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1.
Bioconjug Chem ; 30(10): 2544-2554, 2019 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-31498987

RESUMO

The use of cationic polymer based gene delivery vectors has several limitations such as low transfection efficiency, high toxicity, and inactivation by serum. The present work provides an inorganic based nanocarrier for efficient gene delivery and a method for preparing the same through a facile coprecipitation technique. The vehicle showed high loading capacity of DNA and can release the loaded DNA in a controlled pH-responsive manner. The developed gene delivery vehicle offers remarkable protection against DNase I and also provides protection against thermal damage. This vehicle also demonstrated efficient cellular uptake performance. Transfection and expression of plasmid gene encoding GFP proteins is achieved successfully by this LDH based vehicle. More interestingly, the developed Li-Al LDH efficiently induces GFP-p53 mediated apoptosis in HeLa cells exclusively sparing the normal tissue cells like NIH-3T3. The study demonstrates the potential of the developed inorganic based nanocarrier as a promising nonviral gene vector for tumor treatment.

2.
In Vitro Cell Dev Biol Anim ; 54(10): 736-742, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30324243

RESUMO

Monensin is a metal ionophore used as anticancer agent in many types of cancer cells. In this study, therapeutic potential of monensin was evaluated in TFE3 translocated renal cell carcinoma (RCC) cell line UOK146. UOK146 cells were treated with different concentrations of monensin, and cell death was induced as shown by MTT assay. Autophagy was studied by LC3 western, FACS and LC3 puncta formation after monensin treatment. Mitochondrial potential was studied by staining with TMRM and FACS. Antimetastatic potential of monensin was checked by inhibition of wound closure and MMP7 expression at RNA level. Dead and floating cells after the 10 µM monensin treatment were observed under phase contrast microscope. FACS analysis following TMRM staining showed that mitochondrial membrane gets depolarized after monensin treatment. FACS analysis after acridine orange staining showed increased double positive (green and red) cells, and LC3 upregulation and increased LC3 punta displayed autophagy activation in UOK146 cell line after monensin treatment. These findings showed that monensin acts as antiproliferative agent, activating autophagy and downregulates PRCC-TFE3 fusion transcript in Xp11.2 translocated tumor cell line.


Assuntos
Autofagia/efeitos dos fármacos , Carcinoma de Células Renais/enzimologia , Carcinoma de Células Renais/patologia , Neoplasias Renais/enzimologia , Neoplasias Renais/patologia , Metaloproteinase 7 da Matriz/metabolismo , Monensin/farmacologia , Autofagia/genética , Carcinoma de Células Renais/genética , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Renais/genética , Metaloproteinase 7 da Matriz/genética , Potencial da Membrana Mitocondrial/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
3.
PLoS One ; 13(10): e0205660, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30335789

RESUMO

The IGFN1 (Immunoglobulin-Like And Fibronectin Type III Domain Containing 1) gene has a role in skeletal muscle function and is also involved in metastatic breast cancer, and the isoforms with three N-terminal globular domains are sufficient for its function in skeletal muscle. Two novel splicing isoforms of IGFN1 have been identified in renal cell carcinoma (RCC), one with 5'exon extension and an isoform with a novel exon. The role of G-quadruplex, a non-B DNA, was explored for the splicing alteration of IGFN1 in RCC. G-quadruplexes are the secondary structures acquired by stacking of G-quartets by Hoogsteen hydrogen bonding in DNA and RNA. IGFN1 has intronic potential G-quadruplex forming sequence (PQS) folding into G-quadruplex and is studied for its involvement in aberrant splicing. A PQS in the intron 15 of IGFN1 gene was observed in our in silico analysis by QGRS mapper and non BdB web servers. We observed PQS folds into stable G-quadruplex structure in gel shift assay and circular dichroism (CD) spectroscopy in the presence of G-quadruplex stabilizing agents Pyridostatin (PDS) and KCl, respectively. G-quadruplex formation site with single base resolution was mapped by Sanger sequencing of the plasmid constructs harbouring the cloned PQS and its mutant. This stable G-quadruplex inhibits reverse transcriptase and taq polymerase in reverse transcriptase & PCR stop assays. PDS changes the different splicing isoforms of IGFN1 in UOK146 cell line, displaying involvement of intronic G-quadruplex in IGFN1 splicing. These results lead us to propose that a stable G-quadruplex structure is formed in IGFN1 intron and a reason behind IGFN1 aberrant splicing which could be targeted for therapeutic intervention.


Assuntos
Carcinoma de Células Renais/genética , Proteínas de Transporte/genética , Neoplasias Renais/genética , Processamento de RNA/genética , Aminoquinolinas/farmacologia , Linhagem Celular Tumoral , DNA/genética , Quadruplex G/efeitos dos fármacos , Humanos , Íntrons/genética , Ácidos Picolínicos/farmacologia , Isoformas de Proteínas/genética , RNA/genética , Análise de Sequência de DNA
4.
In Vitro Cell Dev Biol Anim ; 54(4): 295-303, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29556894

RESUMO

Sodium butyrate (SB), a histone deacetylase inhibitor, is emerging as a potent anti-cancer drug for different types of cancers. In the present study, anti-cancer activity of SB in Xp11.2 (TFE3) translocated renal cell carcinoma cell line UOK146 was studied. Anti-proliferative effect of SB in renal cell carcinoma (RCC) cell line UOK146 was evaluated by MTT assay and morphological characteristics were observed by phase contrast microscopy which displayed the cell death after SB treatment. SB induces DNA fragmentation and change in nuclear morphology observed by increased sub-G1 region cell population and nuclear blebbings. Cell cycle arrest at G2/M phase was found after SB treatment. UOK146 cell line shows autophagy mode of cell death as displayed by acridine orange staining and flow cytometry analysis. LC3-II, a protein marker of autophagy, was also found to be upregulated after SB treatment. A tumor suppressor gene DIRAS1 was upregulated after SB treatment, displaying its anti-cancer potential at molecular level. These findings suggest that SB could serve as a novel regulator of tumor suppressors and lead to the discovery of novel therapeutics with better and enhanced anti-cancer activity.


Assuntos
Autofagia/efeitos dos fármacos , Ácido Butírico/farmacologia , Carcinoma de Células Renais/tratamento farmacológico , GTP Fosfo-Hidrolases/genética , Proteínas Supressoras de Tumor/genética , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/patologia , Linhagem Celular , Humanos , Ativação Transcricional/efeitos dos fármacos
5.
Biochim Biophys Acta Gen Subj ; 1862(3): 630-636, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29138008

RESUMO

Transcription Factor E3 (TFE3) translocation is found in a group of different type of cancers and most of the translocations are located in the 5' region of TFE3 which may be considered as Breakpoint Region (BR). In our In silico study by QGRS mapper and non BdB web servers we found a Potential G-quadruplex forming Sequence (PQS) in the intron 2 of TFE3 gene. In vitro G-quadruplex formation was shown by native PAGE in presence of Pyridostatin(PDS), which with inter molecular secondary structure caused reduced mobility to migrate slower. G-quadruplex formation was mapped at single base resolution by Sanger sequencing and Circular Dichroism showed the formation of parallel G-quadruplex. FRET analysis revealed increased and decreased formation of G-quadruplex in presence of PDS and antisense oligonucleotide respectively. PCR stop assay, transcriptional and translational inhibition by PQS showed stable G-quadruplex formation affecting the biological processes. TFE3 minigene splicing study showed the involvement of this G-quadruplex in TFE3 splicing too. Therefore, G-quadruplex is evident to be the reason behind TFE3 induced oncogenesis executed by translocation and also involved in the mRNA splicing.


Assuntos
Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Transformação Celular Neoplásica/genética , Cromossomos Humanos X/genética , DNA de Neoplasias/genética , Quadruplex G , Regulação Neoplásica da Expressão Gênica/genética , Processamento de RNA/genética , Translocação Genética/genética , Aminoquinolinas/farmacologia , Animais , Células COS , Chlorocebus aethiops , Cromossomos Humanos X/ultraestrutura , DNA Recombinante/genética , Quadruplex G/efeitos dos fármacos , Humanos , Íntrons/genética , Oligonucleotídeos Antissenso/farmacologia , Proteínas de Fusão Oncogênica/genética , Proteínas de Fusão Oncogênica/fisiologia , Ácidos Picolínicos/farmacologia , Cloreto de Potássio/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Análise de Sequência de DNA , Transfecção
6.
Gene ; 635: 69-76, 2017 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-28847717

RESUMO

Several studies on experimental animals indicate that the process of organogenesis crucially depends upon the spatiotemporal dose of certain critical bio-molecules. Tooth development is also not an exception. While most of the knowledge regarding the molecular mechanism of tooth development comes from the studies on mouse model, pathogenic variations identified in human tooth agenesis also provide valuable information on mammalian tooth development. Until now five major candidate genes have been identified for tooth agenesis in human. Among them, PAX9 plays the crucial role in tooth development and in non-syndromic congenital tooth agenesis. In this study, microsatellite and SNP based genotyping identifies a disease specific haplotype block, which includes PAX9 gene, segregates with autosomal dominant tooth agenesis phenotype. Direct sequencing of PAX9 identifies a novel heterozygous G to A transition at the third base (c.3G>A) of initiation codon leading to ATG to ATA shift in all affected individuals which is absent in all unaffected relatives and 200 control chromosomes. Further, in vitro functional analysis creating PAX9 minigene construct did apparently show no effect on the splice-site migration. It is therefore proposed that haploinsufficiency of PAX9 is the causal factor for tooth agenesis in this family.


Assuntos
Anodontia/genética , Predisposição Genética para Doença , Fator de Transcrição PAX9/genética , Dente/crescimento & desenvolvimento , Animais , Anodontia/fisiopatologia , Genótipo , Haplótipos , Humanos , Camundongos , Mutação , Organogênese/genética , Polimorfismo de Nucleotídeo Único , Dente/fisiopatologia
7.
J Genet ; 96(6): 885-893, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29321346

RESUMO

Polycystic kidney disease (PKD) is a systemic disorder which adds majority of renal patients to end stage renal disease. Autosomal dominant polycystic kidney disease (ADPKD) is more prevalent and leading cause of dialysis and kidney transplant. Linkage analysis revealed some closely linked loci, two of which are identified as PKD1, PKD2 and an unidentified locus to ADPKD. This study was performed using PCR and automated DNA sequencing in 84 cases and 80 controls to test potential candidature of PKD2 as underlying cause of PKD by in silico and statistical analyses. Two associated symptoms, hypertension (19%) and liver cyst (31%) havemajor contribution to PKD. Gender-based analysis revealed that familial female patients (27%) and familialmale patients (33%) are more hypertensive. Liver cyst, the second major contributing symptom presented by large percentage of sporadic males (46%). Genetic screening of all 15 exons of PKD2 revealed eight pathogenic (c.854_854delG, c.915C>A, c.973C>T, c.1050_1050delC, c.1604_1604delT, c.1790T>C, c.2182_2183delAG, c.2224C>T) and eight likely pathogenic (g.11732A>G, c.646T>C, c.1354A>G, g.39212G>C, c.1789C>A, c.1849C>A, c.2164G>T, c.2494A>G)DNA sequence variants. In our study, 27.38% (23/84) cases shown pathogenic / likely pathogenic variants in PKD2 gene. Some regions of PKD2 prone for genetic variation suggested to be linked with disease pathogenesis. This noticeable hot spot regions hold higher frequency (50%) of pathogenic / likely pathogenic genetic variants constituting single nucleotide variants than large deletion and insertion that actually represents only 41.08% of coding sequence of PKD2. Statistically significant association for IVS3-22AA genotype was observed with PKD, while association of IVS4+62C>T was found insignificant.


Assuntos
Predisposição Genética para Doença , Variação Genética/genética , Rim Policístico Autossômico Dominante/genética , Canais de Cátion TRPP/genética , Grupo com Ancestrais do Continente Asiático/genética , Análise Mutacional de DNA , Éxons/genética , Feminino , Estudos de Associação Genética , Testes Genéticos , Genótipo , Humanos , Índia/epidemiologia , Masculino , Mutação , Rim Policístico Autossômico Dominante/epidemiologia , Análise de Sequência de DNA
8.
Dalton Trans ; 45(16): 7163-77, 2016 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-27009608

RESUMO

Four organometallic complexes [(η(6)-C6H6)RuCl(pmpzdpm)], 1; [(η(6)-C6H6)RuCl(pypzdpm)], 2; [(η(6)-C10H14)RuCl(pmpzdpm)], 3 and [(η(6)-C10H14)RuCl(pypzdpm)], 4 containing 5-(2-pyrimidyl-piperazine)phenyldipyrromethene (pmpzdpm) and 5-(2-pyridylpiperazine)phenyldipyrromethene (pypzdpm) have been designed and synthesized. The complexes 1-4 have been fully characterized by elemental analyses and spectroscopic studies (ESI-MS, IR, (1)H, (13)C NMR, UV-vis). Their electrostatic/intercalative interaction with CT DNA has been investigated by UV-vis and competitive ethidium bromide displacement studies while their protein binding affinity toward bovine serum albumin (BSA) was realized by UV-vis, fluorescence, synchronous and three dimensional (3D) fluorescence studies. The interaction with DNA and protein has further been validated by in silico studies. Cellular uptake, in vitro cytotoxicity and flow cytometric analyses have been performed to determine the mode of cell death against the kidney cancer cell line ACHN. Cell cycle analysis suggested that the complexes cause cell cycle arrest in the subG1 phase and overall results indicated that the in vitro antitumor activity of 1-4 lies in the order of 3 >4 >1 >2 (IC50, 7.0 1; 8.0 2; 2.0 3; 4.0 µM,4 ).


Assuntos
Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , DNA/metabolismo , Compostos Organometálicos/metabolismo , Compostos Organometálicos/farmacologia , Rutênio/química , Soroalbumina Bovina/metabolismo , Animais , Antineoplásicos/química , Apoptose/efeitos dos fármacos , Bovinos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA/química , Humanos , Simulação de Acoplamento Molecular , Conformação de Ácido Nucleico , Compostos Organometálicos/química , Ligação Proteica , Conformação Proteica , Soroalbumina Bovina/química
9.
J Control Release ; 224: 186-198, 2016 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-26774219

RESUMO

Hydrophobic anticancer drug, raloxifene hydrochloride (RH) is intercalated into a series of magnesium aluminum layered double hydroxides (LDHs) with various charge density anions through ion exchange technique for controlled drug delivery. The particle nature of the LDH in presence of drug is determined through electron microscopy and surface morphology. The release of drug from the RH intercalated LDHs was made very fast or sustained by altering the exchangeable anions followed by the modified Freundlich and parabolic diffusion models. The drug release rate is explained from the interactions between the drug and LDHs along with order-disorder structure of drug intercalated LDHs. Nitrate bound LDH exhibits greater interaction with drug and sustained drug delivery against the loosely interacted phosphate bound LDH-drug, which shows fast release. Cell viability through MTT assay suggests drug intercalated LDHs as better drug delivery vehicle for cancer cell line against poor bioavailability of the pure drug. In vivo study with mice indicates the differential tumor healing which becomes fast for greater drug release system but the body weight index clearly hints at damaged organ in the case of fast release system. Histopathological experiment confirms the damaged liver of the mice treated either with pure drug or phosphate bound LDH-drug, fast release system, vis-à-vis normal liver cell morphology for sluggish drug release system with steady healing rate of tumor. These observations clearly demonstrate that nitrate bound LDH nanoparticle is a potential drug delivery vehicle for anticancer drugs without any side effect.


Assuntos
Hidróxido de Alumínio/química , Antineoplásicos/administração & dosagem , Portadores de Fármacos/química , Hidróxido de Magnésio/química , Animais , Ânions/química , Antineoplásicos/química , Preparações de Ação Retardada , Combinação de Medicamentos , Liberação Controlada de Fármacos , Feminino , Células HeLa , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Tamanho da Partícula , Cloridrato de Raloxifeno/administração & dosagem , Cloridrato de Raloxifeno/química , Ratos , Ratos Sprague-Dawley
10.
Neurosci Lett ; 605: 29-33, 2015 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-26282903

RESUMO

Loss-of-function mutation in PINK1 is known for causing autosomal recessive early onset Parkinsonism accounting approximately 6.5% of PD cases. Recently, PINK1 has also been shown to cause Parkinson's disease (PD) in eastern India. Present study is aimed to see its contribution in north-Indian PD patients. A total of 106 PD patients and 60 ethnically matched healthy controls were included in the study. All the patients were screened for mutation in PINK1 by direct DNA sequence analysis of the PCR amplicons covering all exons and exon-intron boundaries. Identified novel variant was reconfirmed by DNA sequencing of 10 randomly selected TA clones containing the variant amplicon. In vitro functional assay of the mutant protein was performed by transfecting COS-7 cell line with wild type and mutant (created by site-directed-mutagenesis) cDNA construct of PINK1 fused to N' terminal GFP followed by western blot analysis. Two potentially pathogenic, one being novel (p.Q267X) and 6 other apparently non-pathogenic variants were identified. Western blot analysis reveals production of truncated PINK1 fusion protein of ∼55kDa in p.Q267X mutant instead of 82/93kDa of wild type PINK1 fusion protein (molecular weight of GFP is ∼27kDa). Our study concludes that PINK1 variants are prevalent for causing Parkinson's disease (PD) in India, as revealed by the occurrence of 1.8% (2/106) in PD patients from north Indian population. The novel homozygous variant of PINK1 (c.799C>T) reported here is the plausible cause for disease manifestation in this patient. Future study, however, would be helpful to understand the functional mechanism how this premature PINK1 protein (p.Q267X) responds to cellular stress leading to the PD pathophysiology.


Assuntos
Doença de Parkinson/genética , Proteínas Quinases/genética , Adulto , Idoso , Sequência de Aminoácidos , Animais , Células COS , Estudos de Casos e Controles , Chlorocebus aethiops , Humanos , Índia , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mutação
11.
Asian Pac J Cancer Prev ; 16(18): 8419-23, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26745095

RESUMO

PURPOSE: Anti-cancer activity evaluation of aqueous extract of CRUEL (herbomineral formulation) capsules on renal cell carcinoma cell lines, and exploration of mechanisms of cell death. MATERIALS AND METHODS: To detect the cytotoxic dose concentration in renal cell carcinoma (RCC) cells, MTT assays were performed and morphological changes after treatment were observed by inverted microscopy. Drug effects against RCC cell lines were assessed with reference to cell cycle distribution (flow cytometry), anti-metastatic potential (wound healing assay) and autophagy(RT-PCR). RESULTS: CRUEL showed anti-proliferative effects against RCC tumor cell lines with an IC50 value of approximately 4mg/mL in vitro, while inducing cell cycle arrest at S-phase of the cell cycle and inhibiting wound healing. LC3 was found to be up-regulated after drug treatment by RT-PCR resulting in an autophagy mode of cell death. CONCLUSIONS: This study provides experimental validation for antitumor activity of CRUEL.


Assuntos
Antineoplásicos/farmacologia , Carcinoma de Células Renais/patologia , Química Farmacêutica , Neoplasias Renais/patologia , Minerais/farmacologia , Preparações de Plantas/farmacologia , Apoptose/efeitos dos fármacos , Western Blotting , Cápsulas/administração & dosagem , Carcinoma de Células Renais/tratamento farmacológico , Carcinoma de Células Renais/metabolismo , Ciclo Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Humanos , Neoplasias Renais/tratamento farmacológico , Neoplasias Renais/metabolismo , Fitoterapia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais Cultivadas
12.
PLoS One ; 9(9): e106811, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25203534

RESUMO

Congenital tooth agenesis in human is characterized by failure of tooth development during tooth organogenesis. 300 genes in mouse and 30 genes in human so far have been known to regulate tooth development. However, candidature of only 5 genes viz. PAX9, MSX1, AXIN2, WNT10A and EDA have been experimentally established for congenitally missing teeth like hypodontia and oligodontia. In this study an Indian family with multiple congenital tooth agenesis was identified. Pattern of inheritance was apparently autosomal dominant type with a rare possibility to be X-linked. Whole genome sequencing of two affected individuals was carried out which revealed 119 novel non-synonymous single nucleotide variations (SNVs) distributed among 117 genes. Out of these only one variation (c.956G>T) located at exon 9 of X-linked EDA gene was considered as pathogenic and validated among all the affected and unaffected family members and unrelated controls. This variation leads to p.Ser319Ile change in the TNF homology domain of EDA (transcript variant 1) protein. In silico analysis predicts that this Ser319 is well conserved across different vertebrate species and a part of putative receptor binding site. Structure based homology modeling predicts that this amino acid residue along with four other amino acid residues nearby, those when mutated known to cause selective tooth agenesis, form a cluster that may have functional significance. Taken together these results suggest that c.956G>T (p.Ser319Ile) mutation plausibly reduces the receptor binding activity of EDA leading to distinct tooth agenesis in this family.


Assuntos
Análise Mutacional de DNA , Ectodisplasinas/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Genômica , Mutação , Anormalidades Dentárias/genética , Adulto , Sequência de Aminoácidos , Animais , Sequência de Bases , Criança , Biologia Computacional , Ectodisplasinas/química , Feminino , Genoma Humano/genética , Humanos , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Linhagem , Conformação Proteica
13.
Asian Pac J Cancer Prev ; 15(5): 1937-41, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24716915

RESUMO

AIMS: To evaluate anti-cancer activity of Asparagus racemosus (AR) leaf extract on UOK146, a renal cell carcinoma cell line, and explore its mechanism of action. MATERIALS AND METHODS: Dried AR leaves were extracted with chloroform and dissolved in DMSO. This extract was applied to UOK146 and cell death was estimated by MTT assay. In addition PRCC-TFE3 fusion transcripts were detected by real time PCR. RESULTS: Extract was found to be cytotoxic with an IC50 of 0.9 mg/ml as estimated by dose response curve. Antitumor activity of the permissible doses of the extract was assessed by the down regulation of PRCC-TFE3 fusion transcript (38%) responsible for oncogenicity of the UOK146 cell line. No increment in the BAX, a proapoptotic marker level was observed. CONCLUSIONS: Evidence of antiproliferative effect, PRCC-TFE3 fusion transcript inhibition and static BAX level clearly indicate that AR extract provides or elicits an apoptosis independent anticancer effect on RCC cells by some specific mechanism of regulation.


Assuntos
Apoptose/efeitos dos fármacos , Asparagus (Planta)/química , Carcinoma de Células Renais/tratamento farmacológico , Morte Celular/efeitos dos fármacos , Neoplasias Renais/tratamento farmacológico , Proteínas de Fusão Oncogênica/genética , Extratos Vegetais/farmacologia , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Carcinoma de Células Renais/genética , Proteínas de Ciclo Celular/genética , Morte Celular/genética , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Humanos , Neoplasias Renais/genética , Proteínas de Neoplasias/genética , Extratos Vegetais/química , Folhas de Planta/química
14.
Int J Low Extrem Wounds ; 13(1): 58-63, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24659626

RESUMO

Diabetes mellitus (DM) is one of the severe metabolic disorders of carbohydrate metabolism worldwide. Developing countries are at higher risk of DM, and there is significant evidence that it is epidemic in many economically developing and newly industrialized countries. Among all other complications associated with DM, delayed wound healing is a major concern in diabetic patients. Wound healing is a natural healing process that starts immediately after injury. This involves interaction of a complex cascade of cellular events that generates resurfacing, reconstitution, and restoration of the tensile strength of injured skin. There are multiple factors responsible for delayed wound healing among which the contribution of DM has been well documented. The wound healing process is also delayed by the metabolic, vascular, neurological, and inflammatory alterations, which are well known in both type 1 and type 2 diabetes. Keratinocytes are crucial for wound re-epithelialization, and defects in directed migration of keratinocytes due to DM are associated with the delayed wound healing process. Many factors responsible for re-epithelialization have been identified, characterized, and well described; however, the genes responsible for the healing process have only partially been illustrated. This article will therefore focus on the efficacy of ANGPTL4 (angiopoietin-like 4) gene, which plays a novel role in keratinocyte migration during wound healing.


Assuntos
Angiopoietinas/genética , Diabetes Mellitus/genética , Regulação da Expressão Gênica , RNA/genética , Pele/lesões , Cicatrização/genética , Ferimentos e Lesões/genética , Proteína 4 Semelhante a Angiopoietina , Angiopoietinas/biossíntese , Diabetes Mellitus/metabolismo , Humanos , Queratinócitos/metabolismo , Queratinócitos/patologia , Ferimentos e Lesões/complicações , Ferimentos e Lesões/metabolismo
15.
Ren Fail ; 36(1): 50-4, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24059749

RESUMO

AIM: Diabetes plays a major role in progression of renal failure. The risk-factor profile changes during the progression of chronic kidney disease (CKD) from mild/moderate to end-stage renal disease. The relationship between glycemic indices, blood pressure, body mass index (BMI) and age at diagnosis in Indians has been less investigated. We assessed association of these risk factors with CKD stages in Indian population. METHODS: This study was carried out on patients (n = 162) who were diagnosed with CKD and normal control group (n = 155). For BMI, National Institutes for Health criteria were used to categorize the patients. RESULT: The mean age of CKD patients were significantly increased with the advancement of stage. BMI, systolic blood pressure (SBP), postprandial sugar level (PP), urea and creatinine were also significantly higher with elevated stages, whereas no differences were observed in diastolic blood pressure (DBP) and fasting blood sugar (FBS). The logistic regression study gave a significant result (p = 0.000) when we compared the group of CKD patients with established/prolonged postprandial blood sugar. It was independently associated with mild CKD [odds ratio (OR) = 5.213, 95% confidence interval (CI) = 2.06-13.21, p = 0.000], moderate CKD (OR = 7.724, 95% CI = 4.05-14.74, p = 0.000) and severe CKD (OR = 7.610, 95% CI = 4.03-14.36, p = 0.000). CONCLUSION: SBP and PP were the best predictors of prevalent nephropathy in this population, while DBP and FBS were found to be less effective. This may have implication for kidney disease risk stratification and protection.


Assuntos
Glicemia , Índice de Massa Corporal , Doenças Cardiovasculares/complicações , Falência Renal Crônica/etiologia , Período Pós-Prandial , Adulto , Idoso , Pressão Sanguínea , Estudos de Casos e Controles , Progressão da Doença , Humanos , Índia , Modelos Logísticos , Pessoa de Meia-Idade
16.
Spectrochim Acta A Mol Biomol Spectrosc ; 70(5): 1109-13, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18061533

RESUMO

Interaction of merbromin with trypsin is of bovine origin has been studied by monitoring the absorption steady-state and time-resolved fluorescence spectral properties of the dye. Studies have been done in media of varying pH at different trypsin concentrations. It has been observed that trypsin brings about a quenching of fluorescence of the dye. The quenching is static in nature and the equilibrium constant of dye-trypsin interaction in the ground-state has been determined from quenching studies. Steady-state anisotropy of the dye increases in presence of trypsin in the medium. Values of micro-viscosity in the vicinity of the fluorophore in media containing trypsin have been determined from measurements of fluorescence anisotropy. Time-resolved fluorescence studies indicate the existence of two decaying states for the dye. The fractional contribution to the time-resolved decay changes with pH. The average lifetime, however, does not depend on the concentration of trypsin.


Assuntos
Merbromina/química , Tripsina/química , Anisotropia , Estrutura Molecular , Espectrometria de Fluorescência , Tripsina/metabolismo
17.
Clin Cancer Res ; 13(24): 7314-21, 2007 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-18094412

RESUMO

PURPOSE: We examined a cohort of patients with alveolar soft part sarcoma (ASPS) treated at our institution and showed the characteristic ASPSCR1-TFE3 fusion transcript in their tumors. Investigation of potential angiogenesis-modulating molecular determinants provided mechanistic and potentially therapeutically relevant insight into the enhanced vascularity characteristic of this unusual tumor. EXPERIMENTAL DESIGN: Medical records of 71 patients with ASPS presenting at the University of Texas M.D. Anderson Cancer Center (1986-2005) were reviewed to isolate 33 patients with formalin-fixed paraffin-embedded material available for study. RNA extracted from available fresh-frozen and formalin-fixed paraffin-embedded human ASPS tumors were analyzed for ASPSCR1-TFE3 fusion transcript expression using reverse transcription-PCR and by angiogenesis oligomicroarrays with immunohistochemical confirmation. RESULTS: Similar to previous studies, actuarial 5- and 10-year survival rates were 74% and 51%, respectively, despite frequent metastasis. ASPSCR1-TFE3 fusion transcripts were identified in 16 of 18 ASPS samples. In the three frozen samples subjected to an angiogenesis oligoarray, 18 angiogenesis-related genes were up-regulated in tumor over adjacent normal tissue. Immunohistochemistry for jag-1, midkine, and angiogenin in 33 human ASPS samples confirmed these results. Comparison with other sarcomas indicates that the ASPS angiogenic signature is unique. CONCLUSION: ASPS is a highly vascular and metastatic tumor with a surprisingly favorable outcome; therapeutically resistant metastases drive mortality. Future molecular therapies targeting overexpressed angiogenesis-promoting proteins (such as those identified here) could benefit patients with ASPS.


Assuntos
Neovascularização Patológica/genética , Sarcoma Alveolar de Partes Moles/genética , Adulto , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Feminino , Humanos , Imuno-Histoquímica , Peptídeos e Proteínas de Sinalização Intracelular , Estimativa de Kaplan-Meier , Masculino , Proteínas de Neoplasias/genética , Proteínas de Fusão Oncogênica/genética , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sarcoma Alveolar de Partes Moles/mortalidade , Sarcoma Alveolar de Partes Moles/patologia , Análise de Sobrevida , Análise Serial de Tecidos
18.
Cancer ; 109(11): 2323-33, 2007 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-17429838

RESUMO

BACKGROUND: p53 is the most commonly mutated gene in cancer, including soft tissue sarcoma (STS). The authors characterized p53 alterations (protein accumulation and gene mutation) in STS to evaluate possible associations with patient outcomes. METHODS: Thirty-one STS specimens (multiple histologies) were analyzed by p53 immunohistochemistry (IHC) and direct DNA sequencing of p53 exons 2-11 and then correlated with outcomes. RESULTS: Direct p53 sequencing detected mutations in 10 of 31 STSs; 7 of 10 were missense mutations, whereas 3 of 10 were either insertions or frameshift mutations, leading to nonfunctional truncated p53; 7 of these p53 mutations have not been previously described. Four p53 exon 4 mutations were identified, a p53 region previously unknown to be mutation prone. Eighteen of the 31 specimens expressed p53 when the authors used the clinical IHC assay of their institution. Interassay concordance of 48% was observed; only 6 of 10 sequencing-identified p53 mutated specimens exhibited nuclear p53 protein expression by IHC, whereas 12 of 18 specimens exhibiting p53 protein expression by IHC harbored sequencing-identified wild-type p53. Decreased survival was observed in STS patients bearing sequencing-identified mutated p53 versus wild-type p53, as was a correlation between IHC-determined nuclear p53 protein expression and decreased survival. CONCLUSIONS: p53 protein stabilization and p53 mutation frequently occur in STS, and both suggest worse outcomes for patients so affected. However, increased p53 protein expression does not necessarily indicate p53 gene mutation. The high incidence of exon 4 mutations found in STS suggests that p53 sequencing should not be limited to the core DNA binding domain.


Assuntos
Mutação/genética , Sarcoma/genética , Proteína Supressora de Tumor p53/genética , Adolescente , Adulto , Idoso , DNA de Neoplasias/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Técnicas Imunoenzimáticas , Incidência , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Prognóstico , Sarcoma/diagnóstico , Sarcoma/metabolismo , Proteína Supressora de Tumor p53/metabolismo
20.
Artigo em Inglês | MEDLINE | ID: mdl-16956787

RESUMO

Electronic absorption and steady state emission properties of a hemicyanine dye [4-[4-(dimethylamino)styryl]-1-docosylpyridinium bromide], have been studied in several pure solvents and two mixed binary solvents (water+ethanol and water+acetonitrile). In pure solvents the band-width of the absorption spectrum correlates well with the Stoke's shift. In mixed aqueous solvents two different molecular forms of the solute, viz. the monomer and the dimer of the solute exists in equilibrium. Thermodynamic parameters (e.g. the Delta G degrees and Delta H degrees ) characterizing the equilibrium have been determined. While the value of Delta G degrees changes very slightly with the composition of the binary mixture, the value of Delta H degrees has been observed to depend significantly with solvent composition.


Assuntos
Compostos de Anilina/química , Carbocianinas/química , Compostos de Piridínio/química , Álcoois , Clorofórmio , Furanos , Cinética , Nitrilos , Distribuição Normal , Soluções , Solventes , Espectrometria de Fluorescência , Espectrofotometria , Água
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