Synthesis and analysis of silica nanocarriers for pectinase immobilization: Enhancing enzymatic stability for continuous industrial applications.

Pectinolytic enzymes are among the important group of industrial enzymes that have wide applications in different food industries. In this study, pectinase-based silica nanocarriers were synthesized using co-precipitation and cross-linking techniques. The resulting silica nanoparticles were investigated using scanning electron microscopy (SEM), energy-dispersive electron microscopy (EDEX), and X-ray diffraction (XRD) for determination of its morphology, elemental composition, and crystalline pattern. Under the optimal immobilization conditions like 1.5 % glutaraldehyde, 3000 IU/mg pectinase concentration, 90 min immobilization time and 40 °C immobilization temperature, pectinase showed maximum immobilization yield. The immobilization of pectinase onto the silica nanocarriers led to enhanced catalytic characteristics, displaying higher enzymatic activity across various temperature and pH levels compared to soluble pectinase. Moreover, the immobilization substantially improved the temperature stability of pectinase, exhibiting 100 % of its initial activity even after 120 h of pre-incubation at 50 °C. Additionally, the silica nanocarrier pectinase retained 100 % of its original activity even after being reused 10 times in a single batch of reactions. These findings indicate that the immobilization of silica nanocarriers effectively enhances pectinase's industrial capabilities, making it economically feasible for industrial use and an efficient system for various biotechnological applications.

Green Synthesis of Low-Glycemic Amylose-Lipid Nanocomposites by High-Speed Homogenization and Formulation into Hydrogel.

In this research, we focused on the production of amylose-lipid nanocomposite material (ALN) through a green synthesis technique utilizing high-speed homogenization. Our aim was to investigate this novel material's distinctive physicochemical features and its potential applications as a low-glycemic gelling and functional food ingredient. The study begins with the formulation of the amylose-lipid nanomaterial from starch and fatty acid complexes, including stearic, palmitic, and lauric acids. Structural analysis reveals the presence of ester carbonyl functionalities, solid matrix structures, partial crystallinities, and remarkable thermal stability within the ALN. Notably, the ALN exhibits a significantly low glycemic index (GI, 40%) and elevated resistance starch (RS) values. The research extends to the formulation of ALN into nanocomposite hydrogels, enabling the evaluation of its anthocyanin absorption capacity. This analysis provides valuable insights into the rheological properties and viscoelastic behavior of the resulting hydrogels. Furthermore, the study investigates anthocyanin encapsulation and retention by ALN-based hydrogels, with a particular focus on the influence of pH and physical cross-link networks on the uptake capacity presenting stearic-acid (SA) hydrogel with the best absorption capacity. In conclusion, the green-synthesized (ALN) shows remarkable functional and structural properties. The produced ALN-based hydrogels are promising materials for a variety of applications, such as medicine administration, food packaging, and other industrial purposes.

PEGylated Graphene Oxide as a Nanodrug Delivery Vehicle for Podophyllotoxin (GO/PEG/PTOX) and In Vitro α-Amylase/α-Glucosidase Inhibition Activities.

This study aims to develop a nanodrug delivery system containing podophyllotoxin (PTOX), a known anticancer drug, loaded on graphene oxide (GO). The system's ability to inhibit α-amylase and α-glucosidase enzymes was also investigated. PTOX was isolated from Podophyllum hexandrum roots with a yield of 2.3%. GO, prepared by Hummer's method, was converted into GO-COOH and surface-mobilized using polyethylene glycol (PEG) (1:1) in an aqueous medium to obtain GO-PEG. PTOX was loaded on GO-PEG in a facile manner with a 25% loading ratio. All the samples were characterized using FT-IR spectroscopy, UV/visible spectroscopy, and scanning electron microscopy (SEM). In FT-IR spectral data, GO-PEG-PTOX exhibited a reduction in acidic functionalities and there was an appearance of the ester linkage of PTOX with GO. The UV/visible measurements suggested an increase of absorbance in 290-350 nm regions for GO-PEG, suggesting the successful drug loading on its surface (25%). GO-PEG-PTOX exhibited a rough, aggregated, and scattered type of pattern in SEM with distinct edges and binding of PTOX on its surface. GO-PEG-PTOX remained potent in inhibiting both α-amylase and α-glucosidase with IC50 values of 7 and 5 mg/mL, closer to the IC50 of pure PTOX (5 and 4.5 mg/mL), respectively. Owing to the 25% loading ratio and 50% release within 48 h, our results are much more promising. Additionally, the molecular docking studies confirmed four types of interactions between the active centers of enzymes and PTOX, thus supporting the experimental results. In conclusion, the PTOX-loaded GO nanocomposites are promising α-amylase- and α-glucosidase-inhibitory agents when applied in vitro and have been reported for the first time.

Effect of sucrose, trehalose, maltose and xylose on rheology, water mobility and microstructure of gluten-free model dough based on high hydrostatic pressure treated starches.

Due to functional and physicochemical properties, starch in its native state has limited range of applications. Simultaneously, information on effects of different sugars and their interactions with modified starch on gluten-free model dough is also limited. To better overcome these restrictions, the effects of sucrose, trehalose, maltose and xylose on rheology, water mobility and microstructure of gluten-free dough prepared with high hydrostatic pressure (HHP) treated maize (MS), potato (PS) and sweet potato starch (SS) were investigated. MS, PS and SS dough with trehalose exhibited a lower degree of dependence of G' on frequency sweep (z'), higher strength (K) and relative elastic part of maximum creep compliance (Je/Jmax), suggesting stable network structure formation. Total gas production (VT) of MS dough with maltose, PS dough with sucrose and SS dough with trehalose was increased from 588 to 1454 mL, 537 to 1498 mL and 637 to 1455 mL respectively. Higher weakly bound water (T22) was found in the dough with trehalose at 60 min of fermentation, suggesting more hydrogen bonds and stable network. Thus, trehalose might be a potential improver in HHP treated starch-based gluten-free products.

Development of Pectinase Based Nanocatalyst by Immobilization of Pectinase on Magnetic Iron Oxide Nanoparticles Using Glutaraldehyde as Crosslinking Agent.

To increase its operational stability and ongoing reusability, B. subtilis pectinase was immobilized on iron oxide nanocarrier. Through co-precipitation, magnetic iron oxide nanoparticles were synthesized. Scanning electron microscopy (SEM) and energy dispersive electron microscopy (EDEX) were used to analyze the nanoparticles. Pectinase was immobilized using glutaraldehyde as a crosslinking agent on iron oxide nanocarrier. In comparison to free pectinase, immobilized pectinase demonstrated higher enzymatic activity at a variety of temperatures and pH levels. Immobilization also boosted pectinase's catalytic stability. After 120 h of pre-incubation at 50 °C, immobilized pectinase maintained more than 90% of its initial activity due to the iron oxide nanocarrier, which improved the thermal stability of pectinase at various temperatures. Following 15 repetitions of enzymatic reactions, immobilized pectinase still exhibited 90% of its initial activity. According to the results, pectinase's catalytic capabilities were enhanced by its immobilization on iron oxide nanocarrier, making it economically suitable for industrial use.

Phytochemical Profiling, Antioxidant, Antimicrobial and Cholinesterase Inhibitory Effects of Essential Oils Isolated from the Leaves of Artemisia scoparia and Artemisia absinthium.

The current studies were focused on the phytochemical profiling of two local wild Artemisia species, Artemisia scoparia and Artemisia absinthium leaves' essential oils, extracted via the hydro distillation method along with evaluation of their antioxidant as well as antimicrobial effects. The constituents of EOs were identified using a combined gas chromatography-mass spectrometric (GC-MS) technique. A total of 25 compounds in A. scoparia essential oil (EOAS) were identified, and 14 compounds with percentage abundance of >1% were tabulated, the major being tocopherol derivatives (47.55%). A total of nine compounds in Artemisia absinthium essential oil (EOAA) were enlisted (% age > 1%), the majority being oleic acid derivatives (41.45%). Strong antioxidant effects were pronounced by the EOAS in DPPH (IC50 = 285 ± 0.82 µg/mL) and in ABTS (IC50 = 295 ± 0.32 µg/mL) free radical scavenging assays. Both the EOs remained potent in inhibiting the growth of bacterial species; Escherichia coli (55−70%) and Shigella flexneri (60−75%) however remained moderately effective against Bacillus subtilis as well as Staphylococcus aureus. Both EOAS and EOAA strongly inhibited the growth of the tested fungal species, especially Aspergillus species (up to 70%). The oils showed anti-cholinesterase potential by inhibiting both Acetylcholinesterase (AChE; IC50 = 30 ± 0.04 µg/mL (EOAS), 32 ± 0.05 µg/mL (EOAA) and Butyrylcholinesterase (BChE; IC50 = 34 ± 0.07 µg/mL (EOAS), 36 ± 0.03 µg/mL (EOAA). In conclusion, the essential oils of A. scoparia and A. absinthium are promising antioxidant, antimicrobial and anticholinergic agents with a different phytochemical composition herein reported for the first time.

In Vitro and In Silico Investigation of Diterpenoid Alkaloids Isolated from Delphinium chitralense.

This study reports the isolation of three new C20 diterpenoid alkaloids, Chitralinine A-C (1-3) from the aerial parts of Delphinium chitralense. Their structures were established on the basis of latest spectral techniques and single crystal X-rays crystallographic studies of chitralinine A described basic skeleton of these compounds. All the isolated Compounds (1-3) showed strong, competitive type inhibition against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in comparison to standard allanzanthane and galanthamine however, chitralinine-C remained the most potent with IC50 value of 11.64 ± 0.08 µM against AChE, and 24.31 ± 0.33 µM against BChE, respectively. The molecular docking reflected a binding free energy of -16.400 K Cal-mol-1 for chitralinine-C, having strong interactions with active site residues, TYR334, ASP72, SER122, and SER200. The overall findings suggest that these new diterpenoid alkaloids could serve as lead drugs against dementia-related diseases including Alzheimer's disease.

Effects of pH and Ionic Salts on the Emulsifying and Rheological Properties of Acorn Protein Isolate.

This study was designed to evaluate the emulsifying and rheological properties of acorn protein isolate (API) in different pH mediums (pH 3, 7 and 9) and in the presence of ionic salts (1 M NaCl and 1 M CaCl2). API shows higher solubility in distilled water at pH 7, while at the same pH, a decrease in solubility was observed for API in the presence of CaCl2 (61.30%). A lower emulsifying activity index (EAI), lower stability index (ESI), larger droplet sizes and slight flocculation were observed for API in the presence of salts at different pHs. Importantly, CaCl2 treated samples showed relevantly higher EAI (252.67 m2/g) and ESI (152.67 min) values at all pH as compared to NaCl (221.76 m2/g), (111.82 min), respectively. A significant increase in interfacial protein concentration (4.61 mg/m2) was observed for emulsion at pH 9 with CaCl2, while the major fractions of API were observed in an interfacial layer after SDS-PAGE analysis. All of the emulsion shows shear thinning behavior (τc > 0 and n < 1), while the highest viscosity was observed for emulsion prepared with CaCl2 at pH 3 (11.03 ± 1.62). In conclusion, API, in the presence of ionic salts at acidic, neutral and basic pH, can produce natural emulsions, which could be substitutes for synthetic surfactants for such formulations.

Neuroprotective Potential of Synthetic Mono-Carbonyl Curcumin Analogs Assessed by Molecular Docking Studies.

Cognitive decline in dementia is associated with deficiency of the cholinergic system. In this study, five mono-carbonyl curcumin analogs were synthesized, and on the basis of their promising in vitro anticholinesterase activities, they were further investigated for in vivo neuroprotective and memory enhancing effects in scopolamine-induced amnesia using elevated plus maze (EPM) and novel object recognition (NOR) behavioral mice models. The effects of the synthesized compounds on the cholinergic system involvement in the brain hippocampus and their binding mode in the active site of cholinesterases were also determined. Compound h2 (p < 0.001) and h3 (p < 0.001) significantly inhibited the cholinesterases and reversed the effects of scopolamine by significantly reducing TLT (p < 0.001) in EPM, while (p < 0.001) increased the time exploring the novel object. The % discrimination index (DI) was significantly increased (p < 0.001) in the novel object recognition test. The mechanism of cholinesterase inhibition was further validated through molecular docking study using MOE software. The results obtained from the in vitro, in vivo and ex vivo studies showed that the synthesized curcumin analogs exhibited significantly higher memory-enhancing potential, and h3 could be an effective neuroprotective agent. However, more study is suggested to explore its exact mechanism of action.

Adolescent health literacy: factors effecting usage and expertise of digital health literacy among universities students in Pakistan.

BACKGROUND: Electronic health tools are of little use if the intended user lacks the skills to effectively engage them. Engaging eHealth requires a skill set, or literacy, of its own. The present study is an effort to probe the relationship of education and Institution (Independent Variables) with the usage and expertise in eHealth literacy (Dependent Variables) among university students. The research is conducted in 16 Higher Education Commission (HEC) Pakistan's recognized universities in Lahore. Both male and female students ranging from BS to PhD programs were the focus of the research. METHODS: Quantitative data was collected through survey method using stratified random sampling technique. There were different kinds of strata in population i.e. general universities, health sciences universities, engineering universities and animal sciences universities etc. The research encompassed a total of 89,664 students in 16 universities, from which sample size of 1513 was drawn through research advisor table (2006). Proportional allocation formula was used to specify the number of respondents from each university. Non-parametric statistics was used since data was not normally distributed. Kruskal-Wallis H test and Mann-Whitney U test were applied to measure the difference of effect of groups of independent variables on the dependent variables. RESULTS: The level of using digital health literacy was not same for all students, as the students of PhD and BS/Masters were significantly different from each other in terms of their usage of digital health literacy. Level of education showed a significant influence on level of expertise in eHealth literacy, confirming that changing the level of education had an effect on level of expertise in digital health literacy, but the size of effect was smaller. MS/MPhil and PhD students were significantly different from each other in their expertise in digital health literacy. CONCLUSION: Results of the study depicted that belonging to different categories of educational levels differently affect the level of usage and that of expertise in digital health literacy among university students.

Effect of protein and oil volume concentrations on emulsifying properties of acorn protein isolate.

This study aims to extract acorn protein isolate (API) from locally abundant waste acorn fruit and investigate its emulsification behavior by mixing different protein (0.1-2% w/v) and oil volume concentrations (5-45% v/v). Significant decrease in emulsifying activity index (EAI) and an increase in emulsifying stability index (ESI) were observed with an increase in API concentrations (P < 0.05). Droplet sizes of emulsions and viscosity were observed to decrease significantly (P < 0.05) with increase in API concentration while the increase was observed in interfacial protein concentration (Г). In contrast, increase in oil volume concentration results in increase of droplet sizes, packing fractions and viscosity, while decrease in Г values was observed. The results reveal that main fractions of API (66.2-14.4 kDa) were migrated to oil-water interface for emulsion stabilization. These results demonstrate the potential application of API in food formulation and development.

Effects of Maillard reaction on physicochemical and functional properties of walnut protein isolate.

In this study, the Maillard reaction (MR) of glucose was applied to improve the physicochemical and functional properties of walnut protein isolate (WNPI). The MR products (MRPs) were prepared with glucose at 0 h (MRP0), 1 h (MRP1), 2 h (MRP2) and 3 h (MRP3) heating at 95 °C. The Infra-Red spectrum showed reduction of amide and S-H functionalities in MRPs with complete intermixing of glucose in MRP3. Scanning electron microscopy indicated changes in the morphology of MRP3 which also exhibited promising antioxidant effect. Significant decrease (P < 0.05) in hydrophobicity values (Ho) and increase (P < 0.05) in emulsifying activity/emulsifying stability indexes values were observed for MRPs. Uniform droplet distribution was observed in microscopy of emulsions while an increase in the interfacial protein concentration ( Г) was obtained for MRPs. These results suggest that MR is useful in improving the utilization of this protein in food product development.

Effect of extraction methods on structural, physiochemical and functional properties of dietary fiber from defatted walnut flour.

The effect of different extraction methods i.e. extraction with alkali (AEDF), enzyme (EEDF) and enzyme plus shear emulsifying hydrolysis (SEDF) on structure, physiochemical as well as the functional characteristics of dietary fiber (DF) from defatted walnut flour were studied. AEDF process showed significantly higher (P < 0.05) amount of water retention capacity (WRC; 5.39 g/g), water swelling capacity (WSC; 3.16 g/mL), and particle size; while, shown lower value of oil adsorption capacity (OAC; 29 g/g) amongst all. Compared to AEDF, no major differences were observed in network except the matrix in EEDF and SEDF was more porous and honey comb like. DF extracted through AEDF, EEDF and SEDF showed good viscosity and emulsifying activity however, less stability indices. The results from this study suggest that AEDF and EEDF and SEDF had specific effects on the structure-functional properties of DF from defatted walnut flour, which has great potential in food applications.

Suppression of Cell Wall Degrading Enzymes and their Encoding Genes in Button Mushrooms (Agaricus bisporus) by CaCl2 and Citric Acid.

Fresh button mushrooms (Agaricus bisporus) were harvested and treated with a solution of 1.5% CaCl2 + 0.5% citric acid and stored for 16 days at 12 °C. The effects of this treatment on firmness, weight, color, cell wall compositions (cellulose and chitin) and cell wall degrading enzymes (cel1ulase, beta-1, 3 glucanase, chitinase and phenylalanine ammonialyase) were investigated during post-harvest storage. The expressions of major genes (Cel1, Glu1, Chi1 and PAL1) involved in cell wall degradation during post-harvest storage were also monitored. The results revealed that the post-harvest chemical treatment maintained better firmness, weight, color and inhibited cellulase, beta-1, 3 glucanase, chitinase and phenylalanine ammonialyase activities. These findings showed that the down-regulation of cell wall degrading enzymes is a possible mechanism that delays the softening of button mushrooms by the application of combined chemical treatment.

Effects of high hydrostatic pressure on emulsifying properties of sweet potato protein in model protein-hydrocolloids system.

The effects of high hydrostatic pressure (HHP) on emulsifying properties of sweet potato protein (SPP) in presence of 0.1%, 0.3% and 0.5% (w/v) of guar gum (GG) and glycerol monostearate (GMS) were investigated. Emulsifying stability index (ESI) of the SPP with GG revealed significant increase (P<0.05) in ESI value at 600MPa treatment, while the stability of SPP-GMS emulsions significantly decreased with increase in GMS concentrations and HHP treatments (200-600MPa). HHP treatment considerably reduced the creaming rate for SPP-GG model while such case was not observed for SPP-GMS model. The flow index for SPP-GG emulsion model was found to decrease with increase in HHP treatment and had non-Newtonian behaviour. The SPP-GMS emulsion models with HHP treatments showed comparatively lower viscosities but had more Newtonian flow character.