. A better understanding of how microvascular lesions depend on ID3 may open new avenues for prevention and treatment of neurological diseases.


Assuntos
Células Endoteliais/metabolismo , Estrogênios/farmacologia , Proteínas Inibidoras de Diferenciação/metabolismo , Proteínas de Neoplasias/metabolismo , Doenças Vasculares/metabolismo , Linhagem Celular , Movimento Celular , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/fisiologia , Endotélio Vascular/efeitos dos fármacos , Endotélio Vascular/metabolismo , Quinase 2 de Adesão Focal/metabolismo , Pontos de Checagem da Fase G2 do Ciclo Celular , Humanos , Proteínas Inibidoras de Diferenciação/genética , Proteínas de Neoplasias/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
15.
Mol Carcinog ; 54(9): 800-9, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24665044

RESUMO

Arsenic (As) induces pre-malignant and malignant dermatological lesions, non-dermatological health effects and cancers in humans. Senescence involves telomere length changes and acquisition of senescence-associated secretory phenotype (SASP), which promotes carcinogenesis. Though in vitro studies have shown that As induces senescence, population based studies are lacking. We investigated the arsenic-induced senescence, telomere length alteration and its contribution towards development of As-induced skin cancer. The study participants included 60 each of As-exposed individuals with skin lesion (WSL), without skin lesions (WOSL) and 60 unexposed controls. Exposure assessment of drinking water and urine was done. SA ß-gal activity, ELISA, and quantification of senescence proteins, alternative lengthening of telomere (ALT) associated proteins and telomerase activity were performed. Relative telomere length (RTL) was determined by qPCR. A significantly higher number of senescent cells, over-expression of p53 and p21 were observed in the As-exposed individuals when compared to unexposed. SASP markers, MMP-1/MMP-3 were significantly higher in the WSL but not IL-6/IL-8. A significant increase of RTL was observed in the WSL group, which was telomerase-independent but exhibited an over-expression of ALT associated proteins TRF-1 and TRF-2 with higher increase in TRF-2. An increased risk for developing As-induced skin lesions was found for individuals having RTL greater than 0.827 (odds ratio, 13.75; 95% CI: 5.66-33.41; P < 0.0001). Arsenic induces senescence in vivo, but the SASP markers are not strictly over-expressed in the As-induced skin lesion group, whereas telomerase-independent elongation of telomere length might be useful for predicting the risk of development of As-induced skin lesions.


Assuntos
Envelhecimento/efeitos dos fármacos , Arsênio/toxicidade , Água Potável/efeitos adversos , Telômero/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Adulto , Carcinogênese/induzido quimicamente , Carcinogênese/metabolismo , Carcinogênese/patologia , Estudos de Casos e Controles , Água Potável/análise , Feminino , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Transdução de Sinais/efeitos dos fármacos , Pele/efeitos dos fármacos , Pele/metabolismo , Pele/patologia , Dermatopatias/induzido quimicamente , Dermatopatias/epidemiologia , Dermatopatias/metabolismo , Dermatopatias/patologia , Telomerase/metabolismo , Telômero/patologia , Proteína Supressora de Tumor p53/metabolismo
16.
PLoS One ; 9(8): e104159, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25090023

RESUMO

Microvascular lesions resulting from endothelial cell dysfunction are produced in the brain, lung, kidney, and retina of patients of complex chronic diseases. The environmental and molecular risk factors which may contribute in the development of microvascular damage are unclear. The mechanism(s) responsible for initiating microvascular damage remain poorly defined, although several inciting factors have been proposed, including environmental toxicants-induced oxidative stress. Enhanced neovascularization has been implicated in either the development or progression of proliferative vascular lesions. Here, we present evidence for how PCB-induced ROS may contribute to the development of a neovascular phenotype with the aim of elucidating the role of environmental toxicants in endothelial dysfunction with a specific focus on the inhibitor of differentiation protein ID3. We used a combination of phenotype and immunohistochemical analysis followed by validating with protein expression and post-translational modifications with Western Blot and MALDI-TOF/TOF analysis. We also looked for a correlation between ID3 expression in vascular tissue. Our results showed that PCB-induced ROS mediated a highly tube branched neovascular phenotype that also depended on ID3 and Pyk2; and PCB153 treatment increased the size of endothelial spheroids under conditions typically used for clonal selection of stem cell spheroids. High ID3 protein expression correlated with a greater degree of malignancy and oxidative DNA damage marker 8-OHdG in blood vessels from human subjects. PCB153 treatment increased both serine and tyrosine phosphorylation of endothelial ID3. Stable ID3 overexpression increased cell survival of human microvascular endothelial cell line hCMEC/D3. In summary, our data provide evidence that ID3 may play a critical role in regulating vascular endothelial cell survival and development of microvascular lesions induced by persistent environmental pollutants such as PCB153. Findings of this study are important because they provide a new paradigm by which PCBs may contribute to the growth of microvascular lesions.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Células Endoteliais/patologia , Proteínas Inibidoras de Diferenciação/biossíntese , Microvasos/patologia , Proteínas de Neoplasias/biossíntese , Bifenilos Policlorados/toxicidade , Poluentes Atmosféricos/toxicidade , Diferenciação Celular/genética , Dano ao DNA/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Quinase 2 de Adesão Focal/biossíntese , Quinase 2 de Adesão Focal/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana , Humanos , Proteínas Inibidoras de Diferenciação/genética , Microvasos/efeitos dos fármacos , Microvasos/metabolismo , Proteínas de Neoplasias/genética , Oxirredução/efeitos dos fármacos
17.
Metallomics ; 6(4): 864-73, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24473091

RESUMO

Arsenic in drinking water is of critical concern in West Bengal, India, as it results in several physiological symptoms including dermatological lesions and cancers. Impairment of the DNA repair mechanism has been associated with arsenic-induced genetic damage as well as with several cancers. ERCC2 (Excision Repair Cross-Complementing rodent repair, complementation group 2), mediates DNA-repair by interacting with Cdk-activating kinase (CAK) complex, which helps in DNA proof-reading during transcription. Arsenic metabolism alters epigenetic regulation; we tried to elucidate the regulation of ERCC2 in arsenic-exposed humans. Water, urine, nails, hair and blood samples from one hundred and fifty seven exposed and eighty eight unexposed individuals were collected. Dose dependent validation was done in vitro using HepG2 and HEK-293. Arsenic content in the biological samples was higher in the exposed individuals compared with the content in unexposed individuals (p < 0.001). Bisulfite-modified methylation specific PCR showed a significant (p < 0.0001) hypomethylation of the ERCC2 promoter in the arsenic-exposed individuals. Densitometric analysis of immunoblots showed a nearly two-fold increase in expression of ERCC2 in exposed individuals, but there was an enhanced genotoxic insult as measured by micronuclei frequency. Immuno-precipitation and western blotting revealed an increased (p < 0.001) association of Cdk7 with ERCC2 in highly arsenic exposed individuals. The decrease in CAK activity was determined by observing the intensity of Ser(392) phosphorylation in p53, in vitro, which decreased with an increase in arsenic dose. Thus we infer that arsenic biotransformation leads to promoter hypomethylation of ERCC2, which in turn inhibits the normal functioning of the CAK-complex, thus affecting DNA-repair; this effect was highest among the arsenic exposed individuals with dermatological lesions.


Assuntos
Arsênio/toxicidade , Quinases Ciclina-Dependentes/metabolismo , Exposição Ambiental/efeitos adversos , Poluentes Químicos da Água/toxicidade , Proteína Grupo D do Xeroderma Pigmentoso/metabolismo , Adulto , Arsênio/análise , Metilação de DNA/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Exposição Ambiental/análise , Feminino , Células HEK293 , Células Hep G2 , Humanos , Masculino , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Poluentes Químicos da Água/análise , Proteína Grupo D do Xeroderma Pigmentoso/genética , Adulto Jovem , Quinase Ativadora de Quinase Dependente de Ciclina
18.
Toxicol Sci ; 135(2): 300-8, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23872714

RESUMO

Over 26 million people in West Bengal, India, are exposed to very high levels of arsenic through drinking water, leading to several deleterious endpoints including cancers. To elucidate the role of promoter methylation in arsenic-induced dermatological and nondermatological health effects, methylation status of p16 and DAPK genes was determined. A case-control study was conducted involving 72 individuals with arsenic-induced skin lesions (cases) and 50 individuals without skin lesions (controls), having similar arsenic exposure through drinking water. Methylation status was determined by bisulfite conversion of genomic DNA and methylation-specific PCR. Expression of the genes was determined by real-time PCR and Western blot analysis. Associations between the promoter methylation status and nondermatological health effects were determined from epidemiological survey data. Significant hypermethylation was found in the promoters of both DAPK and p16 genes in the cases compared with the controls resulting in downregulation of both the genes in the cases. There was a 3.4-fold decrease in the expression of death-associated protein kinase and 2.2-fold decrease in gene expression of p16 in the cases compared to the controls, the lowest expression being in the cancer tissues. Promoter hypermethylation of the genes was also associated with higher risk of developing arsenic-induced skin lesions, peripheral neuropathy, ocular and respiratory diseases. This study for the first time makes an attempt to correlate epigenetic modifications of the tumor suppressor genes with dermatological and nondermatological health outcomes in a population chronically exposed to arsenic.


Assuntos
Arsênio/toxicidade , Proteínas Quinases Associadas com Morte Celular/genética , Epigênese Genética , Genes p16 , Dermatopatias/induzido quimicamente , Adulto , Estudos de Casos e Controles , Metilação de DNA , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Dermatopatias/genética
19.
PLoS One ; 8(5): e61015, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23675407

RESUMO

MicroRNAs have been implicated in many critical cellular processes including apoptosis. We have previously found that apoptosis in pancreatic cancer cells was induced by adamantyl retinoid-related (ARR) molecule 3-Cl-AHPC. Here we report that 3-Cl-AHPC-dependent apoptosis involves regulating a number of microRNAs including miR-150* and miR-630. 3-Cl-AHPC stimulated miR-150* expression and caused decreased expression of c-Myb and IGF-1R in the pancreatic cancer cells. 3-Cl-AHPC-mediated reduction of c-Myb resulted in diminished binding of c-Myb with IGF-1R and Bcl-2 promoters, thereby causing repression of their transcription and protein expression. Over-expression of miR-150* also resulted in diminished levels of c-Myb and Bcl-2 proteins. Furthermore, the addition of the miRNA inhibitor 2'-O-methylated miR-150 blocked 3-Cl-AHPC-mediated increase in miR-150* levels and abrogated loss of c-Myb protein. Knockdown of c-Myb in PANC-1 cells resulted in enhanced apoptosis both in the presence or absence of 3-Cl-AHPC confirming the anti-apoptotic property of c-Myb. Overexpression of miR-630 also induced apoptosis in the pancreatic cancer cells and inhibited target protein IGF-1R mRNA and protein expression. Together these results implicate key roles for miR-150* and miR-630 and their targeting of IGF-1R to promote apoptosis in pancreatic cancer cells.


Assuntos
Apoptose/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias Pancreáticas/genética , Receptor IGF Tipo 1/genética , Adamantano/análogos & derivados , Adamantano/farmacologia , Antígeno CD24/metabolismo , Linhagem Celular Tumoral , Cinamatos/farmacologia , Expressão Gênica , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Genes bcl-2 , Humanos , Receptores de Hialuronatos/metabolismo , MicroRNAs/metabolismo , Células-Tronco Neoplásicas/metabolismo , Neoplasias Pancreáticas/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Proteínas Proto-Oncogênicas c-myb/genética , Proteínas Proto-Oncogênicas c-myb/metabolismo , Receptor IGF Tipo 1/metabolismo , Esferoides Celulares , Células Tumorais Cultivadas
20.
Mutat Res ; 755(1): 1-5, 2013 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-23644288

RESUMO

Prolonged consumption of arsenic-laden water above the threshold limit of 10µg/L causes a plethora of dermatological and non-dermatological multi-organ health problems, including cancer and death. Among several mechanisms of arsenic-induced toxicity and carcinogenicity studied so far, role of arsenic in impairment of immune system is less understood. Epidemiological data, animal model as well as cell line based studies have indicated that arsenic targets immune system and is associated with characteristic immunosupression, which may further adversely affect respiratory function. However, to the best of our knowledge, there is no study with respect to arsenic susceptibility investigating the role of genetic variation having immunological function. Hence, we have recruited a total of 432 arsenic-exposed individuals, of which 219 individuals with characteristic arsenic-induced skin lesions (cases) and 213 individuals without arsenic-induced skin lesion(controls), from arsenic-exposed districts of West Bengal, India. To find any probable association between arsenicism and the exonic single nucleotide polymorphisms (SNPs) in NALP2 gene, an important component of inflammasome complex, we screened the entire coding region (exon) in all the study participants. Among 9 SNPs found in NALP2 gene, the A1052E polymorphism (at least with one minor allele), was significantly overrepresented in controls and hence implies decreased risk toward the development of skin lesions [OR=0.67, 95% CI: 0.46-0.97]. Since, development of non-dermatological health effects are also important factor to properly look into, we have attempted to correlate the genetic variation of NALP2 with the extent of cytogenetic damage as measured by chromosomal aberration assay and adverse health effects including peripheral neuropathy, eye problem and respiratory diseases in the study population. We observed individuals with the protective genotype had less chromosomal aberration (p<0.05), and were also less susceptible toward arsenic-related respiratory diseases [OR=0.47; 95%CI: 0.23-0.89]. These findings suggest that NALP2 A1052E SNP plays an important role toward development of arsenic-induced skin lesions, chromosomal damage and respiratory diseases.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Intoxicação por Arsênico/complicações , Arsênio/efeitos adversos , Oftalmopatias/genética , Doenças do Sistema Nervoso Periférico/genética , Polimorfismo de Nucleotídeo Único/genética , Síndrome do Desconforto Respiratório/genética , Dermatopatias/genética , Adulto , Proteínas Reguladoras de Apoptose , Intoxicação por Arsênico/genética , Estudos de Casos e Controles , Aberrações Cromossômicas , Oftalmopatias/induzido quimicamente , Oftalmopatias/epidemiologia , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Índia/epidemiologia , Masculino , Doenças do Sistema Nervoso Periférico/induzido quimicamente , Doenças do Sistema Nervoso Periférico/epidemiologia , Prognóstico , Síndrome do Desconforto Respiratório/induzido quimicamente , Síndrome do Desconforto Respiratório/epidemiologia , Dermatopatias/induzido quimicamente , Dermatopatias/epidemiologia
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