Simulated Gastrointestinal Fluids Research Articles

Isolation and identification of probiotic Bacillus subtilis AJQ03 from the intestinal tract of Anguilla japonica (Japanese eel).

In recent years, the use of fish-derived probiotics in aquaculture has become more widespread. However, research on Anguilla japonica-derived probiotics is still limited. To evaluate the potential of probiotics for disease control in eel aquaculture, isolates were obtained from the intestinal tract of healthy Anguilla japonica. These isolates were assessed for their adhesion properties, inhibition of pathogen adhesion, and hydrolytic enzyme production. Morphological characteristics and 16S rRNA sequence analysis were used for identification. Results showed that the AJQ03 strain adhered to the intestinal mucus and inhibited common pathogenic bacteria through adhesion inhibition, and further produced amylase, lipase, protease, and cellulase. Based on morphological characteristics and 16S rRNA sequencing, AJQ03 was identified as Bacillus subtilis. The strain demonstrated tolerance to various extreme conditions, as well as survival in simulated gastrointestinal fluids and superior growth in intestinal fluid compared to Luria-Bertani (LB) broth. In vitro safety tests showed that AJQ03 was not resistant to 32 antibiotics and exhibited γ hemolysis on blood plate. In vivo safety tests demonstrated a 100% survival rate for the fish, with stable organ indices, reduced bacterial loads in the liver and spleen, and complete bacterial clearance by day 7 without residue. Intestinal bacterial load results confirmed effective colonization by strain AJQ03. Analysis of the impact of AJQ03 on the gut microbiota of A. japonica revealed a significant increase in the relative abundance of Bacillus at the genus level, corroborating the colonization efficiency of AJQ03. Additionally, the relative abundances of Klebsiella, Pseudomonas, and Aeromonas were significantly lower compared to the controls, indicating that strain AJQ03 effectively reduced harmful bacteria and improved gut microbiota composition. This study confirms that B. subtilis AJQ03, isolated from the intestine of A. japonica, can serve as a probiotic candidate in A. japonica aquaculture.

Enhancement of exercise-induced fatigue alleviation and liver selenium regulation through in situ nanoselenium synthesis by Lactobacillus rhamnosus cells, empowered by Ganoderma lucidum spore loading.

Given the increasing awareness of the negative effects of fatigue on daily activities, mental health, and quality of life, antifatigue supplements are becoming increasingly popular among consumers. Selenium has been found to have antifatigue potential in high dosage, but may cause toxicity effects to the body. In this study, inorganic selenium was first converted to nanoselenium particles via in situ synthesis by Lactobacillus rhamnosus SHA113 (Se-LRS), and then loaded by Ganoderma lucidum spores (GLS). The resulting products were not only assessed for their antioxidant activities, but also the antifatigue potential in mice. As a result, both Se-LRS and the Se-LRS/GLS complex exhibited higher antioxidant and antibacterial activities in simulated gastrointestinal fluids compared to isolated selenium nanoparticles. The Se-LRS/GLS complex demonstrated sustained release of selenium in simulated gastrointestinal fluids and showed significant alleviation of exercise-induced fatigue indicators, but relatively lower liver selenium accumulation in the mice, surpassing the effects of isolated nanoselenium. No toxicity was found to Caco-2 cells for Se-LRS/GLS complex at 2µg/mL. This is a novel approach to enhance the antifatigue potential of selenium without causing extra toxicity.

From Cellulose Solutions to Aerogels and Xerogels: Controlling Properties for Drug Delivery.

A cheap and easy-to-recycle solvent, namely, aqueous NaOH with no additives, was used to dissolve cellulose and make cross-linker-free materials with varying porosity, testing them as drug delivery devices. Cellulose solutions were gelled, coagulated in a nonsolvent (water, ethanol), and dried either using supercritical CO2 (aerogels) or low-vacuum evaporation (named "xerogels"). Aerogels had densities of around 0.1 g/cm3 and specific surface areas (SSAs) of 200-400 m2/g. A significant influence of the first nonsolvent and drying mode on material properties was recorded: when the first nonsolvent was ethanol and low-vacuum drying was performed from ethanol, aerogel-like xerogels were obtained with densities of around 0.2 g/cm3 and SSAs of 200-260 m2/g. Other conditions (under evaporative drying) resulted in cellulose with much lower porosity and SSA. All materials were evaluated as drug delivery devices in simulated gastrointestinal fluids; theophylline was used as a model drug. Materials of high porosity exhibited shrinking and rapid drug release, whereas denser materials swelled and showed slower release. Two release mechanisms were suggested: diffusion through aqueous media in pores and diffusion through swollen pore walls. The results demonstrate a large spectrum of options for tuning the properties of porous cellulose materials for drug release applications.

Colonic targeting insulin-loaded trimethyl chitosan nanoparticles coated pectin for oral delivery: In vitro and In vivo studies

Colon-targeted delivery offers several benefits for oral protein delivery, such as low proteolytic enzyme activity, natural pH, and extended residence time, which improve the bioavailability of the encapsulated protein. Therefore, we hypothesize that developing a novel colonic nanocarrier system based on modified chitosan, which enhances solubility at natural pH and is coated with a colon-degradable polymer, will provide an effective delivery system for oral insulin. This study aims to synthesize insulin-loaded pectin-trimethyl chitosan nanoparticles (Ins-P-TMC-NPs) as an oral insulin delivery system and evaluate their efficacy both in vitro and in streptozotocin-induced diabetic male rats. N-trimethyl chitosan (TMC), synthesized via a methylation method, was used to prepare insulin-TMC nanoparticles coated with pectin via the ionic gelation method. The nanoparticles were characterized for their physicochemical properties, cumulative release profile, and surface morphology. In vitro biological cytotoxicity and cellular uptake of the nanoparticles were evaluated against HT-29 cells using an MTT assay and fluorescent microscopy, respectively. The in vivo blood glucose-lowering effect was assessed in diabetic male Sprague–Dawley rats, with in vivo toxicity evaluated in the liver, spleen, duodenum, pancreas, and kidney through histological studies (H&E staining). The results showed that Ins-P-TMC-NPs were spherical, with an average size of 379.40 ± 40.26 nm, a polydispersity index of 24.10 ± 1.03 %, a zeta potential of +17.20 ± 0.52 mV, and a loading efficiency of 83.21 ± 1.23 %. Compared to uncoated TMC nanoparticles, Ins-P-TMC-NPs reduced insulin loss in simulated gastrointestinal fluid by approximately 67.23 ± 0.97 % and provided controlled insulin release in simulated colonic fluid. In vitro bioactivity studies revealed that Ins-P-TMC-NPs were non-toxic, with cell viability of 91.12 ± 0.91 %, and exhibited high cellular uptake in the HT-29 cell line with a fluorescence intensity of 37.80 ± 2.40. Furthermore, in vivo studies demonstrated a sustained reduction in blood glucose levels after oral administration, peaking after 8 h with a glucose reduction of 87 ± 1.03 %. Histological sections showed no signs of toxicity. Overall, the developed colon-targeted oral insulin delivery system shows great potential as a candidate for oral insulin administration.

Preparation of curcumin-loaded chitosan/lecithin nanoparticles with increased anti-oxidant activity and in vivo bioavailability

As a natural polyphenol, curcumin (Cur) has exhibited a range of bioactive properties, including anti-inflammatory, anti-oxidant, and anti-infection properties. However, the chemical instability and low water solubility of Cur hinder its wide application. Herein, Cur-loaded chitosan/lecithin nanoparticles (CCL NPs) were prepared by the electrostatic self-assembly method. The prepared CCL NPs showed a small particle size (122.86 ± 1.53 nm) with homogeneous distribution (PDI = 0.17 ± 0.01). The high EE (79.34 ± 2.93 %) and LC (9.33 ± 0.34 %) indicated that most of Cur was encapsulated in CCL NPs. Meanwhile, the Cur was released from CCL NPs in a quick and sustained way after being exposed to simulated gastrointestinal fluids. The CCL NPs displayed superior anti-oxidant activity than that of free Cur. Moreover, the in vivo pharmacokinetic studies showed that the CCL NPs could lead to a ~ 2.64-fold increase in oral bioavailability compared with that of free Cur. All these findings indicated that the formation of CCL NPs would be a promising platform to deliver Cur in the food industry.

Locust bean gum glutarate nanocomposite hydrogel microspheres of gliclazide: Optimization by box-Behnken design and preclinical evaluation of anti-diabetic efficacy

In this study, carboxymethyl locust bean gum was synthesized and nanocomposite hydrogel microspheres (GHMs) of gliclazide were produced based on nanosilicate reinforcement and aluminium-ion driven gelation process, followed by covalent crosslinking with glutaric anhydride (GA). The effect of three independent variables (polymer and GA concentration, incubation time) on the drug entrapment efficiency (DEE%) and percent drug release at 8 h, was optimized using Box-Behnken design. The highest DEE (%) and lowest drug release was achieved at the following optimized conditions: polymer (2.43 %), GA (0.34 %), and incubation (27 min). The nanoscilicate-enriched matrix provided a maximum of 75.13 % DEE, 92.33 % gel fraction, and excellent flowability. The optimized microspheres had virtually spherical morphology, according to FE-SEM analysis. FTIR study indicated a hydrogen bonding interaction between the drug and other formulation components. X-ray and DSC measurements suggested that the crystallinity of the drug decreased following integration into the matrix. The optimized GHM demonstrated anomalous diffusion of gliclazide in simulated gastrointestinal fluids for 12 h without disintegration. In streptozotocin (50 mg/kg BW)-induced diabetic rats, the optimized formulation diminished blood glucose level by about 72 % in 12 h. Thus, the nanocomposite GHMs created by combining nanosilicate with GA had an outstanding potential for long-term control of diabetes.

Exploring the probiotic potential of Lactiplantibacillus pentosus SM1: Resistance, anti-microbial activity, anti-biofilm, cytotoxic activity, and safety properties

Lactiplantibacillus pentosus SM1 displayed notable resistance to acidic environments, with over 76% survival after 3 h in low pH conditions, and demonstrated viability in simulated gastrointestinal fluids. The Lpb. pentosus SM1 exhibited surface hydrophobicity, auto-aggregation, co-aggregation, and adhesion values of 59.60%, 38.80%, 44.90%, and 13.10% respectively. The anti-adhesion capability of Lpb. pentosus SM1 against Listeria monocytogenes adhesion to Caco-2 cells was demonstrated by its ability to compete with, inhibit, and displace at rates of 54.10%, 48.80%, and 23.90%, respectively. The Lpb. pentosus SM1 exhibited broad-spectrum anti-microbial activity, particularly against Gram-positive bacteria, and inhibited biofilm formation. The strain also showcased antioxidant activity, effectively scavenging DPPH (57.60%) and ABTS (60.54%) radicals. Notably, it absorbed 51.60% of cholesterol, highlighting its potential for cholesterol-lowering benefits. The cytotoxicity values were measured at 36.57 mg/mL for HT-29 and 38.20 mg/mL for Hela cancer cell lines. Importantly, Lpb. pentosus SM1 was free of hemolytic activity and biogenic amines, demonstrating a favorable safety profile. Lpb. pentosus SM1 exhibited the strongest resistance to ampicillin, while its resistance to erythromycin was the weakest, with inhibition zones measuring 15.50 mm and 26.40 mm, respectively. These findings suggest that Lpb. pentosus SM1 has promising applications as a probiotic candidate for promoting gut health and preventing pathogenic infections.

Composite Hydrogel of Polyacrylamide/Starch/Gelatin as a Novel Amoxicillin Delivery System.

This study investigates the development and characterization of a novel composite hydrogel composed of polyacrylamide (PAAm), starch, and gelatin for use as an amoxicillin delivery system. The optical properties, swelling behavior, and drug release profile of the composite hydrogel's were studied to evaluate its efficacy and potential applications. UV-visible spectroscopy was employed to determine the optical properties, revealing significant transparency in the visible range, which is essential for biomedical applications. The incorporation of starch and gelatin into the polyacrylamide matrix significantly enhanced the hydrogel's swelling capacity and biocompatibility. Studies on drug delivery demonstrated a sustained release profile of amoxicillin in simulated gastrointestinal fluids, which is essential for maintaining therapeutic levels for a prolonged amount of time. The results indicate that the composite hydrogel of PAAm/starch/gelatin has good swelling behavior, appealing optical characteristics, and a promising controlled drug release mechanism. These results point to this hydrogel's considerable potential as a drug delivery method, providing a viable path toward enhancing the medicinal effectiveness of amoxicillin and maybe other medications.

Design and Characterization of Liposomal-Based Carriers for the Encapsulation of Rosa canina Fruit Extract: In Vitro Gastrointestinal Release Behavior.

The increasing demand for natural compounds as an alternative to synthetic antioxidants and conservans has led to the utilization of secondary plant metabolites in the food industry, as these bioactive compounds possess great antioxidative and antimicrobial properties without side effects on human health. Despite this, the sensitivity of plant-derived compounds is a restrictive factor in terms of their full potential. The current research aimed to characterize rosehip-fruit-extract-loaded liposomes (non-treated and UV-irradiated) in terms of their density, surface tension, viscosity, chemical composition (FTIR and HPLC analyses), and thermal behavior. In the storage stability study, the vesicle size, polydispersity index (PDI), zeta potential, conductivity, and mobility of the liposomes were monitored. FTIR analysis confirmed that the plant compounds were successfully loaded within the carrier, while no chemical reaction between the rosehip fruit extract and phospholipids was detected. The results of the HPLC analysis evidence the high potential for liposomal encapsulation to protect sensitive bioactives in the rosehip fruit extract from the degrading effect of UV irradiation. The size of the rosehip-fruit-extract-encapsulated liposomes increased on the seventh day of storage from 250 nm to 300 nm, while the zeta potential values were between -21 mV and -30 mV in the same period and further stabilized over 60 days of monitoring. In Vitro release studies in water and simulated gastrointestinal fluids showed that the presence of enzymes and bile salts (in intestinal fluid) enhanced the rosehip-polyphenol permeability from liposomes (70.3% after 6 h) compared with their release in water after 24 h and in gastric fluid after 4 h (38.9% and 41.4%, respectively). The obtained results indicate that the proliposome method was an effective method for rosehip fruit extract liposomal encapsulation and for the delivery of these plant-derived bioactives in foods.

Prediction, identification, and analysis of major B-cell linear epitopes of bomb m 6 from silkworm pupa

Bomb m 6 is one of the most representative allergens in silkworm pupae. The aim of the present study was to investigate the B-cell linear epitopes of Bomb m 6. Sequence propensity– and machine learning–based strategies were first used to predict 12 and 8 candidate epitopes, respectively. Then, 46 overlapping peptides [P1–P46; length, 15 amino acids (AAs); offset, 5 AAs] spanning the Bomb m 6 sequence were synthesized and tested for IgE/IgG reactivity by dot blotting. Seven IgE-binding epitopes and six IgG-binding epitopes were identified. AA61–80, AA136–150, and AA146–150 were identified as the shared dominant IgE/IgG-binding epitopes. The epitopes contain mainly α-helix and β-sheet structures and high proportions of hydrophobic AAs, and are distributed in charged regions. The major peptides of the epitopes were poorly resistant to simulated gastrointestinal fluid digestion. These findings provide a reference for the prevention of and immunotherapy for silkworm pupa allergy.

In Vitro evaluation of probiotic properties of lactic acid bacteria strains isolated from juçara fruit native to the Atlantic Forest

Lactic acid bacteria (LAB) are the microorganisms most commonly used as probiotics. Since probiotic benefits are strain-dependent, there is a continuous need for research into new cultures with probiotic properties. Fruits such as juçara (Euterpe edulis Martius), a palm species from the Atlantic Forest threatened with extinction, are rich niches for microorganisms, including LAB. This study investigated the probiotic properties of Lactococcus lactis J7 and Leuconostoc pseudomesenteroides JF17 strains isolated from juçara fruits native to the Atlantic Forest. Probiotic characteristics, such as tolerance to simulated gastrointestinal fluids or juices, hydrophobicity, autoaggregation, coaggregation properties, inhibition of pathogenic microorganisms, and technological properties were evaluated. The survival rate of L. lactis J7 and L. pseudomesenteroides JF17 decreased after exposure to simulated gastrointestinal conditions; however, L. lactis J7 was more resistant, maintaining viability at the end of the enteric phase of 6.07 ± 0.16 log CFU mL-1. The J7 strain also exhibited the highest values in hydrophobicity (12.55 ± 0.52%), autoaggregation (25.63 ± 0.75%), and coaggregation capacity with E. coli ATCC 25922 (21.52 ± 0.98%) and S. Enteritidis ATCC 13076 (22.68 ± 1.01%). Both J7 and JF17 strains demonstrated antimicrobial activity, confirmed by the multilayer agar plate system. Additionally, the growth of the strains was temperature-dependent, and they were able to thrive in high concentrations of sodium chloride (6%). Thus, L. lactis J7 is a promising probiotic candidate for the development of functional products. Nevertheless, further studies using animal models are necessary to explore the properties of these probiotic bacteria. Novel strains isolated from fruits should be studied to broaden the application of probiotic microorganisms in the development of foods and medicines.

Structural characterization and Bacteroides proliferation promotion activity of a novel homogeneous arabinoglucuronoxylan from Commelina communis L.

Commelina communis L., a functional food and herbal plant in Asia, has been used against obesity, diabetes, and infections for centuries. A growing body of studies has demonstrated that indigestible polysaccharides are significant in obesity management. However, the structures and bioactivities of homogeneous polysaccharides from C. communis remain unclear. This study presented the structural characterization, simulated digestion, and human gut Bacteroides proliferation promotion activity of a novel homogeneous polysaccharide (CCB-3) from C. communis. The results showed that CCB-3 was an arabinoglucuronoxylan, primarily composed of arabinose, galactose, xylose, glucuronic acid (GlcA), and 4-O-methyl GlcA with a molecular weight (Mw) of 58.8 kDa. Following a 6-hour exposure to simulated gastrointestinal fluid, the Mw of CCB-3 remained unchanged, revealing that CCB-3 was an indigestible polysaccharide. Notably, CCB-3 could promote the proliferation of B. thetaiotaomicron, B. ovatus, and B. cellulosilyticus and produce short-chain fatty acids (SCFAs) and 1,2-propanediol. These findings might shed light on the discovery of polysaccharide-based leading compounds from C. communis against obesity.

Alginate-millet starch composite matrices as novel carriers for controlled delivery of grape seed polyphenols: Preparation, characterization, and in vitro release behaviour

Grape seed polyphenols (GSP) were encapsulated in alginate-modified millet starch composite matrix using internal gelation. Millet starch was modified using ultrasound (US) for 10, 20, 30, and 40 min. The application of US reduced the long-range order and increased the short-range order of starch, consequently increasing its solubility. FESEM analysis revealed that US-treatment induced fissures and grooves on the surface of starch granules and disintegrated the starch particles. Particle size distribution analysis confirmed a significant reduction in mean particle size. US-treatment facilitated enhanced alginate-starch interactions owing to an increased surface area and improved hydrogen bonding, as corroborated by FTIR, XRD, and DSC results. The composite microencapsulates prepared from 40 min US-treated starch demonstrated the highest encapsulation efficiency (89.20 ± 1.10 %), heat and UV-light stability, and delayed GSP release in simulated gastrointestinal fluids. The best fit for the release behaviour of GSP from the composite microencapsulates was observed with Peppas-Sahlin model.

In vivo antihyperglycemic effect of Ptilostemon casabonae (L.) Greuter leaf extract and its liposomal formulation

Prolonged exposure to hyperglycemia is recognized as the primary factor in the pathogenesis of diabetes, a metabolic disease that affects millions of people and one of the top 10 causes of death globally. Ptilostemon casabonae (L.) Greuter is an edible plant whose extract has recently shown inhibitory activity against α-glucosidase and antioxidant activity. This suggests that this plant could be a candidate to develop natural remedies against hyperglycemia. In this study, the in vivo antihyperglycemic effect of an extract from P. casabonae leaves is presented, and a nanoformulation is proposed to possibly enhance such effect, upon oral administration in rats. Liposomes coated with a gastro-resistant polymer, Eudragit, were developed for the loading and delivery of the extract. Nanosized (ca. 90 nm), unilamellar vesicles were produced, as demonstrated by light scattering results and cryo-TEM micrographs. The effects of the loading of the extract and the coating with Eudragit on the liposomes' bilayered structure were described by Small-Angle X-ray Scattering. The physical stability of the P. casabonae Eudragit-coated liposomes was evaluated in storage and in simulated gastrointestinal fluids, results showing that the vesicles’ key features were preserved. The vesicles efficiently entrapped rutin (57.4 % ± 5.7) and apigenin (82 % ± 9.1), main flavonoids in the extract, as indicated by the quali-quantitative characterization via HPLC-PDA. P. casabonae extract was found to exert a significant antihyperglycemic effect in rats subjected to a sucrose load, with a reduction in peak glucose level of 33 and 63 % at doses of 40 and 100 mg/kg, respectively. A 10 mg/kg dose of the extract in Eudragit-coated liposomes produced the same reduction (30 %) as that obtained with a 4-fold higher dose of the extract on its own.

Protective effects of potential probiotics Lacticaseibacillus rhamnosus SN21-1 and Lactiplantibacillus plantarum SN21-2 against Salmonella typhimurium infection in broilers

This study aimed to explore the probiotic characteristics of Lacticaseibacillus rhamnosus SN21-1 and Lactiplantibacillus plantarum SN21-2 by genotype and phenotype analysis, assess their safety in vitro and in vivo, and investigate the effects of L. rhamnosus SN21-1 and L. plantarum SN21-2 on Salmonella typhimurium-infected broilers in an in vivo experiment. L. rhamnosus SN21-1 and L. plantarum SN21-2 showed antimicrobial activity against pathogens, including S. Typhimurium, resistance to simulated gastrointestinal digestive fluid, and adhesion to HT-29 cells. In addition, L. rhamnosus SN21-1 and L. plantarum SN21-2 showed no resistance to most common antimicrobial agents and no haemolysis in vitro. Whole-genome sequence analyses of L. rhamnosus SN21-1 and L. plantarum SN21-2 provided basic genomic information, functional genes underlying the probiotic characteristics, and evidence of safety. Furthermore, feeding with L. rhamnosus SN21-1 or L. plantarum SN21-2 for 28 d had no significant effect on the growth or blood biochemical parameters of the broilers, and hematoxylin-eosin staining revealed no liver, spleen, heart, or kidney damage. Additionally, L. rhamnosus SN21-1 or L. plantarum SN21-2 did not translocate to the blood, liver, spleen, heart, or kidney of the broilers. Moreover, L. rhamnosus SN21-1 and L. plantarum SN21-2 significantly reduced S. Typhimurium counts in the faeces and caecal contents of S. Typhimurium-infected broilers and reduced small intestinal bleeding in S. Typhimurium-infected broilers. Consequently, L. rhamnosus SN21-1 and L. plantarum SN21-2 have excellent probiotic characteristics and are safe for use as anti-S. typhimurium probiotics in broilers.

Probiotics encapsulated by calcium pectin/chitosan-calcium pectin/sodium alginate-pectin-whey through biofilm-based microencapsulation strategy and their preventive effects on ulcerative colitis

Biofilm-based probiotics have emerged as a compelling approach for administering probiotics. Selecting appropriate wall materials for enclosing probiotics in biofilm structure is viewed as an advanced method of microencapsulation. In this study, Lactiplantibacillus paraplanturm LR-1 was enclosed in three different types of wall materials to create calcium pectin beads (CPB), chitosan-calcium pectin beads (C-CPB), and sodium alginate-pectin-whey beads (SPW). The biofilm cells of LR-1 were cultured externally and then encapsulated to create CPB-ex-LR-1, C-CPB-ex-LR-1 and SPW-ex-LR-1. In contrast, LR-1 was cultured within the microcapsules to form biofilms in situ, resulting in CPB-situ-LR-1, C-CPB-situ-LR-1 and SPW-situ-LR-1. The microcapsules displayed a relatively uniform morphology with the water contents below 10%. Nevertheless, there were noticeable edge effects in the distribution of strains within the in situ-type microcapsules. The detection of distinctive peaks unique to Lactiplantibacillus within the microcapsules confirmed the successful encapsulation. Additionally, a decrease in the intensity of the peaks linked to the wall materials indicated the formation of an amorphous structure within the microcapsules. The bacterial concentrations of situ-type microcapsules were found to surpass 10 lg CFU/g. The SPW-situ-LR-1 demonstrated superior tolerance performance, optimal release properties in simulated gastrointestinal fluids, and exceptional stability during the storage period. Additionally, SPW-situ-LR-1 showed similar effectiveness in alleviating colitis compared to fresh Biofilm-LR-1. Both Biofilm-LR-1 and SPW-situ-LR-1 pre-treatments exhibited positive effects on regulation, with SPW-situ-LR-1 administration significantly increasing the presence of Lactobacillus. This study provides valuable insights into the development of the next-generation probiotics.

Poly(allylamine)-adorned heptylcarboxymethyl galactomannan nanocarriers of canagliflozin for controlling type-2 diabetes: Optimization by Box-Behnken design and in vivo performance

In the past three decades, the prevalence of type-2 diabetes has arisen dramatically in countries of all income levels. A novel, most effective nanotechnology-based strategy may reduce the prevalence of diabetes. Recently, the shell-crosslinked polysaccharide-based micellar nanocarriers (MNCs) have shown great promise in terms of stability, controlled drug release, and improved in vivo performance. In this study, heptyl carboxymethyl guar gum was synthesized and characterized by ATR-FTIR, 1HNMR spectroscopy, surface charge, critical micelle concentration (23.9 μg/mL), and cytotoxicity analysis. Box-Behnken design was used to optimize the diameter, zeta potential, drug entrapment efficiency (DEE), and drug release characteristics of poly (allylamine)-crosslinked MNCs containing canagliflozin. The optimized MNCs revealed spherical morphology under TEM and had 149.3 nm diameter (PDI 21.2 %), +53.8 mV zeta potential, and 84 % DEE. The MNCs released about 63 % of the drug in 12 h under varying pH of the simulated gastrointestinal fluid. DSC and x-ray analyses suggested amorphous dispersion of drugs in the MNCs. CAM assay demonstrated the biocompatibility of the MNCs. The MNCs showed hemolysis of <1 %, 85 % mucin adsorption, and stability over three months. The MNCs demonstrated excellent anti-diabetic efficacy in streptozotocin-nicotinamide-induced diabetic rats, continuously lowering blood glucose levels up to 12 h.

Bile salt integrated cerasomes: A potential nanocarrier for enhancement of the oral bioavailability of idarubicin hydrochloride

Cerasomes are a modified form of liposomes containing both inorganic and organic parts and due to their strong polyorganosiloxane surface have remarkably high morphological stability and provide easier functionalization compared with conventional liposomes. To investigate the potential of these nanocarriers for oral delivery, bile salt integrated cerasomes (named bilocerasomes) encapsulating idarubicin hydrochloride (IDA) were prepared and characterized. The optimum formulation showed excellent stability in the simulated gastrointestinal fluids as well as under storage conditions. The oral pharmacokinetics of the IDA solution, empty nanocarrier + drug solution, and IDA-loaded bilocerasome were evaluated. The nanoformulation significantly increased the area under the drug concentration–time curve and the mean residence time (∼14.3- and 9-fold, respectively). The results obtained from cell uptake and chylomicron flow blocking approach revealed that bilocerasomes are absorbed into the intestinal cells via a clathrin/caveolin-independent endocytosis pathway and transported to the systemic circulation extensively via the intestinal lymphatic vessels. Considering the high stability of the prepared bilocerasome, noticeable participation of lymphatic transport in its systemic absorption and marked enhancement in the oral absorption of IDA, bilocerasomes can be introduced as a capable carrier for improving the oral bioavailability of drugs, particularly those that hepatic first-pass metabolism seriously limits their oral absorption.

Intracellular Gelation-Mediated Living Bacteria for Advanced Biotherapeutics in Mouse Models.

Despite its significant potential in various disease treatments and diagnostics, microbiotherapy is consistently plagued by multiple limitations ranging from manufacturing challenges to in vivo functionality. Inspired by the strategy involving nonproliferating yet metabolically active microorganisms, we report an intracellular gelation approach that can generate a synthetic polymer network within bacterial cells to solve these challenges. Specifically, poly(ethylene glycol dimethacrylate) (PEGDA, 700 Da) monomers are introduced into the bacterial cytosol through a single cycle of freeze-thawing followed by the initiation of intracellular free radical polymerization by UV light to create a macromolecular PEGDA gel within the bacterial cytosol. The molecular crowding resulting from intracytoplasmic gelation prohibits bacterial division and confers robust resistance to simulated gastrointestinal fluids and bile acids while retaining the ability to secrete functional proteins. Biocompatibility assessments demonstrate that the nondividing gelatinized bacteria are effective in alleviating systemic inflammation triggered by intravenous Escherichia coli injection. Furthermore, the therapeutic efficacy of gelatinized Lactobacillus rhamnosus in colitis mice provides additional support for this approach. Collectively, intracellular gelation indicates a universal strategy to manufacture next-generation live biotherapeutics for advanced microbiotherapy.

Revolutionizing the probiotic functionality, biochemical activity, antibiotic resistance and specialty genes of Pediococcus acidilactici BCB1H via in-vitro and in-silico approaches.

This study presents a comprehensive genomic exploration, biochemical characterization, and the identification of antibiotic resistance and specialty genes of Pediococcus acidilactici BCB1H strain. The functional characterization, genetic makeup, biological activities, and other considerable parameters have been investigated in this study with a prime focus on antibiotic resistance and specialty gene profiles. The results of this study revealed the unique susceptibility patterns for antibiotic resistance and specialty genes. BCB1H had good invitro probiotic properties, which survived well in simulated artificial gastrointestinal fluid, and exhibited acid and bile salt resistance. BCB1H didn't produce hemolysis and had certain antibiotic sensitivity, making it a relatively safe LAB strain. Simultaneously, it had good self-coagulation characteristics and antioxidant activity. The EPS produced by BCB1H also had certain antioxidant activity and hypoglycemic function. Moreover, the genome with a 42.4 % GC content and a size of roughly 1.92 million base pairs was analyzed in the genomic investigations. The genome annotation identified 192 subsystems and 1,895 genes, offering light on the metabolic pathways and functional categories found in BCB1H. The identification of specialty genes linked to the metabolism of carbohydrates, stress response, pathogenicity, and amino acids highlighted the strain's versatility and possible uses. This study establishes the groundwork for future investigations by highlighting the significance of using multiple strains to investigate genetic diversity and experimental validation of predicted genes. The results provide a roadmap for utilizing P.acidilactici BCB1H's genetic traits for industrial and medical applications, opening the door to real-world uses in industries including food technology and medicine.