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1.
Environ Res ; 204(Pt B): 112067, 2021 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-34543636

RESUMO

COVID-19 positive patients can egest live SARS-CoV-2 virus and viral genome fragments through faecal matter and urine, raising concerns about viral transmission through the faecal-oral route and/or contaminated aerosolized water. These concerns are amplified in many low- and middle-income countries, where raw sewage is often discharged into surface waterways and open defecation is common. Nonetheless, there has been no evidence of COVID-19 transmission via ambient urban water, and the virus viability in such aquatic matrices is believed to be minimal and not a matter of concern. In this manuscript, we attempt to discern the presence of SARS-CoV-2 genetic material (ORF-1ab, N and S genes) in the urban water (lakes, rivers, and drains) of the two Indian cities viz., Ahmedabad (AMD), in western India with 9 wastewater treatment plants (WWTPs) and Guwahati (GHY), in the north-east of the country with no such treatment facilities. The present study was carried out to establish the applicability of environmental water surveillance (E-wat-Surveillance) of COVID-19 as a potential tool for public health monitoring at the community level. 25.8% and 20% of the urban water samples had detectable SARS-CoV-2 RNA load in AMD and GHY, respectively. N-gene > S-gene > ORF-1ab-gene were readily detected in the urban surface water of AMD, whereas no such observable trend was noticed in the case of GHY. The high concentrations of SARS-CoV-2 genes (e.g., ORF-1ab; 800 copies/L for Sabarmati River, AMD and S-gene; 565 copies/L for Bharalu urban river, GHY) found in urban waters suggest that WWTPs do not always completely remove the virus genetic material and that E-wat-Surveillance of COVID-19 in cities/rural areas with poor sanitation is possible.

2.
Sci Total Environ ; 792: 148367, 2021 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-34465041

RESUMO

Following the proven concept, capabilities, and limitations of detecting the RNA of Severe Acute Respiratory Coronavirus 2 (SARS-CoV-2) in wastewater, it is pertinent to understand the utility of wastewater surveillance data on various scale. In the present work, we put forward the first wastewater surveillance-based city zonation for effective COVID-19 pandemic preparedness. A three-month data of Surveillance of Wastewater for Early Epidemic Prediction (SWEEP) was generated for the world heritage city of Ahmedabad, Gujarat, India. In this expedition, 116 wastewater samples were analyzed to detect SARS-CoV-2 RNA, from September 3rd to November 26th, 2020. A total of 111 samples were detected with at least two out of three SARS-CoV-2 genes (N, ORF 1ab, and S). Monthly variation depicted a significant decline in all three gene copies in October compared to September 2020, followed by a sharp increment in November 2020. Correspondingly, the descending order of average effective gene concentration was: November (~10,729 copies/L) > September (~3047 copies/L) > October (~454 copies/L). Monthly variation of SARS-CoV-2 RNA in the wastewater samples may be ascribed to a decline of 20.48% in the total number of active cases in October 2020 and a rise of 1.82% in November 2020. Also, the monthly recovered new cases were found to be 16.61, 20.03, and 15.58% in September, October, and November 2020, respectively. The percentage change in the gene concentration was observed in the lead of 1-2 weeks with respect to the percentage change in the provisional figures of confirmed cases. SWEEP data-based city zonation was matched with the heat map of the overall COVID-19 infected population in Ahmedabad city, and month-wise effective gene concentration variations are shown on the map. The results expound on the potential of WBE surveillance of COVID-19 as a city zonation tool that can be meaningfully interpreted, predicted, and propagated for community preparedness through advanced identification of COVID-19 hotspots within a given city.


Assuntos
COVID-19 , Humanos , Índia/epidemiologia , Pandemias , RNA Viral , SARS-CoV-2 , Águas Residuárias
3.
J Hazard Mater ; 416: 126125, 2021 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-34492919

RESUMO

The ongoing COVID-19 pandemic increases the consumption of antimicrobial substances (ABS) due to the unavailability of approved vaccine(s). To assess the effect of imprudent consumption of ABS during the COVID-19 pandemic, we compare the 2020 prevalence of antidrug resistance (ADR) of Escherichia coli (E. coli) with a similar survey carried out in 2018 in Ahmedabad, India using SARS-CoV-2 gene detection as a marker of ABS usage. We found a significant ADR increase in 2020 compared to 2018 in ambient water bodies, harbouring a higher incidence of ADR E.coli towards non-fluoroquinolone drugs. Effective SARS-CoV-2 genome copies were found to be associated with the ADR prevalence. The prevalence of ADR depends on the efficiency of WWTPs (Wastewater Treatment Plants) and the catchment area in its vicinity. In the year 2018 study, prevalence of ADR was discretely distributed, and the maximum ADR prevalence recorded was ~60%; against the current homogenous ADR increase, and up to 85% of maximum ADR among the incubated E.coli isolated from the river (Sabarmati) and lake (Chandola and Kankaria) samples. Furthermore, wastewater treatment plants showed less increase in comparison to the ambient waters, which eventually imply that although SARS-CoV-2 genes and faecal pollution may be diluted in the ambient waters, as indicated by low Ct-value and E.coli count, the danger of related aftermath like ADR increase cannot be nullified. Also, Non-fluoroquinolone drugs exhibited overall more resistance than quinolone drugs. Overall, this is probably the first-ever study that traces the COVID-19 pandemic imprints on the prevalence of antidrug resistance (ADR) through wastewater surveillance and hints at monitoring escalation of other environmental health parameters. This study will make the public and policyholders concerned about the optimum use of antibiotics during any kind of treatment.


Assuntos
COVID-19 , Escherichia coli/genética , Humanos , Pandemias , SARS-CoV-2 , Águas Residuárias
4.
J Hazard Mater ; 411: 124822, 2021 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-33858073

RESUMO

Cyanobacteria, the first photoautotrophs have remarkable adaptive capabilities against most abiotic stresses, including Cd. A model cyanobacterium, Anabaena sp. PCC 7120 has been commonly used to understand cyanobacterial plasticity under different environmental stresses. However, very few studies have focused on the acute Cd toxicity. In this context, Anabaena was subjected to 100 µM Cd for 48 h (acute Cd stress, ACdS) and then transferred into the fresh medium for post-stress recovery (PSR). We further investigated the dynamics of morpho-ultrastructure, physiology, cytosolic proteome, thylakoidal complexes, chelators, and transporters after ACdS, as well as during early (ER), mid (MR), and late (LR) phases of PSR. The findings revealed that ACdS induced intracellular Cd accumulation and ROS production, altered morpho-ultrastructure, reduced photosynthetic pigments, and affected the structural organization of PSII, which subsequently hindered photosynthetic efficiency. Anabaena responded to ACdS and recovered during PSR by reprogramming the expression pattern of proteins/genes involved in cellular defense and repair; CO2 access, Calvin-Benson cycle, glycolysis, and pentose phosphate pathway; protein biosynthesis, folding, and degradation; regulatory functions; PSI-based cyclic electron flow; Cd chelation; and efflux. These modulations occurred in an integrated and coordinated manner that facilitated Anabaena to detoxify Cd and repair ACdS-induced cellular damage.


Assuntos
Anabaena , Cianobactérias , Anabaena/genética , Anabaena/metabolismo , Proteínas de Bactérias/metabolismo , Cádmio/toxicidade , Cianobactérias/metabolismo , Fotossíntese , Proteoma
5.
J Agric Food Chem ; 69(11): 3371-3379, 2021 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-33688734

RESUMO

Partially acetylated chito-oligosaccharides (paCOSs) are bioactive compounds with potential medical applications. Their biological activities are largely dependent on their structural properties, in particular their degree of polymerization (DP) and the position of the acetyl groups along the glycan chain. The production of structurally defined paCOSs in a purified form is highly desirable to better understand the structure/bioactivity relationship of these oligosaccharides. Here, we describe a newly discovered chitinase from Paenibacillus pabuli (PpChi) and demonstrate by mass spectrometry that it essentially produces paCOSs with a DP of three and four that carry a single N-acetylation at their reducing end. We propose that this specific composition of glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) residues, as in GlcN(n)GlcNAc1, is due to a subsite specificity toward GlcN residues at the -2, -3, and -4 positions of the partially acetylated chitosan substrates. In addition, the enzyme is stable, as evidenced by its long shelf life, and active over a large temperature range, which is of high interest for potential use in industrial processes. It exhibits a kcat of 67.2 s-1 on partially acetylated chitosan substrates. When PpChi was used in combination with a recently discovered fungal auxilary activity (AA11) oxidase, a sixfold increase in the release of oligosaccharides from the lobster shell was measured. PpChi represents an attractive biocatalyst for the green production of highly valuable paCOSs with a well-defined structure and the expansion of the relatively small library of chito-oligosaccharides currently available.


Assuntos
Quitinases , Quitosana , Acetilação , Animais , Quitina/metabolismo , Quitinases/metabolismo , Quitosana/metabolismo , Oligossacarídeos , Paenibacillus
6.
Biomacromolecules ; 22(4): 1600-1613, 2021 04 12.
Artigo em Inglês | MEDLINE | ID: mdl-33749252

RESUMO

Commercial mucin glycoproteins are routinely used as a model to investigate the broad range of important functions mucins fulfill in our bodies, including lubrication, protection against hostile germs, and the accommodation of a healthy microbiome. Moreover, purified mucins are increasingly selected as building blocks for multifunctional materials, i.e., as components of hydrogels or coatings. By performing a detailed side-by-side comparison of commercially available and lab-purified variants of porcine gastric mucins, we decipher key molecular motifs that are crucial for mucin functionality. As two main structural features, we identify the hydrophobic termini and the hydrophilic glycosylation pattern of the mucin glycoprotein; moreover, we describe how alterations in those structural motifs affect the different properties of mucins-on both microscopic and macroscopic levels. This study provides a detailed understanding of how distinct functionalities of gastric mucins are established, and it highlights the need for high-quality mucins-for both basic research and the development of mucin-based medical products.


Assuntos
Glicoproteínas , Mucinas , Animais , Glicoproteínas/metabolismo , Glicosilação , Hidrogéis , Lubrificação , Mucinas/metabolismo , Suínos
7.
J Biosci Bioeng ; 131(5): 491-500, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33610455

RESUMO

Cyanobacteria hold promise as cell factories for the photoautotrophic conversion of carbon dioxide to useful chemicals. For the eventual commercial viability of such processes, cyanobacteria need to be engineered for (i) efficient channeling of carbon flux toward the product of interest and (ii) improved product tolerance, the latter being the focus of this study. We chose the recently reported, fast-growing, high light and CO2 tolerant cyanobacterium Synechococcus elongatus PCC 11801 for adaptive laboratory evolution. In two parallel experiments that lasted over 8400 h of culturing and 100 serial passages, S. elongatus PCC 11801 was evolved to tolerate 5 g/L n-butanol or 30 g/L 2,3-butanediol representing a 100% improvement in concentrations tolerated. The evolved strains retained alcohol tolerance even after being passaged several times without the alcohol stress suggesting that the changes were permanent. Whole genome sequencing of the n-butanol evolved strains revealed mutations in a number of stress responsive genes encoding translation initiation factors, RpoB and an ABC transporter. In 2,3-butanediol evolved strains, genes for ClpC, a different ABC transporter, glyceraldehyde-3-phosphate dehydrogenase and phosphoribulokinase were found to be mutated. Furthermore, the evolved strains showed significant improvement in tolerance toward several other alcohols. Notably, the n-butanol evolved strain could tolerate up to 32 g/L ethanol, thereby making it a promising host for photosynthetic production of biofuels via metabolic engineering.


Assuntos
Evolução Molecular Direcionada , Solventes/farmacologia , Synechococcus/efeitos dos fármacos , Synechococcus/genética , Álcoois/farmacologia , Biocombustíveis , Dióxido de Carbono/metabolismo , Fotossíntese/efeitos dos fármacos , Synechococcus/metabolismo
8.
Sci Total Environ ; 764: 144164, 2021 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-33385648

RESUMO

Soil salinization has become a major global agricultural issue that threatens sustainable development goals related to food security, agriculture, resource conservation, and nutrition. The higher levels of salinity have detrimental effects on soil physico-chemical and biological characteristics and plant metabolism. Also, salinity poses a negative impact on the abundance and distribution of soil microbes and soil-dwelling organisms. Research has always been trying to overcome the salinity issue, but it does not fit well in conventional approaches. This review unravels traditional and modern salinity management techniques. Out of the available salinity management techniques, some are focused on enhancing soil properties (chemical amendments, biochar, earthworms, and their vermicompost, compost, microbial inoculants, electro remediation), some focus on improving plant properties (seed priming, afforestation, crop selection, genetic improvements, agroforestry) and some techniques augment both soil as well as plant properties in a synergic manner. Therefore, it is imperative to find a conclusive solution by integrating traditional and modern methods to find the most effective response to regionally-specific salinity related problems. This review aimed at critical analysis of the salinity problems, its impact on agroecosystem, and different management approaches available to date with a balanced viewpoint that would help to draw a possible roadmap towards the future investigation in this domain for sustainable management of salinity issues around the globe.


Assuntos
Salinidade , Solo , Agricultura , Medição de Risco
9.
Sci Total Environ ; 766: 144389, 2021 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-33429300

RESUMO

A culture-independent DNA metabarcoding analysis of the bacterial communities was carried out throughout a complete vermicomposting cycle of municipal solid waste material using the earthworm Eisenia fetida. 16S rRNA amplicons from the initial material (0 days), an intermediate (42 days), and a final stage (84 days) were sequenced in an Illumina NGS platform and compared. A steady increase in community diversity was observed corresponding to a 2.5-fold higher taxa richness and correspondingly risen values of the Shannon and Simpson ecological indexes and the evenness parameter. A total of 49,665 operational taxonomic units (OTUs) were counted. From the qualitative standpoint, a clear successional shift was observed with an initial community dominated by putatively plant-associated groups belonging to the Rhizobiales order within the Alphaproteobacteria class, regressively leaving the scores of relative abundance (RA) to the Firmicutes phylum and in particular to the Bacilli. Vermistabilization of municipal solid waste (MSW) increased (p < 0.001) the TKN and total P content in the final vermicompost, while pH, TOC, and C/N ratio declined (p < 0.001) in the process. Likewise, a progressive decrease was noticed in ß-glucosidase, acid phosphatase, and urease activity while protease and dehydrogenase showed a slight increase, followed by a steep fall. A strong positive correlation was observed among the canonical functions of physico-chemical attributes and enzyme activities. The canonical correspondence analysis (CCA) revealed that significant families did not change on the temporal scale; however, their abundance was influenced by the abiotic environmental factors. In comparison to prior studies on vermicomposting that used different earthworm species (Eisenia andrei) and different substrates, results reflect a considerable degree of substrate specificity for the earthworm species used. The results offer clues to optimize the vermistabilization of MSW along with its potential use in agriculture, to foster improved levels of the circular economy.


Assuntos
Oligoquetos , Animais , Bactérias/genética , RNA Ribossômico 16S/genética , Solo , Resíduos Sólidos
10.
Front Microbiol ; 11: 571093, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33178154

RESUMO

Many Stramenopile species belonging to oomycetes from the genus Saprolegnia infect fish, amphibians, and crustaceans in aquaculture farms and natural ecosystems. Saprolegnia parasitica is one of the most severe fish pathogens, responsible for high losses in the aquaculture industry worldwide. Most of the molecules reported to date for the control of Saprolegnia infections either are inefficient or have negative impacts on the health of the fish hosts or the environment resulting in substantial economic losses. Until now, the whole proteome of S. parasitica has not been explored for a systematic screening of novel inhibitors against the pathogen. The present study was designed to develop a consensus computational framework for the identification of potential target proteins and their inhibitors and subsequent experimental validation of selected compounds. Comparative analysis between the proteomes of Saprolegnia, humans and fish species identified proteins that are specific and essential for the survival of the pathogen. The DrugBank database was exploited to select food and drug administration (FDA)-approved inhibitors whose high binding affinity to their respective protein targets was confirmed by computational modeling. At least six of the identified compounds significantly inhibited the growth of S. parasitica in vitro. Triclosan was found to be most effective with a minimum inhibitory concentration (MIC100) of 4 µg/ml. Optical microscopy showed that the inhibitors affect the morphology of hyphal cells, with hyper-branching being commonly observed. The inhibitory effects of the compounds identified in this study on Saprolegnia's mycelial growth indicate that they are potentially usable for disease control against this class of oomycete pathogens. Similar approaches can be easily adopted for the identification of potential inhibitors against other plant and animal pathogenic oomycete infections.

11.
Sci Rep ; 10(1): 19125, 2020 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-33154404

RESUMO

The current outbreak of Covid-19 infection due to SARS-CoV-2, a virus from the coronavirus family, has become a major threat to human healthcare. The virus has already infected more than 44 M people and the number of deaths reported has reached more than 1.1 M which may be attributed to lack of medicine. The traditional drug discovery approach involves many years of rigorous research and development and demands for a huge investment which cannot be adopted for the ongoing pandemic infection. Rather we need a swift and cost-effective approach to inhibit and control the viral infection. With the help of computational screening approaches and by choosing appropriate chemical space, it is possible to identify lead drug-like compounds for Covid-19. In this study, we have used the Drugbank database to screen compounds against the most important viral targets namely 3C-like protease (3CLpro), papain-like protease (PLpro), RNA-dependent RNA polymerase (RdRp) and the spike (S) protein. These targets play a major role in the replication/transcription and host cell recognition, therefore, are vital for the viral reproduction and spread of infection. As the structure based computational screening approaches are more reliable, we used the crystal structures for 3C-like main protease and spike protein. For the remaining targets, we used the structures based on homology modeling. Further, we employed two scoring methods based on binding free energies implemented in AutoDock Vina and molecular mechanics-generalized Born surface area approach. Based on these results, we propose drug cocktails active against the three viral targets namely 3CLpro, PLpro and RdRp. Interestingly, one of the identified compounds in this study i.e. Baloxavir marboxil has been under clinical trial for the treatment of Covid-19 infection. In addition, we have identified a few compounds such as Phthalocyanine, Tadalafil, Lonafarnib, Nilotinib, Dihydroergotamine, R-428 which can bind to all three targets simultaneously and can serve as multi-targeting drugs. Our study also included calculation of binding energies for various compounds currently under drug trials. Among these compounds, it is found that Remdesivir binds to targets, 3CLpro and RdRp with high binding affinity. Moreover, Baricitinib and Umifenovir were found to have superior target-specific binding while Darunavir is found to be a potential multi-targeting drug. As far as we know this is the first study where the compounds from the Drugbank database are screened against four vital targets of SARS-CoV-2 and illustrates that the computational screening using a double scoring approach can yield potential drug-like compounds against Covid-19 infection.


Assuntos
Infecções por Coronavirus/tratamento farmacológico , Bases de Dados de Produtos Farmacêuticos , Avaliação Pré-Clínica de Medicamentos/métodos , Terapia de Alvo Molecular , Pneumonia Viral/tratamento farmacológico , COVID-19 , Proteases 3C de Coronavírus , Análise Custo-Benefício , Cisteína Endopeptidases/química , Cisteína Endopeptidases/metabolismo , Avaliação Pré-Clínica de Medicamentos/economia , Humanos , Simulação de Acoplamento Molecular , Pandemias , Conformação Proteica , Proteínas não Estruturais Virais/antagonistas & inibidores , Proteínas não Estruturais Virais/química , Proteínas não Estruturais Virais/metabolismo
12.
Nanomaterials (Basel) ; 10(6)2020 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-32580447

RESUMO

The use of cadmium sulphide quantum dot (CdS QD)-enabled products has become increasingly widespread. The prospect of their release in the environment is raising concerns. Here we have used the yeast model Saccharomyces cerevisiae to determine the potential impact of CdS QD nanoparticles on living organisms. Proteomic analyses and cell viability assays performed after 9 h exposure revealed expression of proteins involved in oxidative stress and reduced lethality, respectively, whereas oxidative stress declined, and lethality increased after 24 h incubation in the presence of CdS QDs. Quantitative proteomics using the iTRAQ approach (isobaric tags for relative and absolute quantitation) revealed that key proteins involved in essential biological pathways were differentially regulated over the time course of the experiment. At 9 h, most of the glycolytic functions increased, and the abundance of the number of heat shock proteins increased. This contrasts with the situation at 24 h where glycolytic functions, some heat shock proteins as well as oxidative phosphorylation and ATP synthesis were down-regulated. It can be concluded from our data that cell exposure to CdS QDs provokes a metabolic shift from respiration to fermentation, comparable to the situation reported in some cancer cell lines.

13.
Front Microbiol ; 11: 521, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32296406

RESUMO

To develop more ecologically sustainable agricultural practices requires that we reduce our reliance on synthetic chemical pesticides for crop protection. This will likely involve optimized biocontrol approaches - the use of beneficial soil microbes to attack potential plant pathogens to protect plants from diseases. Many bacterial species, including strains of Bacillus subtilis, have been explored for their biocontrol properties, as they can control the growth of harmful fungi, often by disrupting the fungal cell wall. A strain that is not often considered for this particular application is Bacillus subtilis natto, primarily known for fermenting soybeans via cell wall degradation in the Japanese probiotic dish "natto." Because deconstruction of the fungal cell wall is considered an important biocontrol trait, we were motivated to explore the possible anti-fungal properties of the B. subtilis natto strain. We show that B. subtilis natto can use complex fungal material as a carbon source for growth, and can effectively deconstruct fungal cell walls. We found degradation of fungal cell wall proteins, and showed that growth on a mix of peptides was very strong. We also found that intact fungal cell walls can induce the secretion of chitinases and proteases. Surprisingly, we could show that chitin, the bulk component of the fungal cell wall, does not permit successful growth of the natto strain or induce the secretion of chitinolytic enzymes, although these were produced during exposure to proteins or to complex fungal material. We have further shown that protease secretion is likely a constitutively enabled mechanism for nutrient scavenging by B. subtilis natto, as well as a potent tool for the degradation of fungal cell walls. Overall, our data highlight B. subtilis natto as a promising candidate for biocontrol products, with relevant behaviors that can be optimized by altering growth conditions. Whereas it is common for bacterial biocontrol products to be supplied with chitin or chitosan as a priming polysaccharide, our data indicate that this is not a useful approach with this particular bacterium, which should instead be supplied with either glucose or attenuated fungal material.

14.
J Environ Manage ; 255: 109914, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32063304

RESUMO

Rapid demographic expansion along with increasing urbanization has aggravated the problem of solid waste management. Therefore, scientists are seeking waste management methods that are eco-friendly, cost effective and produce immediate results. In the developing world, municipal solid waste (MSW) contains mostly organic substances, therefore vermicomposting could be a better and cost-effective option for waste management. In this study, vermicomposting of organic portion of MSW with cow dung (additive) was performed using Eisenia fetida. The results showed significant (p < 0.001) decline in pH (13.17%), TOC (21.70%), C: N (62.53%) and C: P (57.66%) ratios, whilst total N (108.9%), P (84.89%) and K (21.85%) content increased (p < 0.001) in matured vermicompost. Different enzymatic activities declined during termination phase of vermicomposting experiment with maximum decrease of 41.72 (p = 0.002) and 39.56% (p = 0.001) in protease and ß-glucosidase, respectively. FT-IR, TGA, DSC and SEM studies suggested that final vermicompost was more stabilized as compared to initial waste mixture, characterized by reduced levels of aliphatic materials, carbohydrates and increase in aromatic groups possibly due to biosynthesis of humic substances. Both, the conventional (physicochemical and enzyme activity) and advanced techniques depict maturity and stability of the ready vermicompost. However, FT-IR, TGA, DSC and SEM were proved to be more promising, fast and reliable techniques over conventional analyses.


Assuntos
Oligoquetos , Resíduos Sólidos , Animais , Bovinos , Feminino , Substâncias Húmicas , Esterco , Solo , Espectroscopia de Infravermelho com Transformada de Fourier
15.
BMC Res Notes ; 13(1): 60, 2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-32028996

RESUMO

OBJECTIVE: Liverworts possess historical adaptive strategies for abiotic stresses because they were the first plants that shifted from water to land. Proteomics is a state-of-the-art technique that can capture snapshots of events occurring at the protein level in many organisms. Herein, we highlight the comparison and optimization of an effective protein extraction and precipitation protocol for two-dimensional gel electrophoresis (2-DE) of liverworts. RESULTS: We compared three different protein extraction methods, i.e.,1.5 M Tris-HCl (pH 8.8), 50 mM Tris-HCl (pH 7.5), and polyvinylpolypyrrolidone (PVPP) extraction, followed by three precipitation methods, i.e., 80% ethanol, 80% acetone, and 20% tricholoroacetic acid (TCA)-acetone, in a liverwort Dumortiera hirsuta. Among these methods, 50 mM Tris-HCl (pH 7.5) extraction, followed by 20% TCA-acetone precipitation, appeared to be more suitable for 2-DE. Furthermore, we performed modifications during protein washing, re-solubilization in rehydration buffer and isoelectric focusing (IEF). The modifications provided us better results in terms of protein yield, resolution, spot numbers, and intensities for 2-DE gels of D. hirsuta and other two liverworts, i.e., Marchantia paleacea and Plagiochasma appendiculatum. Furthermore, we randomly selected spots from the 2-DE gel of D. hirsuta and identified using mass spectrometry, which confirms the applicability of this protocol for liverworts proteomics.


Assuntos
Eletroforese em Gel Bidimensional/métodos , Hepatófitas/metabolismo , Proteínas de Plantas/isolamento & purificação , Ecossistema , Células Germinativas Vegetais/metabolismo
16.
Environ Monit Assess ; 192(3): 191, 2020 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-32080774

RESUMO

Varanasi, India's historic cultural capital, struggles with efficient waste management practices. This impacts environment and human well-being in terms of waste generation that is estimated around 550-650 TPD with a generation rate of 0.42 kg capita-1 day-1 (n = 117). The present study aims to explore and characterize wastes, current practices, ecological profiling, and phytotoxicity of an abandoned open dumping site, and vermicomposting of organic fraction of municipal solid waste (OFMSW) as sustainable waste management approach. Compositional analysis of waste indicates organic fraction (46.13%) as a major component along with a considerable amount of heavy metals. The calorific value and moisture content of municipal solid waste (MSW) was 2351.4 cal g-1 and 34.72%, respectively. Ecological profiling of the dumping site revealed that floral diversity and ecological species/indicators were negatively affected. Likewise, phytotoxicity results displayed a negative impact on germination and physiology of maize (Zea mays L.) plants grown on dumping site soil. Vermistabilization of OFMSW showed a significant increase in N (56.10-89.48%), P (33.93-82.87%), and K (25.55-50.42%) and a decrease in total organic carbon (15.15-24.81%). Similarly, C/N and C/P ratios decreased by 1.89-2.51 and 1.72-2.18 folds, respectively. A survey of stakeholders suggested that open dumping was the main practice adopted by Varanasi Municipal Corporation (VMC) during 2013-2015. Recently (2017-2018), VMC adopted different methods, such as door-to-door collection and source segregation for effective waste management. Waste characteristics and nutrient profile of the vermicompost explains that vermicomposting could be used for efficient waste management in Varanasi, further reducing the collection, transportation, and disposal costs of waste, which enables to close the loop and move towards a circular economy. Moreover, implications of existing waste management practices and possible management options need to be addressed scientifically. Therefore, this research outcome will help in designing a successful waste management plan for Varanasi and other cities with similar waste characteristics.


Assuntos
Eliminação de Resíduos , Gerenciamento de Resíduos , Cidades , Monitoramento Ambiental , Alimentos , Humanos , Índia , Resíduos Sólidos
17.
Astrobiology ; 19(12): 1442-1458, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31038352

RESUMO

The evolutionarily ancient Aquificales bacterium Sulfurihydrogenibium spp. dominates filamentous microbial mat communities in shallow, fast-flowing, and dysoxic hot-spring drainage systems around the world. In the present study, field observations of these fettuccini-like microbial mats at Mammoth Hot Springs in Yellowstone National Park are integrated with geology, geochemistry, hydrology, microscopy, and multi-omic molecular biology analyses. Strategic sampling of living filamentous mats along with the hot-spring CaCO3 (travertine) in which they are actively being entombed and fossilized has permitted the first direct linkage of Sulfurihydrogenibium spp. physiology and metabolism with the formation of distinct travertine streamer microbial biomarkers. Results indicate that, during chemoautotrophy and CO2 carbon fixation, the 87-98% Sulfurihydrogenibium-dominated mats utilize chaperons to facilitate enzyme stability and function. High-abundance transcripts and proteins for type IV pili and extracellular polymeric substances (EPSs) are consistent with their strong mucus-rich filaments tens of centimeters long that withstand hydrodynamic shear as they become encrusted by more than 5 mm of travertine per day. Their primary energy source is the oxidation of reduced sulfur (e.g., sulfide, sulfur, or thiosulfate) and the simultaneous uptake of extremely low concentrations of dissolved O2 facilitated by bd-type cytochromes. The formation of elevated travertine ridges permits the Sulfurihydrogenibium-dominated mats to create a shallow platform from which to access low levels of dissolved oxygen at the virtual exclusion of other microorganisms. These ridged travertine streamer microbial biomarkers are well preserved and create a robust fossil record of microbial physiological and metabolic activities in modern and ancient hot-spring ecosystems.


Assuntos
Biodiversidade , Extremófilos/fisiologia , Fontes Termais/microbiologia , Microbiota/fisiologia , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Ciclo do Carbono , DNA Bacteriano/isolamento & purificação , Extremófilos/isolamento & purificação , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/metabolismo , Fósseis/microbiologia , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos , Sedimentos Geológicos/microbiologia , Oxirredução , Filogenia , RNA Ribossômico 16S/genética , Enxofre/metabolismo
18.
Cell Surf ; 5: 100030, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32743146

RESUMO

As an obligate biotroph, Blumeria graminis f. sp. hordei (Bgh) cannot be grown in an axenic culture, and instead must be cultivated on its host species, Hordeum vulgare (barley). In this study an in vitro system utilizing n-hexacosanal, a constituent of the barley cuticle and known inducer of Bgh germination, was used to cultivate Bgh and differentiate conidia up to the appressorial germ tube stage for analysis. Transcriptomic and proteomic profiling of the appressorial germ tube stage revealed that there was a significant shift towards energy and protein production during the pre-penetrative phase of development, with an up-regulation of enzymes associated with cellular respiration and protein synthesis, modification and transport. Glycosidic linkage analysis of the cell wall polysaccharides demonstrated that during appressorial development an increase in 1,3- and 1,4-linked glucosyl residues and xylosyl residues was detected along with a significant decrease in galactosyl residues. The use of this in vitro cultivation method demonstrates that it is possible to analyse the pre-penetrative processes of Bgh development in the absence of a plant host.

19.
Front Plant Sci ; 9: 1681, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30510561

RESUMO

Plasmodesmata are channels that link adjacent cells in plant tissues through which molecular exchanges take place. They are involved in multiple processes vital to plant cells, such as responses to hormonal signaling or environmental challenges including osmotic stress, wounding and pathogen attack. Despite the importance of plasmodesmata, their proteome is not well-defined. Here, we have isolated fractions enriched in plasmodesmata from cell suspension cultures of Populus trichocarpa and identified 201 proteins that are enriched in these fractions, thereby providing further insight on the multiple functions of plasmodesmata. Proteomics analysis revealed an enrichment of proteins specifically involved in responses to stress, transport, metabolism and signal transduction. Consistent with the role of callose deposition and turnover in the closure and aperture of the plasmodesmata and our proteomic analysis, we demonstrate the enrichment of callose synthase activity in the plasmodesmata represented by several gene products. A new form of calcium-independent callose synthase activity was detected, in addition to the typical calcium-dependent enzyme activity, suggesting a role of calcium in the regulation of plasmodesmata through two forms of callose synthase activities. Our report provides the first proteomic investigation of the plasmodesmata from a tree species and the direct biochemical evidence for the occurrence of several forms of active callose synthases in these structures. Data are available via ProteomeXchange with identifier PXD010692.

20.
Biomacromolecules ; 19(3): 872-882, 2018 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-29451983

RESUMO

The mucus gel covers the wet epithelia that forms the inner lining of the body. It constitutes our first line of defense protecting the body from infections and other deleterious molecules. Failure of the mucus barrier can lead to the inflammation of the mucosa such as in inflammatory bowel diseases. Unfortunately, there are no effective strategies that reinforce the mucus barrier properties to recover or enhance its ability to protect the epithelium. Herein, we describe a mucus engineering approach that addresses this issue where we physically cross-link the mucus gel with low molar mass chitosan variants to reinforce its barrier functions. We tested the effect of these chitosans on mucus using in-lab purified porcine gastric mucins, which mimic the native properties of mucus, and on mucus-secreting HT29-MTX epithelial cell cultures. We found that the lowest molar mass chitosan variant (degree of polymerization of 8) diffuses deep into the mucus gels while physically cross-linking the mucin polymers, whereas the higher molar mass chitosan variants (degree of polymerization of 52 and 100) interact only superficially. The complexation resulted in a tighter mucin polymer mesh that slowed the diffusion of dextran polymers and of the cholera toxin B subunit protein through the mucus gels. These results uncover a new use for low molar mass mucoadhesive polymers such as chitosans as noncytotoxic mucosal barrier enhancers that could be valuable in the prevention and treatment of mucosal diseases.


Assuntos
Quitosana , Células Epiteliais/metabolismo , Mucosa Intestinal/metabolismo , Mucinas/metabolismo , Animais , Linhagem Celular , Quitosana/farmacocinética , Quitosana/farmacologia , Toxina da Cólera/farmacocinética , Toxina da Cólera/farmacologia , Dextranos/farmacocinética , Dextranos/farmacologia , Células Epiteliais/patologia , Humanos , Mucosa Intestinal/patologia , Suínos
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