RESUMEN
Gene therapy is a transfectant method for the treatment of hereditary disease, which transfers the gene mutation into the cells. In the view of the high prospects of utilization of cationic gemini surfactants as a non-viral vector for the gene transfection, we have made a comprehensive study on the interactions between a recently synthesized series of ester-functionalized cationic Cm-E2O-Cm gemini surfactants (m = 12, 14 and 16) with calf thymus deoxyribonucleic acid (ctDNA) utilizing various techniques. The micellization behavior of gemini surfactants has been altered in the presence of ctDNA. A series of measurements (fluorescence, UV-vis and time-resolved fluorescence) show that the quenching of ctDNA proceeds by a static mechanism. The competitive displacement studies (EB, AO and HO), KI quenching analysis, CD studies and viscosity measurements suggested intercalative binding mode in a stoichiometry ratio of 1:1 with the Kb (binding constant) order being: C16-E2O-C16 > C14-E2O-C14 > C12-E2O-C12. The thermodynamic parameters show that the geminis interacted with ctDNA spontaneously through ionic/electrostatic interactions. Furthermore, the theoretical approaches offer accurate insights about the binding of gemini surfactants with DNA, and are in consistence with the experimental results.
Asunto(s)
ADN/química , Teoría Funcional de la Densidad , Ésteres/química , Tensoactivos/química , Animales , Cationes/química , Bovinos , Dicroismo Circular , Estructura Molecular , Termodinámica , ViscosidadRESUMEN
Anaerobic Ammonium Oxidation ("anammox") is a bacterial process in which nitrite and ammonium are converted into nitrogen gas and water, yielding energy for the cell. Anammox is an important branch of the global biological nitrogen cycle, being responsible for up to 50% of the yearly nitrogen removal from the oceans. Strikingly, the anammox process uniquely relies on the extremely reactive and toxic compound hydrazine as a free intermediate. Given its global importance and biochemical novelty, there is considerable interest in the enzymes at the heart of the anammox pathway. Unfortunately, obtaining these enzymes in sufficiently large amounts for biochemical and structural studies is problematic, given the slow growth of pure cultures of anammox bacteria when high cell densities are required. However, the anammox process is being applied in wastewater treatment to remove nitrogenous waste in processes like DEamMONification (DEMON). In plants using such processes, which rely on a combination of aerobic ammonia-oxidizers and anammox organisms, kilogram amounts of anammox bacteria-containing sludge are readily available. Here, we report a protein isolation protocol starting from anammox cells present in DEMON sludge from a wastewater treatment plan that readily yields pure preparations of key anammox proteins in the tens of milligrams, including hydrazine synthase HZS and hydrazine dehydrogenase (HDH), as well as hydroxylamine oxidoreductase (HAO). HDH and HAO were active and of sufficient quality for biochemical studies and for HAO, the crystal structure could be determined. The method presented here provides a viable way to obtain materials for the study of proteins not only from the central anammox metabolism but also for the study of other exciting aspects of anammox bacteria, such as for example, their unusual ladderane lipids.
Asunto(s)
Oxidación Anaeróbica del Amoníaco , Bacterias/enzimología , Proteínas Bacterianas/metabolismo , Reactores Biológicos/microbiología , Complejos Multienzimáticos/metabolismo , Aguas del Alcantarillado/microbiología , Compuestos de Amonio/metabolismo , Bacterias/clasificación , Bacterias/genética , Proteínas Bacterianas/química , Proteínas Bacterianas/aislamiento & purificación , Cristalografía por Rayos X , Hidrazinas/metabolismo , Cinética , Complejos Multienzimáticos/química , Complejos Multienzimáticos/aislamiento & purificación , Nitritos/metabolismo , Nitrógeno/metabolismo , Nitrosomonas/clasificación , Nitrosomonas/genética , Oxidación-Reducción , Oxidorreductasas/química , Oxidorreductasas/aislamiento & purificación , Oxidorreductasas/metabolismo , FilogeniaRESUMEN
Anaerobic ammonium oxidation (anammox) is a microbial process responsible for significant nitrogen loss from the oceans and other ecosystems. The redox reactions at the heart of anammox are catalyzed by large multiheme enzyme complexes that rely on small cytochrome c proteins for electron shuttling. Among the most highly abundant of these cytochromes is a unique heterodimeric complex composed of class I and class II c-type cytochromes called NaxLS, which has distinctive biochemical and spectroscopic properties. Here, we present the 1.7 Å resolution crystal structure of this complex from the anammox organism Kuenenia stuttgartiensis (KsNaxLS). The structure reveals that the heme irons in each subunit exhibit a rare His/Cys ligation, which, as we show by substitution, causes the observed unusual spectral properties. Unlike its individual subunits, the KsNaxLS complex binds nitric oxide (NO) only at the distal heme side, forming 6cNO adducts. This is likely due to steric immobilization of the proximal heme-binding motifs upon complex formation, a finding that may be of functional relevance, because NO is an intermediate in the central anammox metabolism. Pulldown experiments with K. stuttgartiensis cell-free extract showed that the KsNaxLS complex binds specifically to one of the central anammox enzyme complexes, hydrazine synthase, which uses NO as one of its substrates. It is therefore possible that the KsNaxLS complex plays a role in binding the volatile NO to retain it in the cell for transfer to hydrazine synthase. Alternatively, we propose that KsNaxLS may shuttle electrons to this enzyme complex.
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Bacterias/metabolismo , Proteínas Bacterianas/metabolismo , Citocromos c/metabolismo , Óxido Nítrico/metabolismo , Oxidorreductasas/metabolismo , Secuencias de Aminoácidos , Proteínas Bacterianas/química , Sitios de Unión , Monóxido de Carbono/química , Monóxido de Carbono/metabolismo , Cristalografía por Rayos X , Citocromos c/química , Citocromos c/genética , Dimerización , Simulación de Dinámica Molecular , Mutagénesis , Óxido Nítrico/química , Oxidación-Reducción , Oxidorreductasas/química , Estructura Terciaria de Proteína , Subunidades de Proteína/metabolismoRESUMEN
OBJECTIVE: The current study aimed to delve into the comparative clinical outcomes between external beam radiation therapy (EBRT) and sequential High Dose Rate Intracavitary Brachytherapy (HDRICBT) with or without concurrent cisplatin administration on the day of intracavitary brachytherapy (ICBT) insertion in women with locally advanced cervical cancer. METHODS: In this study, conducted between January 2017 and July 2018 at a leading institute in India, diagnosed and untreated patients of locally advanced carcinoma cervix were randomized into two groups. Arm 1 received concurrent cisplatin before each course of brachytherapy, while Arm 2 underwent brachytherapy alone. The outcomes were compared in terms of acute and late toxicities, treatment response, and follow-up. Data analysis was performed using SPSS 16, with statistical significance set at p < 0.05. RESULTS: Both study arms showed similar complete response (CR) rates of 73.3%, with no significant advantage of concurrent cisplatin before brachytherapy. However, a noteworthy trend emerged during follow-up. In the concurrent cisplatin group, the CR rate increased from 73.3% post 1 month of brachytherapy to 86.7% at 3 months and 83.3% at 6 months. Contrastingly, the control group showed CR rates of 73.3% post 1 month, 80% at 3 months, and 76.6% at 6 months. While not statistically significant, this observation suggests a possible enhancement in response rates with concurrent cisplatin and ICBT. CONCLUSIONS: Future studies focusing on the optimal drug, dosage, scheduling, and combining cisplatin with other agents are recommended to further explore the potential benefits observed in this study.
Asunto(s)
Braquiterapia , Quimioradioterapia , Cisplatino , Centros de Atención Terciaria , Neoplasias del Cuello Uterino , Humanos , Neoplasias del Cuello Uterino/radioterapia , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/terapia , Neoplasias del Cuello Uterino/tratamiento farmacológico , Femenino , Braquiterapia/métodos , Cisplatino/administración & dosificación , Cisplatino/uso terapéutico , India , Persona de Mediana Edad , Quimioradioterapia/métodos , Estudios de Seguimiento , Dosificación Radioterapéutica , Adulto , Antineoplásicos/uso terapéutico , Pronóstico , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/radioterapia , Carcinoma de Células Escamosas/terapia , Resultado del Tratamiento , Terapia CombinadaRESUMEN
Correction for 'Probiotic and prebiotic supplementation ameliorates chronic restraint stress-induced male reproductive dysfunction' by Mohd Akram et al., Food Funct., 2023, 14, 8558-8574, https://doi.org/10.1039/D3FO03153E.
RESUMEN
The implications of surfactant-enzyme/protein interactions in a variety of fields, including biotechnology, cosmetics, paints and pharmaceuticals, have attracted a lot of attention in contemporary studies. Herein, we have employed several in vitro and in silico techniques such as excitation and absorption spectroscopies, circular dichroism and FT-IR spectroscopies, density functional and molecular dynamics simulations to understand the interaction behavior of oxy-diester-based green cationic Gemini surfactants, N1,N1,N14,N14-tetramethyl-2,13-dioxo-N1,N14-dialkyl-3,6,12-tetraoxateradecane-1,14-diaminiumdichloride (abbreviated as Cm-E2O2-Cm, where 'm' stands for alkyl chain length, m = 12 and 14) with one of the main digestive proteins, pepsin. The spectroscopic techniques confirm the static quenching effect of surfactants on pepsin. The calculated physical parameters (Ksv, Kb and ΔG) and their order reveal the distinguished implications for the surfactants' chain lengths. The spontaneity of interaction was also confirmed by negative Gibbs free energy change values. The extrinsic spectroscopic study with pyrene as fluorescence probe, FT-IR and CD techniques indicated a potential conformational change in pepsin induced by the Gemini surfactants. DFT, docking and MD simulations provided the theoretical understanding regarding the quantum mechanical environment, location of binding and stability of the protein-surfactant complexation in energy terms. We believe this study will be a humble addition to our existing knowledge in the field of protein-surfactant interactions.Communicated by Ramaswamy H. Sarma.
Asunto(s)
Ésteres , Pepsina A , Espectroscopía Infrarroja por Transformada de Fourier , Ésteres/química , Dicroismo Circular , Tensoactivos/químicaRESUMEN
Restraint stress (RS) can induce male reproductive deficits by activating the hypothalamic-pituitary-adrenal (HPA) axis and causing oxidative stress. Previous studies have shown that probiotics can alleviate neurological and metabolic disorders induced by stress. However, the effects of probiotics on RS-induced reproductive deficits have not been fully elucidated. This study aimed to investigate whether Lactobacillus rhamnosus NCDC-610 (Probiotic-1) and Lactobacillus fermentum NCDC-400 (Probiotic-2) with prebiotic (fructooligosaccharides (FOS)) could prevent RS-induced reproductive deficits. C57BL6/J mice were subjected to RS for four hours daily before oral administration of probiotics (4 × 109 CFU per mice) either separately or concurrently with FOS. The results showed that oral administration of Probiotic-1 and Probiotic-2 protected against RS-induced sperm deficits, including sperm count, motility, morphology, and histopathology of testes, and improved intestinal health. Furthermore, Probiotic-1 and Probiotic-2 prevented RS-induced changes in testosterone levels by up-regulating the expressions of steroidogenic acute regulatory protein (StAR), cytochrome P450 side-chain cleavage enzyme (P450scc), and 17ß-hydroxysteroid dehydrogenase (17ßHSD) in the testes. Additionally, Probiotic-1 and Probiotic-2 increased the activities of catalase and superoxide dismutase and reduced the fold change of interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-alpha (TNF-α), indicating a protective effect against RS-induced oxidative stress. Oral administration of Probiotic-1 and Probiotic-2, either separately or concurrently with FOS (probiotic dose of 4 × 109 CFU per mice and prebiotic 5% w/v), prevented RS-induced activation of the HPA axis and improved male fertility. These findings suggest that L. rhamnosus NCDC-610 and L. fermentum NCDC-400 are safe and effective probiotics for mitigating stress-induced male reproductive deficits.
Asunto(s)
Prebióticos , Probióticos , Masculino , Animales , Ratones , Sistema Hipotálamo-Hipofisario , Sistema Hipófiso-Suprarrenal , Semen , Ratones Endogámicos C57BLRESUMEN
The primary objective of this study was to investigate the protective effects of ropinirole (ROP) medication given for an extended period following the induction of cognitive decline, oxidative stress, and deterioration of mitochondria in a Wistar rat model by Aß1-42 . This study aimed to examine the neuroprotective efficacy of ROP in a stereotaxis model of AD. The Wistar rats were randomly assigned into four groups. Group I was considered as a sham, group II served as Aß-infusion alone, Group III was Aß1-42 + ROP (5 mg/kg/i.p.), and Group IV was Aß1-42 + ROP (10 mg/kg/i.p.). Our research revealed that ROP (10 mg/kg, b.wt.) attenuates the cognitive deficits caused by Aß1-42 -infused, which also correlates with the barnes maze, where (10 mg/kg, b.w.t.) shows significant improvement in spatial learning and memory. At the same time, ROP was rescued from oxidative damage, decreased lipid peroxidation rates, and inhibited acetylcholinesterase activity caused, demonstrating antioxidant benefits. In addition, a higher dose of ROP restored mitochondrial membrane potential in Aß1-42 rats. Furthermore, histopathological examination showed that ROP treatment reduced neuronal loss, especially in the hippocampus. We conclude that ROP's protective effects in reducing oxidative stress and modulating mitochondrial function might have a propensity in AD pathogenesis.
Asunto(s)
Disfunción Cognitiva , Fármacos Neuroprotectores , Ratas , Animales , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Acetilcolinesterasa/metabolismo , Acetilcolinesterasa/farmacología , Acetilcolinesterasa/uso terapéutico , Roedores/metabolismo , Aprendizaje por Laberinto , Ratas Wistar , Disfunción Cognitiva/inducido químicamente , Disfunción Cognitiva/tratamiento farmacológico , Estrés Oxidativo , Modelos Animales de EnfermedadRESUMEN
Probiotics have been suggested as alternatives to pharmacological drugs in the treatment of a variety of medical problems, including obesity management, which is often linked to low sperm production. Also, probiotic fermented products are known to boost host immune response, immunosenescence, infection tolerance, and redox homeostasis, but their direct role in male fertility has been less investigated. This study assessed the effect of two probiotic strains, L. fermentum NCDC 400 and L. rhamnosus NCDC 610, and fructooligosaccharide (FOS) fermented milk supplementation. We identified the significantly reduced oxidative stress markers in the plasma and liver of HF diet-fed animals. We determined the role of key testicular enzymes of steroidogenic pathway genes StAR, P450scc, and 17ßHSD in maintaining the testosterone concentration and restoring testicular structures. In conclusion, the present work illustrated the ability of both probiotics L. fermentum NCDC 400 and L. rhamnosus NCDC 610 as regulatory agents with beneficial effects on weight loss and endogenous testosterone with substantially improved sperm motility in male diet-induced obesity (DIO) models. Our findings indicate that fermented milk supplementation may be an alternative treatment for preventing obesity and other related metabolic syndromes.
Asunto(s)
Productos Lácteos Cultivados , Microbioma Gastrointestinal/fisiología , Probióticos , Motilidad Espermática/fisiología , Animales , Dieta Alta en Grasa , Masculino , Ratones , Ratones Endogámicos C57BL , Obesidad , Salud ReproductivaRESUMEN
Oxidative damage and mitochondrial dysfunction are two prominent pathological features and gradually understood as important pathogenic events for neurodegenerative diseases, including aging and Alzheimer's disease (AD). The present study was aimed to explore the prolonged treatment of pramipexole (PPX) following amyloid beta (Aß1-42)-induced cognitive impairments , oxidative stress, and mitochondrial dysfunction in a Wistar rat model. We have found that PPX (1.0 mg/kg, b.wt.) improves cognitive impairments of Aß1-42-infused rats in Morris water maze. At the same time, PPX attenuated Aß1-42-induced oxidative damage and increased reduced-glutathione content level, decreased lipid peroxidation rate and suppressed the activity of acetylcholinesterase and shows antioxidant effects. Additionally, PPX treatment has shown inhibition of mitochondrial reactive oxygen species production and restored mitochondrial membrane potential, oxidative phosphorylation, and enhanced ATP levels in Aß1-42 rats. Furthermore, PPX treatment reduced bioenergetics loss and dynamics alterations by upregulating PGC-1α protein level and mitigating translocation of Bax and Drp-1 to mitochondria and cytochrome-c release into the cytoplasm. PPX also increased mitofusin-2 protein expression, a basic element of mitochondrial fusion process. We conclude that remedial role of PPX in mitigating oxidative damage and mitochondrial perturbation that are modulated in Aß1-42 rats may have the propensity in AD pathogenesis.
Asunto(s)
Enfermedad de Alzheimer , Péptidos beta-Amiloides , Acetilcolinesterasa/metabolismo , Adenosina Trifosfato/metabolismo , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Animales , Antioxidantes/farmacología , Biomarcadores/metabolismo , Citocromos/metabolismo , Citocromos/farmacología , Glutatión/metabolismo , Hipocampo , Aprendizaje por Laberinto , Trastornos de la Memoria/inducido químicamente , Trastornos de la Memoria/tratamiento farmacológico , Trastornos de la Memoria/metabolismo , Mitocondrias , Estrés Oxidativo , Fragmentos de Péptidos , Pramipexol/efectos adversos , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Proteína X Asociada a bcl-2/metabolismoRESUMEN
Agroforestry, an integration of farming system with woody perennials leads to the generation of potential agroforestry residues. The conventional treatment of agroforestry waste includes landfilling, thermal management, and decomposition which is accompanied with their own share of disadvantages. The ample amount of residues and products needs effective management to reap the economic and environmental benefits. The channel of waste collection, transportation, and recycle or valorization into products like biofuel, fertilizers, biochar, industrial chemicals is essential to maintain a circular sustainable bioeconomy. Global market value of biowaste to bioenergy (BtB) technology is roughly US $25.32 billion and is projected to enhance to US $40 billion by 2023. Employment of an appropriate pretreatment technology such as fermentation, hydrolysis, gasification etc. is going to elevate the degree of valorization along with surpassing the mobilization barrier. The sustainability assessment of the management process can be achieved with multiple models including technoeconomic analysis, life cycle assessment and multi criteria approach which are dependent on both hard and soft indices. Additionally, the loopholes of the agroforestry sectors would be managed by the introduction of appropriate policies which are undertaken globally by the Orlando and Lugo declarations, food and agriculture organization, Millennium Development Goals, Global Research Alliance and Guidelines for Sustainable Agriculture and Rural Development. The present review envisaged the agroforestry waste management strategy and its sustainability assessment primarily based upon Social, Economic and Environmental parameters without tormenting the future generations.
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Agricultura Forestal , Administración de Residuos , Agricultura , Biocombustibles , ReciclajeRESUMEN
Anaerobic ammonium oxidation (anammox) is a bacterial process in which ammonium and nitrite are combined into dinitrogen gas and water, yielding energy for the cell. This process relies on a series of redox reactions catalyzed by a set of enzymes, with electrons being shuttled to and from these enzymes, likely by small cytochrome c proteins. For this system to work productively, these electron carriers require a degree of specificity toward the various possible redox partners they encounter in the cell. Here, we compare two cytochrome c proteins from the anammox model organism Kuenenia stuttgartiensis. We show that they are highly homologous, are expressed at comparable levels, share the same fold, and display highly similar redox potentials, yet one of them accepts electrons from the metabolic enzyme hydroxylamine oxidase (HAO) efficiently, whereas the other does not. An analysis of the crystal structures supplemented by Monte Carlo simulations of the transient redox interactions suggests that this difference is at least partly due to the electrostatic field surrounding the proteins, illustrating one way in which the electron carriers in anammox could attain the required specificity. Moreover, the simulations suggest a different "outlet" for electrons on HAO than has traditionally been assumed.
RESUMEN
To study the interaction of the series of pyridinium based gemini surfactants (GS) (referred to as m-Py-m, m = 14, 16); 4,4'-(propane-1,3-diyl)bis(1-(2-(tetradecyloxy)-2-oxoethyl) dipyridinium chloride (14-Py-14), and 4,4'-(propane-1,3-diyl) bis(1-(2-(hexadecyloxy)-2-oxoethyl)dipyridinium chloride (16-Py-16) with bovine serum albumin (BSA), various physicochemical and spectroscopic tools such as tensiometry, steady-state fluorescence, synchronous fluorescence, pyrene fluorescence, UV-visible, far-UV circular dichroism (CD) were utilized at physiological pH (7.4) and 298 K in combination with computational molecular modeling analysis. The tensiometric results show significant modifications in interfacial and thermodynamic parameters for m-Py-m GS upon BSA combination, deciphering the gemini surfactant-BSA interaction. Steady-state fluorescence analysis evaluates the structural alterations of BSA with the addition of m-Py-m GS. The plots of Stern-Volmer, modified Stern-Volmer, and thermodynamic parameters were used to determine the binding type of m-Py-m GS to BSA. The synchronous fluorescence spectra state a mild effect of gemini surfactants on the emission intensity of tyrosine (Tyr) residues, on the other hand, tryptophan (Trp) residues showed a significant effect. Post addition of GS, the plot of pyrene fluorescence reveals the mild micropolarity fluctuations via the probe (pyrene) molecules encapsulated in BSA. UV-visible experiments support the complex formation between the BSA and m-Py-m GS. Far-UV CD measurements revealed the modifications in the secondary structure of protein produced by m-Py-m GS. Furthermore, we also used the computational molecular modeling for attaining deep insight into BSA and m-Py-m GS binding and the results are supported with our experimental results.
Asunto(s)
Albúmina Sérica Bovina , Tensoactivos , Sitios de Unión , Dicroismo Circular , Unión Proteica , Estructura Secundaria de Proteína , Albúmina Sérica Bovina/metabolismo , Espectrometría de Fluorescencia , TermodinámicaRESUMEN
The fluoroquinolone antibiotic drug namely ciprofloxacin hydrochloride (CFH) is widely prescribed for the treatment of different bacterial infections. The interaction of CFH with a synthetic polymer, polyvinyl pyrrolidone (PVP), and biopolymer, bovine serum albumin (BSA) was studied by UVvisible and fluorescence spectroscopic methods at different temperatures. The binding constant (K b ) for the CFH-PVP complex was determined from the Benesi-Hildebrand plot. PVP of different molecular weights (MW) (such as 24,000, 40,000, 360,000, and 700,000 g. mole-1) were used for the interaction between CFH and PVP. The gradual increase in K b value and the complexation reaction was found to be much enhanced with the augmentation of the MW of PVP. The values of K b were also found to be increased with increasing temperatures as well as with the increase of electrolyte/acetic acid concentration. The Gibbs free energy of binding (∆G 0) values of the interaction process was negative which indicates the complex formation is thermodynamically spontaneous. The positive values of enthalpy (∆H 0) and entropy (∆S 0) of binding connote that the binding force for CFH-PVP complexation is hydrophobic in nature and the complexation is entropy controlled. The negative intrinsic enthalpy (∆H *,0) values indicate the high stability of CFH-PVP complexes. Molecular docking calculation discloses the existence of similar binding forces between CFH and PVP obtained by the analysis of experimental data from UV-visible spectroscopic method. The binding constant between CFH and BSA (K b ), quenching constant (K sv ), the number of binding sites (n), and the quenching rate constant (K q ) for the CFH-BSA system were also calculated. The values of K sv , K q , and n for the CFH-BSA system are lower in 0.05 mol L-1 urea solution and higher in PVP solutions compared to those of aqueous medium.
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Antibacterianos/química , Biopolímeros/química , Ciprofloxacina/química , Simulación del Acoplamiento Molecular/métodos , Povidona/análogos & derivados , Albúmina Sérica Bovina/química , Análisis Espectral/métodos , Temperatura , Ácido Acético/química , Interacciones Farmacológicas , Electrólitos/química , Peso Molecular , Povidona/química , Soluciones , TermodinámicaRESUMEN
Cytoplasmic male sterility(CMS), a maternally inherited trait, provides a promising means to harness yield gains associated with hybrid vigor. In pigeonpea [Cajanus cajan (L.) Huth], nine types of sterility-inducing cytoplasm have been reported, of which A2 and A4 have been successfully deployed in hybrid breeding. Unfortunately, molecular mechanism of the CMS trait is poorly understood because of limited research invested. More recently, an association between a mitochondrial gene (nad7) and A4 -CMS has been demonstrated in pigeonpea; however, the mechanism underlying A2 -CMS still remains obscure. The current investigation aimed to analyze the differences in A2 -CMS line (ICPL 88039A) and its isogenic maintainer line (ICPL 88039B) at transcriptome level using next-generation sequencing. Gene expression profiling uncovered a set of 505 genes that showed altered expression in response to CMS, of which, 412 genes were upregulated while 93 were downregulated in the fertile maintainer line vs. the CMS line. Further, gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction (PPI) network analyses revealed association of CMS in pigeonpea with four major pathways: glucose and lipid metabolism, ATP production, pollen development and pollen tube growth, and reactive oxygen species (ROS) scavenging. Patterns of digital gene expression were confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) of six candidate genes. This study elucidates candidate genes and metabolic pathways having potential associations with pollen development and male sterility in pigeonpea A2 -CMS. New insights on molecular mechanism of CMS trait in pigeonpea will be helpful to accelerate heterosis utilization for enhancing productivity gains in pigeonpea.
Asunto(s)
Infertilidad Masculina , Infertilidad Vegetal , Citoplasma , Infertilidad Masculina/metabolismo , Fitomejoramiento , Infertilidad Vegetal/genética , TranscriptomaRESUMEN
Nitrate is an abundant nutrient and electron acceptor throughout Earth's biosphere. Virtually all nitrate in nature is produced by the oxidation of nitrite by the nitrite oxidoreductase (NXR) multiprotein complex. NXR is a crucial enzyme in the global biological nitrogen cycle, and is found in nitrite-oxidizing bacteria (including comammox organisms), which generate the bulk of the nitrate in the environment, and in anaerobic ammonium-oxidizing (anammox) bacteria which produce half of the dinitrogen gas in our atmosphere. However, despite its central role in biology and decades of intense study, no structural information on NXR is available. Here, we present a structural and biochemical analysis of the NXR from the anammox bacterium Kuenenia stuttgartiensis, integrating X-ray crystallography, cryo-electron tomography, helical reconstruction cryo-electron microscopy, interaction and reconstitution studies and enzyme kinetics. We find that NXR catalyses both nitrite oxidation and nitrate reduction, and show that in the cell, NXR is arranged in tubules several hundred nanometres long. We reveal the tubule architecture and show that tubule formation is induced by a previously unidentified, haem-containing subunit, NXR-T. The results also reveal unexpected features in the active site of the enzyme, an unusual cofactor coordination in the protein's electron transport chain, and elucidate the electron transfer pathways within the complex.
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Bacterias/enzimología , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Oxidorreductasas/química , Oxidorreductasas/metabolismo , Bacterias/química , Bacterias/genética , Proteínas Bacterianas/genética , Dominio Catalítico , Microscopía por Crioelectrón , Cristalografía por Rayos X , Cinética , Complejos Multiproteicos/química , Complejos Multiproteicos/genética , Complejos Multiproteicos/metabolismo , Nitratos/metabolismo , Nitritos/metabolismo , Oxidación-Reducción , Oxidorreductasas/genéticaRESUMEN
This paper presents the results of a kinetic study performed between ninhydrin and a Ni(II) dipeptide complex under various conditions. The rate of formation of the imine adduct was measured spectrophotometrically both in plain aqueous media and in aqueous micellar media in which CTAB (cetyltrimethylammonium bromide) is used as the surfactant. These studies were carried out at pH 5 and over a temperature a range of 50 to 70°C. Studies were also conducted to elucidate the effect of some organic sodium salts on the rate of this reaction. In these studies, it was found that the formation of imine adduct followed a first-order kinetics with respect to [Ni(II)-Gly-Leu]+ in both aqueous and micellar medium. A fractional-order kinetics was observed with respect to ninhydrin, again in both aqueous and micellar media. Increase in the total concentration of CTAB from 0 to 40×10-3 mol dm-3 resulted in approximately two folds increase in the pseudo-first-order rate constant (kψ). The rate constant (kΨ) in micellar medium first increased with increase in CTAB concentration, reached a maximum value, and finally, with further increase in CTAB concentration, a decreasing effect was observed. Quantitative kinetic analysis of kψ-[CTAB] data was performed on the basis of the pseudo-phase model of the micelles. The rate profile in presence of CTAB suggests a cooperative effect in the enhanced formation of the imine adduct as is commonly found in enzyme catalyzed reactions. Addition of organic sodium salts (such as benzoate, salicylate and tosylate) enhanced the rate at lower concentrations but rates start to decrease at higher concentrations. This suggests that tightly binding organic counter-anions were the most effective. Viscosity of the reaction media seems to affect the kinetic behavior in micellar media.
Asunto(s)
Cetrimonio/química , Dipéptidos/química , Iminas/química , Metales/química , Ninhidrina/química , Tensoactivos/química , Aniones/química , Benzoatos/química , Catálisis , Concentración de Iones de Hidrógeno , Micelas , Fenómenos Químicos Orgánicos , Salicilatos/química , Espectrofotometría , Temperatura , Viscosidad , Agua/químicaRESUMEN
Accumulation of different protein-surfactant mixtures affords further knowledge about the structure-property interactions of biomacromolecules. They will help design suitable surfactants, which, in turn, can enhance the utilization of protein-surfactant complexes in biotechnologies, cosmetics, and food industry realms. Owing to their adaptable and remarkably notable properties, we are describing herein the interaction of C m -E2O-C m gemini surfactants (m = 12, 14, and 16) with α-CHT by employing various spectroscopic techniques including with molecular docking and density functional theory (DFT) method. Results have revealed complex formation, unfolding, and a static quenching mechanism in the interaction of gemini surfactants with α-CHT. The Stern-Volmer constant (K SV), quenching constant (k q), the number of binding sites (n), and binding constant (K b) were interrogated by utilizing the fluorescence quenching method, UV-vis, synchronous, 3-D, and resonance Rayleigh scattering fluorescence studies. The data perceive the α-CHT-C m -E2O-C m complex formation along with conformational alterations induced in α-CHT. The contribution of aromatic residues to a nonpolar environment is illustrated by pyrene fluorescence. Fourier transform infrared spectroscopy and circular dichroism outcomes reveal conformational modifications in the secondary structure of α-CHT with the permutation of gemini surfactants. The computational calculations (molecular docking and DFT) further corroborate the complex formation between α-CHT and C m -E2O-C m gemini surfactants and the contribution of electrostatic/hydrophobic interaction forces therein.
RESUMEN
Glutamatergic N-methyl-D-aspartate (NMDA) receptors have critical roles in several neurological and psychiatric diseases. Dizocilpine (MK-801) is a ligand at phencyclidine recognition sites that is associated with NMDA receptor-coupled cation channels, where it acts as a potent noncompetitive antagonist of central glutamate receptors. In this study, we investigate the effect of clozapine on MK-801-induced neurochemical and neurobehavioral alterations in the prefrontal cortex of mice. Acute administration of NMDA noncompetitive antagonist MK-801 impairs motor coordination, grip strength, and locomotor activity. Clozapine is the only medication that is indicated for treating refractory schizophrenia, due to its superior efficacy among all antipsychotic agents; however, its mechanism is not well understood. To understand its mechanism, we investigated the effects of clozapine on motor coordination, locomotor activity, and grip strength in mice against the NMDA receptor antagonist MK-801. MK-801 induced elevations in acetylcholinesterase (AChE) activity, monoamine oxidase (MAO) activity, and c-fos expression. The administration of clozapine inhibited the effects caused by MK-801 (0.2 mg/kg body weight). Motor coordination and grip strength paradigms that had been altered by MK-801 were restored by clozapine. Moreover, clozapine also ameliorated MK-801-induced elevation in AChE and MAO activity. Our immunostaining results demonstrated that clozapine treatment reduced overexpression of the neuronal activity marker c-fos in cortices of the brain. Results of the current study determine that clozapine ameliorated cognition in MK-801-treated mice via cholinergic and neural mechanisms. These findings show that clozapine possesses the potential to augment cognition in diseases such as schizophrenia.
Asunto(s)
Clozapina/farmacología , Maleato de Dizocilpina/toxicidad , Aprendizaje/efectos de los fármacos , Memoria/efectos de los fármacos , Actividad Motora/efectos de los fármacos , Esquizofrenia/tratamiento farmacológico , Antagonistas de la Serotonina/farmacología , Animales , Antipsicóticos/farmacología , Antagonistas de Aminoácidos Excitadores/toxicidad , Masculino , Ratones , Corteza Prefrontal/efectos de los fármacos , Corteza Prefrontal/fisiopatología , Esquizofrenia/inducido químicamenteRESUMEN
It is expected that in 2050, there will be more than 20% of senior citizens aged over 60 years worldwide. Such alarming statistics require immediate attention to improve the health of the aging population. Since aging is closely related to the loss of antioxidant defense mechanisms, this situation eventually leads to numerous health problems, including fertility reduction. Furthermore, plant extracts have been used in traditional medicine as potent antioxidant sources. Although many experiments had reported the impact of various bioactive compounds on aging or fertility, there is a lack of review papers that combine both subjects. In this review, we have collected and discussed various bioactive compounds from 26 different plant species known to affect both longevity and fertility. These compounds, including phenolics and terpenes, are mostly involved in the antioxidant defense mechanisms of diverse organisms such as rats, mites, fruit flies, roundworms, and even roosters. A human clinical trial should be considered in the future to measure the effects of these bioactive compounds on human health and longevity. Ultimately, these plant-derived compounds could be developed into health supplements or potential medical drugs to ensure a healthy aging population.