RESUMEN
RAD51 and the breast cancer suppressor BRCA2 have critical functions in DNA double-strand (dsDNA) break repair by homologous recombination and the protection of newly replicated DNA from nucleolytic degradation. The recombination function of RAD51 requires its binding to single-stranded DNA (ssDNA), whereas binding to dsDNA is inhibitory. Using reconstituted MRE11-, EXO1-, and DNA2-dependent nuclease reactions, we show that the protective function of RAD51 unexpectedly depends on its binding to dsDNA. The BRC4 repeat of BRCA2 abrogates RAD51 binding to dsDNA and accordingly impairs the function of RAD51 in protection. The BRCA2 C-terminal RAD51-binding segment (TR2) acts in a dominant manner to overcome the effect of BRC4. Mechanistically, TR2 stabilizes RAD51 binding to dsDNA, even in the presence of BRC4, promoting DNA protection. Our data suggest that RAD51's dsDNA-binding capacity may have evolved together with its function in replication fork protection and provide a mechanistic basis for the DNA-protection function of BRCA2.
Asunto(s)
ADN de Cadena Simple , Recombinasa Rad51 , Proteína BRCA2/genética , Proteína BRCA2/metabolismo , ADN/genética , Roturas del ADN de Doble Cadena , Reparación del ADN , Replicación del ADN , ADN de Cadena Simple/genética , Recombinasa Rad51/genética , Recombinasa Rad51/metabolismoRESUMEN
Tellimagrandin-I (TL) and camptothin A (CA) are ellagitannins widely found in diverse plant species. Numerous studies demonstrated their significant biological activities, which include antitumor, antioxidant, and hepatoprotective properties. Despite this protective profile, the effects of TL and CA on DNA have not been comprehensively investigated. Thus, the aim of this study was to determine the mutagenic and antimutagenic effects attributed to TL and CA exposure on Salmonella enterica serovar Typhimurium strains using the Ames test. In addition, the cytotoxic and genotoxic effects were examined on human lymphocytes, employing both trypan blue exclusion and CometChip assay. The antigenotoxic effect was determined following TL and CA exposure in the presence of co-treatment with doxorubicin (DXR). Our results from the Ames test indicated that TL or CA did not display marked mutagenic activity. However, TL or CA demonstrated an ability to protect DNA against the damaging effects of the mutagens 4-nitroquinoline-1-oxide and sodium azide, thereby exhibiting antimutagenic properties. In relation to human lymphocytes, TL or CA did not induce significant cytotoxic or genotoxic actions on these cells. Further, these ellagitannins exhibited an ability to protect DNA from damage induced by DOX during co-treatment, indicating their potential beneficial usefulness as antigenotoxic agents. In conclusion, the protective effects of TL or CA against mutagens, coupled with their absence of genotoxic and cytotoxic effects on human lymphocytes, emphasize their potential therapeutic value in chemopreventive strategies.
Asunto(s)
Antimutagênicos , Salmonella enterica , Humanos , Salmonella typhimurium/genética , Salmonella enterica/genética , Taninos Hidrolizables/farmacología , Serogrupo , Pruebas de Mutagenicidad , Mutágenos/toxicidad , Antimutagênicos/farmacología , Extractos Vegetales/farmacología , Carcinógenos/farmacología , ADN/farmacología , LinfocitosRESUMEN
Polysaccharide from Asarum sieboldii Miq (ASP) was extracted and five phosphorylation polysaccharides with different degree of substitution were obtained, namely ASPP1, ASPP2, ASPP3, ASPP4, and ASPP5 (ASPPs). The physical and chemical structure and biological activities were studied. The results suggested that the carbohydrate and protein content were reduced while uronic acid was increased after phosphorylation modification. The molecular weight of ASPPs was significantly lower than that of ASP. ASPPs were acidic heteropolysaccharides mainly composed of galacturonic acid, galactose, glucose, fructose, and arabinose. The UV-vis spectrum indicated that the polysaccharides did not contain nucleic acid or protein after modification. The Fourier transform infrared spectrum demonstrated that ASPPs contained characteristic absorption peaks of P=O and P-O-C near 1270 and 980â cm-1 . ASPPs presented a triple helix conformation, but it was not presented in ASP. The scanning electron microscopy analysis showed that the surface topography and particle structure of ASP were different after modification. Compared with ASP, ASPPs enhanced the activity to scavenge DPPH and ABTS free radicals and possessed more protective ability to DNA oxidation caused by OHâ , GSâ , and AAPH free radicals. These results suggest that chemical modification is beneficial for the exploitation and utilization of natural polysaccharides.
Asunto(s)
Antioxidantes , Asarum , Antioxidantes/farmacología , Antioxidantes/química , Fosforilación , Polisacáridos/farmacología , Polisacáridos/química , Radicales Libres , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Xanthorrhizol, an important marker of Curcuma xanthorrhiza, has been recognized for its different pharmacological activities. A green strategy for selective xanthorrhizol extraction is required. Herein, natural deep eutectic solvents (NADESs) based on glucose and organic acids (lactic acid, malic acid, and citric acid) were screened for the extraction of xanthorrhizol from Curcuma xanthorrhiza. Ultrasound-assisted extraction using glucose/lactic acid (1:3) (GluLA) gave the best yield of xanthorrhizol. The response surface methodology with a Box-Behnken Design was used to optimize the interacting variables of water content, solid-to-liquid (S/L) ratio, and extraction to optimize the extraction. The optimum conditions of 30% water content in GluLA, 1/15 g/mL (S/L), and a 20 min extraction time yielded selective xanthorrhizol extraction (17.62 mg/g) over curcuminoids (6.64 mg/g). This study indicates the protective effect of GluLA and GluLA extracts against oxidation-induced DNA damage, which was comparable with those obtained for ethanol extract. In addition, the stability of the xanthorrhizol extract over 90 days was revealed when stored at -20 and 4 °C. The FTIR and NMR spectra confirmed the hydrogen bond formation in GluLA. Our study reported, for the first time, the feasibility of using glucose/lactic acid (1:3, 30% water v/v) for the sustainable extraction of xanthorrhizol.
Asunto(s)
Antioxidantes , Curcuma , Fenoles , Extractos Vegetales , Rizoma , Curcuma/química , Antioxidantes/química , Antioxidantes/farmacología , Antioxidantes/aislamiento & purificación , Rizoma/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Fenoles/química , Fenoles/aislamiento & purificación , Fenoles/farmacología , Disolventes Eutécticos Profundos/química , Ondas UltrasónicasRESUMEN
Nowadays, olive leaf polyphenols have been at the center of scientific interest due to their beneficial effects on human health. The most abundant polyphenol in olive leaves is oleuropein. The biological properties of oleuropein are mainly due to the hydroxytyrosol moiety, a drastic catechol group, whose biological activity has been mentioned many times in the literature. Hence, in recent years, many nutritional supplements, food products, and cosmetics enriched in hydroxytyrosol have been developed and marketed, with unexpectedly positive results. However, the concentration levels of hydroxytyrosol in olive leaves are low, as it depends on several agricultural factors. In this study, a rapid and easy methodology for the production of hydroxytyrosol-enriched extracts from olive leaves was described. The proposed method is based on the direct acidic hydrolysis of olive leaves, where the extraction procedure and the hydrolysis of oleuropein are carried out in one step. Furthermore, we tested the in vitro bioactivity of this extract using cell-free and cell-based methods, evaluating its antioxidant and DNA-protective properties. Our results showed that the hydroxytyrosol-enriched extract produced after direct hydrolysis of olive leaves exerted significant in vitro antioxidant and geno-protective activity, and potentially these extracts could have various applications in the pharmaceutical, food, and cosmetic industries.
Asunto(s)
Glucósidos Iridoides , Olea , Alcohol Feniletílico/análogos & derivados , Humanos , Antioxidantes/farmacología , Grecia , Hidrólisis , Hojas de la Planta , Extractos Vegetales/farmacologíaRESUMEN
Peroxiredoxin 1 is a member of the typical 2-Cys peroxiredoxin family, which serves diverse functions in gene expression, immune and inflammatory responses, and tumor progression. In this study, we aimed to analyze the structural, functional, and immunomodulatory properties of peroxiredoxin 1 from Epinephelus akaara (EaPrx1). The open reading frame of EaPrx1 is 597 base pairs in length, encoding 198 amino acids, with a molecular weight of approximately 22 kDa. The in silico analysis revealed that EaPrx1 shares a conserved thioredoxin fold and signature motifs that are critical for its catalytic activity and oligomerization. Further, EaPrx1 is closely related to Epinephelus lanceolatus Prx1 and clustered in the Fishes group of the vertebrate clade, revealing that EaPrx1 was conserved throughout evolution. In terms of tissue distribution, a high level of EaPrx1 expression was observed in the spleen, brain, and blood tissues. Likewise, in immune challenge experiments, significant transcriptional modulations of EaPrx1 upon lipopolysaccharide, polyinosinic:polycytidylic acid, and nervous necrosis virus injections were noted at different time points, indicating the immunological role of EaPrx1 against pathogenic infections. In the functional analysis, rEaPrx1 exhibited substantial DNA protection, insulin disulfide reduction, and tissue repair activities, which were concentration-dependent. EaPrx1/pcDNA™ 3.1 (+)-transfected fathead minnow cells revealed high cell viability upon arsenic toxicity, indicating the heavy metal detoxification activity of EaPrx1. Taken together, the transcriptional and functional studies imply critical roles of EaPrx1 in innate immunity, redox regulation, apoptosis, and tissue-repair processes in E. akaara.
Asunto(s)
Lubina , Enfermedades de los Peces , Animales , Peroxirredoxinas/genética , Peroxirredoxinas/química , Lubina/genética , Lubina/metabolismo , Inmunidad Innata/genética , Antioxidantes/metabolismo , Oxidación-Reducción , Filogenia , Regulación de la Expresión Génica , Proteínas de Peces/químicaRESUMEN
Herbicides are one of the main parts of pesticides used today. Due to the high efficiency and widespread use of glyphosate-based herbicides, the search for substances reducing their genotoxicity is an important interdisciplinary task. One possible approach for solving the problem of herbicide toxicity is to use compounds that can protect DNA from damage by glyphosate derivatives. For the first time, a method for developing DNA-protecting measures against glyphosate isopropylamine salt (GIS) damage was presented and realized, based on low-toxicity water-soluble pillar[5]arene derivatives. Two- and three-component systems based on pillar[5]arene derivatives, GIS, and model DNA from salmon sperm, as well as their cytotoxicity, were studied. The synthesized pillar[5]arene derivatives do not interact with GIS, while GIS is able to bind DNA from salmon sperm with lgKa = 4.92. The pillar[5]arene betaine derivative containing fragments of L-phenylalanine and the ester derivative with diglycine fragments bind DNA with lgKa = 5.24 and lgKa = 4.88, respectively. The study of the associates (pillar[5]arene-DNA) with GIS showed that the interaction of GIS with DNA is inhibited only by the betaine pillar[5]arene containing fragments of L-Phe (lgKa = 3.60). This study has shown a possible application of betaine pillar[5]arene derivatives for nucleic acid protection according to its competitive binding with biomacromolecules.
Asunto(s)
Herbicidas , Ácidos Nucleicos , Masculino , Humanos , Betaína/farmacología , Herbicidas/farmacología , Herbicidas/química , Semen , ADN , Cloruro de Sodio , Cloruro de Sodio Dietético , GlifosatoRESUMEN
We studied the influence of epiphysectomy and administration of melatonin to epiphysectomized outbred white rats on the level and daily dynamics of damage to the genetic material of developing male germ cells. Epiphysectomy leads to an increase in the level of damage in the DNA structure and the disappearance of the circadian rhythm of the activity of repair enzyme PARP-1 and the apoptosis-inducing enzyme caspase-3. The administration of melatonin to animals after epiphysectomy reduces the level of DNA damage, restores the circadian rhythm of activity of PARP-1 and caspase-3. These fundings suggest that melatonin can indirectly protect DNA of maturing male gametes.
Asunto(s)
Melatonina , Ratas , Animales , Masculino , Melatonina/farmacología , Caspasa 3/genética , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Ritmo Circadiano , ADNRESUMEN
Glyphosate is a controversial herbicide. Its genotoxicity and presence in various ecosystems have been reported. The use of ascorbic acid and resveratrol could protect different organisms from glyphosate-induced genetic damage. In the present study, specific genetic damage induced by glyphosate was evaluated in erythrocytes of Oreochromis niloticus, Ambystoma mexicanum and human lymphocytes. Simultaneously, the antigenotoxic capacity of various concentrations of ascorbic acid and resveratrol was evaluated by means of pretreatment and simultaneous treatment protocols. The 0.03, 0.05 and 0.07 mM concentrations of glyphosate induced significant genotoxic activity (p < 0.05) in human lymphocytes and in erythrocytes of the species studied, and could cause genomic instability in these populations. The reduction in genetic damage observed in human lymphocytes exposed to high concentrations of glyphosate is only apparent: excessive genetic damage was associated with undetectable excessive tail migration length. A significant (p < 0.05) antigenotoxic effect of ascorbic acid and resveratrol was observed in all concentrations, organisms and protocols used. Both ascorbic acid and resveratrol play an important role in maintaining the integrity of DNA. Ascorbic acid in Oreochromis niloticus, Ambystoma mexicanum reduced glyphosate-induced genetic damage to a basal level. Therefore, our data indicate that these antioxidants could help preserve the integrity of the DNA of organisms exposed to glyphosate. The consumption of antioxidants is a useful tool against the genotoxicity of glyphosate.
RESUMEN
The roles of two interrelated DNA protection protein in starved cells (Dps)-putative Dps Dgeo_0257 and Dgeo_0281-as orthologous proteins to DrDps1 for DNA binding, protection, and metal ion sensing were characterised in a Deinococcus geothermalis strain. Dgeo_0257 exhibited high DNA-binding affinity and formed a multimeric structure but lacked the conserved amino acid sequence for ferroxidase activity. In contrast, the Dgeo_0281 (DgDps1) protein was abundant in the early exponential phase, had a lower DNA-binding activity than Dgeo_0257, and was mainly observed in its monomeric or dimeric forms. Electrophoretic mobility shift assays demonstrated that both purified proteins bound nonspecifically to DNA, and their binding ability was affected by certain metal ions. For example, in the presence of ferrous and ferric ions, neither Dgeo_0257 nor Dgeo_0281 could readily bind to DNA. In contrast, both proteins exhibited more stable DNA binding in the presence of zinc and manganese ions. Mutants in which the dps gene was disrupted exhibited higher sensitivity to oxidative stress than the wild-type strain. Furthermore, the expression levels of each gene showed an opposite correlation under H2O2 treatment conditions. Collectively, these findings indicate that the putative Dps Dgeo_0257 and DgDps1 from D. geothermalis are involved in DNA binding and protection in complementary interplay ways compared to known Dps.
Asunto(s)
Deinococcus , Peróxido de Hidrógeno , Peróxido de Hidrógeno/metabolismo , Proteínas Bacterianas/metabolismo , Proteínas de Unión al ADN/metabolismo , Deinococcus/metabolismo , Metales/metabolismo , ADN/metabolismoRESUMEN
Genomes of all organisms are persistently threatened by endogenous and exogenous assaults. Bacterial mechanisms of genome maintenance must provide protection throughout the physiologically distinct phases of the life cycle. Spore-forming bacteria must also maintain genome integrity within the dormant endospore. The nucleoid-associated proteins (NAPs) influence nucleoid organization and may alter DNA topology to protect DNA or to alter gene expression patterns. NAPs are characteristically multifunctional; nevertheless, Dps, HU and CbpA are most strongly associated with DNA protection. Archaea display great variety in genome organization and many inhabit extreme environments. As of yet, only MC1, an archaeal NAP, has been shown to protect DNA against thermal denaturation and radiolysis. ssDNA are intermediates in vital cellular processes, such as DNA replication and recombination. Single-stranded binding proteins (SSBs) prevent the formation of secondary structures but also protect the hypersensitive ssDNA against chemical and nuclease degradation. Ionizing radiation upregulates SSBs in the extremophile Deinococcus radiodurans.
Asunto(s)
Proteínas de Unión al ADN , Deinococcus , Animales , Proteínas Bacterianas/metabolismo , ADN/metabolismo , Replicación del ADN , ADN de Cadena Simple/metabolismo , Proteínas de Unión al ADN/metabolismo , Deinococcus/genética , Deinococcus/metabolismo , Estadios del Ciclo de VidaRESUMEN
The long-term use of Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) in treatment of different chronic inflammatory disorders is strongly restricted by their serious gastrointestinal adverse effects. Therefore, there is still an urgent need to search for new, safe, and efficient anti-inflammatory agents. Previously, we have reported the Mannich base-type derivatives of pyrrolo[3,4-d]pyridazinone which strongly inhibit cyclooxygenase, have better affinity to COX-2 isoenzyme and exert promising anti-oxidant activity. These findings encouraged us to perform further optimization of that structure. Herein, we present the design, synthesis, molecular docking, spectroscopic, and biological studies of novel pyrrolo[3,4-d]pyridazinone derivatives bearing 4-aryl-1-(1-oxoethyl)piperazine pharmacophore 5a,b-6a,b. The new compounds were obtained via convenient, efficient, one-pot synthesis. According to in vitro evaluations, novel molecules exert no cytotoxicity and act as selective COX-2 inhibitors. These findings stay in good correlation with molecular modeling results, which additionally showed that investigated compounds take a position in the active site of COX-2 very similar to Meloxicam. Moreover, all derivatives reduce the increased level of reactive oxygen and nitrogen species and prevent DNA strand breaks caused by oxidative stress. Finally, performed spectroscopic and molecular docking studies demonstrated that new compound interactions with bovine serum albumin (BSA) are moderate, formation of complexes is in one-to-one ratio, and binding site II (subdomain IIIA) is favorable.
Asunto(s)
Antioxidantes/química , Antioxidantes/farmacología , Inhibidores de la Ciclooxigenasa 2/química , Inhibidores de la Ciclooxigenasa 2/farmacología , Diseño de Fármacos , Bases de Mannich/química , Oxadiazoles/química , Piridazinas/farmacología , Pirroles/farmacología , Antioxidantes/síntesis química , Dominio Catalítico , Línea Celular , Supervivencia Celular/efectos de los fármacos , Dicroismo Circular , Ciclooxigenasa 2/metabolismo , Inhibidores de la Ciclooxigenasa 2/síntesis química , Femenino , Fibroblastos/metabolismo , Humanos , Concentración 50 Inhibidora , Persona de Mediana Edad , Simulación del Acoplamiento Molecular , Estructura Molecular , Piridazinas/síntesis química , Piridazinas/química , Pirroles/síntesis química , Pirroles/química , Albúmina Sérica Bovina/química , Relación Estructura-ActividadRESUMEN
The surface of the Earth is exposed to harmful ultraviolet radiation (UVR: 280-400 nm). Prolonged skin exposure to UVR results in DNA damage through oxidative stress due to the production of reactive oxygen species (ROS). Mycosporine-like amino acids (MAAs) are UV-absorbing compounds, found in many marine and freshwater organisms that have been of interest in use for skin protection. MAAs are involved in photoprotection from damaging UVR thanks to their ability to absorb light in both the UV-A (315-400 nm) and UV-B (280-315 nm) range without producing free radicals. In addition, by scavenging ROS, MAAs play an antioxidant role and suppress singlet oxygen-induced damage. Currently, there are over 30 different MAAs found in nature and they are characterised by different antioxidative and UV-absorbing capacities. Depending on the environmental conditions and UV level, up- or downregulation of genes from the MAA biosynthetic pathway results in seasonal fluctuation of the MAA content in aquatic species. This review will provide a summary of the MAA antioxidative and UV-absorbing features, including the genes involved in the MAA biosynthesis. Specifically, regulatory mechanisms involved in MAAs pathways will be evaluated for controlled MAA synthesis, advancing the potential use of MAAs in human skin protection.
Asunto(s)
Aminoácidos/química , Protectores Solares/química , Rayos Ultravioleta/efectos adversos , Animales , Antioxidantes/metabolismo , Vías Biosintéticas/efectos de los fármacos , Humanos , Especies Reactivas de Oxígeno/metabolismo , Piel/efectos de los fármacos , Protectores Solares/farmacologíaRESUMEN
Dried flower buds of Lonicera japonica and L. macranthoides have long been used as herbs in numerous Chinese traditional medicines. Comparisons of three phenolic fractions (i.e., free, esterified, and insoluble-bound phenolics) in three different organs (i.e., flower, leaf, and stem) of the two species revealed that the free phenolics were the highest in terms of total phenol and total flavonoid content, composed of the most numerous phenolics and flavonoids; thus, they exhibited the most excellent antioxidant activities (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonate) (ABTS), and oxygen radical absorbance capacity (ORAC)), as well as protective effects on DNA damage induced by free radicals. In identical free and esterified phenolics of a same organ, higher contents and bioactivities were observed in L. macranthoides than in L. japonica. Phenolics identified by ultra-performance liquid chromatography with a diode array detector, alongside tandem mass spectrometry coupled with a quadrupole time-of-flight mass spectrometer (UPLC-DADâ»QTOF-MS/MS) mainly included chlorogenic acid and its five derivatives, three flavonoids that were only found in the free phenolic fraction and closely correlated with its bioactivity, and caffeic acid that was the major contributor to antioxidant activity of the esterified and insoluble-bound phenolic fractions. It was, thus, concluded that, like L. japonica, L. macranthoides, which was underestimated since being separately listed by the 2010 edition of the Chinese Pharmacopoeia, is also a good (and better) herbal medicine.
Asunto(s)
Lonicera/química , Fenoles/química , Antioxidantes/química , Compuestos de Bifenilo/química , Cromatografía Líquida de Alta Presión , Daño del ADN/genética , Picratos/químicaRESUMEN
BACKGROUND: Flavonoids possess a rich polypharmacological profile and their biological role is linked to their oxidation state protecting DNA from oxidative stress damage. However, their bioavailability is hampered due to their poor aqueous solubility. This can be surpassed through encapsulation to supramolecular carriers as cyclodextrin (CD). A quercetin- 2HP-ß-CD complex has been formerly reported by us. However, once the flavonoid is in its 2HP-ß-CD encapsulated state its oxidation potential, its decomplexation mechanism, its potential to protect DNA damage from oxidative stress remained elusive. To unveil this, an array of biophysical techniques was used. METHODS: The quercetin-2HP-ß-CD complex was evaluated through solubility and dissolution experiments, electrochemical and spectroelectrochemical studies (Cyclic Voltammetry), UV-Vis spectroscopy, HPLC-ESI-MS/MS and HPLC-DAD, fluorescence spectroscopy, NMR Spectroscopy, theoretical calculations (density functional theory (DFT)) and biological evaluation of the protection offered against H2O2-induced DNA damage. RESULTS: Encapsulation of quercetin inside the supramolecule's cavity enhanced its solubility and retained its oxidation profile. Although the protective ability of the quercetin-2HP-ß-CD complex against H2O2 was diminished, iron serves as a chemical stimulus to dissociate the complex and release quercetin. CONCLUSIONS: We found that in a quercetin-2HP-ß-CD inclusion complex quercetin retains its oxidation profile similarly to its native state, while iron can operate as a chemical stimulus to release quercetin from its host cavity. GENERAL SIGNIFICANCE: The oxidation profile of a natural product once it is encapsulated in a supramolecular carrier was unveiled as also it was discovered that decomplexation can be triggered by a chemical stimilus.
Asunto(s)
Ciclodextrinas/metabolismo , Daño del ADN/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Hierro/metabolismo , Quercetina/metabolismo , Disponibilidad Biológica , Ciclodextrinas/química , Humanos , Hierro/química , Células Jurkat , Oxidantes/farmacología , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Quercetina/químicaRESUMEN
Notwithstanding the enormous reproductive potential encapsulated within a mature mammalian oocyte, these cells present only a limited window for fertilization before defaulting to an apoptotic cascade known as post-ovulatory oocyte aging. The only cell with the capacity to rescue this potential is the fertilizing spermatozoon. Indeed, the union of these cells sets in train a remarkable series of events that endows the oocyte with the capacity to divide and differentiate into the trillions of cells that comprise a new individual. Traditional paradigms hold that, beyond the initial stimulation of fluctuating calcium (Ca2+) required for oocyte activation, the fertilizing spermatozoon plays limited additional roles in the early embryo. While this model has now been drawn into question in view of the recent discovery that spermatozoa deliver developmentally important classes of small noncoding RNAs and other epigenetic modulators to oocytes during fertilization, it is nevertheless apparent that the primary responsibility for oocyte activation rests with a modest store of maternally derived proteins and mRNA accumulated during oogenesis. It is, therefore, not surprising that widespread post-translational modifications, in particular phosphorylation, hold a central role in endowing these proteins with sufficient functional diversity to initiate embryonic development. Indeed, proteins targeted for such modifications have been linked to oocyte activation, recruitment of maternal mRNAs, DNA repair and resumption of the cell cycle. This review, therefore, seeks to explore the intimate relationship between Ca2+ release and the suite of molecular modifications that sweep through the oocyte to ensure the successful union of the parental germlines and ensure embryogenic fidelity.
Asunto(s)
Calcio/metabolismo , Oocitos/metabolismo , Cigoto/metabolismo , Animales , Cationes Bivalentes/metabolismo , Reparación del ADN , Desarrollo Embrionario , Epigénesis Genética , Fertilización , Humanos , Oocitos/citología , Fosforilación , Procesamiento Proteico-Postraduccional , Cigoto/citologíaRESUMEN
Culture broth of Ganoderma lucidum was determined for antioxidant, antibacterial and α-amylase inhibitory activities. The culture broth contained protein as determined by Bradford method equaled to 0.2 mg/ml and total phenol content as 0.078 mg GAE/mg protein (0.557 mg GAE/g extract). It exhibited radicals scavenging activities against ABTS+· and DPPH· radicals with a half maximal inhibitory concentration (IC50) less than 1.70 ± 0.02 and 2.28 ± 0.02 µg protein/ml, respectively and reducing power equaled to 4.38 ± 0.02 µmol Trolox/µg protein as investigated by ferric ion reducing antioxidant power method. The culture broth experimented into two approached; (1) treated with pronase and (2) filtered through a membrane with 10 kDa molecular weight cut-off (MWCO). The pronase-treated culture broth exhibited insignificant lower antioxidant activities, but the retained culture broth 10 kDa MWCO resulted in significant decrease in antioxidant activities suggesting that the small proteins might play the key role in antioxidant activity. The culture broth could protect DNA damage from hydroxyl radicals (·OH) generated by Fenton's reaction. This culture broth showed antibacterial activity towards pathogenic strains Staphylococcus epidermidis and Pseudomonas aeruginosa and also had an interesting α-amylase inhibitory activity. This study suggested that apart from the fruiting bodies and the mycelial of G. lucidum, its culture broth also had potential applications as a value-added ingredient in the product such as in cosmetics and in nutraceuticals.
RESUMEN
Apple pomace (AP), the residue that remains after the extraction of juice from apple accounts for ~25 % of total apple weight. Current study is aimed at identification of phytochemicals and utilization of Dehydrated apple pomace (DAP) in the preparation of bakery products with potential health benefits. DAP was prepared by drying the pomace obtained by crushing peeled apple fruits. DAP was incorporated into bakery products such as bun, muffin and cookies for value addition. Bioactivity such as free radical scavenging, cyto/DNA protectivity was evaluated in these products. DAP contained 17 g/100 g starch, 49.86 g/100 g fructose and 37 g/100 g dietary fibre. The phenolics and flavonoids content was 1.5 mg/g and 3.92 mg/g, respectively. Increase in DAP resulted in decreased volume and enhanced firmness of buns and muffins. DAP at 15 % in buns, 30 % in muffins and 20 % in cookies were found to be acceptable. DAP blended products exhibited better free radical scavenging as well as cyto/DNA protective properties suggesting the retention of bioactivity after baking. Addition of DAP potentially enhanced the bioactivity of the products evaluated.
RESUMEN
In liver, the major site of iron storage, iron overload is associated with oxidative damage of protein, lipid, and DNA and causes protein oxidation, lipid peroxidation, and rupture of hepatocytes, leading to cell death. Serum ferritin and liver iron content are the main forecasters of moderate to severe iron overload in the liver. The sequels of excess iron deposition in the liver are fibrosis and enhanced levels of serum enzymes and bilirubin markers. Emblica officinalis (EO) fruit extract was found efficient in lessening intraperitoneally injected iron dextran-induced liver toxicity in Swiss albino mice. Mice administered with different doses of 70% methanol extract of EO (50, 100, and 200 mg kg(-1) body weight) showed significant decrease in liver iron, serum ferritin, and serum enzyme levels, along with the decrease in lipid peroxidation, protein oxidation, and collagen content. The activity was further supported by its considerable iron chelation with half-maximal inhibitory concentration of 70.24 ± 2.74 µg ml(-1) and the protection on ferrous ion-mediated DNA breakdown with 50% protection ([P]50) of 1.04 ± 0.01 µg ml(-1). Simultaneously, the extract effectively induced the antioxidant enzyme levels and also exhibited the potential activity of reductive release of ferritin iron. These findings suggest that the EO extract may be used as a potent drug for the treatment of pathological sequences arisen in the iron overload-induced liver damage.
Asunto(s)
Antioxidantes/farmacología , Quelantes del Hierro/farmacología , Sobrecarga de Hierro/tratamiento farmacológico , Hígado/efectos de los fármacos , Phyllanthus emblica/metabolismo , Animales , Catalasa/análisis , Cromatografía Líquida de Alta Presión , Ferritinas/sangre , Glutatión Transferasa/análisis , Hierro/metabolismo , Hígado/patología , Masculino , Metanol , Ratones , Superóxido Dismutasa/análisisRESUMEN
The acute genoprotective effect of Panax quinquefolius (American ginseng) has been investigated. The experiment was carried out to explore the DNA protective effect after a single dose of American ginseng tea bag infusion. Fourteen subjects (6 males and 8 females) were recruited in this study. Seven of them (3 males and 4 females) were asked to drink a cup of freshly prepared American ginseng infusions. Water was taken by the remaining subjects as the control group. Blood samples of both groups were taken before and 2 h post-ingestion. The blood samples were challenged with ultraviolet B irradiation followed by using comet assay. Completed slides were stained with Giemsa stain and DNA damage was assessed. Results showed a significant decrease in comet score after American ginseng supplementation and no change in the control group. The current study demonstrated a cup of American ginseng infusion could protect cellular DNA from oxidative stress at least within 2 h.