Exploiting Apical Sodium-Dependent Bile Acid Transporter (ASBT)-Mediated Endocytosis with Multi-Functional Deoxycholic Acid Grafted Alginate Amide Nanoparticles as an Oral Insulin Delivery System.

OBJECTIVE: Oral administration of insulin is a potential candidate for managing diabetes. However, it is obstructed by the gastrointestinal tract barriers resulting in negligible oral bioavailability. METHODS: This investigation presents a novel nanocarrier platform designed to address these challenges. In this regard, the process involved amination of sodium alginate by ethylene diamine, followed by its conjugation with deoxycholic acid. RESULTS: The resulting DCA@Alg@INS nanocarrier revealed a significantly high insulin loading content of 63.6 ± 1.03% and encapsulation efficiency of 87.6 ± 3.84%, with a particle size of 206 nm and zeta potentials of -3 mV. In vitro studies showed sustained and pH-dependent release profiles of insulin from nanoparticles. In vitro cellular studies, confocal laser scanning microscopy and flow cytometry analysis confirmed the successful attachment and internalization of DCA@Alg@INS nanoparticles in Caco-2 cells. Furthermore, the DCA@Alg@INS demonstrated a superior capacity for cellular uptake and permeability coefficient relative to the insulin solution, exhibiting sixfold and 4.94-fold enhancement, respectively. According to the uptake mechanism studies, the results indicated that DCA@Alg@INS was mostly transported through an energy-dependent active pathway since the uptake of DCA@Alg@INS by cells was significantly reduced in the presence of NaN3 by ~ 92% and at a low temperature of 4°C by ~ 94%. CONCLUSIONS: Given the significance of administering insulin through oral route, deoxycholic acid-modified alginate nanoparticles present a viable option to surmount various obstacles presented by the gastrointestinal.

Alginate/whey protein isolate-based emulgel as an alternative margarine replacer in processed cheese: Impact on rheological, mechanical, nutritional, and sensory characteristics.

The effects of partial or full replacement of margarine by alginate/whey protein isolate-based olive oil emulgel on nutritional, physicochemical, mechanical, and rheological properties of processed cheese (PC) were investigated in this work. All formulated samples had the same amount of total fat, DM, and pH. According to the results of the fatty acids profile, the PC sample in which the margarine was fully replaced by the emulgel (EPC100) had the highest (49.84%) oleic acid content and showed a reduction of 23.7% in SFA compared with the control sample (EPC0; formulated just with margarine). In addition, EPC0 had the highest hardness among various cheese samples, which was also confirmed by its compact microstructure. Dynamic oscillatory measurements revealed that EPC100 had the highest crossover strain (or resistance to deformation). The high rigidity of this sample was related to the 3-dimensional structure of emulgel. According to the creep test results, EPC100 showed the lowest relative recovery (flowability). A high temperature dependency of viscoelastic moduli was observed in EPC0 at 42°C. No significant differences were observed between the color attributes and sensory properties of the various cheese samples. Alginate/whey protein isolate-based olive oil emulgel can be considered as a healthy margarine replacer in PC.

Comparative phylogenetic analysis of SARS-CoV-2 spike protein-possibility effect on virus spillover.

Coronavirus disease 2019 has developed into a dramatic pandemic with tremendous global impact. The receptor-binding motif (RBM) region of the causative virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), binds to host angiotensin-converting enzyme 2 (ACE2) receptors for infection. As ACE2 receptors are highly conserved within vertebrate species, SARS-CoV-2 can infect significant animal species as well as human populations. An analysis of SARS-CoV-2 genotypes isolated from human and significant animal species was conducted to compare and identify mutation and adaptation patterns across different animal species. The phylogenetic data revealed seven distinct phylogenetic clades with no significant relationship between the clades and geographical locations. A high rate of variation within SARS-CoV-2 mink isolates implies that mink populations were infected before human populations. Positions of most single-nucleotide polymorphisms (SNPs) within the spike (S) protein of SARS-CoV-2 genotypes from the different hosts are mostly accumulated in the RBM region and highlight the pronounced accumulation of variants with mutations in the RBM region in comparison with other variants. These SNPs play a crucial role in viral transmission and pathogenicity and are keys in identifying other animal species as potential intermediate hosts of SARS-CoV-2. The possible roles in the emergence of new viral strains and the possible implications of these changes, in compromising vaccine effectiveness, deserve urgent considerations.

Chimeric Hepatitis B core virus-like particles harboring SARS-CoV2 epitope elicit a humoral immune response in mice.

BACKGROUND: Virus-like particles are an interesting vector platform for vaccine development. Particularly, Hepatitis B virus core antigen has been used as a promising VLP platform. It is highly expressed in different recombinant expression systems, such as E. coli, and self-assembled in vitro. It effectively improves the immunogenicity of foreign antigenic epitopes on its surface. Various foreign antigens from bacteria, viruses, and protozoa can be genetically inserted into such nanoparticles. The effective immunogenicity due to VLP vaccines has been reported. However, no research has been performed on the SARS-CoV2 vaccine within this unique platform through genetic engineering. Considering the high yield of target proteins, low cost of production, and feasibility of scaling up, E. coli is an outstanding expression platform to develop such vaccines. Therefore, in this investigation, we planned to study and develop a unique HBc VLP-based vaccine against SARS-Cov2 utilizing the E. coli expression system due to its importance. RESULTS: Insertion of the selected epitope was done into the major immunodominant region (MIR) of truncated (149 residues) hepatitis B core capsid protein. The chimeric protein was constructed in PET28a+ and expressed through the bacterial E. coli BL21 expression system. However, the protein was expressed in inclusion body forms and extracted following urea denaturation from the insoluble phase. Following the extraction, the vaccine protein was purified using Ni2 + iminodiacetic acid (IDA) affinity chromatography. SDS-PAGE and western blotting were used to confirm the protein expression. Regarding the denaturation step, the unavoidable refolding process was carried out, so that the chimeric VLP reassembled in native conformation. Based on the transmission electron microscopy (TEM) analysis, the HBC VLP was successfully assembled. Confirming the assembled chimeric VLP, we explored the immunogenic effectivity of the vaccine through mice immunization with two-dose vaccination with and without adjuvant. The utilization of adjuvant was suggested to assess the effect of adjuvant on improving the immune elicitation of chimeric VLP-based vaccine. Immunization analysis based on anti-spike specific IgG antibody showed a significant increase in antibody production in harvested serum from immunized mice with HBc-VLP harboring antigenic epitope compared to HBc-VLP- and PBS-injected mice. CONCLUSIONS: The results approved the successful production and the effectiveness of the vaccine in terms of humoral IgG antibody production. Therefore, this platform can be considered a promising strategy for developing safe and reasonable vaccines; however, more complementary immunological evaluations are needed.

Microencapsulation of Lacticaseibacillus rhamnosus GG and L. plantarum 299 V by reverse spherification: a promising method to improve the survival of probiotics.

This research reports the first application of the reverse spherification (RVS) method for encapsulation of two probiotics (Lacticaseibacillus rhamnosus GG and L. plantarum 299 V) compared to the basic spherification (BS). These probiotics were encapsulated in different solutions encompassing various contents of alginate, gelatin, and gellan gum. The RVS bead diameters was about 1.5 times bigger and hardness was 70%-80% lower than BS samples. As determined by Raman spectral mapping, the RVS beads had two calcium alginate walls but the BS beads had only one. The inner wall of the RVS beads was more than three times thicker than outer wall. The encapsulation yields of gelatin/gellan gum and gelatin beads prepared by both methods were >1.5% alginate beads. All the RVS-prepared beads were resistant to stomach acid and showed no significant reduction in the intestine. Furthermore, the incorporation of gelatin and gellan gum into alginate led to higher cell protection. For 1.5% alginate beads, <67% survival was achieved after acid exposure but in others, >77% survival was observed; RVS beads were about 1 log above than BS ones. The proposed novel microencapsulation method efficiently increased the viability of probiotic bacteria compared to the conventional approaches.

Sublethal chlorine stress promotes the biofilm-forming ability of Salmonella enterica serovars enteritidis and expression of the related genes.

Chlorine treatment is the most common disinfection method in food-related environments. In addition to being simple and inexpensive, this method is very effective if used properly. However, insufficient chlorine concentrations only cause a sublethal oxidative stress in the bacterial population and may alter the growth behavior of stressed cells. In the present study, the effect of sublethal chlorine stress on the biofilm formation characteristics of Salmonella Enteritidis was evaluated. Our results demonstrated that, sublethal chlorine stress (350 ppm total chlorine) activates the biofilm (csgD, agfA, adrA and bapA) and quorum-sensing (sdiA and luxS) related genes in planktonic cells of S. Enteritidis. The higher expression of these genes illustrated that the chlorine stress induced the initiation of the biofilm formation process in S. Enteritidis. Results of the initial attachment assay confirmed this finding. In addition, the number of chlorine-stressed biofilm cells was significantly higher than non-stressed biofilm cells after 48 h incubation at 37 °C. In S. Enteritidis ATCC 13076 and S. Enteritidis KL19, the number of chlorine-stressed biofilm cells were 6.93 ± 0.48 and 7.49 ± 0.57 log CFU/cm2, while the number of non-stressed biofilm cells were 5.12 ± 0.39 and 5.63 ± 0.51 log CFU/cm2, respectively. These findings were confirmed by measurements of the major components of biofilm, i.e., eDNA, protein and carbohydrate. The amount of these components in 48-h biofilms was higher when the cells were initially subjected to sublethal chlorine stress. However, the up-regulation of the biofilm and quorum sensing genes was not observed in 48-h biofilm cells, indicating that the effect of chlorine stress had vanished in the subsequent generations of Salmonella. In total, these results revealed that sublethal chlorine concentrations can promote the biofilm-forming ability of S Enteritidis.

Dual-band wide-angle perfect absorber based on the relative displacement of graphene nanoribbons in the mid-infrared range.

In this paper, a novel graphene-based dual-band perfect electromagnetic absorber operating in the mid-infrared regime has been proposed. The absorber has a periodic structure which its unit cell consists of a sliver substrate and two graphene nanoribbons (GNRs) of equal width separated with a dielectric spacer. Two distinct absorption peaks at 10 and 11.33 µm with absorption of 99.68% and 99.31%, respectively have been achieved due to a lateral displacement of the GNRs. Since graphene surface conductivity is tunable, the absorption performance can be tuned independently for each resonance by adjusting the chemical potential of GNRs. Also, it has been proved that performance of the proposed absorber is independent of the incident angle and its operation is satisfactory when the incident angle varies from normal to ±75°. To simulate and analyze the spectral behavior of the designed absorber, the semi-analytical method of lines (MoL) has been extended. Also, the finite element method (FEM) has been applied in order to validate and confirm the results.

Betanin alleviates oxidative stress through the Nrf2 signaling pathway in the liver of STZ-induced diabetic rats.

BACKGROUND: Continuing hyperglycemia causes and exacerbate oxidative stress. Betanin as the principal pigment of red beet root has antioxidant, anti-inflammatory, and anti-diabetic properties. The purpose of this study was to investigate the potency of betanin on antioxidant defense in STZ-induced diabetic rats' livers. METHODS: STZ at a single dose of 60 mg/kg body weight was intraperitoneally injected and betanin (10, 20, and 40 mg/kg/day) was administered orally for 28 days. Malondialdehyde (MDA), total antioxidant capacity (TAC), protein carbonyl (PC) levels, and the enzyme activity of superoxide dismutase (SOD), catalases and glutathione peroxidases (GPx) were evaluated in the liver. Furthermore, gene expression of Nrf2 and mentioned antioxidant enzymes were measured by Real-time PCR. RESULTS: Betanin (10 and 20 mg/kg) significantly reduced PC levels and increased antioxidant enzyme activity in diabetic rats compared to the control diabetic group (P < 0.01). In comparison to the diabetic control group, all studied genes expression in diabetic rats were increased significantly with betanin at doses of 10 and 20 mg/kg (P < 0.02). The increase in gene expression at 20 mg/kg of betanin was significantly stronger than others (P < 0.015) except for the catalase (P = 0.201), that was almost the same. Moreover, treatment of diabetic rats with 20 mg/kg of betanin could significantly increase TAC levels (P < 0.05) and decrease MDA levels (P < 0.001) compared to diabetic control group. CONCLUSIONS: Betanin could increase the antioxidant capacity of liver tissue associated with the Nrf2-mediated pathway in a dose-dependent manner.

Plant-derived VLP: a worthy platform to produce vaccine against SARS-CoV-2.

After its emergence in late 2019 SARS-CoV-2 was declared a pandemic by the World Health Organization on 11 March 2020 and has claimed more than 2.8 million lives. There has been a massive global effort to develop vaccines against SARS-CoV-2 and the rapid and low cost production of large quantities of vaccine is urgently needed to ensure adequate supply to both developed and developing countries. Virus-like particles (VLPs) are composed of viral antigens that self-assemble into structures that mimic the structure of native viruses but lack the viral genome. Thus they are not only a safer alternative to attenuated or inactivated vaccines but are also able to induce potent cellular and humoral immune responses and can be manufactured recombinantly in expression systems that do not require viral replication. VLPs have successfully been produced in bacteria, yeast, insect and mammalian cell cultures, each production platform with its own advantages and limitations. Plants offer a number of advantages in one production platform, including proper eukaryotic protein modification and assembly, increased safety, low cost, high scalability as well as rapid production speed, a critical factor needed to control outbreaks of potential pandemics. Plant-based VLP-based viral vaccines currently in clinical trials include, amongst others, Hepatitis B virus, Influenza virus and SARS-CoV-2 vaccines. Here we discuss the importance of plants as a next generation expression system for the fast, scalable and low cost production of VLP-based vaccines.

A short overview of CRISPR-Cas technology and its application in viral disease control.

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs) together with CRISPR-associated (Cas) proteins have catalysed a revolution in genetic engineering. Native CRISPR-Cas systems exist in many bacteria and archaea where they provide an adaptive immune response through sequence-specific degradation of an invading pathogen's genome. This system has been reconfigured for use in genome editing, drug development, gene expression regulation, diagnostics, the prevention and treatment of cancers, and the treatment of genetic and infectious diseases. In recent years, CRISPR-Cas systems have been used in the diagnosis and control of viral diseases, for example, CRISPR-Cas12/13 coupled with new amplification techniques to improve the specificity of sequence-specific fluorescent probe detection. Importantly, CRISPR applications are both sensitive and specific and usually only require commonly available lab equipment. Unlike the canonical Cas9 which is guided to double-stranded DNA sites of interest, Cas13 systems target RNA sequences and thus can be employed in strategies directed against RNA viruses or for transcriptional silencing. Many challenges remain for these approach, including issues with specificity and the requirement for better mammalian delivery systems. In this review, we summarize the applications of CRISPR-Cas systems in controlling mammalian viral infections. Following necessary improvements, it is expected that CRISPR-Cas systems will be used effectively for such applications in the future.

Utilization of the human gamma-satellite insulator for the enhancement of anti-PCSK9 monoclonal antibody expression in Chinese hamster ovary cells.

Monoclonal antibodies (mAbs) are widely employed as invaluable therapeutics for a vast number of human disorders. Several approaches have been introduced for the improvement of mAb production in Chinese hamster ovary (CHO) cells due to the increasing demand for these products. In this regard, various chromatin-modifying elements such as insulators have been incorporated in the expression vectors to augment mAb expression. In this study, human gamma-satellite insulator containing vectors were utilized for the expression of an anti-proprotein convertase subtilisin/kexin type 9 (PCSK9) mAb in CHO-K1 cells. To this aim, dual expression vectors encoding the antibody light chain (LC) and heavy chain (HC) with or without the insulator element were constructed, and mAb expression was evaluated in transient and stable expression. Based on the results, mAb expression significantly increased in the stable cell pool, and clonal cells developed using the human gamma-satellite insulator. In contrast, transient antibody expression was not affected by the insulator element. Finally, the enhancement of LC and HC mRNA levels was found in the insulator containing stable cell pools using the quantitative real-time-polymerase chain reaction (qRT-PCR). Our findings showed the positive effect of the human gamma-satellite insulator on the stable expression of an anti-PCSK9 immunoglobulin G1 (IgG1) mAb in CHO-K1 cells using dual expression vectors.

Natural flavor biosynthesis by lipase in fermented milk using in situ produced ethanol.

ABSTRACT: Many flavoring agents on the market are extracted from natural sources or synthesized chemically. Due to the disadvantages of both methods, biotechnology is becoming a promising alternative. In this study, short chain ethyl esters with fruity notes were biosynthesized in UHT whole milk via coupling ethanolic fermentation with lipase (Palatase®) transesterification. Kluyveromyces marxianus, Lactobacillus fermentum and Lb. paracasei were used for fermentation. Milk fat was esterified with in situ produced ethanol by adding lipase at 0, 8 and 24 h of fermentation. Viable cell counts and pH were monitored during 48 h fermentation period. Flavor active ethyl esters, ethanol and free fatty acids were analyzed using headspace SPME-GC. Free fatty acid levels were lower in K. marxianus samples than lactobacilli. K. marxianus produced higher amounts of ethanol and esters than lactic acid bacteria. Viable cell counts decreased after lipase application at 0 and 8 h, possibly due to fatty acid production. Addition of lipase at 24 h resulted in improved cell counts as well as ethanol and ester production in the case of K. marxianus. This study demonstrated that fermenting milk with alcohol producing cultures in conjunction with lipase application can be an alternative to artificial flavorings in fermented milks.

Muscle mass and function are related to respiratory function in chronic obstructive pulmonary disease.

Background: Chronic obstructive pulmonary disease (COPD), as an airway limitation condition, is accompanied by alteration of muscle mass and function. We aimed to determine the relationship between disease severity and body composition, muscle function, and nutritional status in COPD patients. Methods: This cross-sectional study was conducted on 129 COPD participants. Muscle strength, body composition, and calf circumference (CC) were measured using a hydraulic hand dynamometer, bioelectrical impedance analysis (BIA), and a tape measure, respectively. Furthermore, fat-free mass index (FFMI), body mass index (BMI) and muscle mass value were calculated by equations. Forced expiratory volume in one second (FEV1) was assessed as well. Nutritional status was also evaluated by subjective global assessment (SGA) questionnaire. SPSS software (version 21 ) was used, chi-square, fisher's exact test, univariate and multivariate linear regression models were used for statistical analysis. P-values less than 0.05 were considered significant. Results: Based on FEV1 classification, 52.7% of the patients had severe conditions. The reports indicated that the prevalence of low CC was 54.2%, low muscle mass 38.7%, low FFMI 34.8%, low right handgrip strength 61.2% and low left handgrip strength 64.3%. Furthermore, there was an increasing trend based on FEV1 in low CC (p=0.032), low muscle mass (p=0.005), low FFMI (p=0.002), low right handgrip strength (p=0.004) and low left handgrip strength (p=0.014). The results of univariate analysis showed muscle mass (p=0.036), total protein (p=0.043), FFM (p=0.047), FFMI (p=0.007), SGA (p=0.029), right handgrip strength (p=0.004) and left hand grip strength (p=0.023) were associated with FEV1. In addition, the results of multivariate analysis demonstrated low values of FFMI (p=0.005) and right handgrip strength (p=0.042) were the main detrimental factors for FEV1. The results of multivariate analysis were confirmed by stepwise model. Conclusion: Low values of muscle mass and function are prevalent among COPD patients. The present study revealed that low FFMI and handgrip strength were closely related to disease severity.

Fortified whey beverage for improving muscle mass in chronic obstructive pulmonary disease: a single-blind, randomized clinical trial.

BACKGROUND: The development of effective nutritional supports for patients with chronic obstructive pulmonary diseases (COPD) is still challenging. This study was conducted to investigate the efficacy of daily consumption of fortified whey on inflammation, muscle mass, functionality, and quality of life in patients with moderate-to-severe COPD. METHODS: A single-blind, randomized trial study was performed on patients with COPD (n = 46). Participants in the intervention group (n = 23) daily received 250 ml of whey beverage fortified with magnesium and vitamin C for 8 weeks. Any changes in inflammatory cytokines (including interleukin- 6 (IL-6) and tumor necrosis factor (TNFα)) were the primary outcomes and the secondary outcomes were fat-free mass, handgrip strength, malnutrition, glutathione and malondialdehyde serum concentrations, and health-related quality of life (HRQoL). Body composition and muscle strength were measured by Bioelectrical Impedance Analysis (BIA) and hydraulic hand dynamometer, respectively. Fat-free mass index (FFMI) was also calculated. RESULTS: At the end of the study, 44 patients were analyzed. There were significant decreases in IL-6 concentrations in the intervention group compared to the control group. Also, FFMI, body protein, and handgrip strength increased significantly in the intervention group with significant changes between two groups. Moreover, improvement in health-related quality of life was observed in the intervention group compared to the control group. There were no significant changes in other study variables. CONCLUSIONS: This novel nutritional intervention decreased inflammatory cytokines levels, improved indices of skeletal muscle mass and muscle strength, and ultimately, increased HRQoL in patients with moderate-to-severe COPD. Thus, it is suggested to do further studies to assess the effects of nutrition intervention on COPD progression. TRIAL REGISTRATION: IR.SUMS.REC.1396.85 ( https://www.irct.ir/ ).

Incorporation of pomegranate rind powder extract and pomegranate juice into frozen burgers: oxidative stability, sensorial and microbiological characteristics.

This study was aimed to evaluate the antibacterial and antioxidant characteristics of incorporated pomegranate juice (PJ) and pomegranate rind powder extract (PRPE) into meat burgers. The peroxide value, thiobarbituric acid reactive substances, and metmyoglobin content for different burgers during 90 days storage at - 18 °C were evaluated. Total anthocyanin content, total phenolic content (TPC) and free radical scavenging activity (RSA or IC50) for PJ and PRPE were measured as 18.90 (mg/mL), 4380 ppm, 0.136 (mg/mL) and 0.40 (mg/mL), 5598 ppm, 0.084(mg/mL), respectively. Incorporation of PRPE with a high concentration of TPC resulted in less oxidation of lipid in comparison with other formulations. The highest and lowest scores in the sensory analysis and total acceptance at the 90th day corresponded to burgers containing PJ and control, respectively. Butylated hydroxytoluene may be substituted in whole or part with PJ and PRPE due to their desired effects on burgers' properties.

Combined CD44, c-MET, and EGFR expression in p16-positive and p16-negative head and neck squamous cell carcinomas.

PURPOSE/OBJECTIVE(S): To examine the association between CD44 and c-MET expression in relation to p16 and EGFR in patients with head and neck squamous cell carcinoma (HNSCC). MATERIALS/METHODS: Immunohistochemical staining of CD44, p16, EGFR, and c-MET was performed on 105 locally advanced HNSCC patients treated with chemoradiation. CD44 expression was correlated with c-MET, EGFR, and p16, locoregional control (LRC), distant metastases (DM), disease-free survival (DFS) and overall survival (OS). RESULTS: High CD44 expression was present in 33% of patients and was associated with non-oropharynx primaries (P < 0.001), high c-MET expression (P < 0.001), p16-negative (P < 0.001) and EGFR-positive tumors (P < 0.001). Fifty-seven percent of CD44 high expressing tumors had high c-MET expression compared to 21% of CD44 low expressing tumors (P < 0.001). High CD44 expression predicted for worse LRC (HR: 2.44; 95% CI: 1.16-5.13; P = 0.018), DFS (HR: 2.61; 95% CI: 1.46-4.67; P = 0.001), and OS (HR: 2.52; 95% CI: 1.30-4.92; P = 0.007) but not DM (P = 0.57) on univariate analysis. Patients with both high CD44 and c-MET expression had a poor prognosis with a 2-year DFS of 30% compared to 70% in the rest of the cohort (P = 0.003). On multivariable analysis, after adjusting for site, T-stage, smoking history, and EGFR status, high c-MET (P = 0.039) and negative p16 status (P = 0.034) predicted for worse DFS, while high CD44 expression did not (P = 0.43). CONCLUSIONS: High CD44 expression is associated with high c-MET expression, p16-negative tumors, and EGFR-positive tumors. The combination of these markers predicts for poor prognosis in HNSCC patients treated with chemoradiation.

Protective effects of synbiotic diets of Bacillus coagulans, Lactobacillus plantarum and inulin against acute cadmium toxicity in rats.

BACKGROUND: Cadmium is a heavy metal that causes oxidative stress and has toxic effects in humans. The aim of this study was to investigate the influence of two probiotics along with a prebiotic in preventing the toxic effects of cadmium in rats. METHODS: Twenty-four male Wistar rats were randomly divided into four groups namely control, cadmium only, cadmium along with Lactobacillus plantarum (1 × 109 CFU/day) and inulin (5% of feedstuff) and cadmium along with Bacillus coagulans (1 × 109 spore/day) and inulin (5% of feedstuff). Cadmium treated groups received 200 µg/rat/day CdCl2 administered by gavage. During the 42-day experimental period, they were weighed weekly. For evaluation of changes in oxidative stress, the levels of some biochemicals and enzymes of serum including SOD, GPX, MDA, AST, ALT, total bilirubin, BUN and creatinine, and also SOD level of livers were measured at day 21 and 42 of treatment. The cadmium content of kidney and liver was determined by using atomic absorption mass spectrophotometry. Data were analyzed using analysis of variance (ANOVA) followed by Duncan's post hoc test. RESULTS: Treatment of cadmium induced rats with synbiotic diets significantly improved the liver enzymes and biochemical parameters that decreased AST, ALT, total bilirubin, BUN and metal accumulation in the liver and kidney and increased body weight, serum and liver SOD values in comparison with the cadmium-treated group. No significant differences were observed with MDA and GPX values between all groups (p > 0.05). CONCLUSIONS: This study showed that synbiotic diets containing probiotics (L. plantarum and B. coagulans) in combination with the prebiotic (inulin) can reduce the level of cadmium in the liver and kidney, preventing their damage and recover antioxidant enzymes in acute cadmium poisoning in rat.

Effects of microbial transglutaminase on physicochemical properties, electrophoretic patterns and sensory attributes of veggie burger.

The main objective of this study was to investigate the effects of microbial-transglutaminase (MTGase 0-0.75%)/sodium-caseinate (SC 0-2%) as crosslinker agents on proximate analysis, binding properties (expressible moisture and shrinkage), texture analysis, electrophoretic patterns, instrumental color, and sensory properties of veggie burgers. Addition of SC and MTGase positively affected shrinkage and expressible moisture. It also increased hardness, springiness, chewiness, and cutting-force of burgers. Presence of SC had no effects on cohesiveness of burgers. Total protein and ash of samples were increased by treatment with SC. The lightness (L*) of samples was significantly decreased by 0.75% MTGase. No significant influence of SC on samples color parameters was observed. The results indicated that distinct protein bands were not formed on the SDS-PAGE of burger samples and resulted in a smearing pattern on the gel. When soy-protein was incubated with MTGase, a progressive decrease in the intensity of the bands corresponding to the subunits 7S and 11S globulins was observed concomitant with disappearance of A3 and B3 bands. Electrophoresis pattern of gluten was slightly changed after MTGase treatment. There were significant differences in color, taste, appearance, mouth feel, and overall acceptability between treated and control samples. Results suggest that production of veggie burgers using MTGase alone or in combination with SC brings about covalent cross-linking between homologous and heterologous proteins to form high-molecular weight polymers, thereby improving the mechanical properties of veggie burgers and profoundly increases the acceptability of the end product.

Intradialytic Oral Protein Supplementation and Nutritional and Inflammation Outcomes in Hemodialysis: A Randomized Controlled Trial.

BACKGROUND: Malnutrition is a common finding in hemodialysis patients and can increase oxidative stress and inflammation levels. STUDY DESIGN: A randomized, controlled, nonblinded, parallel trial. SETTING & PARTICIPANTS: 92 hemodialysis patients from a single center with malnutrition according to subjective global assessment (SGA) score (SGA score > 7). INTERVENTION: 3 treatment groups (23 patients each) received 220mL of fermented vitamin E-fortified whey beverage (15g of whey protein concentrate + 600IU of vitamin E) or 220mL of fermented whey beverage (15g of whey protein concentrate) or vitamin E (600IU) 3 times a week for 8 weeks. The control group (23 patients) received no intervention. OUTCOME & MEASUREMENTS: Primary outcomes were change in SGA score and malnutrition-inflammation score (MIS) from baseline to the end of the trial. RESULTS: At the end of the study, 83 patients were analyzed (2, 3, 1, and 3 patients left the study in the vitamin E-fortified whey beverage, whey beverage, vitamin E, and control groups, respectively). Changes in SGA scores were -3.48 (95% CI, -4.90 to -2.00), -3.22 (95% CI, -4.13 to -2.30), -1.70 (95% CI, -3.20 to -0.24), and 1.56 (95% CI, 0.60 to 2.50) for the vitamin E-fortified whey beverage, whey beverage, vitamin E, and control groups, respectively (overall P<0.001; P≤0.001 for each treatment group vs control). Changes in MISs were -3.17 (95% CI, -4.40 to -1.90), -1.83 (95% CI, -2.50 to -1.10), -2.30 (95% CI, -3.50 to -1.10), and 1.48 (95% CI, 0.65 to 2.30) for the vitamin E-fortified whey beverage, whey beverage, vitamin E, and control groups, respectively (overall P<0.001; P<0.001 for each treatment group vs control). Few adverse effects were reported in any group. LIMITATIONS: Lack of blinding, small sample size, and short duration. CONCLUSIONS: Whey protein in the form of a new fermented whey beverage and vitamin E supplementation may improve SGA score and MIS in the short term.

Mitochondrial oxidative stress and dysfunction induced by isoniazid: study on isolated rat liver and brain mitochondria.

Isoniazid (INH or isonicotinic hydrazide) is used for the treatment and prophylaxis of tuberculosis. Liver and brain are two important target organs in INH toxicity. However, the exact mechanisms behind the INH hepatotoxicity or neurotoxicity have not yet been completely understood. Considering the mitochondria as one of the possible molecular targets for INH toxicity, the aim of this study was to evaluate the mechanisms of INH mitochondrial toxicity on isolated mitochondria. Mitochondria were isolated by differential ultracentrifugation from male Sprague-Dawley rats and incubated with different concentrations of INH (25-2000 µM) for the investigation of mitochondrial parameters. The results indicated that INH could interact with mitochondrial respiratory chain and inhibit its activity. Our results showed an elevation in mitochondrial reactive oxygen species (ROS) formation, lipid peroxidation and mitochondrial membrane potential collapse after exposure of isolated liver mitochondria in INH. However, different results were obtained in brain mitochondria. Noteworthy, significant glutathione oxidation, adenosine triphosphate (ATP) depletion and lipid peroxidation were observed in higher concentration of INH, as compared to liver mitochondria. In conclusion, our results suggest that INH may initiate its toxicity in liver mitochondria through interaction with electron transfer chain, lipid peroxidation, mitochondrial membrane potential decline and cytochrome c expulsion which ultimately lead to cell death signaling.