RESUMO
Calcineurin is a phosphatase whose primary targets in T cells are NFAT transcription factors, and inhibition of calcineurin activity by treatment with cyclosporin A (CsA) or FK506 is a cornerstone of immunosuppressive therapies. Here we found that calcineurin was recruited to the T cell antigen receptor (TCR) signaling complex, where it reversed inhibitory phosphorylation of the tyrosine kinase Lck on Ser59 (LckS59). Loss of calcineurin activity impaired phosphorylation of Tyr493 of the tyrosine kinase ZAP-70 (ZAP-70Y493), as well as some downstream pathways in a manner consistent with signaling in cells expressing LckS59A (Lck that cannot be phosphorylated) or LckS59E (a phosphomimetic mutant). Notably, CsA inhibited integrin-LFA-1-dependent and NFAT-independent adhesion of T cells to the intercellular adhesion molecule ICAM-1, with little effect on cells expressing mutant Lck. These results provide new understanding of how widely used immunosuppressive drugs interfere with essential processes in the immune response.
Assuntos
Calcineurina/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Linfócitos T/imunologia , Proteína-Tirosina Quinase ZAP-70/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Ciclosporina/farmacologia , Humanos , Imunossupressores/farmacologia , Molécula 1 de Adesão Intercelular/metabolismo , Células Jurkat , Ativação Linfocitária/efeitos dos fármacos , Antígeno-1 Associado à Função Linfocitária/metabolismo , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/genética , Camundongos , Camundongos Transgênicos , Fatores de Transcrição NFATC/genética , Fatores de Transcrição NFATC/metabolismo , Fosforilação/efeitos dos fármacos , Fosforilação/genética , Ligação Proteica , Transdução de Sinais , Linfócitos T/efeitos dos fármacos , Tacrolimo/farmacologiaRESUMO
Natural antioxidants have become vital to minimize macromolecular damage caused by Reactive Oxygen Species (ROS). This study investigated the antioxidant property of ß-cryptoxanthin (ß-CRX) extracted from Kocuria marina DAGII and its protective effect against macromolecular damages by generating ROS via two models: UV radiation and the Fenton reaction. ß-cryptoxanthin exhibited the highest scavenging activity towards hydrogen peroxide radicals with an IC50 value of 38.30 ± 1.13 µg/ml, favoring the hydrogen atom transfer mechanism. The total antioxidant capacity value of 872.0101 ± 1.84 µg BHT/mg ß-CRX indicated the cumulative ROS scavenging ability of ß-cryptoxanthin. ß-cryptoxanthin could protect against ROS-induced lipid peroxidation, protein oxidation, and DNA damage. The highest lipid peroxidation and protein oxidation inhibition values of ß-cryptoxanthin against ROS were 99.371 ± 0.51% and 78.19 ± 0.15%, respectively. ß-cryptoxanthin also showed a protective effect in maintaining DNA intactness against ROS-mediated DNA damage. Allium cepa test showed the non-genotoxic nature of ß-cryptoxanthin and its protective effect against ROS genotoxic effects. A photo-stability study of ß-cryptoxanthin toward UVA and UVB radiation showed a rapid bleaching result of UVB obeying pseudo-zero order kinetics with an average R2 value of 0.9897 and a higher k value (-6.3 × 10-11 ± 0.2 M/s) than UVA (k value -3.1 × 10-11 ± 0.17 M/s), signifying that UVB is more potent toward photo-degradation. The good SPF value of 23.1737 ± 0.15 showed the UV protection capability of ß-cryptoxanthin. Thus, the present study suggests that ß-cryptoxanthin could be a valuable antioxidant to protect against ROS-induced various macromolecular damages and act as a good UV protectant.
Assuntos
Antioxidantes , beta-Criptoxantina , Antioxidantes/farmacologia , Espécies Reativas de Oxigênio , Dano ao DNA , HidrogênioRESUMO
ZAP-70 is a tyrosine kinase that is essential for initiation of T cell antigen receptor (TCR) signaling. We have found that T cell p38 MAP kinase (MAPK), which is directly phosphorylated and activated by ZAP-70 downstream of the TCR, in turn phosphorylates Thr-293 in the interdomain B region of ZAP-70. Mutant T cells expressing ZAP-70 with an alanine substitution at this residue (ZAP-70T293A) had enhanced TCR proximal signaling and increased effector responses. Lack of ZAP-70T293 phosphorylation increased association of ZAP-70 with the TCR and prolonged the existence of TCR signaling microclusters. These results identify a tight negative feedback loop in which ZAP-70-activated p38 reciprocally phosphorylates ZAP-70 and destabilizes the signaling complex.
Assuntos
Genes Codificadores dos Receptores de Linfócitos T/fisiologia , Proteína-Tirosina Quinase ZAP-70/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Sequência de Aminoácidos , Regulação da Expressão Gênica , Humanos , Células Jurkat , Fosforilação , Transdução de Sinais , Proteína-Tirosina Quinase ZAP-70/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
In the present investigation, growth kinetics of Kocuria marina DAGII during batch production of ß-Cryptoxanthin (ß-CRX) was studied by considering the effect of glucose and maltose as a single and binary substrate. The importance of mixed substrate over single substrate has been emphasised in the present study. Different mathematical models namely, the Logistic model for cell growth, the Logistic mass balance equation for substrate consumption and the Luedeking-Piret model for ß-CRX production were successfully implemented. Model-based analyses for the single substrate experiments suggested that the concentrations of glucose and maltose higher than 7.5 and 10.0 g/L, respectively, inhibited the growth and ß-CRX production by K. marina DAGII. The Han and Levenspiel model and the Luong product inhibition model accurately described the cell growth in glucose and maltose substrate systems with a R 2 value of 0.9989 and 0.9998, respectively. The effect of glucose and maltose as binary substrate was further investigated. The binary substrate kinetics was well described using the sum-kinetics with interaction parameters model. The results of production kinetics revealed that the presence of binary substrate in the cultivation medium increased the biomass and ß-CRX yield significantly. This study is a first time detailed investigation on kinetic behaviours of K. marina DAGII during ß-CRX production. The parameters obtained in the study might be helpful for developing strategies for commercial production of ß-CRX by K. marina DAGII.
Assuntos
beta-Criptoxantina/biossíntese , Micrococcaceae/crescimento & desenvolvimento , Modelos Biológicos , CinéticaRESUMO
T cells are critical to immunity against pathogenic Salmonella including Salmonella Typhi which causes systemic infection, typhoid, in humans. The strategies that this pathogen employs to keep T-cell mediated immune responses in check during establishment of systemic infection are not completely understood. Here, we show that the virulence polysaccharide Vi, which distinguishes S. Typhi from localized gastroenteritis-producing nontyphoidal Salmonella serovars, is a potent inhibitor of T-cell activation. Vi released by S. Typhi interacts with the membrane prohibitin complex and inhibits IL-2 secretion from T cells stimulated through the T-cell receptor (TCR) but does not affect PMA-activated interleukin 2 (IL-2) secretion. Treatment with Vi suppresses early activation events including TCR down-regulation, actin polymerization, and phosphorylation of ERK. Coadministration of Vi with anti-CD3 Ab reduces secretion of IL-2 and interferon γ in mice. Our findings reveal a mechanism by which S. Typhi may target T-cell immunity during establishment of typhoid.
Assuntos
Interações Hospedeiro-Patógeno , Interleucina-2/antagonistas & inibidores , Ativação Linfocitária/efeitos dos fármacos , Polissacarídeos Bacterianos/metabolismo , Proteínas Repressoras/antagonistas & inibidores , Salmonella typhi/imunologia , Linfócitos T/efeitos dos fármacos , Animais , Membrana Celular/efeitos dos fármacos , Membrana Celular/imunologia , Células Cultivadas , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Proibitinas , Linfócitos T/imunologiaRESUMO
In this study, we fabricated a novel biodegradable functional film using natural polysaccharides by adding jujube seed powder as an active ingredient. Scanning electron microscopy analysis showed agglomerate formation in the film with increasing concentration of seed powder. Fourier transform-infrared spectroscopy study demonstrated an electrostatic interaction between pectin and chitosan. The water solubility and swelling degree significantly decreased from 55.5 to 47.7 % and 66.0 to 41.9 %, respectively, depicting the film's water resistance properties. Higher opacity and lower transmittance value of the film indicated its protective effect towards light-induced oxidation of food. It was observed that the fabricated active film biodegraded to 82.33 % in 6 days. The DPPH radical scavenging activity of 98.02 % was observed for the functional film. The film showed antifungal activity against B. cinerea and P. chrysogenum. The highest zone of inhibition was obtained against food spoiling bacteria B. subtilis followed by S. aureus, P. aeruginosa and E. coli. Genotoxicity studies with the fabricated film showed a mitotic index of 8 % compared to 3 % in the control film. We used the fabricated film to preserve grapefruits, and the result showed that it could preserve grapes for ten days with an increase in antioxidant activity and polyphenolic content.
Assuntos
Quitosana , Frutas , Extratos Vegetais , Ziziphus , Pós/farmacologia , Embalagem de Alimentos , Staphylococcus aureus , Escherichia coli , Quitosana/química , Polissacarídeos/farmacologia , Água/farmacologia , Pectinas/farmacologiaRESUMO
The serine/threonine phosphatase calcineurin is a component of the T cell receptor (TCR) signalosome, where it promotes T cell activation by dephosphorylating LckS59. Using small interfering RNA (siRNA)-mediated knockdown and CRISPR-Cas9-targeted genetic disruption of the calcineurin A chain α and ß isoforms, we find that calcineurin also functions as an adaptor in TCR-signaled human T cells. Unlike inhibition of its phosphatase activity, in the absence of calcineurin A, TCR signaling results in attenuated actin rearrangement, markedly reduced TCR-Lck microcluster formation and recruitment of the adaptor RhoH, and diminished phosphorylation of critical targets downstream of Lck such as TCRζ and ZAP-70. Reconstitution of deficient T cells with either calcineurin Aα or Aß restores TCR microcluster formation and signaling, as does reconstitution with a phosphatase-inactive Aα chain. These results assign a non-enzymatic adaptor function to calcineurin in the formation and stabilization of a functional TCR signaling complex.
Assuntos
Calcineurina , Receptores de Antígenos de Linfócitos T , Transdução de Sinais , Calcineurina/metabolismo , Humanos , Receptores de Antígenos de Linfócitos T/metabolismo , Células Jurkat , Linfócitos T/metabolismo , Fosforilação , Proteína Tirosina Quinase p56(lck) Linfócito-Específica/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteína-Tirosina Quinase ZAP-70/metabolismoRESUMO
Perishable foods like fruits and vegetables, meat, fish, and dairy products have short shelf-life that causes significant postharvest losses, which poses a major challenge for food supply chains. Biopolymers have been extensively studied as sustainable alternatives to synthetic plastics, and pectin is one such biopolymer that has been used for packaging and preservation of foods. Pectin is obtained from abundantly available low-cost sources such as agricultural or food processing wastes and by products. This review is a complete account of pectin extraction from agro-wastes, development of pectin-based composite films and coatings, their characterizations, and their applications in food packaging and preservation. Compared to conventional chemical extraction, supercritical water, ultrasound, and microwave assisted extractions are a few examples of modern and more efficient pectin extraction processes that generate almost no hazardous effluents, and thus, such extraction techniques are more environment friendly. Pectin-based films and coatings can be functionalized with natural active agents such as essential oils and other phytochemicals to improve their moisture barrier, antimicrobial and antioxidant properties. Application of pectin-based active films and coatings effectively improved shelf-life of fresh cut-fruits, vegetables, meat, fish, poultry, milk, and other food perishable products.
Assuntos
Embalagem de Alimentos , Pectinas , Embalagem de Alimentos/métodos , Conservação de Alimentos/métodos , Antioxidantes/farmacologia , Antioxidantes/química , Manipulação de AlimentosRESUMO
Dietzia maris NIT-D, a canthaxanthin producer, was isolated during routine screening of pigment-producing bacteria. Response surface methodology was applied for statistical designing of process parameters for biomass and canthaxanthin production. The effects of four process parameters (considered as independent variables), namely temperature (10-30 °C), pH (4.75-5.75), shaker speed (75-135 rpm) and percentage inoculum (0.5-2.5 %) on the biomass and canthaxanthin yield (considered as dependent variables) were studied. As much as 122 mg L(-1) of canthaxanthin was obtained when Dietzia maris NIT-D was incubated for 120 h at 25 °C and 120 rpm, initial pH and percentage inoculum being 5.5 and 2 % respectively. The pigment yield is the highest reported till date, with Dietzia maris as the test organism. The maximum biomass yield was 7.39 g L(-1) under optimized process parameters. The predicted values were also verified by validation experiments in 5-day fermentation. Different mathematical models were used to describe growth and production, considering the effect of glucose in batch mode. The kinetic constants were calculated by fitting the experimental data to the models. Cell growth was inhibited beyond a glucose concentration of 15 g L(-1). Andrews' model gave the best fit with a R (2) value of 0.9993. During the exponential growth phase, the specific growth rate was found to remain fairly constant with respect to time. There was no inhibitory effect due to intracellular product accumulation for all concentrations of glucose. This observation is the first of its kind, as previous studies have reported that increasing accumulation of intracellular carotenoid exerts greater degree of inhibition on growth.
Assuntos
Cantaxantina/biossíntese , Corynebacterium/crescimento & desenvolvimento , Corynebacterium/metabolismo , Modelos Biológicos , Concentração de Íons de HidrogênioRESUMO
BACKGROUND: Mangiferin is a C-glycoside xanthone molecule having a wide range of therapeutic properties. Hence, the present study aims to understand the efficacy of mangiferin against colorectal cancer (CRC) and to elucidate the mechanisms of action of mangiferin on colorectal cancer. METHOD: The molecular mechanism of mangiferin against colorectal cancer was studied using Autodock Vina software. Pharmacophore analysis of mangiferin concerning five COX-2 inhibitor drugs was carried out using the PharmaGist server to analyze the possibility of using mangiferin as a COX-2 inhibitor. In vitro analysis of Mangiferin against various cancer cell lines was performed. RESULTS: The molecular mechanism of action of mangiferin against CRC was assessed by docking with multiple target proteins involved in the progression of CRC. Docking studies showed good binding scores (kcal/mol) ranging from - 10.3 to - 6.7. Mangiferin showed a good affinity towards enzymes like COX-2 and LA4H involved in Arachidonic acid (AA) metabolism with a binding score(kcal/mol) of - 10.1 and - 10.3 respectively. The pharmacophore feature assessment of mangiferin was done for COX-2 inhibitor drugs, which further confirmed that mangiferin poses the same pharmacophore feature as that of COX-2 inhibitor drugs. Furthermore, the binding affinity of mangiferin was compared with five COX-2 inhibitor drugs to prove its efficacy as an inhibitor. Mangiferin also had a cytotoxic effect against colorectal cancer (HT 29), cervical cancer (HeLa), and breast cancer (MCF 7) cell lines. The study could establish that Mangiferin might be a promising candidate for the treatment of colorectal cancer. CONCLUSION: In short, these studies exploited the possibility of mangiferin as a lead molecule to develop anticancer/anti-inflammatory drugs for the treatment of CRC.
RESUMO
Prohibitin is a highly conserved ubiquitously expressed protein involved in several key cellular functions. Targeting of this protein in the membrane by the virulence polysaccharide, Vi, of human typhoid-causing pathogen, Salmonella enterica serovar Typhi (S. Typhi), results in suppression of IL-2 secretion from T cells activated through the T-cell receptor (TCR). However, the mechanism of this suppression remains unclear. Here, using Vi as a probe, we show that membrane prohibitin associates with the src-tyrosine kinase, p56lck (Lck), and actin in human model T cell line, Jurkat. Activation with anti-CD3 antibody brings about dissociation of this complex, which coincides with downstream ERK activation. The trimolecular complex reappears towards culmination of proximal TCR signaling. Engagement of cells with Vi prevents TCR-triggered activation of Lck and ERK by inhibiting dissociation of the former from prohibitin. These findings suggest a regulatory role for membrane prohibitin in Lck activation and TCR signaling.
Assuntos
Membrana Celular/metabolismo , Complexos Multiproteicos/metabolismo , Proibitinas/metabolismo , Salmonella typhi/patogenicidade , Linfócitos T/fisiologia , Actinas/metabolismo , Humanos , Terapia de Imunossupressão , Células Jurkat , Ativação Linfocitária , Polissacarídeos Bacterianos/imunologia , Ligação Proteica , Receptores de Antígenos de Linfócitos T/metabolismo , Transdução de Sinais , Fatores de Virulência/imunologiaRESUMO
Sulfur is a growth-limiting and secondary macronutrient as well as an indispensable component for several cellular components of crop plants. Over the years various scientists have conducted several experiments on sulfur metabolism based on different aspects of plants. Sulfur metabolism in seeds has immense importance in terms of the different sulfur-containing seed storage proteins, the significance of transporters in seeds, the role of sulfur during the time of seed germination, etc. The present review article is based on an overview of sulfur metabolism in seeds, in respect to source to sink relationships, S transporters present in the seeds, S-regulated seed storage proteins and the importance of sulfur at the time of seed germination. Sulfur is an essential component and a decidable factor for seed yield and the quality of seeds in terms of oil content in oilseeds, storage of qualitative proteins in legumes and has a significant role in carbohydrate metabolism in cereals. In conclusion, a few future perspectives towards a more comprehensive knowledge on S metabolism/mechanism during seed development, storage and germination have also been stated.
RESUMO
Immunization of human volunteers with a single dose of pneumococcal surface protein A (PspA) stimulates broad cross-reactive Abs to heterologous PspA molecules that, when transferred, protect mice from fatal infection with Streptococcus pneumoniae. In this study, we report the molecular characterization of 36 mouse mAbs generated against the extracellular domain of PspA (PspA(3-286)) from strain R36A. Abs to PspA(3-286) were encoded by diverse V(H) and V(kappa) families/genes. The H chain CDR3 and L chain CDR3 lengths were 3-13 (7.8 +/- 0.5) and 8-9 (8.7 +/- 0.2) codons, respectively. Unexpectedly, seven hybridomas expressed H chains that lack D(H) gene-derived amino acids. Nontemplate-encoded addition(s) were observed in the H chain expressed in six of these seven hybridomas; Palindromic addition(s) were absent. Absence of D(H) gene-derived amino acids did not prevent anti-PspA(3-286) mAbs from attaining average relative avidity. Avidity maturation occurred during primary IgG anti-PspA(3-286) polyclonal Ab response in PspA(3-286)- and R36A-immunized mice. Compared with PspA(3-286)-immunized mice, the relative avidity of the primary polyclonal IgG Abs was higher in R36A immunized mice on days 72, 86, and 100. Two pairs of clonally related hybridomas were observed. D(H) genes expressed in the majority (75.9%) of the hybridomas used reading frame 3. Analysis of replacement/silent mutation ratio in the CDR and framework regions provided evidence for Ag-driven selection in 11 mAbs. Based on epitope localization experiments, the mAbs were classified into 12 independent groups. ELISA additivity assay indicated that members within a group recognized topographically related epitopes. This study provides molecular insights into the biology of D(H)-less Abs.
Assuntos
Anticorpos Antibacterianos/biossíntese , Afinidade de Anticorpos/genética , Diversidade de Anticorpos/genética , Proteínas de Bactérias/imunologia , Deleção de Genes , Rearranjo Gênico de Cadeia Pesada de Linfócito B/genética , Cadeias Pesadas de Imunoglobulinas/genética , Streptococcus pneumoniae/imunologia , Sequência de Aminoácidos , Animais , Anticorpos Antibacterianos/genética , Anticorpos Antibacterianos/metabolismo , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/metabolismo , Sequência de Bases , Epitopos de Linfócito B/metabolismo , Feminino , Hibridomas , Cadeias Pesadas de Imunoglobulinas/metabolismo , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/metabolismo , Proteínas de Membrana/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos CBA , Dados de Sequência Molecular , Família Multigênica/imunologiaRESUMO
Inhibitors of calcineurin phosphatase activity (CNIs) such as cyclosporin A (CsA) are widely used to treat tissue transplant rejection and acute graft-versus-host disease (aGVHD), for which inhibition of gene expression dependent on nuclear factor of activated T cells (NFAT) is the mechanistic paradigm. We recently reported that CNIs inhibit TCR-proximal signaling by preventing calcineurin-mediated dephosphorylation of LckS59, an inhibitory modification, raising the possibility of another mechanism by which CNIs suppress immune responses. Here we used T cells from mice that express LckS59A, which cannot accept a phosphate at residue 59, to initiate aGVHD. Although CsA inhibited NFAT-dependent gene upregulation in allo-aggressive T cells expressing either LckWT or LckS59A, it was ineffective in treating disease when the T cells expressed LckS59A. Two important NFAT-independent T cell functions were found to be CsA-resistant in LckS59A T cells: upregulation of the cytolytic protein perforin in tissue-infiltrating CD8+ T cells and antigen-specific T/DC adhesion and clustering in lymph nodes. These results demonstrate that effective treatment of aGVHD by CsA requires NFAT-independent inhibition of TCR signaling. Given that NFATs are widely expressed and off-target effects are a major limitation in CNI use, it is possible that targeting TCR-associated calcineurin directly may provide effective therapies with less toxicity.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Inibidores de Calcineurina/farmacologia , Ciclosporina/farmacologia , Doença Enxerto-Hospedeiro/tratamento farmacológico , Fatores de Transcrição NFATC/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Transdução de Sinais/imunologia , Doença Aguda , Animais , Doença Enxerto-Hospedeiro/genética , Doença Enxerto-Hospedeiro/imunologia , Camundongos , Camundongos Knockout , Fatores de Transcrição NFATC/genética , Receptores de Antígenos de Linfócitos T/genética , Transdução de Sinais/genéticaRESUMO
Oxidative stress is considered to be the leading cause of many chronic and degenerative diseases leading to death and disability. Antioxidant therapy could be efficient in preventing oxidative stress-induced damages in cells. In the present study the efficacy of mangiferin-chitosan nanoparticles (MCNs) as a therapeutic agent against the oxidative stress, since mangiferin (MGF), a polyphenol protects and attenuates oxidative stress against various diseases. However, the bioactivity of MGF does not result in vivo biological effect owing to its low bioavailability and poor solubility. Mangiferin nanoparticles (NPs) were developed to improve the solubility and bioavailability of mangiferin. Nanoparticles had significant free radical scavenging activity in the cell-free system and had a synergistic action with phase II antioxidant enzymes such as catalases and peroxidases. A nephropathic system was developed to investigate the attenuative role of mangiferin-chitosan nanoparticles against induced oxidative stress, on normal kidney epithelial (NKE) cells. Pre-treatment with the nanoparticles exclusively prevented the induction of cytotoxicity induced by NaF and maintained the level of intracellular antioxidant enzyme in the cells. Nanoparticles had significant lipid peroxidation and protein oxidation inhibition activities. These nanoparticles can be used in food and pharmaceutical industries as a therapeutic agent to prevent the oxidative stress-induced health disorders.
Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Quitosana/química , Nanopartículas/química , Estresse Oxidativo/efeitos dos fármacos , Xantonas/química , Sobrevivência Celular , Células Cultivadas , Espaço Intracelular/metabolismo , Rim/citologia , Rim/efeitos dos fármacos , Rim/metabolismo , Modelos Biológicos , Oxirredução , Espécies Reativas de Oxigênio/metabolismo , Análise EspectralRESUMO
The successful translation of nanostructure-based bioimaging and/or drug delivery system needs extensive in vitro and in vivo studies on biocompatibility, biodistribution, clearance, and toxicity for its diagnostic applications. Herein, we have investigated the in vitro cyto-hemocompatibility, in vivo biodistribution, clearance, and toxicity in mice after systemic administration of GdF3 nanoparticles loaded PEGylated mesoporous carbon capsule (GdF3-PMCC)-based theranostic system. In vitro cyto-hemocompatibility study showed a very good biocompatibility up to concentration of 500 µg/ml. Biodistribution studies carried out from 1 h to 8 days showed that GdF3-PMCC was found in major organs, such as liver, kidney, spleen, and muscle till 4th day and it was negligible in any tissue after 8th day. The clearance study was carried out for a period of 8 days and it was observed that the urinary system is the main route of excretion of GdF3-PMCC. The tissue toxicity study was done for 15 days and histopathological analysis indicated that the GdF3-PMCC based theranostic system does not have any adverse effect in tissues. Thus, PMCCs are nontoxic and can be applied as theranostic agents in contrast to the other carbon-based systems (PEGylated carbon nanotubes and PEGylated graphene oxide) which showed significant toxicity.
Assuntos
Antineoplásicos/administração & dosagem , Materiais Biocompatíveis/química , Carbono/química , Portadores de Fármacos/química , Compostos de Flúor/administração & dosagem , Gadolínio/química , Nanopartículas/administração & dosagem , Polietilenoglicóis/química , Nanomedicina Teranóstica , Animais , Antineoplásicos/farmacocinética , Antineoplásicos/farmacologia , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Materiais Biocompatíveis/administração & dosagem , Coagulação Sanguínea/efeitos dos fármacos , Cápsulas , Carbono/administração & dosagem , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/administração & dosagem , Sistemas de Liberação de Medicamentos , Compostos de Flúor/farmacocinética , Compostos de Flúor/toxicidade , Gadolínio/administração & dosagem , Gadolínio/efeitos adversos , Gadolínio/farmacocinética , Gadolínio/toxicidade , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Taxa de Depuração Metabólica , Camundongos , Músculos/efeitos dos fármacos , Músculos/metabolismo , Nanopartículas/química , Polietilenoglicóis/administração & dosagem , Espécies Reativas de Oxigênio , Baço/efeitos dos fármacos , Baço/metabolismo , Distribuição TecidualRESUMO
Clear identification among early-stage cancer patients of those at highest risk of having metastatic disease would be of great benefit in treatment planning and management. Considerable additional benefit would accrue to high-risk patients if their responses to specific therapeutic alternatives could be predicted. Molecular biomarkers in the form of gene expression profiles are proving to be more effective tools for both prognostic and predictive patient stratification than more traditional methods such as patient demographics and histopathology indicators. Such biomarkers must be clinically validated before they can be effectively used to manage patients in clinical studies or clinical practice. This can be most efficiently accomplished by analyzing archived clinical samples with well-characterized clinical outcomes. Doing studies of this type requires reoptimization of traditional molecular expression profiling techniques to analyze RNA from fixed paraffin-embedded tissues. We have modified molecular methods for RNA extraction, RNA quantification, reverse transcription, and quantitative PCR to work optimally in archived clinical samples in order to develop a clinically validated assay for breast cancer prognosis and prediction of patient response to hormonal and chemotherapy.
Assuntos
Biomarcadores Tumorais/análise , Formaldeído/metabolismo , Perfilação da Expressão Gênica/métodos , Inclusão em Parafina/métodos , RNA/genética , Fixação de Tecidos/métodos , Desoxirribonuclease I/metabolismo , Endopeptidase K/metabolismo , Humanos , Reação em Cadeia da Polimerase , RNA/isolamento & purificação , RNA/metabolismo , Transcrição Reversa , SoluçõesRESUMO
The total carotenoid content and textural properties of pumpkin pieces thermally treated at 55 degrees C, 65 degrees C, 75 degrees C, 85 degrees C and 95 degrees C for different blanch times (3 min, 5 min, and 10 min) and subsequently kept for storage at 0 degrees C, -18 degrees C and -40 degrees C for a period of 80 days were measured. The total carotenoids content increased in the thermally treated pumpkin pieces, the increase being greater at 55 degrees C than at 95 degrees C. Storing the pumpkin pieces caused the total carotenoids content to decrease by 35-40% at 0 degrees C, by 15-20% at -18 degrees C and by 25-30% at -40 degrees C. A low-temperature blanch gave a firmer texture to pumpkin in comparison with high-temperature blanching. The texture of thermally treated pumpkins degraded when kept for storage, the losses being greater at 0 degrees C than at -18 degrees C and -40 degrees C. A split plot analysis was performed to study the effect of the factors on the retention of carotenoids.
Assuntos
Carotenoides/metabolismo , Cucurbita/metabolismo , Armazenamento de Alimentos/métodos , Congelamento , Frutas/metabolismo , Temperatura , Frutas/normasRESUMO
The dairy industry produces enormous amount of cheese whey containing the major milk nutrients, but this remains unutilized all over the globe. The present study investigates the production of ß-cryptoxanthin (ß-CRX) by Kocuria marina DAGII using cheese whey as substrate. Response surface methodology (RSM) and an artificial neural network (ANN) approach were implemented to obtain the maximum ß-CRX yield. Significant factors, i.e. yeast extract, peptone, cheese whey and initial pH, were the input variables in both the optimizing studies, and ß-CRX yield and biomass were taken as output variables. The ANN topology of 4-9-2 was found to be optimum when trained with a feed-forward back-propagation algorithm. Experimental values of ß-CRX yield (17.14 mg l-1) and biomass (5.35 g l-1) were compared and ANN predicted values (16.99 mg l-1 and 5.33 g l-1, respectively) were found to be more accurate compared with RSM predicted values (16.95 mg l-1 and 5.23 g l-1, respectively). Detailed kinetic analysis of cellular growth, substrate consumption and product formation revealed that growth inhibition took place at substrate concentrations higher than 12% (v/v) of cheese whey. The Han and Levenspiel model was the best fitted substrate inhibition model that described the cell growth in cheese whey with an R2 and MSE of 0.9982% and 0.00477%, respectively. The potential importance of this study lies in the development, optimization and modelling of a suitable cheese whey supplemented medium for increased ß-CRX production.
RESUMO
Various natural polymers with hydrophilic properties have been used to form hydrogels for the encapsulation and delivery of nutrients and drugs in food and pharmaceutical industries. Among them, chitosan (ChiHG)- and alginate (AlgHG)- based hydrogels have been extensively explored for delivery of several nutraceuticals in recent years. Release of natural canthaxanthin (CX) obtained from Dietzia maris NITD (accession number: HM151403) has been investigated with emphasis on biomedical applications. Significant changes (P < 0.05) in degree of swelling ( % ) and moisture content (% dry basis) were found after encapsulation of bacterial canthaxanthin (BCX), but the gel content remained unchanged. BCX encapsulation efficiency was calculated to be 55.92% and 60.45% in ChiHG and AlgHG, respectively. A noticeable change in heat of fusion ( Δ H m ) d melting point ( T m ) was recorded in ChiHG and AlgHG after BCX encapsulation. Swelling and BCX release from gel matrix was performed under two different pH (1.2 and 7.4). The results showed that swelling of hydrogel and BCX release was facilitated at higher pH (7.4) than acidic pH (1.2). With regard to the release kinetics data, it was found that BCX is released from both ChiHG and AlgHG in a non - Fickian diffusion transport method. In addition, antioxidant activity of BCX encapsulated hydrogels was found significantly higher (P < 0.001) in terms of DPPH, ABTS, nitrite, hydroxyl radical scavenging and reducing power assay. These results indicated that BCX can be successfully encapsulated into a polymeric hydrogel to obtain a dynamic biomaterial that may be used in drug delivery applications in future.