RESUMO
Breast Cancer (BC) is one of the most common and challenging cancers among females worldwide. Conventional treatments for oral cancer rely on the use of radiology and surgery accompanied by chemotherapy. Chemotherapy presents many side effects, and the cells often develop resistance to this chemotherapy. It will be urgent to adopt alternative or complementary treatment strategies that are new and more effective without these negative effects to improve the well-being of patients. A substantial number of epidemiological and experimental studies reported that many compounds are derived from natural products such as curcumin and their analogs, which have a great deal of beneficial anti-BC activity by inducing apoptosis, inhibiting cell proliferation, migration, and metastasis, modulating cancer-related pathways, and sensitizing cells to radiotherapy and chemotherapy. In the present study, we investigated the effect of the curcumin-analog PAC on DNA repair pathways in MCF-7 and MDA-MB-231 human breast-cancer cell lines. These pathways are crucial for genome maintenance and cancer prevention. MCF-7 and MDA-MB-231 cells were exposed to PAC at 10 µM. MTT and LDH assays were conducted to evaluate the effects of PAC on cell proliferation and cytotoxicity. Apoptosis was assessed in breast cancer cell lines using flow cytometry with annexin/Pi assay. The expression of proapoptotic and antiapoptotic genes was determined by RT-PCR to see if PAC is active in programming cell death. Additionally, DNA repair signaling pathways were analyzed by PCR arrays focusing on genes being related and confirmed by quantitative PCR. PAC significantly inhibited breast-cancer cell proliferation in a time-dependent manner, more on MDA-MB-231 triple-negative breast cancer cells. The flow cytometry results showed an increase in apoptotic activity. These data have been established by the gene expression and indicate that PAC-induced apoptosis by an increased Bax and decreased Bcl-2 expression. Moreover, PAC affected multiple genes involved in the DNA repair pathways occurring in both cell lines (MCF-7 and MDA-MB231). In addition, our results suggest that PAC upregulated more than twice 16 genes (ERCC1, ERCC2, PNKP, POLL, MPG, NEIL2, NTHL1, SMUG1, RAD51D, RAD54L, RFC1, TOP3A, XRCC3, XRCC6BP1, FEN1, and TREX1) in MDA-MB-231, 6 genes (ERCC1, LIG1, PNKP, UNG, MPG, and RAD54L) in MCF-7, and 4 genes (ERCC1, PNKP, MPG, and RAD54L) in the two cell lines. In silico analysis of gene-gene interaction shows that there are common genes between MCF-7 and MDA-MB-321 having direct and indirect effects, among them via coexpression, genetic interactions, pathways, predicted and physical interactions, and shared protein domains with predicted associated genes indicating they are more likely to be functionally related. Our data show that PAC increases involvement of multiple genes in a DNA repair pathway, this certainly can open a new perspective in breast-cancer treatment.
Assuntos
Antineoplásicos , Neoplasias da Mama , Curcumina , Neoplasias de Mama Triplo Negativas , Feminino , Humanos , Curcumina/uso terapêutico , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/genética , Neoplasias de Mama Triplo Negativas/patologia , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Apoptose , Proliferação de Células , Expressão Gênica , Reparo do DNA , Antineoplásicos/farmacologia , Proteína Grupo D do Xeroderma Pigmentoso/genética , Proteína Grupo D do Xeroderma Pigmentoso/metabolismo , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Enzimas Reparadoras do DNA/genéticaRESUMO
In this study, we have explored the possible role of ascorbic acid (ASC) and glutathione (GSH) in alleviating arsenate (AsV ) toxicity in brinjal roots. Moreover, we have also focused our attention on the possible involvement of endogenous nitric oxide (NO) in accomplishing this task. AsV treatment negatively impacts the length and fresh weight of roots and shoots as well as the dry weight and fitness of roots, and this was accompanied by greater As accumulation in roots and shoots of brinjal. AsV treatment also declined the endogenous NO level by inhibiting Nitric Oxide Synthase-like (NOS-like) activity. Furthermore, AsV stimulated oxidative stress markers, caused protein damage by their carbonylation due to downregulation in antioxidants [particularly ascorbate (AsA)-GSH cycle], leading to disturbed cellular redox status. This, collectively, led to root cell death in brinjal. However, the addition of either ASC or GSH rescued brinjal roots from the toxic effects of AsV in. Interestingly, lycorine (an inhibitor of ASC biosynthesis) further increased AsV toxicity, while ASC rescued its effects. Moreover, buthionine sulphoximine (BSO, an inhibitor of GSH biosynthesis) interestingly increased further AsV toxicity, while GSH rescued the plant from the As toxic effects. An interesting notion of this study was that GSH rescued the toxic effect of lycorine, while ASC rescued the toxic effect of BSO, though the AsV toxicity mediated by either ASC or GSH was always accompanied by high endogenous NO level and NOS-like activity. All together, these results suggest that ASC and GSH independently mitigate AsV toxicity in brinjal roots, but both might be dependent on endogenous NO for accomplishing the AsV toxicity alleviatory tasks.
Assuntos
Glutationa , Solanum melongena , Antioxidantes , Arseniatos/toxicidade , Ácido Ascórbico , Glutationa/metabolismo , Óxido Nítrico , Estresse Oxidativo , Raízes de Plantas/metabolismoRESUMO
Metal toxicity in crop plants is a matter of scientific concern. Therefore, in recent years efforts have been made to minimize metal toxicity in crop plants. Out of various strategies, priming of seedlings with certain chemicals, like e.g. donors of signaling molecules, nutrients, metabolites or plant hormones has shown encouraging results. However, mechanisms related with the priming-induced mitigation of metal toxicity are still poorly known. Hence, we have tested the potential of 2-oxoglutarate (2-OG) priming in enhancing the arsenate (AsV ) toxicity tolerance in tomato seedlings along with deciphering the probable role of nitric oxide (NO) in accomplishing this task. Arsenate decreased growth, endogenous NO and nitric oxide synthase-like activity but enhanced the accumulation of As, which collectively led to root cell death. Arsenate toxicity also decreased some photosynthetic characteristics (i.e. Fv /Fm, qP, Fv /F0 and Fm /F0 , and total chlorophyll content) but enhanced NPQ. However, priming with 2-OG alleviated the toxic effect of AsV on growth, endogenous NO, cell death and photosynthesis. Moreover, arsenate inhibited the activities of enzymes of nitrogen metabolism (i.e. nitrate reductase, nitrite reductase, glutamine synthetase and glutamine 2-oxoglutarate aminotransferase) but increased the activity of glutamate dehydrogenase and NH4 + content. Superoxide radicals, hydrogen peroxide, lipid peroxidation, protein oxidation and membrane damage increased upon AsV exposure, but the antioxidant enzymes (i.e. superoxide dismutase, catalase and glutathione-S-transferase) showed differential responses. Overall, our results showed that 2-OG is capable of alleviating AsV toxicity in tomato seedlings but the involvement of endogenous NO is probably required.
Assuntos
Arsênio , Solanum lycopersicum , Antioxidantes , Ácidos Cetoglutáricos , Óxido Nítrico , Estresse Oxidativo , Fotossíntese , PlântulaRESUMO
KEY MESSAGE: Exogenous potassium (K+) and endogenous hydrogen sulfide (H2S) synergistically alleviate drought stress through regulating H+-ATPase activity, sugar metabolism and redox homoeostasis in tomato seedlings. Present work evaluates the role of K+ in the regulation of endogenous H2S signaling in modulating the tolerance of tomato (Solanum lycopersicum L. Mill.) seedlings to drought stress. The findings reveal that exposure of seedlings to 15% (w/v) polyethylene glycol 8000 (PEG) led to a substantial decrease in leaf K+ content which was associated with reduced H+-ATPase activity. Treatment with sodium orthovanadate (SOV, PM H+-ATPase inhibitor) and tetraethylammonium chloride (TEA, K+ channel blocker) suggests that exogenous K+ stimulated H+-ATPase activity that further regulated endogenous K+ content in tomato seedlings subjected to drought stress. Moreover, reduction in H+-ATPase activity by hypotaurine (HT; H2S scavenger) substantiates the role of endogenous H2S in the regulation of H+-ATPase activity. Elevation in endogenous K+ content enhanced the biosynthesis of H2S through enhancing the synthesis of cysteine, the H2S precursor. Synergistic action of H2S and K+ effectively neutralized drought stress by regulating sugar metabolism and redox homoeostasis that resulted in osmotic adjustment, as witnessed by reduced water loss, and improved hydration level of the stressed seedlings. The integrative role of endogenous H2S in K+ homeostasis was validated using HT and TEA which weakened the protection against drought stress induced impairments. In conclusion, exogenous K+ and endogenous H2S regulate H+-ATPase activity which plays a decisive role in the maintenance of endogenous K+ homeostasis. Thus, present work reveals that K+ and H2S crosstalk is essential for modulation of drought stress tolerance in tomato seedlings.
Assuntos
Antioxidantes/metabolismo , Desidratação , Sulfeto de Hidrogênio/metabolismo , Potássio/metabolismo , Solanum lycopersicum/fisiologia , Anidrases Carbônicas/metabolismo , Clorofila/metabolismo , Secas , Enzimas/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , ATPases Translocadoras de Prótons/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Plântula/fisiologia , Açúcares/metabolismoRESUMO
Earthquake prediction is a popular topic among earth scientists; however, this task is challenging and exhibits uncertainty therefore, probability assessment is indispensable in the current period. During the last decades, the volume of seismic data has increased exponentially, adding scalability issues to probability assessment models. Several machine learning methods, such as deep learning, have been applied to large-scale images, video, and text processing; however, they have been rarely utilized in earthquake probability assessment. Therefore, the present research leveraged advances in deep learning techniques to generate scalable earthquake probability mapping. To achieve this objective, this research used a convolutional neural network (CNN). Nine indicators, namely, proximity to faults, fault density, lithology with an amplification factor value, slope angle, elevation, magnitude density, epicenter density, distance from the epicenter, and peak ground acceleration (PGA) density, served as inputs. Meanwhile, 0 and 1 were used as outputs corresponding to non-earthquake and earthquake parameters, respectively. The proposed classification model was tested at the country level on datasets gathered to update the probability map for the Indian subcontinent using statistical measures, such as overall accuracy (OA), F1 score, recall, and precision. The OA values of the model based on the training and testing datasets were 96% and 92%, respectively. The proposed model also achieved precision, recall, and F1 score values of 0.88, 0.99, and 0.93, respectively, for the positive (earthquake) class based on the testing dataset. The model predicted two classes and observed very-high (712,375 km2) and high probability (591,240.5 km2) areas consisting of 19.8% and 16.43% of the abovementioned zones, respectively. Results indicated that the proposed model is superior to the traditional methods for earthquake probability assessment in terms of accuracy. Aside from facilitating the prediction of the pixel values for probability assessment, the proposed model can also help urban-planners and disaster managers make appropriate decisions regarding future plans and earthquake management.
RESUMO
Melatonin, a natural agent, has multiple functions in animals as well as in plants. However, its possible roles in plants under abiotic stress are not clear. Nowadays, soil salinity is a major threat to global agriculture because a high soil salt content causes multiple stresses (hyperosmotic, ionic, and oxidative). Therefore, the aim of the present study was to explore: (1) the involvement of melatonin in biosynthesis of photosynthetic pigments and in regulation of photosynthetic enzymes, such as carbonic anhydrase (CA) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco); (2) the role of melatonin in osmoregulation by proline and carbohydrate metabolism; and (3) the function of melatonin in the antioxidant defense system under salinity. Outcomes of the study reveal that under non-saline conditions, application of melatonin (20 and 50 µM) improved plant growth, viz. shoot length, root length, shoot fresh weight (FW), root FW, shoot dry weight (DW), root DW and leaf area and physio-biochemical parameters [chlorophyll (Chl) a and b, proline (Pro) and total soluble carbohydrates (TSC) content, and increased the activity of CA and Rubisco]. However, tomato seedlings treated with NaCl exhibited enhanced Chl degradation, electrolyte leakage (EL), malondialdehyde (MDA) and reactive oxygen species (ROS; superoxide and hydrogen peroxide). ROS were detected in leaf and root. Interestingly, application of melatonin improved plant growth and reduced EL, MDA and ROS levels through upregulation of photosynthesis enzymes (CA, Rubisco), antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase and ascorbate reductase) and levels of non-enzymatic antioxidants [ascorbate (ASC) and reduced glutathione (GSH)], as well as by affecting the ASC-GSH cycle. Additionally, exogenous melatonin also improved osmoregulation by increasing the content of TSC, Pro and Δ¹-pyrroline-5-carboxylate synthetase activity. These results suggest that melatonin has beneficial effects on tomato seedlings growth under both stress and non-stress conditions. Melatonin's role in tolerance to salt stress may be associated with the regulation of enzymes involved in photosynthesis, the antioxidant system, metabolism of proline and carbohydrate, and the ASC-GSH cycle. Also, melatonin could be responsible for maintaining the high ratios of GSH/GSSG and ASC/DHA.
Assuntos
Antioxidantes/metabolismo , Metabolismo dos Carboidratos/efeitos dos fármacos , Melatonina/farmacologia , Prolina/metabolismo , Plântula/metabolismo , Cloreto de Sódio/toxicidade , Solanum lycopersicum/metabolismo , Ácido Ascórbico/metabolismo , Anidrases Carbônicas/metabolismo , Clorofila/metabolismo , Eletrólitos/metabolismo , Glutationa/metabolismo , Peróxido de Hidrogênio/metabolismo , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/enzimologia , Solanum lycopersicum/crescimento & desenvolvimento , Malondialdeído/metabolismo , Proteínas de Plantas/metabolismo , Ribulose-Bifosfato Carboxilase/metabolismo , Salinidade , Plântula/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Solubilidade , Estresse Fisiológico/efeitos dos fármacos , Superóxidos/metabolismoRESUMO
Arsenic (As) poses a significant environmental threat as a metalloid toxin, adversely affecting the health of both plants and animals. Strigolactones (SL) and nitric oxide (NO) are known to play crucial roles in plant physiology. Therefore, the present experiment was designed to investigate the potential cumulative role of SL (GR24-0.20 µM) and NO (100 µM) in mitigating the adverse effect of AsV (53 µM) by modulating physiological mechanisms in two genotypes of tomato (Riogrand and Super Strain 8). A sample randomized design with four replicates was used to arrange the experimental pots in the growth chamber. 45-d old both tomato cultivars under AsV toxicity exhibited reduced morphological attributes (root and shoot length, root and shoot fresh weight, and root and shoot dry weight) and physiological and biochemical characteristics [chlorophyll (Chl) a and b content, activity of δ-aminolevulinic acid dehydratase activity (an enzyme responsible for Chl biosynthesis), and carbonic anhydrase activity (an enzyme responsible for photosynthesis), and enhanced Chl degradation, overproduction of reactive oxygen species (ROS) and lipid peroxidation due to enhanced malondialdehyde (MDA) content. However, the combined application of SL and NO was more effective in enhancing the tolerance of both varieties to AsV toxicity compared to individual application. The combined application of SL and NO improved growth parameters, biosynthesis of Chls, NO and proline. However, the combined application significantly suppressed cellular damage by inhibiting MDA and overproduction of ROS in leaves and roots, as confirmed by the fluorescent microscopy study and markedly upregulated the antioxidant enzymes (catalase, peroxidase, superoxide dismutase, ascorbate dismutase and glutathione reductase) activity. This study provides clear evidence that the combined application of SL and NO supplementation significantly improves the resilience of tomato seedlings against AsV toxicity. The synergistic effect of SL and NO was confirmed by the application of cPTIO (an NO scavenger) with SL and NO. However, further molecular studies could be imperative to conclusively validate the simultaneous role of SL and NO in enhancing plant tolerance to abiotic stress.
Assuntos
Arsênio , Compostos Heterocíclicos com 3 Anéis , Lactonas , Resiliência Psicológica , Solanum lycopersicum , Antioxidantes/metabolismo , Plântula/metabolismo , Óxido Nítrico/metabolismo , Arsênio/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Estresse Oxidativo , Peróxido de Hidrogênio/metabolismoRESUMO
Adoption of plant-derived compounds for the management of oral cancer is encouraged by the scientific community due to emerging chemoresistance and conventional treatments adverse effects. Considering that very few studies investigated eugenol clinical relevance for gingival carcinoma, we ought to explore its selectivity and performance according to aggressiveness level. For this purpose, non-oncogenic human oral epithelial cells (GMSM-K) were used together with the Tongue (SCC-9) and Gingival (Ca9-22) squamous cell carcinoma lines to assess key tumorigenesis processes. Overall, eugenol inhibited cell proliferation and colony formation while inducing cytotoxicity in cancer cells as compared to normal counterparts. The recorded effect was greater in gingival carcinoma and appears to be mediated through apoptosis induction and promotion of p21/p27/cyclin D1 modulation and subsequent Ca9-22 cell cycle arrest at the G0/G1 phase, in a p53-independent manner. At these levels, distinct genetic profiles were uncovered for both cell lines by QPCR array. Moreover, it seems that our active component limited Ca9-22 and SCC-9 cell migration respectively through MMP1/3 downregulation and stimulation of inactive MMPs complex formation. Finally, Ca9-22 behaviour appears to be mainly modulated by the P38/STAT5/NFkB pathways. In summary, we can disclose that eugenol is cancer selective and that its mediated anti-cancer mechanisms vary according to the cell line with gingival squamous cell carcinoma being more sensitive to this phytotherapy agent.
Assuntos
Apoptose , Carcinoma de Células Escamosas , Proliferação de Células , Eugenol , Neoplasias Gengivais , Humanos , Eugenol/farmacologia , Eugenol/uso terapêutico , Neoplasias Gengivais/tratamento farmacológico , Neoplasias Gengivais/patologia , Neoplasias Gengivais/metabolismo , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Quimioterapia Adjuvante/métodosRESUMO
Contamination of soils with chromium (Cr) jeopardized agriculture production globally. The current study was planned with the aim to better comprehend how melatonin (Mel) and hydrogen sulfide (H2S) regulate antioxidant defense system, potassium (K) homeostasis, and nitrogen (N) metabolism in tomato seedlings under Cr toxicity. The data reveal that application of 30 µM Mel to the seedlings treated with 25 µM Cr has a positive effect on H2S metabolism that resulted in a considerable increase in H2S. Exogenous Mel improved phytochelatins content and H+-ATPase activity with an associated increase in K content as well. Use of tetraethylammonium chloride (K+-channel blocker) and sodium orthovanadate (H+-ATPase inhibitor) showed that Mel maintained K homeostasis through regulating H+-ATPase activity under Cr toxicity. Supplementation of the stressed seedlings with Mel substantially scavenged excess reactive oxygen species (ROS) that maintained ROS homeostasis. Reduced electrolyte leakage and lipid peroxidation were additional signs of Mel's ROS scavenging effects. In addition, Mel also maintained normal functioning of nitrogen (N) metabolism and ascorbate-glutathione (AsA-GSH) system. Improved level of N fulfilled its requirement for various enzymes that have induced resilience during Cr stress. Additionally, the AsA-GSH cycle's proper operation maintained redox equilibrium, which is necessary for the biological system to function normally. Conversely, 1 mM hypotaurine (H2S scavenger) abolished the Mel-effect and again Cr-induced impairment on the above-mentioned parameters was observed even in presence of Mel. Therefore, based on the observed findings, we concluded that Mel needs endogenous H2S to alleviate Cr-induced impairments in tomato seedlings.
Assuntos
Sulfeto de Hidrogênio , Melatonina , Melatonina/farmacologia , Melatonina/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sulfeto de Hidrogênio/metabolismo , Sulfeto de Hidrogênio/farmacologia , Cromo/toxicidade , Cromo/metabolismo , Ácido Ascórbico/farmacologia , Ácido Ascórbico/metabolismo , Glutationa/metabolismo , Antioxidantes/metabolismo , Plântula , Nitrogênio/metabolismoRESUMO
Chromium (Cr), a highly toxic redox-active metal cation in soil, seriously threatens global agriculture by affecting nutrient uptake and disturbing various physio-biochemical processes in plants, thereby reducing yields. Here, we examined the effects of different concentrations of Cr alone and in combination with hydrogen sulfide (H2S) application on the growth and physio-biochemical performance of two mungbeans (Vigna radiata L.) varieties, viz. Pusa Vishal (PV; Cr tolerant) and Pusa Ratna (PR; Cr sensitive), growing in a pot in hydroponics. Plants were grown in the pot experiment to examine their growth, enzymatic and non-enzymatic antioxidant levels, electrolyte balance, and plasma membrane (PM) H+-ATPase activity. Furthermore, root anatomy and cell death were analysed 15 days after sowing both varieties in hydroponic systems. The Cr-induced accumulation of reactive oxygen species caused cell death and affected the root anatomy and growth of both varieties. However, the extent of alteration in anatomical features was less in PV than in PR. Exogenous application of H2S promoted plant growth, thereby improving plant antioxidant activities and reducing cell death by suppressing Cr accumulation and translocation. Seedlings of both cultivars treated with H2S exhibited enhanced photosynthesis, ion uptake, glutathione, and proline levels and reduced oxidative stress. Interestingly, H2S restricted the translocation of Cr to aerial parts of plants by improving the nutrient profile and viability of root cells, thereby relieving plants from oxidative bursts by activating the antioxidant machinery through triggering the ascorbate-glutathione cycle. Overall, H2S application improved the nutrient profile and ionic homeostasis of Cr-stressed mungbean plants. These results highlight the importance of H2S application in protecting crops against Cr toxicity. Our findings can be utilised to develop management strategies to improve heavy metal tolerance among crops.
Assuntos
Sulfeto de Hidrogênio , Vigna , Antioxidantes/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Plântula/metabolismo , Vigna/metabolismo , Sulfeto de Hidrogênio/farmacologia , Sulfeto de Hidrogênio/metabolismo , Cromo/toxicidade , Estresse Oxidativo , Glutationa/metabolismo , Produtos Agrícolas/metabolismoRESUMO
This study determined the full length sequence of glucose-6-phosphate dehydrogenase cDNA (G6PD) from the Arabian camel Camelus dromedarius using reverse transcription polymerase chain reaction. The C. dromedarius G6PD has an open reading frame of 1545 bp, and the cDNA encodes a protein of 515 amino acid residues with a molecular weight of 59.0 KDa. The amino acid sequence showed the highest identity with Equus caballus (92%) and Homo sapiens (92%). The G6PD cDNA was cloned and expressed into Escherichia coli as a fusion protein and was purified in a single chromatographic step using nickel affinity gel column. The purity and the molecular weight of the enzyme were checked on SDS-PAGE and the purified enzyme showed a single band on the gel with a molecular weight of 63.0 KDa. The specific activity of G6PD was determined to be 289.6 EU/mg protein with a fold purification of 95.45 and yield of 56.8%.
Assuntos
Camelus/genética , Escherichia coli/genética , Glucosefosfato Desidrogenase/biossíntese , Glucosefosfato Desidrogenase/genética , Proteínas Recombinantes de Fusão/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Eletroforese em Gel de Poliacrilamida , Escherichia coli/química , Escherichia coli/enzimologia , Glucosefosfato Desidrogenase/química , Glucosefosfato Desidrogenase/isolamento & purificação , Immunoblotting , Masculino , Modelos Moleculares , Dados de Sequência Molecular , Conformação Proteica , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/isolamento & purificação , Alinhamento de Sequência , Análise de Sequência de ProteínaRESUMO
Use of untreated municipal wastewater (WW) contains toxic trace elements that pose a serious threat to the soil-plant-human continuum. The use of biochar (BC) is a promising approach to minimize trace element induced toxicity in the ecosystem. Therefore, the present study aims to evaluate the efficacy of BC derived from wheat straw and iron oxide nanoparticles doped biochar (IO-BC) to reduce trace element buildup in soil and plants that consequently affect tomato plant growth and physiological activity under WW irrigation. The BC and IO-BC were applied at four levels (0, 0.5, 1, and 1.5%) in WW irrigated soils. The results indicated that the addition of WW + BC and WW + IO-BC resulted in significant reduction in trace element mobility in soil. Interestingly, the application of WW + IO-BC (1.5%) was more effective in reducing trace element mobility and bioavailability in soil by 78% (As), 58% (Cr), 46% (Pb) and 50% (Cd) compared to WW irrigation, and thus reduced trace element accumulation and toxicity in plants. Results revealed that WW irrigation negatively affected tomato growth, fruit yield, physiology and antioxidative response. Addition of WW + BC and WW + IO-BC ameliorated the oxidative stress (up to 65% and 58% in H2O2 and MDA) and increased plant tolerance (up to 49% in POD and APX activity). The risk indices also showed minimum human health risk (H1 < 1) from tomato after the addition of BC or IO-BC in WW irrigated soils. It is concluded that IO-BC addition in WW irrigated soil could assist in reducing trace elements accumulation and toxicity in tomato and associated human health risks.
Assuntos
Poluentes do Solo , Solanum lycopersicum , Oligoelementos , Carvão Vegetal , Ecossistema , Humanos , Peróxido de Hidrogênio , Nanopartículas Magnéticas de Óxido de Ferro , Solo , Poluentes do Solo/análise , Poluentes do Solo/toxicidadeRESUMO
PURPOSE: Congenital fibrosis of the extraocular muscles type I (CFEOM1), the most common CFEOM worldwide, is characterized by bilateral ptotic hypotropia, an inability to supraduct above the horizontal midline, horizontal strabismus (typically exotropia), and ophthalmoplegia with abnormal synkinesis. This distinct non-syndromic phenotype is considered autosomal dominant and is virtually always from heterozygous missense mutations in kinesin family member 21A (KIF21A). However, there are occasional KIF21A-negative cases, opening the possibility for a recessive cause. The objective of this study is to explore this possibility by assessing CFEOM1 patients exclusively from consanguineous families, who are the most likely to have recessive cause for their phenotype if a recessive cause exists. METHODS: Ophthalmic examination and candidate gene direct sequencing (KIF21A, paired-like homeobox 2A [PHOX2A], tubulin beta-3 [TUBB3]) of CFEOM1 patients from consanguineous families referred for counseling from 2005 to 2010. RESULTS: All 5 probands had classic CFEOM1 as defined above. Three had siblings with CFEOM. None of the probands had mutations in KIF21A, PHOX2A, or TUBB3. CONCLUSIONS: The lack of KIF21A mutations in CFEOM1 patients exclusively from consanguineous families, most of whom had siblings with CFEOM, is strong evidence for a recessive form of CFEOM1. Further studies of such families will hopefully uncover the specific locus(loci).
Assuntos
Cinesinas/genética , Mutação , Músculos Oculomotores/patologia , Oftalmoplegia/genética , Adolescente , Blefaroptose/genética , Criança , Pré-Escolar , Consanguinidade , Primers do DNA/genética , Saúde da Família , Feminino , Genes Dominantes , Genes Recessivos , Heterozigoto , Humanos , Masculino , Mutação de Sentido Incorreto , Oftalmoplegia/patologia , Fenótipo , Arábia SauditaRESUMO
Heat shock proteins are ubiquitous, induced under a number of environmental and metabolic stresses, with highly conserved DNA sequences among mammalian species. Camelus dromedaries (the Arabian camel) domesticated under semi-desert environments, is well adapted to tolerate and survive against severe drought and high temperatures for extended periods. This is the first report of molecular cloning and characterization of full length cDNA of encoding a putative stress-induced heat shock HSPA6 protein (also called HSP70B') from Arabian camel. A full-length cDNA (2417 bp) was obtained by rapid amplification of cDNA ends (RACE) and cloned in pET-b expression vector. The sequence analysis of HSPA6 gene showed 1932 bp-long open reading frame encoding 643 amino acids. The complete cDNA sequence of the Arabian camel HSPA6 gene was submitted to NCBI GeneBank (accession number HQ214118.1). The BLAST analysis indicated that C. dromedaries HSPA6 gene nucleotides shared high similarity (77-91%) with heat shock gene nucleotide of other mammals. The deduced 643 amino acid sequences (accession number ADO12067.1) showed that the predicted protein has an estimated molecular weight of 70.5 kDa with a predicted isoelectric point (pI) of 6.0. The comparative analyses of camel HSPA6 protein sequences with other mammalian heat shock proteins (HSPs) showed high identity (80-94%). Predicted camel HSPA6 protein structure using Protein 3D structural analysis high similarities with human and mouse HSPs. Taken together, this study indicates that the cDNA sequences of HSPA6 gene and its amino acid and protein structure from the Arabian camel are highly conserved and have similarities with other mammalian species.
Assuntos
Camelus/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Animais , Sequência de Bases , Clonagem Molecular , DNA Complementar/química , DNA Complementar/metabolismo , Proteínas de Choque Térmico HSP70/química , Proteínas de Choque Térmico HSP70/genética , Humanos , Masculino , Camundongos , Modelos Moleculares , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Filogenia , Estrutura Terciária de Proteína , RNA/química , RNA/metabolismo , Alinhamento de Sequência , Análise de Sequência de RNA , Homologia de Sequência de AminoácidosRESUMO
A study has been carried out to define the effect of drastic temperature changes on the performance of lab-scale hollow-fibre MBR in treating municipal wastewater at a flux of 10 L m(-2) h(-1) (LMH). The objectives of the study were to estimate the activated sludge properties, the removal efficiencies of COD and NH(3)-N and the membrane fouling tendency under critical conditions of drastic temperature changes (23, 33, 42 & 33 °C) and MLSS concentration ranged between 6,382 and 8,680 mg/L. The study exhibited that the biomass reduction, the low sludge settleability and the supernatant turbidity were results of temperature increase. The temperature increase led to increase in SMP carbohydrate and protein, and to decrease in EPS carbohydrate and protein. The BRE of COD dropped from 80% at 23 °C to 47% at 42 °C, while the FRE was relatively constant at about 90%. Both removal efficiencies of NH(3)-N trended from about 100% at 33 °C to less than 50% at 42 °C. TMP and BWP ascended critically with temperature increase up to 336 and 304 mbar respectively by the end of the experiment. The values of suspended solids (SS) and the turbidity in the final effluent were negligible. The DO in the mixed liquor was varying with temperature change, while the pH was within the range of 6.7-8.3.
Assuntos
Temperatura , Purificação da Água/métodos , Biomassa , Reatores Biológicos , Concentração de Íons de Hidrogênio , Oxigênio/metabolismo , Esgotos/química , Eliminação de Resíduos Líquidos/métodosRESUMO
Hydrogen sulfide (H2S) has emerged as a potential gasotransmitter in plants with a beneficial role in stress amelioration. Despite the various known functions of H2S in plants, not much information is available to explain the associative role of molybdenum (Mo) and hydrogen sulfide (H2S) signaling in plants under arsenic toxicity. In view to address such lacunae in our understanding of the integrative roles of these biomolecules, the present work attempts to decipher the roles of Mo and H2S in mitigation of arsenate (AsV) toxicity in faba bean (Vicia faba L.) seedlings. AsV-stressed seedlings supplemented with exogenous Mo and/or NaHS treatments (H2S donor) showed resilience to AsV toxicity manifested by reduction of apoptosis, reactive oxygen species (ROS) content, down-regulation of NADPH oxidase and GOase activity followed by upregulation of antioxidative enzymes in leaves. Fluorescent localization of ROS in roots reveals changes in its intensity and spatial distribution in response to MO and NaHS supplementation during AsV stress. Under AsV toxicity conditions, seedlings subjected to Mo + NaHS showed an increased rate of nitrogen metabolism evident by elevation in nitrate reductase, nitrite reductase and glutamine synthetase activity. Furthermore, the application of Mo and NaHS in combination positively upregulates cysteine and hydrogen sulfide biosynthesis in the absence and presence of AsV stress. Mo plus NaHS-supplemented seedlings exposed to AsV toxicity showed a substantial reduction in oxidative stress manifested by reduced ELKG, lowered MDA content and higher accumulation of proline in leaves. Taken together, the present findings provide substantial evidence on the synergetic role of Mo and H2S in mitigating AsV stress in faba bean seedlings. Thus, the application of Mo and NaHS reveals their agronomic importance to encounter heavy metal stress for management of various food crops.
Assuntos
Arsênio , Sulfeto de Hidrogênio , Vicia faba , Arsênio/toxicidade , Cisteína , Molibdênio/toxicidade , Nitrogênio , PlântulaRESUMO
Present investigation reports the role of calcium (Ca2+) and hydrogen sulfide (H2S) crosstalk associated with Vigna radiata seedlings subjected to K+ deficient conditions under short-term (24 h) and long-term (72 h) NaCl stress. Perusal of the data reveals that under short-term NaCl stress an initial decline in K+ level led to the elevation in Ca2+ and H2S levels along with improvement in antioxidant system and reduction in reactive oxygen species (ROS) production. Under long-term NaCl stress a further decline in K+ content was deleterious that led to a lower K+/Na+ ratio. This was followed by reduction in antioxidant system along with excessive accumulation of ROS and methylglyoxal content, and increased membrane damage. However, supplementation of the seedling roots with Ca2+ enhanced biosynthesis of H2S through enhancing cysteine pool. The present findings suggest that synergistic action of Ca2+ and H2S induced the activity of H+-ATPase that created H+ gradient which in turn induced Na+/H+ antiport system that accelerated K+ influx and Na+ efflux. All of these together contributed to a higher K+/Na+ ratio, activation of antioxidative defense system, and maintenance of redox homeostasis and membrane integrity in Ca2+-supplemented stressed seedlings. Role of Ca2+ and H2S in the regulation of Na+/H+ antiport system was validated by the use of sodium orthovanadate (plasma membrane H+-ATPase inhibitor), tetraethylammonium chloride (K+ channel blocker), and amiloride (Na+/H+ antiporter inhibitor). Application of Ca2+-chelator EGTA (ethylene glycol-bis(b-aminoethylether)-N,N,N',N'-tetraacetic acid) and H2S scavenger hypotaurine abolished the effect of Ca2+, suggesting the involvement of Ca2+ and H2S in the alleviation of NaCl stress. Moreover, use of EGTA and HT also substantiates the downstream functioning of H2S during Ca2+-mediated regulation of plant adaptive responses to NaCl stress. To sum up, present findings reveal the association of Ca2+ and H2S signaling in the regulation of ion homeostasis and antioxidant defense during K+-deficient NaCl stress.
Assuntos
Cálcio , Sulfeto de Hidrogênio , Raízes de Plantas , Vigna , Antioxidantes/metabolismo , Cálcio/metabolismo , Sulfeto de Hidrogênio/metabolismo , Transporte de Íons , Raízes de Plantas/fisiologia , Potássio/metabolismo , Estresse Salino/fisiologia , Cloreto de Sódio/farmacologia , Trocadores de Sódio-Hidrogênio/metabolismo , Vigna/fisiologiaRESUMO
Blister agents damage the skin, eyes, mucous membranes and subcutaneous tissues. Other toxic effects may occur after absorption. The response of the Scientific Advisory Board (SAB) of the Organisation for the Prohibition of Chemical Weapons (OPCW) to a request from the OPCW Director-General in 2013 on the status of medical countermeasures and treatments to blister agents is updated through the incorporation of the latest information. The physical and toxicological properties of sulfur mustard and clinical effects and treatments are summarised. The information should assist medics and emergency responders who may be unfamiliar with the toxidrome of sulfur mustard and its treatment.
Assuntos
Substâncias para a Guerra Química/intoxicação , Gás de Mostarda/intoxicação , Animais , Humanos , Contramedidas MédicasRESUMO
We investigated rock varnish, a thin, manganese- and iron-rich, dark surface crust, on basaltic lava flows and petroglyphs in the Owens and Rose Valleys (California) by portable X-ray fluorescence (pXRF) and femtosecond laser-ablation inductively-coupled-plasma mass spectrometry (fs-LA-ICPMS). The major element composition of the varnish was consistent with a mixture of Mn-Fe oxyhydroxides and clay minerals. As expected, it contained elevated concentrations of elements that are typically enriched in rock varnish, e.g., Mn, Pb, Ba, Ce, and Co, but also showed unusually high enrichments in U, Cu, and Th. The rare earth and yttrium (REY) enrichment pattern revealed a very strong positive cerium (Ce) anomaly and distinct negative europium (Eu) and Y anomalies. The light rare earth elements (REE) were much more strongly enriched than the heavy REY. These enrichment patterns are consistent with a formation mechanism by leaching of Mn and trace elements from aeolian dust, reprecipitation of Mn and Fe as oxyhydroxides, and scavenging of trace elements by these oxyhydroxides. We inferred accumulation rates of Mn and Fe in the varnish from their areal densities measured by pXRF and the known ages of some of the lava flow surfaces. The areal densities of Mn and Fe, as well as their accumulation rates, were comparable to our previous results from the desert of Saudi Arabia. There was a moderate dependence of the Mn areal density on the inclination of the rock surfaces, but no relationship to its cardinal orientation. We attempted to use the degree of varnish regrowth on the rock art surfaces as an estimate of their age. While an absolute dating of the petroglyphs was not possible because of the lack of suitable calibration surfaces and a considerable amount of variability, the measured degree of varnish regrowth on the various petroglyphs was consistent with chronologies based on archeological and other archaeometric techniques. In particular, our results suggest that rock art creation in the study area continued over an extended period of time, possibly starting around the Pleistocene/Holocene transition and extending into the last few centuries.
Assuntos
Argila/química , Hidróxidos/análise , Ferro/análise , Manganês/análise , California , Meio Ambiente , Monitoramento Ambiental , Espectrometria de Massas , Metais Terras Raras/análise , Espectrometria por Raios X , Propriedades de Superfície , Erupções Vulcânicas , Ítrio/análiseRESUMO
Hydrogen sulfide (H2S) and nitric oxide (NO) have been known to affect vast number of processes in plants under abiotic stresses. Also, calcium (Ca) works as a second messenger in plants, which underpins the abiotic stress-induced damage. However, the sequence of action of these signaling molecules against cadmium (Cd)-induced cellular oxidative damage remains unidentified. Therefore, we studied the synergistic actions and/or relationship of signaling molecules and Ca-dependent activation of tolerance mechanisms in Vigna radiata seedlings under Cd stress. The present study shows that exogenous Ca supplemented to Cd-stressed V. radiata seedlings reduced Cd accumulation and improved the activity of nitrate reductase, and L/D-cysteine desulfhydrase (LCD/DCD) that resulted in improved synthesis of NO and H2S content. Application of Ca also elevated the level of cysteine (Cys) by upregulating the activity of Cys-synthesizing enzymes serine acetyltransferase and O-acetylserine(thiol)lyase in Cd-stressed seedlings. Maintenance of Cys pool under Cd stress contributed to improved H2S content which together with Ca and NO improved antioxidant enzymes and components of ascorbate-glutathione (AsA-GSH) cycle. All these collectively regulated the activity of NADPH oxidase and glycolate oxidase, resulting in the inhibition of Cd-induced generation of reactive oxygen species. The elevated level of Cys also assisted the Cd-stressed seedlings in maintaining GSH pool which retained normal functioning of AsA-GSH cycle and led to enhanced content of phytochelatins coupled with reduced Cd content. The positive effect of these events manifested in an enhanced rate of photosynthesis, carbohydrate accumulation, and growth attributes of the plants. On the contrary, addition of NO scavenger cPTIO [2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide], H2S scavenger HT (Hypotaurine) and Ca-chelator EGTA (Ethylene glycol-bis(b-aminoethylether)-N,N,N',N'-tetraacetic acid) again developed a condition similar to stress and positive effect of the signaling molecules was abolished. The findings of the study postulate that Ca in association with NO and H2S mitigates Cd-induced impairment and enhances the tolerance of the V. radiata plants against Cd stress. The results of the study also substantiate that Ca acts both upstream as well as downstream of NO signals whereas, H2S acts downstream of Ca and NO during Cd-stress responses of the plants.