Broad-spectrum pH functional chitosan-phosphatase beads for the generation of plant-available phosphorus: utilizing the insoluble P pool.

Utilization of organic phosphates and insoluble phosphates for the gradual generation of plant-available phosphorus (P) is the only sustainable solution for P fertilization. Enzymatic conversions are one of the best sustainable routes for releasing P to soil. Phosphatase enzyme aids in solubilizing organic and insoluble phosphates to plant-available P. We herein report the preparation of highly functional chitosan beads co-immobilized with acid phosphatase and alkaline phosphatase enzymes via a glutaraldehyde linkage. The dual enzyme co-immobilized chitosan beads were characterized using Fourier-transform infrared (FTIR), thermogravimetric (TGA), and scanning electron microscopy-energy dispersive x-ray (SEM-EDX) analyses to confirm the immobilization. The co-immobilized system was found to be active for a broader pH range of ∼4-10 than the individually bound enzymes and mixed soluble enzymes. The bound matrix exhibited pH optima at 6 and 9, respectively, for acid and alkaline phosphatase and a temperature optimum at 50°C. The phosphate-solubilizing abilities of the chitosan-enzyme derivatives were examined using insoluble tri-calcium phosphate (TCP) for wide pH conditions of 5.5, 7, and 8.5 up to 25 days. The liberation of phosphate was highest (27.20 mg/mL) at pH 5.5 after the defined period. The residual soil phosphatase activity was also monitored after 7 days of incubation with CBE for three different soils of pH ∼5.5, 7, and 8.5. The residual phosphatase activity increased for all the soils after applying the CBE. The germination index of the Oryza sativa (rice) plant was studied using different pH buffer media upon the application of the CBE in the presence of tri-calcium phosphate as a phosphate source. Overall, the dual-enzyme co-immobilized chitosan beads were highly effective over a wide pH range for generating plant-available phosphates from insoluble phosphates. The chitosan-enzyme derivative holds the potential to be used for sustainable phosphorus fertilization with different insoluble and organic phosphorus sources.

Design, Synthesis, Biological Evaluation and in Silico Studies of Curcumin Pyrrole Conjugates.

Curcumin conjugated heterocyclic compounds are potent candidates with drug likeness against various bacterial pathogens. A set of curcumin-based pyrrole conjugates (CPs) were synthesized and characterized by FT-IR, 1H and 13C NMR and HR-MS techniques. The results of free radical scavenging activity of the synthesized CPs, evaluated by FRAP and CUPRAC assays, showed the potency of these compounds as effective antioxidants. CP3 exhibits the highest antioxidant activity amongst the CPs. The bactericidal efficacy of CPs was screened against ESKAP bacterial pathogens, and CPs were found to possess better antibacterial property than curcumin, specifically against staphylococcus aureus bacteria. In addition, serum albumin (BSA and HSA) binding interaction of these CPs were determined by UV-visible and fluorescence spectrophotometric techniques. In-silico molecular docking study was performed to determine the binding patterns of molecular targets against Staphylococcus aureus tyrosyl tRNA synthetase, and serum albumin proteins. The structure-activity relationship showed that the presence of multiple phenolic hydroxyl groups, and electron withdrawing groups on the structure of CP molecule, enhances its antioxidant and antibacterial activity, respectively.

Pollution indices of selected metals in tea (Camellia sinensis L.) growing soils of the Upper Assam region divulge a non-trifling menace of National Highway.

The study investigated the influence of a National Highway (NH) traversing tea estates (TEs) on heavy metal (HM) contamination in the top soils of Upper Assam, India. The dispersion and accumulation of six HMs, viz. cadmium (Cd), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), and zinc (Zn), within tea-growing soils were assessed using diverse indices: contamination factor (CF), degree of contamination (DC), enrichment factor (EF), geo-accumulation index (Igeo), modified degree of contamination (MDC), Nemerow pollution index (PINemerow), pollution load index (PLI), potential ecological risk factor (Eri), and potential ecological risk index (RI). The order of HM prevalence was Fe > Mn > Zn > Ni > Cu > Cd. Elevated Cd levels near the NH prompted immediate attention, while Cd and Zn showed moderate pollution in CF, EF, and RI. The remaining metals posed minimal individual risk (Eri< 40), resulting in an overall contamination range of "nil to shallow," signifying slight contamination from the studied metals. From MDC values for investigated metals, it was found to be "zero to very low degree of contamination" at all locations except the vicinity of NH. Soil pollution, as determined by PLI, indicated unpolluted soils in both districts, yet PINemerow values indicated slight pollution. The statistical analysis revealed that there is a significant decrease in most of the indices of HM as the distance from NH increases. The application of multivariate statistical techniques namely Principal Component Analysis and Cluster Analysis showed the presence of three distinct homogenous groups of distances based on different indices. This investigation underscores NH-associated anthropogenic effects on TE soil quality due to HM deposition, warranting proactive mitigation measures.

Intercalation vs Adsorption Strategies of Myo-Inositol Hexakisphosphate into Zn-Fe Layered Double Hydroxide: A Tiff between Anion Exchange and Coprecipitation.

Myo-inositol hexakisphosphates (IHPs) or phytates are the most abundant organic phosphates having the potential to serve as a phosphorus reserve in soil. Understanding the fate of IHP interaction with soil minerals tends to be crucial for its efficient storage and utilization as a slow-release organic phosphate fertilizer. We have systematically compared the effective intercalation strategy of a phytate onto Zn-Fe layered double hydroxide (LDH) acting as storage/carrier material through coprecipitation and anion exchange. Powder X-ray diffraction, X-ray photoelectron spectroscopy, elemental analysis, thermogravimetric analysis, FTIR spectra, and molecular modeling demonstrated the formation of phytate-intercalated Zn-Fe LDH through coprecipitation with a maximum loading of 41.34% (w/w) in the pH range of ∼9-10 in a vertical alignment through monolayer formation. No intercalation product was obtained from the anion exchange method, which was concluded based on the absence of shifting in the XRD (003) peak. A change in the zeta potential values from positive to negative and subsequent increase in solution pH, with decreasing phytate concentration, are suggestive of adsorption of IHP onto the LDH surface. The batch adsorption data were best fitted with Langmuir isotherm equation and followed the pseudo-second-order kinetic model. The maximum adsorption capacity was found to be 45.87 mg g-1 at a temperature of 25 ± 0.5 °C and pH 5.63.

The Level of Selected Metals in Made Tea and Tea Infusion from the Roadside Tea Plants and Health Risk Assessment.

The effects of human activities are becoming clearer every year, with multiple reports of struggling and eroded ecosystems resulting in new threats of plant and animal extinctions throughout the world. It has been speculated that roadside tea-growing soils impact on metal dynamics from soil to tea plants and subsequently to tea infusion which may be threatened by increasingly unpredictable and dangerous surroundings. Furthermore, heavy metals released from vehicles on the national highway (NH) could be a source of metal contamination in roadside tea soils and tea plants. This study was articulated to realize the effect of NH on a buildup of selected metals (Cu, Cd, Fe, Mn, Ni, and Zn) in made tea along with repeated tea infusion. In general, metal concentration was found significantly higher in made tea prepared from the young shoots collected from the vicinity of NH. The results also showed that distance from the NH and infusion process significantly influenced to content of the analysed metal in tea infusions. The mean average daily intake (ADI) and hazard quotient (HQ) values of analysed tea samples were found in the orderMn˃Fe˃Zn˃Cu˃Ni˃Cd and Mn˃Cu˃Zn˃Fe˃Ni˃Cd, respectively. The HQ values of all analysed metals were found << 1, indicating that ingestion of tea infusion with analysed heavy metals should not cause a danger to human health. However, this study further demonstrates the consumption of tea infusion prepared from made tea around the vicinity of NH may contribute to a significantly higher quantity of metal intake in the human body. From the hierarchical cluster analysis, it has been observed that there are three homogenous groups of analysed heavy metals.

Insights into the effects of tea pruning litter biochar on major micronutrients (Cu, Mn, and Zn) pathway from soil to tea plant: An environmental armour.

A field study was conducted from 0 to 360 days to investigate the effect of tea pruning litter biochar (TPLBC) on the accumulation of major micronutrients (copper: Cu, manganese: Mn, and zinc: Zn) in soil, their uptake by tea plant (clone: S.3 A/3) and level of contamination in soil due to TPLBC. To evaluate the level of contamination due to TPLBC, a soil pollution assessment was carried out using the geo-accumulation index (Igeo), enrichment factor (EF), contamination factor (CF), potential ecological risk factor (PERF), individual contamination factor (ICF), and risk assessment code (RAC). The total content of Cu, Mn, and Zn gradually increased with increasing doses of TPLBC at 0D, and then decreased with time. The fractionation of the three micronutrients in soil changed after the application of TPLBC. The contamination risk assessment of soil for Cu, Mn, and Zn based on the Igeo, EF, CF, PERF,ICF, and RAC suggested that the application of TPLBC does not have any adverse effect on soil. Except for Mn, the bioconcentration and translocation factors were less than one for Cu and Zn. Results from this study revealed that the application of 400 kg TPLBC ha-1 is significantly better than the other treatments for Cu, Mn, and Zn at a 5% level of significance.

Tea pruning litter biochar amendment in soil reduces arsenic, cadmium, and chromium in made tea (Camellia sinensis L.) and tea infusion: A safe drink for tea consumers.

Effect of tea pruning litter biochar (TPLBC) on arsenic (As), cadmium (Cd) and chromium (Cr) content in made tea and successive tea infusions were investigated in a greenhouse experiment with two tea cultivars (TV23 and S.3A/3). Made tea prepared from TV23 and S.3A/3 clone, a decrease in the concentration of As, Cd, and Cr by 36.73%, 16.22%, 13.96%, and 36.63%, 27.78%, 10.54%, respectively over control on the application of the highest dose of TPLBC (500 kg TPLBC ha-1). Irrespective of treatments, studied element concentrations were significantly higher (p ≤ 0.05) in the first infusion and lower in the third. Considering Ten g made tea consumption per person per day, the maximum average daily intakes of As, Cd and Cr in a higher dose of TPLBC were far below the tolerable weekly intake prescribed by the World Health Organization. As hazard quotient values of selected elements were ≪ 1, no significant adverse health consequences are expected for tea consumers.

Boosting nitrogen fertilization by a slow releasing nitrate-intercalated biocompatible layered double hydroxide-hydrogel composite loaded with Azospirillum brasilense.

Indiscriminate use of chemical fertilizers leads to soil environmental disbalance and therefore, preparation and application of environment-friendly slow-release multifunctional fertilizers are of paramount importance for sustainable crop production in the present scenario. In this study, we propose a slow-release multifunctional composite nitrogen (N) fertilizer, which possesses the ability to supply plant accessible N in the form of ammonium (NH4 +) and nitrate (NO3 -) to improve nitrate assimilation coupled with zinc (Zn, a major micronutrient for plants in the soil) after its degradation. For this purpose, NO3 --intercalated zinc-aluminum (Zn-Al) layered double hydroxide (LDH) was synthesized using a co-precipitation protocol. The prepared LDH was added as 25.45% of total polymer weight to a sodium carboxymethyl cellulose/hydroxyethyl cellulose citric acid (NaCMC/HEC-CA) biodegradable hydrogel. A. brasilense, commonly used nitrogen-fixing bacteria in soils, was added to the LDH-hydrogel composite along with LDH alone to augment the availability of NH4 + and NO3 -. Adjusting the pH under acidic (pH 5.25) and neutral (pH 7) conditions, the release pattern of NO3 - from LDH and the composite was monitored for 30 days at normal temperature. The pH was selected based on the soil analysis data of North East India. The LDH-composite released 90% (w/w) and 85.45% (w/w) of intercalated NO3 - at pH 5.25 and 7.00 respectively in 30 days. However, 100% (w/w) and 87% (w/w) of intercalated NO3 - at pH 5.25 and 7.00 respectively were released in 30 days when only LDH was applied, which indicated the lower performance of LDH alone in comparison to the LDH-composite for the nitrate holding pattern. The pH of the bacteria-loaded system was observed to be acidic (pH = 5-6) during the study of nitrate assimilation and Zn2+ release. A. brasilense improved nitrate assimilation and increased the NH4 + ion concentration in the studied system. A significant increase in Zn2+ release was observed from day 5 in the presence of A. brasilense in the LDH-composite compared with that in the absence of A. brasilense. In conclusion, the prepared LDH-hydrogel-A. brasilense composite fertilizer system increases the availability of plant accessible N form (both NO3 - and NH4 +) and can potentially improve soil fertility with the addition of Zn and bacteria to the soil in the extended course.

Silica Nanoparticle-Endothelial Interaction: Uptake and Effect on Platelet Adhesion under Flow Conditions.

Silica nanoparticles are extensively used in biomedical applications and consumer products. Little is known about the interaction of these NPs with the endothelium and effect on platelet adhesion under flow conditions in circulation. In this study, we investigated the effect of silica nanoparticles on the endothelium and its inflammation, and subsequent adhesion of flowing platelets in vitro. Platelet counts adhered onto the surface of endothelial cells in the presence of nanoparticles increased at both low and high concentrations of nanoparticles. Preincubation of endothelial cells with nanoparticles also increased platelet adhesion. Interestingly, platelet adhesion onto TNF-α-treated endothelial cells decreased in the presence of nanoparticles at different concentrations as compared with the absence of nanoparticles. We monitored the expression of different endothelial proteins, known to initiate platelet adhesion, in the presence and absence of silica nanoparticles. We found that silica nanoparticles caused changes in the endothelium such as overexpression of PECAM that promoted platelet adhesion to the endothelial cell.

Redox-Responsive Polysulfide-Based Biodegradable Organosilica Nanoparticles for Delivery of Bioactive Agents.

Design and development of silica nanoparticles (SiO2 NPs) with a controlled degradation profile promises effective drug delivery with a predetermined carrier elimination profile. In this research, we fabricated a series of redox-responsive polysulfide-based biodegradable SiO2 NPs with low polydispersity and with variations in size (average diameters of 58 ± 7, 108 ± 11, 110 ± 9, 124 ± 9, and 332 ± 6 nm), porosity, and composition (disulfide vs tetrasulfide bonds). The degradation kinetics of the nanoparticles was analyzed in the presence of 8 mM glutathione (GSH), mimicking the intracellular reducing condition. Results indicate that porosity and core composition play the predominant roles in the degradation rate of these nanoparticles. The 108 nm mesoporous disulfide-based nanoparticles showed the highest degradation rate among all the synthesized nanoparticles. Transmission electron microscopy (TEM) reveals that nonporous nanoparticles undergo surface erosion, while porous nanoparticles undergo both surface and bulk erosion under reducing environment. The cytotoxicity of these nanoparticles in RAW 264.7 macrophages was evaluated. Results show that all these nanoparticles with the IC50 values ranging from 233 ± 42 to 705 ± 17 µg mL-1 do not have cytotoxic effect in macrophages at concentrations less than 125 µg mL-1. The degradation products of these nanoparticles collected within 15 days did not show cytotoxicity in the same macrophage cell line after 24 h of incubation. In vitro doxorubicin (DOX) release was examined in 108 nm mesoporous disulfide-based nanoparticles in the absence and presence of 8 mM GSH. It was shown that drug release depends on intracellular reducing conditions. Due to their ease of synthesis and scale up, robust structure, and the ability to control size, composition, release, and elimination, biodegradable SiO2 NPs provide an alternative platform for delivery of bioactive and imaging agents.

Differential Protein Adsorption and Cellular Uptake of Silica Nanoparticles Based on Size and Porosity.

Slight alterations in nanoparticles' surface properties can significantly influence the corona composition which may alter their interaction with the biological milieu. Size and porosity of silica nanoparticles (SNPs) are known to be predominant factors influencing their dose-dependent toxicity. Little is known however about the extent and type of protein adsorption on SNPs as a function of physicochemical properties and the role this might play on mechanisms of cellular uptake and toxicity. In this work we investigated the influence of size and porosity of SNPs on protein adsorption, cellular uptake, and toxicity in RAW 264.7 macrophages. Toxicity of the SNPs was found to be concentration dependent, and the formation of the protein corona mitigated toxicity for all particles. Detailed analysis of the amount of proteins recovered from each nanoparticle revealed similarities in the protein adsorption profile as a function of size and porosity. The mechanism of uptake was highly dependent on size rather than porosity or the adsorbed proteins.

Correlating enzyme density, conformation and activity on nanoparticle surfaces in highly functional bio-nanocomposites.

The biological activity of the immobilized enzyme is crucial for the performance of different nanoparticle mediated enzymatic assays, where enzymatic conversion can be used for label-free analyte detection. In this article we have addressed two significant aspects of enzyme-nanoparticle interactions. First, we have developed copper sulfide (CuS) nanoparticles with an average diameter of 25 nm as a potential enzyme-interface using trypsin protease as a model enzyme. CuS nanoparticles showed high trypsin immobilization capacity of about 14.0 mg m(-2) with the significant retention of native enzymatic activity (75-98%) at room temperature, even beyond the calculated tightly packed monolayer coverage (which is around 4.1 mg m(-2)). Second, we report a quantitative correlation between the structure-functional relationship and the density of immobilized trypsin on a nanoparticle surface. The in situ conformation of immobilized trypsin could be efficiently analyzed by fluorescence, circular dichroism and FT-IR spectroscopic measurements because of the small size of the nanoparticles. Trypsin molecules appear to retain their close-native tertiary and secondary structural features (with a small loss of 1-2% of helical content) in the entire surface density range (2.0-14.0 mg m(-2)) on the CuS nanoparticles. However, interestingly, at a low surface coverage (2.0 mg m(-2)), immobilized trypsin retains almost 98% of its native enzymatic activity, leading to a highly functional bio-nanocomposite. However, at higher surface coverages, the enzyme activity decreases to 77%, indicating the influence of steric crowding. Furthermore, the high functionality of the immobilized trypsin at low surface density on CuS nanoparticle was also confirmed by determining the kinetic parameters of enzymatic activity.

Magnetic nanoparticles for selective capture and purification of an antimicrobial peptide secreted by food-grade lactic acid bacteria.

Biocompatible iron oxide nanoparticles (IONPs) were utilized for charge-based selective capture of the cationic bacteriocin pediocin secreted by food-grade lactic acid bacteria (LAB), resulting in the generation of a nanocomposite, which could be readily separated from other secreted metabolites. Interestingly, pediocin activity was conserved and a membrane-directed antibacterial activity typically associated with pediocin was manifested in the nanocomposite. Efficient sequestration of pediocin was also achieved through facile magnetic separation of IONP-pediocin composites and following desorption at pH 2.0, the recovered IONPs could be recycled for a subsequent round of pediocin adsorption. The steady state adsorption isotherm of pediocin with IONPs followed a Langmuir isotherm model. Following the reversible adsorption-desorption process with IONPs, 16-fold purification of pediocin could be achieved. The HPLC profile of the desorbed pediocin revealed a similar retention time to pediocin purified by an established cell-adsorption method and the HPLC eluted fraction also displayed the signature membrane-directed pediocin activity. The facile capture of pediocin, magnetic separation and the possibility of salvaging IONPs for reuse, accompanied by high retention of pediocin activity during the purification process, enhance the merit of IONPs as robust and effective purification tools for a potentially therapeutic antimicrobial peptide.

Interpreting the adsorption of serum albumin and lactoglobulin onto ZnS nanopaticles: effect of conformational rigidity of the proteins.

The work we have undertaken is to investigate the adsorption of two different proteins (BSA and BLG) having near same IEP and differing in their conformational flexibility, onto the surface of ZnS nanoparticles (ZnS NPs). BSA and BLG both have an IEP value around pH~5. BSA is more prone to conformational deformation and considered "soft" while BLG holds the conformational rigidity and considered as "hard" protein. To ascertain the differences in surface coverage and conformation of the protein onto ZnS surface (PZC ~ 3.7), we have evaluated the adsorption profile at pH 7, where the entire surface behaves negatively. An integrated approach was taken by incorporating zeta (ζ) potential, fluorescence and CD for analyzing the adsorption process. In both systems, an increase in protein surface coverage was observed with the increase in free protein concentration in the solution and ζ values approaching that of native protein at high surface coverage. An alteration in the tertiary structure was observed for both BSA and BLG. The CD spectra analysis reveals that the secondary structure of the BSA was more deviated from the native protein structure, accommodating the increased adsorption value. For BLG no such prominent structural alteration was observed. These findings help us to understand better, how adjustment of the protein adsorption amount can be achieved onto the surface of nanoparticles having like charges.

Efficient removal of chromate and arsenate from individual and mixed system by malachite nanoparticles.

Malachite nanoparticles of 100-150 nm have been efficiently and for the first time used as an adsorbent for the removal of toxic arsenate and chromate. We report a high adsorption capacity for chromate and arsenate on malachite nanoparticle from both individual and mixed solution in pH ∼4-5. However, the adsorption efficiency decreases with the increase of solution pH. Batch studies revealed that initial pH, temperature, malachite nanoparticles dose and initial concentration of chromate and arsenate were important parameters for the adsorption process. Thermodynamic analysis showed that adsorption of chromate and arsenate on malachite nanoparticles is endothermic and spontaneous. The adsorption of these anions has also been investigated quantitatively with the help of adsorption kinetics, isotherm, and selectivity coefficient (K) analysis. The adsorption data for both chromate and arsenate were fitted well in Langmuir isotherm and preferentially followed the second order kinetics. The binding affinity of chromate is found to be slightly higher than arsenate in a competitive adsorption process which leads to the comparatively higher adsorption of chromate on malachite nanoparticles surface.