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Land use change from native vegetation to cropping can significantly affect the quantity and quality of soil organic matter (SOM). However, it remains unclear how the chemical composition of SOM is affected by such changes. This study employed a sequential chemical extraction to partition SOM from an Oxisol into several distinct fractions: water-soluble fractions (ultrapure water (W)), organometal complexes (sodium pyrophosphate (PP)), short-range ordered (SRO) oxides (hydroxylamine-HCl (HH)), and well-crystalline oxides (dithionite-HCl (DH)). Coupled with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), the impact of land use change on the molecular composition of different OM fractions was investigated. Greater amounts of OM were observed in the PP and HH fractions compared to other fractions, highlighting their importance in SOM stabilization. The composition of different OM fractions varied based on extracted phases, with lignin-like and tannin-like compounds being prevalent in the PP and HH fractions, while aliphatic-like compounds dominated in the DH fraction. Despite changes in the concentration of each OM fraction from native vegetation to cropping, there was little influence of land use change on the molecular composition of OM associated with different mineral phases. No significant selective loss or preservation of organic carbon compounds was observed, indicating the composition of SOM remained unchanged.
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Suelo , Suelo/química , Compuestos Orgánicos/análisisRESUMEN
Candida species is the causative agent in approximately 80% of invasive mycoses and drug-resistant Candida albicans is among the four strains of 'critical priority group' framed by WHO. Lichens are endowed with some rare phytochemicals and a plethora of therapeutics viz. antifungal capacities of Roccella montagnei. Biosynthesis of silver nanoparticles (AgNPs) using lichen could offer an eco-friendly, and cost-effective alternative against emerging 'microbial resistance.' Therefore, the objective was to biosynthesize silver nanoparticles (Rm-AgNPs) using a Hydro-alcoholic (1:1) extract of R. montagnei to develop a potent anticandidal agent against Fluconazole-resistant C. albicans NBC099. UV-Spectroscopy identified AgNPs specific-peak of Rm-AgNPs at 420-440 nm and FTIR revealed the presence of amines, alcohol, aromatic compounds, and acids. SEM and TEM analysis indicated that Rm-AgNPs are spherical shaped with a size range of 10-50 nm. Zetasizer analysis indicated that particles are highly stable and have a mean hydrodynamic diameter of 116 nm with a zeta potential charge of - 41 mV. XRD analysis suggested face centered cubic crystal lattice structure. Results indicated that Rm-AgNPs strongly inhibited the growth of NBC099 at a minimum inhibitory concentration (IC50) of ≤ 15 µg. C. albicans culture treated with Rm-AgNPs at concentrations below IC50, down-regulates the production of different virulence factors in NBC099, viz. hyphal formation (> 85%), biofilms production (> 80%), phospholipase, esterase, proteinase activity. The apoptosis assay demonstrated the Rm-AgNPs induced apoptosis in NBC099 cells via oxidative stress. Interestingly, Rm-AgNPs showed negligible cytotoxicity (< 6%) in murine RAW 246.7 macrophage cells at a concentration above 15 µg/mL. Therefore, Rm-AgNPs have been offered as an anti-candida alternative that can be utilized to improve the efficacy of already available medications.
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Ascomicetos , Candida albicans , Nanopartículas del Metal , Animales , Ratones , Fluconazol/farmacología , Plata/farmacología , CandidaRESUMEN
The quantitative assessment of micro-structure and load-induced damages in Al-SiC metal matrix composites (MMC) is important for its design optimization, performance evaluation and structure-property correlation. X-ray Phase contrast micro-tomography is potentially used for evaluation of its 3 dimensional micro-structure manifested in the form of voids, cracks, embedded particles, and load-induced damages. However, the contrast between Al matrix and SiC particles is insufficient for their clear morphological identification and quantitative assessment. In the present study, we have proposed and applied single image-based phase retrieval as a pre-processing step to micro-tomography reconstruction for improved assessment of micro-structure and cohesion-induced damages in Al-SiC MMC. The advantages of applying different phase retrieval techniques in the enhancement of image quality and morphological quantification of SiC particles, pores and cohesion damages are discussed. It is observed that the Paganin method offers the best improvement in contrast to noise ratio for the measurement of SiC particles embedded in the Al matrix.
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BACKGROUND: India is hyperendemic to dengue and over 50% of adults are seropositive. There is limited information on the association between neutralizing antibody profiles from prior exposure and viral RNA levels during subsequent infection. METHODS: Samples collected from patients with febrile illness was used to assess seropositivity by indirect ELISA. Dengue virus (DENV) RNA copy numbers were estimated by quantitative RT-PCR and serotype of the infecting DENV was determined by nested PCR. Focus reduction neutralizing antibody titer (FRNT) assay was established using Indian isolates to measure the levels of neutralizing antibodies and also to assess the cross-reactivity to related flaviviruses namely Zika virus (ZIKV), Japanese encephalitis virus (JEV) and West Nile virus (WNV). RESULTS: In this cross-sectional study, we show that dengue seropositivity increased from 52% in the 0-15 years group to 89% in >45 years group. Antibody levels negatively correlate with dengue RNAemia on the day of sample collection and higher RNAemia is observed in primary dengue as compared to secondary dengue. The geometric mean FRNT50 titers for DENV-2 is significantly higher as compared to the other three DENV serotypes. We observe cross-reactivity with ZIKV and significantly lower or no neutralizing antibodies against JEV and WNV. The FRNT50 values for international isolates of DENV-1, DENV-3 and DENV-4 is significantly lower as compared to Indian isolates. CONCLUSIONS: Majority of the adult population in India have neutralizing antibodies to all the four DENV serotypes which correlates with reduced RNAemia during subsequent infection suggesting that antibodies can be considered as a good correlate of protection.
India is one of the hotspots of dengue infection. The objective of the study was to assess whether having previous exposure to dengue virus could influence how the body will respond to repeat infections with dengue virus. Here, we analysed samples from febrile patients to measure the amount of dengue virus genetic material in the blood, the type of virus and the amount of antibodies, which are proteins produced by the host in response to dengue virus infection. The majority of patient samples demonstrated the capability to restrict all four types of dengue virus in circulation within the country, but reduced capacity to restrict when it comes to international dengue virus types. These data will help to inform future dengue vaccine design and clinical studies in India.
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The engineering of virus-like particles (VLPs) is a viable strategy for the development of vaccines and for the identification of therapeutic targets without using live viruses. Here, we report the generation and characterization of quadruple-antigen SARS-CoV-2 VLPs. VLPs were generated by transient transfection of two expression cassettes in adherent HEK293T cellsâone cassette containing Mpro for processing of three structural proteins (M, E, and N), and the second cassette expressing the Spike protein. Further characterization revealed that the VLPs retain close morphological and antigenic similarity with the native virus and also bind strongly to the SARS-CoV-2 receptor hACE-2 in an in vitro binding assay. Interestingly, the VLPs were found to internalize into U87-MG cells through cholesterol-rich domains in a dynamin-dependent process. Finally, our results showed that mice immunized with VLPs induce robust humoral and cellular immune responses mediated by enhanced levels of IL-4, IL-17, and IFNγ. Taken together, our results demonstrate that VLPs mimic the native virus and induce a strong immune response, indicating the possible use of these particles as an alternative vaccine candidate against SARS-CoV-2. VLPs can also be effective in mapping the initial stages of virus entry and screening inhibitors.
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COVID-19 , SARS-CoV-2 , Animales , COVID-19/prevención & control , Células HEK293 , Humanos , Interleucina-17 , Interleucina-4 , Ratones , Glicoproteína de la Espiga del Coronavirus/genética , Internalización del VirusRESUMEN
The interactions of plastics and soil organisms are complex and inconsistent observations on the effects of plastics on soil organisms have been made in published studies. In this study, we assessed the effects of plastic exposure on plants, fauna and microbial communities, with a meta-analysis. Using a total of 2936 observations from 140 publications, we analysed how responses in plants, soil fauna and microorganisms depended on the plastic concentration, size, type, species and exposure media. We found that overall plastics caused substantial detrimental effects to plants and fauna, but less so to microbial diversity and richness. Plastic concentration was one of the most important factors explaining variations in plant and faunal responses. Larger plastics (>1 µm) caused unfavourable changes to plant growth, germination and oxidative stress, while nanoplastics (NPs; ≤ 1 µm) only increased oxidative stress. On the contrary, there was a clear trend showing that small plastics adversely affected fauna reproduction, survival and locomotion than large plastics. Plant responses were indifferent to plastic type, with most studies conducted using polyethylene (PE) and polystyrene (PS) plastics, but soil fauna were frequently more sensitive to PS than to PE exposure. Plant species played a vital role in some parameters, with the effects of plastics being considerably greater on vegetable plants than on cereal plants.
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Plásticos , Suelo , Ecotoxicología , Plantas , PoliestirenosRESUMEN
Many adults in India have received at least one dose of COVID-19 vaccine with or without a prior history SARS-CoV-2 infection. However, there is limited information on the effect of prior immunity on antibody response upon vaccination in India. As immunization of individuals continues, we aimed to assess whether pre-existing antibodies are further boosted by a single dose of BBV152, an inactivated SARS-CoV-2 vaccine, and, if these antibodies can neutralize SARS-CoV-2 Delta and Omicron variants. Here we show that natural infection during the second wave in 2021 led to generation of neutralizing antibodies against other lineages of SARS-CoV-2 including the Omicron variant, albeit at a significantly lower level for the latter. A single dose of BBV152 boosted antibody titers against the Delta and the Omicron variants but the antibody levels remained low against the Omicron variant. Boosting of antibodies showed negative correlation with baseline neutralizing antibody titers.
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COVID-19 , Vacunas Virales , Anticuerpos Neutralizantes , Anticuerpos Antivirales , COVID-19/prevención & control , Vacunas contra la COVID-19 , Humanos , SARS-CoV-2 , VacunaciónRESUMEN
BACKGROUND: Deficiency of Zn is a major soil constraint in rice plant growth and yield. Edaphic factors such as Zn deficiency in soil in relation to plant performance are still poorly understood. Here, we report promising quantitative trait loci (QTL) conferring tolerance to Zn deficiency, which were identified through biparental mapping. The experiment was conducted using the 236 F7 recombinant inbred line mapping population derived from the cross of Kinandang Patong (Zn deficiency sensitive) and A69-1 (Zn deficiency tolerant). RESULTS: A total of six QTLs (qLB-2B, qLB-4B, qPM-4B, qPM-6B, qRZC-4B, qSZC-4B) on chromosomes 2, 4 and 6 were identified for environment 1, whereas five QTLs (qLB-2 N, qLB-4 N, qPM-4 N, qRZC-4 N, qSZC-4 N) on chromosomes 2 and 4 were detected for environment 2. Among these, five major (51.30, 48.70, 28.60, 56.00, 52.00 > 10 R2 ) and one minor (5.40 < 10 R2 ) QTLs for environment 1 and four major (51.48, 50.20, 53.00, 48.00 > 10 R2 ) and one minor (4.44 < 10) QTLs for environment 2 for Zn deficiency tolerance with a logarithm of odd threshold value higher than 3 were identified. The QTLs (qLB-4B, qPM-4B, qRZC-4B, qSZC-4B, qLB-4 N, qPM-4 N, qRZC-4 N, qSZC-4 N) for leaf bronzing, plant mortality root zinc concentration and shoot zinc concentration identified on chromosome 4 were found to be the most promising and highly reproducible across the locations that explained phenotypic variation from 48.00% to 56.00% with the same marker interval RM6748-RM303. CONCLUSION: The new QTLs and its linked markers identified in the present study can be utilized for Zn deficiency tolerance in elite cultivars using marker-assisted backcrossing. © 2022 Society of Chemical Industry.
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Oryza , Sitios de Carácter Cuantitativo , Oryza/genética , Fenotipo , Suelo , ZincRESUMEN
Pulmonary tuberculosis is one of the common diseases with high prevalence of mortality and morbidity in developing countries. It is one of the rare pulmonary infections which can induce hyponatremia and it is important to recognise hyponatremia because of its potential hazards. OBJECTIVES: To assess the serum sodium levels in patients with pulmonary tuberculosis and to establish a relation between the serum sodium levels and its effect on illness in these patients. MATERIAL: This is a single-center prospective observational Study conducted on patients with Pulmonary Tuberculosis, irrespective of treatment, in the department of General Medicine, B. R. Ambedkar Medical College, Bangalore from August 2019 to June 2021after obtaining ethical clearance. A total of 100 patients were enrolled in the study who met the predefined inclusion criteria of age more than 18 years and having been diagnosed with active tuberculosis. The collected data was analysed using student's T test and Chi-Square Test and the analysis was done using SPSS software version 24.0. OBSERVATION: The mean age was 46.46+/- 15.69, with majority of participants in the age group 46-60 years. Gender wise there was male preponderance in our study with 60%. The average serum sodium concentration in our study was 134.20 +/- 5.59 mmol/l, with 44% prevalence of hyponatremia and 4% SIADH Conclusion: Although mild hyponatremia was seen in more than 50% of patients, we found out that hyponatremia in pulmonary tuberculosis is detected in 44% of our patients with male preponderance. The predominant mechanism of hyponatremia was syndrome of inappropriate anti- diuretic hormone secretion (SIADH), which was present in 65% of cases with hyponatremia. Early detection and treatment of underlying electrolyte abnormality can potentially reduce mortality and morbidity associated with tuberculosis and reduce duration of hospitalization. Further research into the prevalence of potassium, magnesium and chloride abnormalities can add to the lacunae of knowledge.
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Hiponatremia , Síndrome de Secreción Inadecuada de ADH , Tuberculosis Pulmonar , Adolescente , Adulto , Humanos , Hiponatremia/epidemiología , Hiponatremia/etiología , Síndrome de Secreción Inadecuada de ADH/complicaciones , Síndrome de Secreción Inadecuada de ADH/diagnóstico , Síndrome de Secreción Inadecuada de ADH/epidemiología , India/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Sodio , Tuberculosis Pulmonar/complicaciones , Tuberculosis Pulmonar/epidemiologíaRESUMEN
The high flux density of synchrotron white beam offers several advantages in X-ray imaging such as higher resolution and signal-to-noise ratio in 3D/4D micro-tomography, higher frame rate in real-time imaging of transient phenomena, and higher penetration in thick and dense materials especially at higher energies. However, these advantages come with additional challenges to beamline optics, camera and sample due to increased heat load and radiation damage, and to personal safety due to higher radiation dose and ozone gas hazards. In this work, a white beam imaging facility at imaging beamline BL-4, Indus-2, has been developed, while taking care of various instrumental and personal safety challenges. The facility has been tested to achieve 1.5â µm spatial resolution, increased penetration depth up to 900â µm in steel, and high temporal resolutions of â¼10â ms (region of interest 2048 × 2048 pixels) and 70â µs (256 × 2048 pixels). The facility is being used successfully for X-ray imaging, non-destructive testing and dosimetry experiments.
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The combined response of exclusion of solar ultraviolet radiation (UV-A+B and UV-B) and static magnetic field (SMF) pre-treatment of 200 mT for 1 h were studied on soybean (Glycine max) leaves using synchrotron imaging. The seeds of soybean with and without SMF pre-treatment were sown in nursery bags kept in iron meshes where UV-A+B (280-400 nm) and UV-B (280-315 nm) from solar radiation were filtered through a polyester filters. Two controls were planned, one with polythene filter controls (FC)- which allows all the UV (280-400 nm); the other control had no filter used (open control-OC). Midrib regions of the intact third trifoliate leaves were imaged using the phase-contrast imaging technique at BL-4, Indus-2 synchrotron radiation source. The solar UV exclusion results suggest that ambient UV caused a reduction in leaf growth which ultimately reduced the photosynthesis in soybean seedlings, while SMF treatment caused enhancement of leaf growth along with photosynthesis even under the presence of ambient UV-B stress. The width of midrib and second-order veins, length of the second-order veins, leaf vein density, and the density of third-order veins obtained from the quantitative image analysis showed an enhancement in the leaves of plants that emerged from SMF pre-treated seeds as compared to untreated ones grown in open control and filter control conditions (in the presence of ambient UV stress). SMF pre-treated seeds along with UV-A+B and UV-B exclusion also showed significant enhancements in leaf parameters as compared to the UV excluded untreated leaves. Our results suggested that SMF-pretreatment of seeds diminishes the ambient UV-induced adverse effects on soybean.
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Glycine max/efectos de la radiación , Campos Magnéticos , Hojas de la Planta/efectos de la radiación , Sincrotrones , Rayos Ultravioleta , Hojas de la Planta/anatomía & histología , Estomas de Plantas/anatomía & histología , Estomas de Plantas/fisiología , Estomas de Plantas/efectos de la radiación , Haz Vascular de Plantas/anatomía & histología , Haz Vascular de Plantas/efectos de la radiación , Espectrofotometría UltravioletaRESUMEN
Thermoplastic composite materials are emerging rapidly due to the flexibility of attaining customized mechanical and melt flow properties. Due to high ductility, toughness, recyclability, and thermal and electrical conductivity, there is ample scope of using copper particles in thermoplastics for 3d printing applications. In the present study, an attempt was made to investigate the Melt Flow Index (MFI), tensile strength, and electrical and thermal conductivity of nylon 6 and ABS (acrylonitrile butadiene styrene) thermoplastics reinforced with copper particles. Thus, the experiments were conducted by adding different-sized copper particles (100 mesh, 200 mesh, and 400 mesh) in variable compositions (0% to 10%) to ABS and nylon 6 matrix. The impact of single, double, and triple particle-sized copper particles on MFI was experimentally investigated followed by FTIR and SEM analysis. Also, the tensile, electrical, and thermal conductivity testing were done on filament made by different compositions. In general, higher fluidity and mechanical strength were obtained while using smaller particles even with higher concentrations (up to 8%) due to improved bonding strength and adhesion between the molecular chains. Moreover, thermal and electrical conductivity was improved with an increase in concentration of copper particles.
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The newly emerged novel coronavirus, SARS-CoV-2, the causative agent of COVID-19 has proven to be a threat to the human race globally, thus, vaccine development against SARS-CoV-2 is an unmet need driving mass vaccination efforts. The receptor binding domain of the spike protein of this coronavirus has multiple neutralizing epitopes and is associated with viral entry. Here we have designed and characterized the SARS-CoV-2 spike protein fragment 330-526 as receptor binding domain 330-526 (RBD330-526) with two native glycosylation sites (N331 and N343); as a potential subunit vaccine candidate. We initially characterized RBD330-526 biochemically and investigated its thermal stability, humoral and T cell immune response of various RBD protein formulations (with or without adjuvant) to evaluate the inherent immunogenicity and immunomodulatory effect. Our result showed that the purified RBD immunogen is stable up to 72 h, without any apparent loss in affinity or specificity of interaction with the ACE2 receptor. Upon immunization in mice, RBD generates a high titer humoral response, elevated IFN-γ producing CD4+ cells, cytotoxic T cells, and robust neutralizing antibodies against live SARS-CoV-2 virus. Our results collectively support the potential of RBD330-526 as a promising vaccine candidate against SARS-CoV-2.
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Anticuerpos Antivirales/sangre , Vacunas contra la COVID-19/administración & dosificación , Inmunidad Humoral/efectos de los fármacos , Inmunogenicidad Vacunal , Fragmentos de Péptidos/administración & dosificación , Glicoproteína de la Espiga del Coronavirus/administración & dosificación , Células TH1/efectos de los fármacos , Adyuvantes Inmunológicos/administración & dosificación , Animales , Biomarcadores/sangre , Vacunas contra la COVID-19/inmunología , Chlorocebus aethiops , Estabilidad de Medicamentos , Glicosilación , Células HEK293 , Humanos , Inmunización , Interferón gamma/sangre , Masculino , Ratones Endogámicos C57BL , Fragmentos de Péptidos/inmunología , Dominios y Motivos de Interacción de Proteínas , Estabilidad Proteica , Glicoproteína de la Espiga del Coronavirus/inmunología , Células TH1/inmunología , Células TH1/metabolismo , Vacunas de Subunidad/administración & dosificación , Vacunas de Subunidad/inmunología , Células VeroRESUMEN
BACKGROUND: In the last few decades, the availability of synchrotron sources has initiated revolutionary advances in X-ray imaging. AIMS: The study aimed to evaluate the incidence of apical transportation after root canal preparation with ProTaper Gold (PTG), Hyflex electro discharge machining (HEDM), Reciproc (RPC), and WaveOne Gold (WOG) using synchrotron radiation-based micro-computed tomographic (SR-µCT) analysis. MATERIALS AND METHODS: Forty mandibular molars were assigned to four experimental groups (n = 10) according to the file system used for the root canal preparation: Group 1: PTG (25/0.08), Group 2: HEDM (25/0.08), Group 3: RPC (25/0.08), and Group 4: WOG (25/0.07). The specimens were scanned on SR-µCT system before and after the root canal preparation. Apical transportation was assessed at 1, 2, 3, 4, and 5 mm section. STATISTICAL ANALYSIS: Two-way analysis of variance and Wilcoxon Mann-Whitney test was used. RESULTS: No significant difference was found between the groups. Transportation in the mesial direction was of greater magnitude than distal transportation for all the files systems. CONCLUSION: SR-µCT can be used as a reliable diagnostic tool for further implications.
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Níquel , Titanio , Cavidad Pulpar/diagnóstico por imagen , Humanos , Preparación del Conducto Radicular , Sincrotrones , Microtomografía por Rayos XRESUMEN
KEY MESSAGE: WRKY transcription factors are among the largest families of transcriptional regulators. In this review, their pivotal role in modulating various signal transduction pathways during biotic and abiotic stresses is discussed. Transcription factors (TFs) are important constituents of plant signaling pathways that define plant responses against biotic and abiotic stimuli besides playing a role in response to internal signals which coordinate different interacting partners during developmental processes. WRKY TFs, deriving their nomenclature from their signature DNA-binding sequence, represent one of the largest families of transcriptional regulators found exclusively in plants. By modulating different signal transduction pathways, these TFs contribute to various plant processes including nutrient deprivation, embryogenesis, seed and trichome development, senescence as well as other developmental and hormone-regulated processes. A growing body of research suggests transcriptional regulation of WRKY TFs in adapting plant to a variety of stressed environments. WRKY TFs can regulate diverse biological functions from receptors for pathogen triggered immunity, modulator of chromatin for specific interaction and signal transfer through a complicated network of genes. Latest discoveries illustrate the interaction of WRKY proteins with other TFs to form an integral part of signaling webs that regulate several seemingly disparate processes and defense-related genes, thus establishing their significant contributions to plant immune response. The present review starts with a brief description on the structural characteristics of WRKY TFs followed by the sections that present recent evidence on their roles in diverse biological processes in plants. We provide a comprehensive overview on regulatory crosstalks involving WRKY TFs during multiple stress responses in plants and future prospects of WRKY TFs as promising molecular diagnostics for enhancing crop improvement.
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Proteínas de Plantas/fisiología , Estrés Fisiológico/fisiología , Factores de Transcripción/fisiología , Productos Agrícolas/genética , Productos Agrícolas/fisiología , Familia de Multigenes , Reguladores del Crecimiento de las Plantas/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Fenómenos Fisiológicos de las Plantas , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas/genética , Transducción de Señal , Factores de Transcripción/químicaRESUMEN
This article investigates a novel data fusion method to predict clay content and cation exchange capacity using visible near-infrared (visNIR) spectroscopy, portable X-ray fluorescence (pXRF), and X-ray diffraction (XRD) techniques. A total of 367 soil samples from two study areas in regional Australia were analyzed and intra- and interarea calibration options were explored. Cubist models were constructed using information from each device independently and in combination. pXRF produced the most accurate predictions of any individual device. Models based on fused data significantly improved the accuracy of predictions compared with those based on individual devices. The combination of pXRF and visNIR had the greatest performance. Overall, the relative increase in Lin's concordance correlation coefficient ranged from 1% to 12% and the corresponding decrease in root-mean-square error (RMSE) ranged from 10% to 46%. Provision of XRD data resulted in a decrease in observed RMSE values, although differences were not significant. Validation metrics were less promising when models were calibrated in one study area and then transferred to the other. Observed RMSE values were â¼2 to 3 times larger under this model transfer scenario and independent use of XRD was found to have the best overall performance.
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Contaminantes del Suelo , Suelo , Australia , Cationes , Arcilla , Monitoreo del Ambiente , Contaminantes del Suelo/análisis , Espectroscopía Infrarroja Corta , Difracción de Rayos X , Rayos XRESUMEN
Phage display antibody (PDA) libraries, allows the rapid isolation and characterization of high specificity monoclonal antibodies for therapeutic and diagnostic applications. However, selection of positive binding clones from synthetic and semi-synthetic libraries has an inherent bias towards clones containing randomly generated amber stop codons, complicating the identification of high affinity binding antibodies. We screened Tomlinson I and J library against receptor binding domain (RBD) of SARS CoV2, eight clones which showed positive binding in phage ELISA, contained one or more amber stop codons in their single-chain antibody fragment (scFv) gene sequences. The presence of amber stop codons within the antibody sequence causes the premature termination of soluble form of scFv expression in nonsuppressor Escherichia coli strain. In the present study, we have used a novel strategy that allows soluble expression of scFvs having amber stop codon in their gene sequences (without phage PIII protein fusion), in the suppressor strain. This strategy of introduction of Ochre (TAA) codon at the junction of scFv and PIII gene, speeds up the initial screening process which is critical for selecting the right scFvs for further studies. Present strategy leads to the identification of a scFv, B8 that binds specifically with nanomolar affinity toward SARS CoV 2 RBD, which otherwise lost in terms of traditional methodology.
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Bacteriófagos/genética , Codón de Terminación/genética , Anticuerpos de Cadena Única/genética , Anticuerpos Monoclonales/genética , COVID-19/virología , Humanos , Región Variable de Inmunoglobulina/genética , Biblioteca de Péptidos , Unión Proteica/genética , ARN Viral/genética , SARS-CoV-2/genéticaRESUMEN
This study investigates the effect of static magnetic field (SMF) pre-treatment in ameliorating arsenic (As) toxicity in soybean plants in relation to growth, photosynthesis and water transport through leaf venation. Soybean (Glycine max variety JS-9560) seeds pre-treated with SMF (200 mT for 1 h) were grown in four levels of arsenate-polluted soil (As(V); 0, 5, 10 and 50 mg kg-1 ) in order to find out the impact of magnetopriming on plant tolerance against As toxicity. Quantitative image analysis of soybean leaf venation showed a narrowing in the width of midrib with increasing As(V) contamination in non-primed seeds. The morphological variations are also supported by the physiological parameters such as reduction in efficiency of photosystem II, plant performance index, stomatal conductance and photosynthetic rate in the presence of As(V) for non-primed seeds. However, remarkable increase was observed in all the measured parameters by SMF pre-treatment at all the concentrations of As(V) used. Even for the highest concentration of As(V) (50 mg kg-1 soil), SMF pre-treatment caused significant enhancement in plant height (40%), area of third trifoliate leaves (40%), along with increase in width of the midrib (17%) and minor vein (13%), contributing to increase in the water uptake, that resulted in higher primary photochemistry of PSII (12%), performance index (50%), stomatal conductance (57%) and photosynthetic rate (33%) as compared to non-primed ones. Consequently, magnetopriming of dry seeds can be effectively used as pretreatment for reduction of As toxicity in soybean plants.
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Arsénico , Glycine max , Arsénico/toxicidad , Clorofila , Fotosíntesis , Hojas de la Planta , SincrotronesRESUMEN
In present work, a correlationship among quantitative drug-polymer miscibility, molecular relaxation and phase behavior of the dipyridamole (DPD) amorphous solid dispersions (ASDs), prepared with co-povidone (CP), hydroxypropyl methylcellulose phthalate (HPMC P) and hydroxypropyl methylcellulose acetate succinate (HPMC AS) has been investigated. Miscibility predicted using melting point depression approach for DPD with CP, HPMC P and HPMC AS at 25 °C was 0.93% w/w, 0.55% w/w and 0.40% w/w, respectively. Stretched relaxation time (τß) for DPD ASDs, measured using modulated differential scanning calorimetry (MDSC) at common degree of undercooling, was in the order of DPD- CP > DPD-HPMC P > DPD-HPMC AS ASDs. Phase behavior of 12 months aged (25 ± 5 °C and 0% RH) spray dried 60% w/w ASDs was tracked using MDSC. Initial ASD samples had homogeneous phase revealed by single glass transition temperature (Tg) in the MDSC. MDSC study of aged ASDs revealed single-phase DPD-CP ASD, amorphous-amorphous and amorphous-crystalline phase separated DPD-HPMC P and DPD-HPMC AS ASDs, respectively. The results were supported by X-ray micro computed tomography and confocal laser scanning microscopy studies. This study demonstrated a profound influence of drug-polymer miscibility on molecular mobility and phase behavior of ASDs. This knowledge can help in designing "physical stable" ASDs.