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Microplastics (MPs), small synthetic particles, have emerged as perilous chemical pollutants in aquatic habitats, causing grave concerns about their disruptive effects on ecosystems. The fauna and flora inhabiting these specific environments consume these MPs, unwittingly introducing them into the intricate web of the food chain. In this comprehensive evaluation, the current methods of identifying MPs are amalgamated and their profound impacts on marine and freshwater ecosystems are discussed. There are many potential risks associated with MPs, including the dangers of ingestion and entanglement, as well as internal injuries and digestive obstructions, both marine and freshwater organisms. In this review, the merits and limitations of diverse identification techniques are discussed, including spanning chemical analysis, thermal identification, and spectroscopic imaging such as Fourier-transform infrared spectroscopy (FTIR), Raman spectroscopy, and fluorescent microscopy. Additionally, it discusses the prevalence of MPs, the factors that affect their release into aquatic ecosystems, as well as their plausible impact on various aquatic ecosystems. Considering these disconcerting findings, it is imperative that appropriate measures should be taken to assess the potential risks of MP pollution, protect aquatic life and human health, and foster sustainable development.
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Microplásticos , Poluentes Químicos da Água , Humanos , Plásticos/análise , Ecossistema , Poluentes Químicos da Água/análise , Monitoramento Ambiental/métodosRESUMO
Recent evidence has prompted the notion of gut-microbial signatures as an indirect marker of aging and aging-associated decline in humans. However, the underlying host-symbiont molecular interactions contributing to these signatures remain poorly understood. In this study, we address this gap using cheminformatic analyses to elucidate potential gut microbial metabolites that may perturb the longevity-associated NAD+ metabolic network. In silico ADMET, KEGG interaction analysis, molecular docking, molecular dynamics simulation, and molecular mechanics calculation predict a large number of safe and bioavailable microbial metabolites to be direct and/or indirect activators of NAD+-dependent sirtuin proteins. Our simulation results suggest dihydropteroate, phenylpyruvic acid, indole-3-propionic acid, phenyllactic acid, all-trans-retinoic acid, and multiple deoxy-, methyl-, and cyclic nucleotides from intestinal microbiota as the best-performing regulators of NAD+ metabolism. Retracing these molecules to their source microorganisms also suggest commensal Escherichia, Bacteroides, Bifidobacteria, and Lactobacilli to be associated with the highest number of pro-longevity metabolites. These findings from our early-stage study, therefore, provide an informatics-based context for previous evidence in the area and grant novel insights for future clinical investigation intersecting anti-aging drug discovery, probiotics, and gut microbial signatures.
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Microbioma Gastrointestinal , Longevidade , NAD/metabolismo , Algoritmos , Simulação por Computador , Humanos , Ligantes , Simulação de Acoplamento Molecular , Simulação de Dinâmica MolecularRESUMO
Neutrophils form sticky web-like structures known as neutrophil extracellular traps (NETs) as part of innate immune response. NETs are decondensed extracellular chromatin filaments comprising nuclear and cytoplasmic proteins. NETs have been implicated in many gastrointestinal diseases including colorectal cancer (CRC). However, the regulatory mechanisms of NET formation and potential pharmacological inhibitors in the context of CRC have not been thoroughly discussed. In this review, we intend to highlight roles of NETs in CRC progression and metastasis as well as the potential of targeting NETs during colon cancer therapy.
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Neoplasias Colorretais/imunologia , Neoplasias Colorretais/patologia , Armadilhas Extracelulares/imunologia , Neutrófilos/imunologia , Neutrófilos/patologia , Animais , Progressão da Doença , Armadilhas Extracelulares/fisiologia , Humanos , Metástase Neoplásica/imunologiaRESUMO
Disease fatality associated with Ebola, SARS-CoV and dengue infections in humans is attributed to a cytokine storm that is triggered by excessive pro-inflammatory responses. Interleukin (IL)-6 acts as a mediator between pro- and anti-inflammatory reactivity by initiating trans- and classical-signaling, respectively. Hence, IL-6 is assumed to provide a target for a broad range of antiviral agents. Available immunosuppressive antivirals are directed to control an often exaggerated pro-inflammatory response that gives rise to complex clinical conditions such as lymphocytopenia. It is known that IL-6, via its soluble receptor (sIL-6R), initiates a pro-inflammatory response while an anti-inflammatory response is triggered by the membrane-bound IL-6 receptor (IL-6R). Future antivirals should thus aim to target the mechanism that regulates switching between IL-6 trans- and classical-signaling. In this review, we propose that the tumour necrosis factor-α converting enzyme ADAM-17 could be the master molecule involved in regulating IL-6 class switching and through this in controlling pro- and anti-inflammatory responses to viral antigenic stimuli. Therefore, ADAM-17 should be considered as a potential target molecule for novel antiviral drug discovery that would regulate host reactivity to infection and thereby limit or prevent fatal outcomes.
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Antivirais/farmacologia , Antivirais/uso terapêutico , Proteína ADAM17/metabolismo , Animais , Humanos , Imunossupressores/farmacologia , Imunossupressores/uso terapêutico , Inflamação/metabolismo , Interleucina-6/metabolismo , Receptores de Interleucina-6/metabolismo , Transdução de Sinais/efeitos dos fármacosRESUMO
BACKGROUND: Recent epidemiological and experimental studies suggest that cadmium and diabetes-related hyperglycemia may act synergistically to worsen metabolic regulation. The present study aims to evaluate the potential effects of Enhydra fluctuans extract in diabetes and dyslipidemia in cadmium (CdCl2) induced- normal and type 2 diabetic model rats. METHOD: Forty-eight Long-Evans rats were divided equally into the following six groups: Normal Control (N-C), Normal treated with CdCl2 (N-Cd), Normal treated with plant extract (N-P), Normal treated with both plant extract and CdCl2 (N-PCd), Diabetic treated with plant extract (DM-P) and Diabetic treated with both plant extract and CdCl2 (DM-PCd). Blood glucose and other biochemical parameters were estimated by the enzymatic colorimetric method. Histological analysis of liver and heart was done by the hematoxylin-eosin (H & E) method. RESULTS: Twenty-one days treatment of E. fluctuans extracts at a dose of 200 mg/kg significantly reduced blood glucose level in N-PCd and DM-PCd (p < 0.05), and DM-P (p < 0.01) group. The plant extract had no direct effects on total blood lipids but, it had beneficial effects on TG/HDL-C ratio in N-P and DM-PCd groups (p < 0.05). Cd induction significantly reduced body weight [(N-Cd, N-PCd, DM-PCd) (p < 0.01)], and induced liver [N-Cd (p < 0.05), N-PCd, p < 0.001] and renal impairment [N-Cd (p < 0.05)]. In bi-variate association, a significant positive correlation between serum glucose and SGPT (p < 0.05) as well as SGPT and TG/HDL ratio (p = 0.019) was found in DM-P and in the merged group. The histology of liver and heart showed severe damages including inflammation, nuclear pyknosis, loss of myocardial fibers, necrosis and fibrosis in the Cd treated groups compared to plant treated groups. CONCLUSION: E. fluctuans seems to have potent antihyperglycemic effects in diabetes and Cd toxicity along with partial antidyslipidemic properties in euglycemic and diabetic rats. Our study suggests a novel oral antihyperglycemic agent in the present environmental context.
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Asteraceae/química , Cádmio/toxicidade , Diabetes Mellitus Tipo 2/tratamento farmacológico , Hiperglicemia/tratamento farmacológico , Hipoglicemiantes/administração & dosagem , Hipolipemiantes/administração & dosagem , Extratos Vegetais/administração & dosagem , Animais , Glicemia/metabolismo , Cádmio/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Hiperglicemia/etiologia , Hiperglicemia/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Masculino , Ratos , Ratos Long-EvansRESUMO
Heritiera fomes Buch.-Ham., a mangrove plant from the Sundarbans, has adapted to a unique habitat, muddy saline water, anaerobic soil, brackish tidal activities, and high microbial competition. Endophytic fungal association protects this plant from adverse environmental conditions. This plant is used in Bangladeshi folk medicine, but it has not been extensively studied phytochemically, and there is hardly any report on investigation on endophytic fungi growing on this plant. In this study, endophytic fungi were isolated from the surface sterilized cladodes and leaves of H. fomes. The antimicrobial activities were evaluated against two Gram-positive and two Gram-negative bacteria and the fungal strain, Candida albicans. Extracts of Pestalotia sp. showed activities against all test bacterial strains, except that the ethyl acetate extract was inactive against Escherichia coli. The structures of the purified compounds, oxysporone and xylitol, were elucidated by spectroscopic means. The anti-MRSA potential of the isolated compounds were determined against various MRSA strains, that is, ATCC 25923, SA-1199B, RN4220, XU212, EMRSA-15, and EMRSA-16, with minimum inhibitory concentration values ranging from 32 to 128 µg/ml. This paper, for the first time, reports on the anti-MRSA property of oxysporone and xylitol, isolation of the endophyte Pestalotia sp. from H. fomes, and isolation of xylitol from a Pestalotia sp.
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Coriolaceae/química , Dioxanos/química , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Extratos Vegetais/química , Xilitol/química , HumanosRESUMO
Developing countries with predominantly agricultural economies are particularly vulnerable to short-term climate variability and long-term climate change adversities. This provides the context for adopting climate-smart agricultural (CSA) practices to ensure consistent agricultural product supply and food security. Although concepts of CSA tools and strategies are well-established, their adoption trends in developing regions are still undesirable, with little to no empirical understanding of factors contributing to these trends. The current research aims to address this gap of knowledge by assessing the nature of climate change impacts, adaptation strategies, and socio-economic characteristics of the farmers influencing the adoption of CSA techniques in a rural cohort in Southwestern Bangladesh. Systematically designed interview schedules revealed that the most common and devastating consequences of climate change in the study area were increased infestation of pests and diseases. Other overlooked hazards including water-logging, cyclones and cold waves, are posing new threats to regional agriculture by changing conventional patterns. Contrary to the status quo, many CSA technologies, especially indigenous approaches, are widely and effectively adopted by farmers in the studied area to cope with climate change. These adoption tendencies and innovations in CSA were found to be significantly driven by factors such as extension and training programs, education of farmers, and dependence on on-farming income. The findings of the current study provide a preliminary analysis of the attitude, knowledge and trends of CSA technology adoption by farmers in a less-saline floodplain niche of a predominantly salinity-impacted coastal region. The study acts as a model for future relevant investigations and implementations for stakeholders with supply of inputs, technologies and expertise in finding sustainable solutions and appropriate extension for communities around the world confronted with similar challenges.
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Silver nanoparticles (AgNPs) prepared by green synthesis have a lot of potentials in various fields. Among them, as an antioxidant, antibacterial agent, and nanoprobe for the colorimetric detection of mercury (Hg2+) ions is thought to be the most important. The antibacterial, antioxidant, and colorimetric sensing potential of the greenly produced AgNPs utilizing Piper chaba stem extract are all predicted in this investigation. By using the disc diffusion method, the antibacterial activity of greenly produced AgNPs are assessed, and the findings are measured from the zone of inhibition (ZOI). It is revealed that the Staphylococcus aureus, Micrococcus spp., Escherichia coli, and Pseudomonas aeruginosa bacterial strains are significantly resisted by the greenly produced AgNPs. The antioxidant activity test of AgNPs reveals a considerable impact on free radical scavenging having the inhibitory concentration (IC 50) is 1.13 mL (equivalent to 0.45 mg mL-1). Also, with a low limit of detection of 28 ppm, the resulting AgNPs are used as highly selective and economical colorimetric sensors for Hg2+ detection. The study's findings support the hypothesis that Piper chaba stems can serve as a source for the production of AgNPs with high antibacterial and antioxidant activity and usefulness for simple colorimetric readings of Hg2+.
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A metabolic network of energy-sensing molecular pathways drives the biological ageing process. Regulating certain network elements can help decelerate the ageing process and ameliorate ageing associated disorders. Bioactive phytopeptides are a prospective avenue for anti-ageing therapeutics and rejuvenation biotechnology. The present study investigates the potential of therapeutic plant peptides against cellular senescence by targeting three key proteins in the ageing network - target of rapamycin (mTOR), adenosine monophosphate-activated protein kinase (AMPK) and sirtuin 1 (SIRT1). This investigation screened a library of reported bioactive peptides using standard cheminformatic methods including in-silico ADMET, molecular docking, molecular dynamics simulation and molecular mechanics calculation. The retrieved simulation data predict 25 diverse phytopeptides as potential safe and drug-like anti-ageing biologics with half-lives >20 h and bioavailability scores >0.40. The best docked peptide, Cycloleonuripeptide B, exhibited strong binding affinity and stable complex formation with mTOR (-17.5 kCal/mol), SIRT1 (-28.54 kCal/mol) and two active sites in AMPK (-41.8 kCal/mol; -36.0 kCal/mol) during molecular dynamics simulations. The computational study acts as a foundation for future laboratory and clinical research into the potential of repurposing therapeutic phytopeptides against cellular senescence and associated pathophysiology. Communicated by Ramaswamy H. Sarma.
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Proteínas Quinases Ativadas por AMP , Sirtuína 1 , Proteínas Quinases Ativadas por AMP/metabolismo , Simulação de Acoplamento Molecular , Simulação de Dinâmica Molecular , Peptídeos/química , Peptídeos/farmacologia , Estudos Prospectivos , Sirtuína 1/metabolismo , Serina-Treonina Quinases TOR/metabolismoRESUMO
A marked increase in the global prevalence of ischemic heart disease demands focused research for novel and more effective therapeutic strategies. At present, atherosclerotic cardiovascular disease (ACVD) is the leading cause of the global incidence of heart attacks and a major contributor to many peripheral cardiac diseases. Decades of research have unearthed the complex and multidimensional pathophysiology of ACVD encompassing oxidative stress, redox imbalance, lipid peroxidation, pro-inflammatory signaling, hyperglycemic stress and diabetes mellitus, chronic low-grade inflammation and aging, immune dysregulation, vascular dysfunction, loss of hemostasis, thrombosis, and fluid shear stress. However, the scientific basis of therapeutic interventions using conventional understandings of the disease mechanisms has been subject to renewed scrutiny with novel findings in recent years. This critical review attempts to revise the pathophysiological mechanisms of atherosclerosis using a recent body of literature, with a focus on lipid metabolism and associated cellular and biochemical processes. The comprehensive study encompasses different molecular perspectives in the development and progression of coronary atherosclerosis. The review also summarizes currently prescribed small molecule therapeutics in inflammation and ACVD, and overviews prospective management measures under development including peptides and microRNA therapeutics. The study provides updated insights into the current knowledge of coronary atherosclerosis, and highlights the need for effective prevention, management and development of novel intervention approaches to overcome this chronic epidemic.
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Heritiera fomes is a mangrove plant with a rich history of ethnomedicinal usage against chronic inflammation. Biochemical analyses of H. fomes have exposed a plethora of bioactive phytochemicals that contribute to this therapeutic effect by perturbing enzymes of a complex inflammatory network mediated by arachidonic acid (AA) metabolism. This study is the first instance of utilizing cheminformatic approaches to elucidate a molecular linkage between these phytochemical interventions and the multi-enzyme AA metabolic network regulation. Analysis of the simulations reflects H. fomes as a functional reservoir of multiple safe and potent natural anti-inflammatory compounds. The investigation suggests two phytocompounds extracted from the plant: a sesquiterpene lactone and a flavone glycoside, as candidate inhibitors of multiple catalytic checkpoints of the inflammatory network. The outcomes of this research act as a primary guideline for future laboratory and clinical testing of anti-inflammatory potentials of H. fomes as an exploitable source of safe and potent drug-like molecules.Communicated by Ramaswamy H. Sarma.
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Coriolaceae , Compostos Fitoquímicos , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Ácido Araquidônico , Redes e Vias Metabólicas , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Extratos Vegetais/química , Extratos Vegetais/farmacologiaRESUMO
Cells undergo a controlled and systematic cycle of growth, replication and death. However, the integrity of this process gradually declines, leading to accumulation of senescent cells, a major hallmark of biological ageing. Dietary algae, particularly marine algae, have been long reported to exert anti-ageing benefits as cosmeceuticals and nutraceuticals with limited understanding of the molecular mechanisms underlying their activity. In this study, we have incorporated 1,202 previously reported bioactive small phycocompounds and subjected them to cheminformatic queries to assess these interactions. In-silico ADMET, 2-phase docking, metabolic pathway interaction and molecular dynamics simulations reveal multiple marine phycocompounds to have safe and effective senolytic potentials. We employed a novel deep convolutional neural network driven screening approach to identify (2R*, 3S*, 6R*, 7S*, 10R*, 13R*)-7,13-Dihydroxy-2,6-cyclo-1(9),14-xenicadiene-18,19-dial derived from Dilophus Fasciola, Laurendecumenyne A from Laurencia decumbens and 4-Bromo-3-ethyl-9-[(2E)-2-penten-4-yn-1-yl]-2,8-dioxabicyclo[5.2.1]decan-6-ol from Laurencia sp. to be potent inhibitors of multiple target senescent-cell anti-apoptotic pathway proteins. We simulated the best overall target inhibitors, specific protein inhibitors and molecular pathway regulators with each target protein and found stable interactions with minimum deviations (mean RMSD = 0.17 ± 0.01 nm) and gyrations (mean Rg = 1.64 ± 0.16 nm) of the simulated protein-compound complexes. Finally, molecular mechanics calculation suggests potent (mean ΔG = -69.56 ± 27.19 kCal/mol) and frequent hydrophobic interactions between the top performing marine phycocompounds and target proteins.
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Simulação de Dinâmica Molecular , Senoterapia , Simulação de Acoplamento MolecularRESUMO
Municipal solid waste (MSW) represents an inevitable by-product of human activity and a major crisis for communities across the globe. In recent times, the recycling of MSW has drawn attention as the process can add value through resources from the recovered waste materials and facilitates the process of circular economy. However, during the unprecedented coronavirus (COVID-19) outbreak, the risk of infection with the highly contagious virus has proven detrimental to the continuation of MSW as a valuable resource. The volume of waste, especially household waste, is higher; face masks, PPE (personal protective equipment), and hazardous materials such as batteries and empty chlorine bottles are examples of extra waste that have arisen during the pandemic. Various countries have set up initiatives for MSW management, including safety measurements for employees in the MSW management sector. The use of disinfectant prior to sorting waste, as well as storing waste for 9 days, may help to inactivate the COVID-19 virus, ensuring an appropriate safety level for MSW management. This work aimed at studying different MSW management strategies, specific challenges, and possible solutions for better understanding for those involved in waste management, in addition to providing a possible management strategy during and post-COVID-19 pandemic.
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COVID-19 , Eliminação de Resíduos , Gerenciamento de Resíduos , Humanos , Pandemias , SARS-CoV-2 , Resíduos Sólidos/análiseRESUMO
Healthcare waste comprises the waste generated by healthcare facilities, medical laboratories and biomedical research facilities. Improper treatment of this waste poses serious risks of disease transmission to waste pickers, waste workers, health workers, patients, and the community in general through exposure to infectious agents. Poor management of the waste emits harmful and deleterious contaminants into society. However, contamination of highly contagious agents such as the COVID-19 virus has created enormous instability in healthcare waste handling and subsequent recycling because of the volume of the waste generated and its contagious nature. Several countries have adopted safety measures to combat this contamination and manage healthcare waste; however, these measures are insufficient and vary depending on the context of the country. In addition, the WHO has set out guidelines for management of healthcare waste. These guidelines are helping to manage the highly contagious healthcare waste resulting from the current pandemic. Proper healthcare waste management may add value by reducing the spread of the COVID-19 virus and increasing the recyclability of materials instead of sending them to landfill. Disinfecting and sorting out healthcare waste facilitates sustainable management and allows their utilization for valuable purposes. This review discusses the different healthcare solid waste management strategies practiced in different countries, the challenges faced during this management, and the possible solutions for overcoming these challenges. It also provides useful insights into healthcare solid waste management scenarios during the COVID-19 pandemic and a possible way forward.
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COVID-19 , Eliminação de Resíduos , Gerenciamento de Resíduos , Atenção à Saúde , Humanos , Pandemias , Reciclagem , SARS-CoV-2 , Resíduos Sólidos/análiseRESUMO
The process of biological aging or senescence refers to the gradual loss of homeostasis and subsequent loss of function - leading to higher chances of mortality. Many mechanisms and driving forces have been suggested to facilitate the evolution of a molecular circuit acting as a trade-off between survival and proliferation, resulting in senescence. A major observation on biological aging and longevity in humans and model organisms is the prevalence of significant sexual divergence in the onset, mechanisms and effects of aging associated processes. In the current account, we describe possible mechanisms by which aging, sex and reproduction are evolutionarily intertwined in order to maintain systemic energy homeostasis. We also interrogate existing literature on the sexual dimorphism of genetic, cellular, metabolic, endocrine and epigenetic processes driving cellular and systemic aging. Subsequently, based on available evidence, we propose a hypothetic model of sex-limited decoupling of female longevity from sirtuins, a major family of regulator proteins of the survival-proliferation trade-off. We also provide necessary considerations to be made in order to test the hypothesis and explore the physiological and therapeutic implications of this decoupling event in male and female longevity after reaching reproductive maturity. HYPOTHESIS STATEMENT: Sirtuins provide survival benefits in a sex-nonspecific manner but the dependency on sirtuins in driving metabolic networks after reaching reproductive maturity is evolutionarily decoupled from female longevity.
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Envelhecimento/genética , Envelhecimento/fisiologia , Metabolismo Energético/fisiologia , Caracteres Sexuais , Sirtuínas/metabolismo , Animais , Evolução Biológica , Feminino , Masculino , Sirtuínas/genéticaRESUMO
Phytol (PYT) is a diterpene member of the long-chain unsaturated acyclic alcohols. PYT and some of its derivatives, including phytanic acid (PA), exert a wide range of biological effects. PYT is a valuable essential oil (EO) used as a fragrance and a potential candidate for a broad range of applications in the pharmaceutical and biotechnological industry. There is ample evidence that PA may play a crucial role in the development of pathophysiological states. Focusing on PYT and some of its most relevant derivatives, here we present a systematic review of reported biological activities, along with their underlying mechanism of action. Recent investigations with PYT demonstrated anxiolytic, metabolism-modulating, cytotoxic, antioxidant, autophagy- and apoptosis-inducing, antinociceptive, anti-inflammatory, immune-modulating, and antimicrobial effects. PPARs- and NF-κB-mediated activities are also discussed as mechanisms responsible for some of the bioactivities of PYT. The overall goal of this review is to discuss recent findings pertaining to PYT biological activities and its possible applications.
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Óleos Voláteis/farmacologia , Fitol/farmacologia , Óleos de Plantas/farmacologia , Adjuvantes Imunológicos/farmacologia , Analgésicos/farmacologia , Animais , Ansiolíticos/farmacologia , Anti-Infecciosos/farmacologia , Anti-Inflamatórios/farmacologia , Anticonvulsivantes/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Biotecnologia , Indústria Farmacêutica , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Testes de Sensibilidade Microbiana , Receptores Ativados por Proliferador de Peroxissomo/efeitos dos fármacosRESUMO
The successful construction of an immunosensor depends on having an effective procedure for immobilising the bio-recognition element to the transducer surface. In the present study, an amino-terminated 4-aminothiophenol (ATP) self-assembled monolayer (SAM) was modified with heterobifunctional crosslinker sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate to couple reduced anti-myoglobin half-antibody fragments. The disulphide groups present in the hinge region of IgG molecules were selectively cleaved by 2-mercaptoethylamine to produce reduced half-antibody fragments with free sulphydryl groups. The maleimide terminated 4-ATP SAM modified surface was coupled to these reduced antibody fragments to produce highly oriented immobilization of the half-antibody via its Fc domain and to allow free access to the Fv bindings sites. This represents an improvement by comparison with biotin/avidin mediated IgG attachment which is essentially randomly oriented. Functional immunosensors were able to detect myoglobin in both phosphate buffered saline and whole serum over the range of concentrations from 10(-13)M to 10(-6)M, and order of magnitude better than avidin/biotin linked immunosensors. In addition, atomic force microscopy (AFM) was carried out to elucidate the nanotopology of the immunosensor surface at different stages of fabrication; the images demonstrate that half antibodies bind as described and show structural changes on subsequent antigen binding.