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1.
Cell Biol Int ; 44(6): 1312-1330, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32068317

RESUMO

Hyaluronan-binding protein 1 (HABP1), a multi-compartmental, multi-functional protein has a wide range of functions, which can be attributed to its ability to associate with a variety of cellular ligands. Earlier we have reported that HABP1 overexpression in rat normal fibroblasts (F-HABP07) shows chronic generation of reactive oxygen species (ROS), induction of autophagy, and apoptosis. However, a significant proportion of cells remained viable after the majority went through apoptosis from 60 to 72 h. In this study, an attempt has been made to delineate the cellular events in the declined population of surviving cells. It has been elucidated here that, these cells at later time points of growth, that is, 72 and 84 h, not only appeared to shrink but also are devoid of autophagic vacuoles and displayed polyploidy. F-HABP07 cells exhibited an altered cytoskeletal structure from their parental cell line F111, assumed to be caused upon inhibition of actin polymerization and decrease in IQ motif-containing GTPase activating protein 1 (IQGAP1), a key protein associated with maintenance of cytoskeletal integrity. Enhanced expression and nuclear localization of AKT observed in F-HABP07 cells appears to be contributing toward the maintenance of high ROS levels in these cells and also potentially modulating the IQGAP1 activity. These observations, in fact have been considered to result in sustained DNA damage, which then leads to increased expression of p53 and activation of p21 and carry out the cellular events responsible for senescence. Subsequent assessment of the presence of positive ß-gal staining and enhanced expression of p16INK4a in F-HABP07, confirmed that HABP1 overexpressing fibroblasts undergo senescence.


Assuntos
Proteínas de Transporte/fisiologia , Senescência Celular , Fibroblastos/citologia , Proteínas Mitocondriais/fisiologia , Animais , Apoptose , Autofagia , Proteínas de Transporte/genética , Linhagem Celular , Humanos , Ácido Hialurônico/metabolismo , Proteínas Mitocondriais/genética , Ratos , Espécies Reativas de Oxigênio/metabolismo
2.
Cell Biol Int ; 43(7): 820-834, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30958601

RESUMO

The pathological mechanism underlying glaucoma has always been a complex aspect of this permanently blinding disease but proteomic studies have been helpful in elucidating it to a great extent in several studies. This study was designed to evaluate the expression and to get an idea about the function of two novel markers (ligatin and fibulin-7) identified in human aqueous humor (hAH) in relation to glaucomatous progression. A significant increase in the protein content of glaucomatous hAH compared to that of non-glaucomatous controls (NG-Ctrls) was observed. Ligatin, fibulin-7, and its proteolysis were revealed in hAH of primary open angle glaucoma (POAG), primary angle closure glaucoma (PACG) and NG-Ctrls. Quantification confirmed no significant difference in expression of ligatin, whereas fibulin-7 was significantly (P < 0.05) low in hAH of PACG in comparison to NG-Ctrls and POAG. Importantly the immunohistochemical assay for both indicated their possible involvement in the maintenance of the appropriate structure of TM in vivo. Since oxidative stress is a major contributor to glaucomatous pathogenesis, in vitro analysis of nuclear and cytoplasmic fractions indicated intracellular changes in localization and expression of ligatin upon oxidative insult of human trabecular meshwork (TM) cells. While no such changes were found for fibulin-7 expression. This was also corroborated with the immunocytochemical assay. Though a study with a small sample size, this is the first report which confirms the presence of ligatin and fibulin-7 in hAH, quantified their differential expression, and indicated the possibility of their involvement in the maintenance of the TM structure.


Assuntos
Humor Aquoso/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Fator de Iniciação 2 em Eucariotos/metabolismo , Glaucoma de Ângulo Fechado/metabolismo , Glaucoma de Ângulo Aberto/metabolismo , Proteínas de Membrana/metabolismo , Malha Trabecular/metabolismo , Idoso , Biomarcadores/metabolismo , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Progressão da Doença , Feminino , Glaucoma de Ângulo Fechado/patologia , Glaucoma de Ângulo Aberto/patologia , Humanos , Pessoa de Meia-Idade , Estresse Oxidativo , Proteólise , Proteômica
3.
Cell Biol Int ; 40(7): 821-31, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27109893

RESUMO

Cancer cells exhibit various degrees of mitochondrial metabolic alterations. Owing to their multiple roles, mitochondria are attractive target for cancer therapy. Cancerous cells have high glucose (HG) requirements for their growth. Depriving them of glucose has been an approach used in many studies to restrict their perpetuation. However, such deprivation can negatively affect the surrounding normal cells in vivo. Keeping this in view, we treated HeLa cells with only physiological glucose (PG, 5.5 mM) and a combination of physiological glucose with a very low dose (1 nM) of rotenone (PGT), taking high glucose (HG, 25 mM)-treated HeLa cells as normal. We demonstrated that HeLa cells under PG condition mainly exhibited growth arrest. The PGT combination induced apoptosis in HeLa cells by generation of ROS, decrease in ATP production even with around 1.89-fold increase in glucose consumption, cell cycle arrest at S-phase and substantial increase in sub-diploid (Sub-D) population. The oxidative stress generated in both PG and PGT conditions stabilised p53 by localising it in the nuclei of HeLa cells, which would have otherwise undergone HPV-mediated inactivation. Pre-mature senescence induced due to limited glucose availability was found to be regulated by nuclear translocated p53 which, in turn, induced p21, pAkt and pERK. The cyto-toxic effect of rotenone on glucose deprived HeLa cells, synergistically activated NFκB, caspase-3 and Bax along with reduced expression of hyaluronan, a ROS scavenging molecule on their cell surface. Thus, our finding might be a valuable approach to specifically target cancerous cells in a more physiologically feasible condition and can serve as a relevant biochemical basis to gain new insights into cancer therapy.


Assuntos
Carcinogênese/efeitos dos fármacos , Transformação Celular Neoplásica/efeitos dos fármacos , Complexo I de Transporte de Elétrons/antagonistas & inibidores , Glucose/deficiência , Rotenona/farmacologia , Apoptose/efeitos dos fármacos , Carcinogênese/metabolismo , Caspase 3/metabolismo , Caspases/metabolismo , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Senescência Celular/efeitos dos fármacos , Complexo I de Transporte de Elétrons/metabolismo , Glucose/administração & dosagem , Células HeLa , Humanos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos
4.
Cell Biol Int ; 40(1): 107-20, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26524696

RESUMO

This study aimed to determine whether the prolonged exposure of the human trabecular meshwork (HTM) cell line to a low dose (1 nM) of rotenone could simulate a glaucomatous-like condition and serve as a cellular model for its etiological analysis. Under 1-nM rotenone exposure for 24-72 h, HTM cells showed a decrease in cell viability as assessed by an MTT assay and showed mitochondrial dysfunction as assessed by measuring H2 DCFDA fluorescence; a decrease in ATP level was also observed. Flow cytometric analysis showed an increase in cellular size and granularity. Elevated AF showed initial senescence. LF staining with SBB and its spectrofluorometric quantification confirmed growth arrest. An accumulation of cytoplasmic myocilin, IL-6, and MMP-9 at 72 h of exposure supported glaucomatous induction. TEM revealed morphological changes in mitochondria and nuclei of treated cells. Signaling cascades were assessed by immunoblotting and immunocytochemical analysis. This study showed a shift in status of the cells from initial senescence to induction of apoptosis in the HTM cell line due to continuous low-dose exposure to rotenone; however, at 72 h, both senescence and apoptotic features are apparent in these cells. This is the first report that reveals the potential of a prolonged low-dose exposure of rotenone to simulate senescence in the HTM cell line to cause a glaucomatous condition.


Assuntos
Glaucoma/etiologia , Rotenona/farmacologia , Malha Trabecular/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Senescência Celular/efeitos dos fármacos , Proteínas do Citoesqueleto/metabolismo , Relação Dose-Resposta a Droga , Proteínas do Olho/metabolismo , Glaucoma/induzido quimicamente , Glaucoma/metabolismo , Glicoproteínas/metabolismo , Humanos , Interleucina-6/metabolismo , Mitocôndrias/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Transdução de Sinais , Malha Trabecular/citologia , Malha Trabecular/metabolismo
5.
J Biol Chem ; 287(23): 19750-64, 2012 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-22451658

RESUMO

Overexpression of the mature form of hyaluronan-binding protein 1 (HABP1/gC1qR/p32), a ubiquitous multifunctional protein involved in cellular signaling, in normal murine fibroblast cells leads to enhanced generation of reactive oxygen species (ROS), mitochondrial dysfunction, and ultimately apoptosis with the release of cytochrome c. In the present study, human liver cancer cell line HepG2, having high intracellular antioxidant levels was chosen for stable overexpression of HABP1. The stable transformant of HepG2, overexpressing HABP1 does not lead to ROS generation, cellular stress, and apoptosis, rather it induced enhanced cell growth and proliferation over longer periods. Phenotypic changes in the stable transformant were associated with the increased "HA pool," formation of the "HA cable" structure, up-regulation of HA synthase-2, and CD44, a receptor for HA. Enhanced cell survival was further supported by activation of MAP kinase and AKT-mediated cell survival pathways, which leads to an increase in CYCLIN D1 promoter activity. Compared with its parent counterpart HepG2, the stable transformant showed enhanced tumorigenicity as evident by its sustained growth in low serum conditions, formation of the HA cable structure, increased anchorage-independent growth, and cell-cell adhesion. This study suggests that overexpression of HABP1 in HepG2 cells leads to enhanced cell survival and tumorigenicity by activating HA-mediated cell survival pathways.


Assuntos
Proteínas de Transporte/biossíntese , Proliferação de Células , Ciclina D1/metabolismo , Ácido Hialurônico/biossíntese , Proteínas Mitocondriais/biossíntese , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Proteínas de Transporte/genética , Adesão Celular/genética , Sobrevivência Celular/genética , Ciclina D1/genética , Ativação Enzimática/genética , Glucuronosiltransferase/genética , Glucuronosiltransferase/metabolismo , Células Hep G2 , Humanos , Receptores de Hialuronatos/genética , Receptores de Hialuronatos/metabolismo , Ácido Hialurônico/genética , Proteínas Mitocondriais/genética , Proteínas Proto-Oncogênicas c-akt/genética , Coelhos , Regulação para Cima/genética
6.
Appl Biochem Biotechnol ; 195(1): 172-195, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36070165

RESUMO

Tupistra nutans Wall. ex Lindl. is a medicinal plant found in the Eastern Himalayan region. Besides being used as a folk medicine for pain and high blood sugar, its inflorescence is consumed as a vegetable. However, its medicinal properties have not been proven in vitro and in vivo till now. Therefore, in this study, we reported the phytochemicals present in the methanolic extract of Tupistra nutans Wall. ex Lindl. inflorescence (METNI) and its comparative effect in liver carcinoma HepG2 cells against non-cancerous murine fibroblast F111 cells. Phytochemical profiling by gas chromatography-mass spectrometry (GC-MS) analysis showed that METNI was rich in unsaturated fatty acids, vitamin E, and anticancer compounds like diosgenin, linoleic acid, and palmitoleic acid. METNI was found to have in vitro antioxidant property as determined by DPPH and pyrogallol methods, and UV protection property as investigated by fluorescence-based and spectrophotometric methods. MTT assay revealed METNI caused significantly more cell proliferation inhibition in HepG2 (IC50 = 138 µg/ml) compared to F111 (IC50 = 347 µg/ml) cells. Although in both HepG2 and F111 cells METNI showed significant antioxidant activity, it led to intracellular ROS generation and cell cycle alteration at higher exposure. The obtained results suggest that Tupistra nutans can be a promising application for anticancer drug and skin care product development, but can be harmful if overconsumed.


Assuntos
Antioxidantes , Extratos Vegetais , Camundongos , Animais , Antioxidantes/farmacologia , Antioxidantes/química , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Inflorescência , Metanol , Fibroblastos , Compostos Fitoquímicos/farmacologia
7.
3 Biotech ; 13(10): 339, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37705865

RESUMO

Cancerous cells display metabolic engineering through enhanced utilization of nutrients to support their increased requirements for proliferation, bioenergetics, biosynthesis, redox homeostasis, and cell signaling. To investigate the extent to which malignant cells rely on glycolysis and glutaminolysis, the effects of differential deprivation of nutrients such as d-glucose, l-glutamine, and pyruvate on proliferation, morphology, cell cycle, oxidative stress, mitochondrial function, autophagic vacuole formation, and migration in MDA-MB-231, HepG2, and HeLa cells were investigated in this study. Cell viability assay,  cell morphology, and ATP assay showed higher dependence of MDA-MB-231 and HepG2 cells on glucose and glutamine, respectively, for cell survival, growth, ATP production, and proliferation, while HeLa cells were equally dependent on both. However, the combination of all three nutrients displayed maximum proliferation. Differential deprivation of glucose in the absence of glutamine resulted in G0/G1 plus G2/M arrest in MDA-MB-231, whereas G0/G1 arrest in HepG2 and S-phase arrest in HeLa cells occurred at 48 h. Although the differential withdrawal of nutrients revealed a varying degree of effect dependent on cell type, nutrient type, nutrient concentrations, and deprivation time, a general trend of increased oxidative stress, loss of mitochondrial membrane potential, and ATP and antioxidant (GSH) depletion led to mitochondrial dysfunction in all three cell lines and inhibition of cell migration in MDA-MB-231 and HeLa cells at 48 h. Extreme deprivation of nutrients formed autophagic vacuoles. Importantly, normal cells (HEK293) remained unaffected under most of the nutrient-deprived conditions examined. This study enhances our understanding of the impact of differential nutrient deprivation on critical characteristics of cancer cells, contributing to the development of metabolism-based effective anticancer strategies. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03759-w.

8.
J Mater Chem B ; 12(1): 202-221, 2023 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-38073612

RESUMO

This study delves into investigating alternative methodologies for anti-microbial therapy by focusing on the mechanistic assessment of carbon dots (CDs) synthesized from F. benghalensis L. extracts. These biogenic CDs have shown remarkable broad-spectrum anti-bacterial activity even against multi-drug resistant (MDR) bacterial strains, prompting a deeper examination of their potential as novel anti-microbial agents. The study highlights the significant detrimental impact of CDs on bacterial cells through oxidative damage, which disrupts the delicate balance of ROS control within the cells. Notably, even at low doses, the anti-bacterial activity of CDs against MDR strains of P. aeruginosa and E. cloacae is highly effective, demonstrating their promise as potent antimicrobial agents. The research sheds light on the capacity of CDs to generate ROS, leading to membrane lipid peroxidation, loss of membrane potential, and rupture of bacterial cell membranes, resulting in cytoplasmic leakage. SEM and TEM analysis revealed time-dependent cell surface, morphological, and ultrastructural changes such as elongation of the cells, irregular surface protrusion, cell wall and cell membrane disintegration, internalization, and aggregations of CDs. These mechanisms offer a comprehensive explanation of how CDs exert their anti-bacterial effects. We also determined the status of plasma membrane integrity and evaluated live (viable) and dead cells upon CD exposure by flow cytometry. Furthermore, comet assay, biochemical assays, and SDS PAGE identify DNA damage, carbohydrate and protein leakage, and distinct differences in protein expression, adding another layer of understanding to the mechanisms behind CDs' anti-bacterial activity. These findings pave the way for future research on managing ROS levels and developing CDs with enhanced anti-bacterial properties, presenting a breakthrough in anti-microbial therapy.


Assuntos
Anti-Infecciosos , Carbono , Espécies Reativas de Oxigênio/metabolismo , Carbono/química , Anti-Infecciosos/farmacologia , Estresse Oxidativo , Membrana Celular/metabolismo , Bactérias
9.
Cell Biol Int ; 36(11): 1021-7, 2012 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-22775755

RESUMO

Oxidative stress induced by serum starvation and H2O2 exposure, both triggers apoptosis in retinal neuronal cell line RGC-5 (retinal ganglion cell-5). We have examined whether, despite excess generation of ROS (reactive oxygen species) and apoptosis induction, there is any dissimilarity in nuclear morphology and apoptotic signalling pathway in RGC-5 under these conditions. Sub-confluent cells were treated either with H2O2 or maintained in SFM (serum-free medium). ROS level was detected along with nuclear morphology and ultrastructural analysis. Generation of excess intracellular ROS, nuclear localization of Bax and caspase 3 activation along with decrease of cellular viability, confirmed apoptosis induction in RGC-5 by 72 h serum starvation and 500 M H2O2 exposure for 1 h. Nuclear swelling as supported by nuclear cytoplasmic ratio and conspicuous black spots with nuclear remodelling were observed only upon SFM, but not with H2O2 treatment. Serum starvation did not alter JNK1 (c-Jun N-terminal kinase 1) expression, although nuclear translocation and higher level of pJNK (phospho-JNK) was evident. Conversely, H2O2 exposure blocked the expression and activation of JNK1 to phospho-JNK as a negligible level of pJNK was present in the cytoplasm. Despite similar ROS generation in both the conditions, difference in nuclear morphology and JNK1 expression leads to the hypothesis that RGC-5 cells may follow different signalling pathways when challenged with serum starvation and H2O2.


Assuntos
Apoptose , Núcleo Celular/ultraestrutura , Proteína Quinase 8 Ativada por Mitógeno/metabolismo , Estresse Oxidativo , Neurônios Retinianos/citologia , Transporte Ativo do Núcleo Celular , Animais , Caspase 3/metabolismo , Diferenciação Celular , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/enzimologia , Forma do Núcleo Celular , Proliferação de Células , Sobrevivência Celular , Reprogramação Celular , Meios de Cultura Livres de Soro/metabolismo , Citoplasma/enzimologia , Citoplasma/metabolismo , Ativação Enzimática , Peróxido de Hidrogênio/efeitos adversos , Sistema de Sinalização das MAP Quinases , Microscopia Eletrônica de Varredura , Fosforilação , Ratos , Espécies Reativas de Oxigênio/metabolismo , Neurônios Retinianos/efeitos dos fármacos , Neurônios Retinianos/enzimologia , Soro/metabolismo , Proteína X Associada a bcl-2/metabolismo
10.
Saudi J Biol Sci ; 29(5): 3838-3847, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35844432

RESUMO

Current studies are focusing on the anti-cancerous properties of natural bioactive compounds, primarily those included in the human diet. These compounds have the potential to alter the redox balance that can hinder cancer cell's growth. In cancer cells, an abnormal rate of ROS production is balanced with higher antioxidant activities, which if not maintained, results in cancer cells being prone to cell death due to oxidative stress. Here, we have analyzed the effects of Chrysin and Capsaicin on the HeLa cells viability and cellular redox signaling. Both these compounds stimulate cellular and mitochondrial ROS overproduction that perturbs the cellular redox state and results in mitochondrial membrane potential loss. Apart from this, these compounds induce cell cycle arrest and induce premature senescence, along with the overexpression of p21, p53, and p16 protein at lower concentration treatment of Chrysin or Capsaicin. Moreover, at higher concentration treatment with these compounds, pro-apoptotic activity was observed with the high level of Bax and cleaved caspase-3 along with suppression of the Bcl-2 protein levels. In-Silico analysis with STITCH v5 also confirms the direct interaction of Chrysin and Capsaicin with target protein p53. This suggests that Chrysin and Capsaicin trigger an increase in mitochondrial ROS, and p53 interaction leading to premature senescence and apoptosis in concentration dependent manner and have therapeutic potential for cancer treatment.

11.
Nanoscale ; 14(29): 10399-10417, 2022 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-35819245

RESUMO

Cancer cells use nutrients like D-glucose (Glc) and L-glutamine (Q) more efficiently for their development. This increased nutritional dependency of malignant cells has been commonly employed in various in vitro and in vivo models of anticancer therapies. This study utilized a combination of a low dose (25 µg mL-1) of S2, a phytosynthesized gold nanoparticle (AuNP) that was previously proven to be non-toxic, and deprivation of extracellular glutamine as an anticancer strategy in the human cervical cancer cell line HeLa. We discovered that 24 h Q deprivation led to a less significant decrease in the viability of HeLa cells while a low dose of S2 caused a non-significant reduction in the viability of HeLa cells. However, combining these two treatments resulted in highly significant inhibition of cell growth, as measured by the MTT test and morphological examination. Glutamine starvation in HeLa cells was found to induce cellular uptake of S2 via clathrin-mediated endocytosis, thus facilitating the improved antitumor effects of the combined treatment. Flow cytometry-based assays using fluorescent probes H2DCFDA and MitoSOX Red confirmed that this combination therapy involved the development of oxidative stress conditions owing to a surplus of cytosolic reactive oxygen species (cytoROS) and mitochondrial superoxide (mtSOX) generation. Furthermore, the investigated combinatorial treatment also indicated mitochondrial inactivity and disintegration, as evidenced by the drop in the mitochondrial membrane potential (Δψm) and the decrease in the mitochondrial mass (mtMass) in a flow-cytometric assay utilizing the probes. Tetramethylrhodamine ethyl ester and MitoTracker Green FM, respectively. Cell cycle arrest in the G0/G1 phase, induction of cell death via apoptosis/necrosis, and inhibition of migration capacities of HeLa cells were also seen after the combined treatment. Thus, this research provides insight into a new combinatorial approach for reducing the dose of nanoparticles and increasing their efficacy to better inhibit the growth of human cervical cancer cells by leveraging their extracellular glutamine dependence.


Assuntos
Nanopartículas Metálicas , Neoplasias do Colo do Útero , Apoptose , Pontos de Checagem do Ciclo Celular , Sobrevivência Celular , Feminino , Pontos de Checagem da Fase G1 do Ciclo Celular , Glutamina/metabolismo , Glutamina/farmacologia , Glutamina/uso terapêutico , Ouro/metabolismo , Células HeLa , Humanos , Potencial da Membrana Mitocondrial , Mitocôndrias/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Neoplasias do Colo do Útero/tratamento farmacológico
12.
Int J Biol Macromol ; 181: 180-192, 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-33771548

RESUMO

Fucoidan is a sulfated polysaccharide obtained from marine algae and known for various pharmacological activities. In this study, we investigated the effect of Fucoidan on cell viability, redox balance, cytoskeletal component F-actin, HDAC inhibition, autophagy, and senescence phenomenon in human cervical cancer HeLa cell line in comparison to positive control suberoylanilide hydroxamic acid by flow cytometry, fluorescence microscopy, and western blotting. Our observations revealed that Fucoidan exposure induces cytotoxicity in HeLa cells via ROS and mitochondrial superoxide generation and loss of ATP. Colorimetrical studies suggested that Fucoidan impairs the function of HDAC expression. Fucoidan treatment also contributes to the change in the granularity of cells, senescence-associated heterochromatin foci formation that leads to senescence in HeLa cells. Moreover, we visualize that Fucoidan exhibits autophagosomes formation with monodansylcadaverine, and flow cytometry analysis by acridine orange further substantiates that Fucoidan triggers autophagy in HeLa cells. Additionally, the changes in the expression of proteins p21, p16, BECN1, and HDAC1 were seen as markers of senescence, autophagy, and HDAC inhibition by FACS and immunoblotting. Molecular docking study validates Fucoidan-HDAC1 association in corroboration with the experimental data. Collectively, these mechanistic studies demonstrated that Fucoidan could be a therapeutic molecule for targeting HDACs in cervical cancer.


Assuntos
Epigênese Genética , Polissacarídeos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Acetilação/efeitos dos fármacos , Acetilcisteína/farmacologia , Actinas/metabolismo , Autofagossomos/efeitos dos fármacos , Autofagossomos/metabolismo , Autofagia/efeitos dos fármacos , Autofagia/genética , Proteína Beclina-1/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Senescência Celular/efeitos dos fármacos , Senescência Celular/genética , Regulação para Baixo/efeitos dos fármacos , Epigênese Genética/efeitos dos fármacos , Células HeLa , Histona Desacetilases/metabolismo , Histonas/metabolismo , Humanos , Lisina/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Estresse Oxidativo , Superóxidos/metabolismo , Vorinostat/farmacologia
13.
Environ Pollut ; 282: 117003, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-33848911

RESUMO

Microbial communities are considered as vital members to reflect the health of a riverine system. Among them, pathogenic and fecal indicators imply health risks involved with potability of river water. The present study explores the diverse microbial communities, distribution pattern of potential pathogens, and fecal indicators between the geographically distinct Himalayan and Peninsular river systems of India. It also inquires into the environmental factors associated with community variance and distribution pattern of microbial indicators. The application of high-throughput amplicon sequencing approach unveiled significant demarcation (p < 0.004, Anosim R = 0.62) of samples suggesting unique microbial diversities in these two river sediments. Random forest analysis revealed Desulfobulbulus, PSB_M_3, and Opitutus in Himalayan, while DA101, Bacillus, and Streptomyces in the Peninsular as significant contributors to develop overall dissimilarity between the river systems. Permutational multivariate analysis of variance and co-occurrence network analysis were used to study the relationships between microbial taxa and environmental factors. Amongst the various studied environmental parameters, pH, K, Ca, Mg, Ba, and Al in the Himalayan and salinity, Na, temperature, and Th in the Peninsular significantly influenced shaping of distinct microbial communities. Furthermore, the potential pathogenic genera, including Flavobacterium, Clostridium, Arcobacter, Pseudomonas, and Bacillus were highly prevalent in both the river systems. Arcobacter, Clostridium, Acinetobacter, Bacteroides, and Caloramator were the prominent fecal indicators in these river systems. Our findings provide salient information about the crucial role and interplay between various environmental factors and anthropogenic influences in framing the microbiome of the distinct river systems in India. Moreover, assessing potential pathogenic and fecal indicators suggest the public health risk associated with untreated sewage discharge into these water sources. The detection of various F/S indicators and potentially pathogenic bacteria in Himalayan and Peninsular river systems emphasize the urgent need for future monitoring and management of major riverine systems in India.


Assuntos
Metagenômica , Rios , Monitoramento Ambiental , Fezes , Índia , Prevalência
14.
Genome Announc ; 6(1)2018 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-29301875

RESUMO

The Yamuna River is the backbone of domestic, irrigation, and industrial activities in Delhi, India, yet the complex dynamics of its microbes and their contribution to biogeochemical cycles in a polluted environment remain elusive. This is an introductory report describing the microbial community in the Yamuna River, using high-throughput metagenomics.

15.
Genome Announc ; 6(11)2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29545300

RESUMO

We report here targeted deep-sequencing metagenomic data that reveal a high level of diversity in the microbiota residing in the sediment of the Periyar River in a reserve forest of the Western Ghats. Of the 4,674 operational taxonomic units discovered, the dominant phyla represented were Proteobacteria (33.12%), Actinobacteria (14.58%), Acidobacteria (12.81%), and Bacteroidetes (9.89%).

16.
J Androl ; 27(4): 604-10, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16582418

RESUMO

The proprotein form of hyaluronan binding protein 1 (HABP1) has been reported to be present in the pachytene spermatocytes and the round spermatids of the adult testis. To explore the role of HABP1 proprotein in spermatogenesis, its expression in the testes of adult rats was compared with that in the testes of developing rats and that in the testes of adult rats that received estriadiol to halt spermatogenesis. Immunoblotting revealed that the mature form of HABP1 was consistently present in the testis, but its precursor form was not found in the testis of animals aged 7, 14, 21, and 28 days. However, immunohistochemical analysis revealed the presence of the proprotein form in the pachytene spermatocytes and the round spermatids of testes from rats aged 21 and the 28 days, the appearance of which correlated well with the appearance of these cells during spermatogenesis. Reverse-transcriptase polymerase chain reaction revealed transcriptional upregulation of HABP1 in the testes of adult rats, compared with the testes of developing rats. Finally, loss of HABP1 proprotein expression from the pachytene spermatocytes and round spermatids was observed in the testes from rats in which spermatogenesis was arrested. Collectively, these findings demonstrate the appearance of HABP1 proprotein in the pachytene spermatocytes and the round spermatids during the initial stages of postnatal testis development and suggest that this expression may be crucial for spermatogenesis.


Assuntos
Receptores de Hialuronatos/análise , Precursores de Proteínas/análise , Espermátides/química , Espermatócitos/química , Espermatogênese/fisiologia , Animais , Benzoatos/farmacologia , Estradiol/análogos & derivados , Estradiol/farmacologia , Masculino , Proteínas Mitocondriais , Ratos , Ratos Sprague-Dawley , Espermatogênese/efeitos dos fármacos , Testículo/crescimento & desenvolvimento
17.
Sci Rep ; 6: 31492, 2016 08 16.
Artigo em Inglês | MEDLINE | ID: mdl-27526963

RESUMO

The steady rise in antimicrobial resistance poses a severe threat to global public health by hindering treatment of an escalating spectrum of infections. We have previously established the potent activity of α-MSH, a 13 residue antimicrobial peptide, against the opportunistic pathogen Staphylococcus aureus. Here, we sought to determine whether an increase in cationic charge in α-MSH could contribute towards improving its staphylocidal potential by increasing its interaction with anionic bacterial membranes. For this we designed novel α-MSH analogues by replacing polar uncharged residues with lysine and alanine. Similar to α-MSH, the designed peptides preserved turn/random coil conformation in artificial bacterial mimic 1,2-dimyristoyl-sn-glycero-3-phosphocholine:1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (7:3, w/w) vesicles and showed preferential insertion in the hydrophobic core of anionic membranes. Increased cationic charge resulted in considerable augmentation of antibacterial potency against MSSA and MRSA. With ~18-fold better binding than α-MSH to bacterial mimic vesicles, the most charged peptide KKK-MSH showed enhanced membrane permeabilization and depolarization activity against intact S. aureus. Scanning electron microscopy confirmed a membrane disruptive mode of action for KKK-MSH. Overall, increasing the cationic charge improved the staphylocidal activity of α-MSH without compromising its cell selectivity. The present study would help in designing more effective α-MSH-based peptides to combat clinically relevant staphylococcal infections.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Cátions/metabolismo , Metabolismo dos Lipídeos , Staphylococcus aureus/efeitos dos fármacos , alfa-MSH/química , alfa-MSH/farmacologia , Antibacterianos/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/ultraestrutura , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Ligação Proteica , Relação Estrutura-Atividade , alfa-MSH/metabolismo
18.
J Trace Elem Med Biol ; 30: 184-93, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25617233

RESUMO

Accumulated evidence over the years indicate that cadmium (Cd) may be a possible etiological factor for neurodegenerative diseases. This may possibly be linked to excessive generation of free radicals that damages the organs in the body depending on their defence mechanism. Since Cd is a toxic agent that affect several cell types, the aim of this study was to shed light on the effect of Cd and its consequences on different organs of the mice body. To test the hypothesis of concentration dependent Reactive Oxygen Species (ROS) generation and DNA damage, observations were done in the serum of 4-5 weeks old male Swiss albino mice by treating with cadmium chloride (CdCl2) in drinking water for 30 days. The expression of Bcl-2-associated X protein (Bax) an apoptotic marker protein was two times higher in brain compared to liver at an exposure level of 0.5mgL(-1) CdCl2. Furthermore the correlation and linkage data analysis of antioxidant defence system revealed a rapid alteration in the brain, compared to any other organs considered in this study. We report that even at low dose of Cd, it impaired the brain due to lipid peroxidase sensitivity which favoured the Cd-induced oxidative injury in the brain.


Assuntos
Encéfalo/patologia , Cloreto de Cádmio/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Ensaio Cometa , Dano ao DNA , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Camundongos , Proteínas Mitocondriais/metabolismo , Especificidade de Órgãos/efeitos dos fármacos , Oxirredução/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Análise de Sobrevida , Proteína X Associada a bcl-2/metabolismo
19.
J Reprod Immunol ; 53(1-2): 45-54, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11730903

RESUMO

Hyaluronan binding protein 1 (HABP1) was reported to be present on human sperm surface and its involvement in fertilization has already been elucidated (Mol. Repro. Dev. 38 (1994) 69). In the present communication, we report a significant reduction in the level of this protein in sperms from asthenozoospermic and oligozoospermic patients as compared to normozoospermic one. Further evidence of the absence of HABP1 in sperms, having motility <20% is documented, which again is a determining factor for fertilization. HABP1 was quantitatively determined using anti-HABP1 antibody from sperm extracts isolated from semen samples of both the fertile and infertile groups demonstrating low sperm motility. Sperm samples with low motility revealed a significant reduction in the level of HABP1 in immunoblot detection as well as immunolocalization experiment. It suggests that decreased HABP1 level may be associated with low motility of sperms, which in turn might cause infertility in the patient. Thus, the sperm surface HABP1 level can be correlated with the degree of sperm motility, an important criteria for fertilization.


Assuntos
Receptores de Hialuronatos/metabolismo , Infertilidade Masculina/metabolismo , Motilidade dos Espermatozoides/fisiologia , Estudos de Casos e Controles , Fertilidade/fisiologia , Humanos , Masculino , Oligospermia/metabolismo
20.
DNA Cell Biol ; 23(5): 301-10, 2004 May.
Artigo em Inglês | MEDLINE | ID: mdl-15169609

RESUMO

The gene encoding Hyaluronan binding protein 1 (HABP1) and its homologs have been reported across eukaryotes, from yeast to human. We have reported the presence of processed pseudogenes in several human chromosomes, along with the location of the HABP1 gene on chromosome 17p12-p13. In this study, we report not only the presence of HABP1 pseudogene in other animal species, but also the presence of a homologous sequence in Methanosarcina barkeri, an ancient life form. This sequence has 44.8% homology to the human HABP1 cDNA and 45.3% homology with the HABP1 pseudogene in human chromosome 21. This sequence has a high G + C content (57%), characteristic of archaea, a family to which M. barkeri belongs. The presence of this HABP1 cDNA like fragment in M. barkeri might enable us to shed light on the evolution of the HABPl gene and whether it was present in a common ancestral organism before the lineages separated.


Assuntos
Evolução Molecular , Receptores de Hialuronatos/genética , Methanosarcina barkeri/genética , Pseudogenes/genética , Animais , Sequência de Bases , Proteínas de Transporte , Cromossomos Humanos Par 21 , Genes Arqueais , Humanos , Proteínas Mitocondriais , Dados de Sequência Molecular , Filogenia , Homologia de Sequência do Ácido Nucleico
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