Management of Cardiovascular Diseases by Short-Chain Fatty Acid Postbiotics.

PURPOSE OF REVIEW: Global health concerns persist in the realm of cardiovascular diseases (CVDs), necessitating innovative strategies for both prevention and treatment. This narrative review aims to explore the potential of short-chain fatty acids (SCFAs)-namely, acetate, propionate, and butyrate-as agents in the realm of postbiotics for the management of CVDs. RECENT FINDINGS: We commence our discussion by elucidating the concept of postbiotics and their pivotal significance in mitigating various aspects of cardiovascular diseases. This review centers on a comprehensive examination of diverse SCFAs and their associated receptors, notably GPR41, GPR43, and GPR109a. In addition, we delve into the intricate cellular and pharmacological mechanisms through which these receptors operate, providing insights into their specific roles in managing cardiovascular conditions such as hypertension, atherosclerosis, heart failure, and stroke. The integration of current information in our analysis highlights the potential of both SCFAs and their receptors as a promising path for innovative therapeutic approaches in the field of cardiovascular health. The idea of postbiotics arises as an optimistic and inventive method, presenting new opportunities for preventing and treating cardiovascular diseases.

The synergistic effect of titanium dioxide nanoparticles and yeast isolated from fermented foods in reduction of aflatoxin B1.

The presence of aflatoxins in food products can lead to health risks in human societies. Therefore, in the present study, the effect of yeast strains isolated from fermented products and titanium dioxide nanoparticles (TiO2-NPs) was studied on aflatoxin reduction. Yeast strains were isolated from fermented products such as sweet fruits and dairy products and identified using biochemical, ascospore (testing by culture medium optimization V8 which is called V8NLF), and molecular methods. The probiotic activity of four selected yeasts was evaluated. Then, the effect of selected yeast isolates and TiO2-NPs on reducing aflatoxin B1 (AFB1) in the medium was studied by measuring AFB1 using ELISA and HPLC. The results of biochemical and molecular identification experiments indicate that the selected strain (Y1) is Saccharomyces cerevisiae. The selected strains showed good tolerance to different concentrations of bile salt, pH, and NaCl, indicating appropriate probiotic activity. It also showed antimicrobial activity against Escherichia coli, Shigella dysenteriae, and Salmonella typhimurium. Selected strain and TiO2-NPs showed AFB1 reducing activity in the medium and when combined, showed synergistic effects in reducing AFB1. TiO2-NPs in combination with selected yeast strains have a high ability to remove AFB1 from the medium and, therefore, can be used for future studies.

Hierarchically-structured laccase@Ni3(PO4)2 hybrid nanoflowers for antibiotic degradation: Application in real wastewater effluent and toxicity evaluation.

Laccase@Ni3(PO4)2 hybrid nanoflowers (HNFs) were prepared by the anisotropic growth of biomineralized nickel phosphate. The immobilization yield was 77.5 ± 3.6 %, and the immobilized enzyme retained 50 % of its initial activity after 18 reusability cycles. The immobilized and free enzymes lost 80 % of their activity after 18 and 6 h incubation in municipal wastewater effluent (MWWE), respectively. The increase in α-helix content (8 %) following immobilization led to a more rigid enzyme structure, potentially contributing to its improved stability. The removal of ciprofloxacin from MWWE by laccase@Ni3(PO4)2·HNFs/p-coumaric acid oxidation system was optimized using a Box-Behnken design. Under the optimized conditions [initial laccase activity (0.05 U mL-1), the concentration of p-coumaric acid (2.9 mM), and treatment time (4.9 h)], the biocatalyst removed 90 % of ciprofloxacin (10 mg L-1) from MWWE. The toxicity of ciprofloxacin against some G+ and G- bacteria was reduced by 35-70 %, depending on their strain. The EC50 of ciprofloxacin for the alga Raphidocelis subcapitata reduced from 3.08 to 1.07 mg L-1 (p-value <0.05) after the bioremoval. Also, the acute and chronic toxicity of identified biodegradation products was lower than ciprofloxacin at three trophic levels, as predicted by ECOSAR software.

Developing a new biologic toxin binder for reducing AFB1 toxicity in laying hens.

Aflatoxin (AFs)-contaminated diet in feeding domestic animals is one of the biggest health concerns for humans. Therefore, various methods have been developed to detoxify AFs. In the present study, adding Saccharomyces cerevisiae probiotic yeast and titanium dioxide nanoparticles (TiO2-NPs) reduces the toxicity of AF B1 (AFB1) in laying hens was studied. After preparing the laying hens, they were fed with a diet containing AFB1 for 14 days and supplemented with S. cerevisiae and TiO2-NPs. Weight changes, serum levels of albumin, globulin, total protein, aspartate transaminase (AST), and alanine transaminase (ALT) were measured over 14 days. Also, on day 14, after killing the animals, their liver tissue was extracted, and the AFB1 content was measured by high-performance liquid chromatography (HPLC) and studied histopathologically using hematoxylin-eosin staining. The results showed that adding S. cerevisiae strain and TiO2-NPs to the diet of chicks with aflatoxicosis prevented weight loss, detoxified the liver, increased total protein, decreased albumin, and globulin content. Histopathological images showed damage to the liver tissue of laying hens fed diets containing AFB1. However, S. cerevisiae and TiO2-NPs were able to prevent liver damage. In general, it was concluded that adding S. cerevisiae along with TiO2-NPs could be a good optiofor reducing AFB1 toxicity in laying hens.

A new probiotic Lactobacillus plantarum strain isolated from traditional dairy together with nanochitosan particles shows the synergistic effect on aflatoxin B1 detoxification.

The present study aimed to evaluate the effects of new Lactobacillus plantarum strain isolated from dairy products as well as chitosan nanoparticles on reducing aflatoxin B1 (AFB1) toxicity In vitro. After collection and preparation of yogurt, cheese, milk, and whey products, lactic acid bacteria (LABs) were isolated and identified using biochemical and molecular methods. pH, bile, and salt tolerance tests were used to measure probiotic activity. Then, the antimicrobial activity of LABs against gastrointestinal pathogens was studied. The strain isolated from cheese (C1) was selected as the appropriate strain and antibiotic susceptibility test was performed for this strain. Then, the effect of C1 isolate and chitosan nanoparticles on reducing aflatoxin B1 (AFB1) in the medium was studied by measuring AFB1 using the enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC). The results of biochemical evaluations indicated the separation of different strains of L. plantarum. Antimicrobial activity test showed extensive antimicrobial activity of C1 isolate. The results showed that this strain has good probiotic activities. This strain was shown to be resistant to erythromycin, fusidic acid, gentamicin, kanamycin, nalidixic acid, neomycin, ofloxacin, and vancomycin antibiotics. C1 strain together with chitosan nanoparticles was able to reduce AFB1 in the medium and, when both were used simultaneously, a synergistic effect was seen in reducing AFB1 from the medium. Based on the findings, it can be concluded that the new C1 L. plantarum strains together with chitosan nanoparticles had synergistic effects on reducing AFB1 toxin in food products.

Nanobiosensor Based on Sugar Code-AuNPs Aggregation: A Key to Opening New Gates in Rapid Diagnosis of Streptococcal Pharyngitis.

Streptococcal pharyngitis is mainly caused by Streptococcus pyogenes (GAS), which if left untreated can lead to rheumatic heart disease. The accurate diagnosis of streptococcal pharyngitis is a challenge for clinicians because several symptoms of streptococcal pharyngitis are similar to viral pharyngitis. There are some commercially available biosensors for the rapid diagnosis of streptococcal pharyngitis. Nevertheless, they are not widely used by physicians, mainly because of their high price and dependence on the instrument. Serotype M1 GAS is the most prevalent cause of streptococcal pharyngitis and binds to H-1 antigen, a sugar code found on oral epithelial cells. Here, we present a nanobiosensor based on aggregation of H-1 antigen-conjugated gold nanoparticles for the rapid, qualitative, and quantitative detection of M1 GAS, which is inspired by the sugar code-lectin interaction. It is noteworthy that M1 GAS was detected in a wide concentration range (1 × 103-1×106 CFU/ml) with a linear response and a short detection time of 20 min. Good reproducibility, easy-to-use, and relatively low production cost are among other attractive features of this nanobiosensor. This work provides a strategic roadmap for developing a new generation of biosensors via targeting the sugar code-lectin interaction in future studies.

A Simplified Process for Purification and Refolding of Recombinant Human Interferon-α2b

Background: Interferon α-2b is a vital biotherapeutic produced through the recombinant DNA technology in E. coli. The recombinant IFN-α2b normally appears as intercellular IBs, which requires intensive refolding and purification steps. Method: Purification of IFN-α2b from solubilized IB was performed using two-phase extraction. To optimize refolding conditions, the effects of pH and different additives, including cysteine, cystine, urea, glycerol, Triton X-100, NaCl, and arginine, were investigated. Optimal refolding buffer (0.64 mM of urea, 5.57 mM of cysteine , and 1.8 mM of cystine) was obtained using RSM. The refolding process was performed by an optimized refolding buffer in the dilution and fed-batch refolding method at different protein concentrations (25-1000 µg/mL). Result: At a final protein concentration of 500 µg/mL, the fed-batch refolding method yielded in a biological activity of 2.24 × 108 IU/mg, which was nearly twice that of dilution method. Conclusion: Fed-batch refolding method resulted in the biologically active IFN-α2b with high purity, which can be used for research and industrial purposes.

Bacteriocin activity of various iranian honey-associated bacteria and development of a simple medium for enhanced bacteriocin activity.

PURPOSE: Honey is a promising source of bacterial strains producing metabolites with antimicrobial activity. There is a great variety in the antimicrobial activity of honey from different areas of nature. Therefore, the aim of present study was to investigate the antibacterial activity of Iranian honey from different regions and to optimize the culture condition for the highly potent bacterial isolate. METHODS: Honey samples were collected from ten different regions of Iran and were screened for bacteriocin-producing bacteria. The best bacteriocin-producing strain was characterized and identified by 16S rDNA analysis. One-factor-at-a-time method was used for optimization of culture medium and the yield and time-course of bacteriocin production were compared in both shake flask and bio-reactor. RESULTS: The Bacillus subtilis SB1 that was isolated from Sabalan honey showed potent antibacterial activity with prominent thermal stability. The optimum medium for the bacteiocin production was a yeast extract-based medium. The optimum incubation temperature for bacteriocin production was 34 °C. Bacteriocin production was higher near neutral pH conditions than that produced at acidic or alkaline environment. The results of cell growth and bacteriocin assays revealed that the exponential phase of growth and antibacterial compounds production was started rapidly in bioreactor than flask. CONCLUSIONS: Findings of this study supported the folkloric application of honey against some infectious diseases. B.subtilis SB1 that isolated from Sabalan honey was a potential source for bacteriocins-like compounds. Our studies suggested a simple buffered nitrogen-based medium for SB1 growth and bacteriocin activity in both shake flask and bioreactor.

Efficacy of probiotics for managing infantile colic due to their anti-inflammatory properties: a meta-analysis and systematic review.

BACKGROUND: Infantile colic (IC) is excessive crying in otherwise healthy children. Despite vast research efforts, its etiology remains unknown. PURPOSE: Most treatments for IC carry various side effects. The collection of evidence may inform researchers of new strategies for the management and treatment of IC as well as new clues for understanding its pathogenesis. This review and meta-analysis aimed to evaluate the efficacy and possible mechanisms of probiotics for mananaging IC. METHODS: Ten papers met the study inclusion and exclusion criteria, and the meta-analysis was conducted using Review Manager (RevMan) software and a random-effects model. RESULTS: This meta-analysis revealed that probiotics are effective for treating infantile colic, while the review showed that this efficacy may be due to their anti-inflammatory effects. CONCLUSION: Probiotics may be an important treatment option for managing infantile colic due to their anti-inflammatory properties.

Comparative effects of probiotic and paraprobiotic addition on microbiological, biochemical and physical properties of yogurt.

Paraprobiotics are inactivated probiotics that exert various health and technological benefits making them suitable for production of functional yogurt. In the present study, probiotic yogurt containing Lactobacillus acidophilus ATCC SD 5221 and Bifidobacterium lactis BB-12 and paraprobiotic yogurt containing inactivated form of the mentioned bacteria were produced and were compared regarding microbiological, biochemical, and physical properties during 28 days of storage at refrigerated temperature. Results revealed that the greatest mean pH drop rate, mean acidity increase rate, mean redox potential increase rate, final acidity and final redox potential were observed in yogurt containing inactivated L. acidophilus added before fermentation. The highest lactic acid after 28 days of storage was obtained in samples prepared by addition of paraprobiotic form of L. acidophilus after fermentation. Yogurt samples with B. lactis and L. acidophilus added after fermentation showed the highest and lowest acetic acid level, respectively after 28 days of storage. The samples containing L. acidophilus and B. lactis had the highest acetaldehyde on day 0 while on day 28, L. acidophilus had more impact on acetaldehyde generation in yogurts. Addition of paraprobiotics increased viability of starter cultures. In addition, incorporation of inactivated probiotic cells into yogurt resulted in lower syneresis and the higher WHC compared to probiotic yogurt samples. Regarding color parameters, it was observed that color parameters (a*, b* and L*) were not influenced by paraprobiotic in probiotic and paraprobiotic yogurts. Overall, it can be concluded that incorporation of paraprobiotics into yogurt involves less technological challenges and can be considered as a suitable appropriate alternative for probiotics in development of functional yogurt.

Development of an enzyme-enhancer system to improve laccase biological activities.

The present investigation reports an in-vitro study using combination of laccase and an enhancer capable of inhibiting the growth of pathogenic microorganisms, preventing biofilm formation, and whitening teeth. Laccase-cinnamic acid system remarkably inhibited the growth of Aggregatibacter actinomycetemcomitans, Candida albicans, S. aureus, and Streptococcus mutans whilst showed no significant effects on Gram-negative bacteria. Data presented that cinnamic acid (10 mM) with laccase (0.125 U ml-1) led to a maximum decrease of about 90%, in S. mutans biofilm formation. The confocal laser scanning microscopy showed considerable detachment of S. mutans cells from glass substratum. The combined laccase-cinnamic acid system could remove teeth discoloration caused by coffee. SEM of the teeth surface exhibited no damages such as surface cracking or fracture. Liquid chromatography-tandem mass spectrometry (LC-MS) and cyclic voltammetry (CV) studies showed that laccase can catalyze the one-electron oxidation of cinnamic acid to the respective radical. This radical can then undergo several fates, including recombination with another radical to form a dimeric species, dismutation of the radical back to cinnamic acid or decarboxylation to give various reduced oxygen species. Therefore, the redox potential values of phenolic monomers/oligomers are related with their biological activities.

Interferon ß-1b in treatment of severe COVID-19: A randomized clinical trial.

In this study, efficacy and safety of interferon (IFN) ß-1b in the treatment of patients with severe COVID-19 were evaluated. Among an open-label, randomized clinical trial, adult patients (≥18 years old) with severe COVID-19 were randomly assigned (1:1) to the IFN group or the control group. Patients in the IFN group received IFN ß-1b (250 mcg subcutaneously every other day for two consecutive weeks) along with the national protocol medications while in the control group, patients received only the national protocol medications (lopinavir/ritonavir or atazanavir/ritonavir plus hydroxychloroquine for 7-10 days). The primary outcome of the study was time to clinical improvement. Secondary outcomes were in-hospital complications and 28-daymortality. Between April 20 and May 20, 2020, 80 patients were enrolled and finally 33 patients in each group completed the study. Time to clinical improvment in the IFN group was significantly shorter than the control group ([9(6-10) vs. 11(9-15) days respectively, p = 0.002, HR = 2.30; 95% CI: 1.33-3.39]). At day 14, the percentage of discharged patients was 78.79% and 54.55% in the IFN and control groups respectively (OR = 3.09; 95% CI: 1.05-9.11, p = 0.03). ICU admission rate in the control group was significantly higher than the IFN group (66.66% vs. 42.42%, p = 0.04). The duration of hospitalization and ICU stay were not significantly different between the groups All-cause 28-day mortality was 6.06% and 18.18% in the IFN and control groups respectively (p = 0.12). IFN ß-1b was effective in shortening the time to clinical improvement without serious adverse events in patients with severe COVID-19. Furthermore, admission in ICU and need for invasive mechanical ventilation decreased following administration of IFN ß-1b. Although 28-day mortality was lower in the IFN group, further randomized clinical trials with large sample size are needed for exact estimation of survival benefit of IFN ß-1b.

Solubilization of Human Interferon ß-1b Inclusion Body Proteins by Organic Solvents.

Purposes: Solubilization of inclusion bodies expressed in E. coli is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high purity and yield. The aim of this study was to examine the benefit of organic solvents such as alcohols in solubilization of recombinant interferon ß-1b inclusion bodies. Methods: Effect of important parameters inclusion pH, concentration and type of denaturant and concentration of alcoholic solvents were optimized to formulate a suitable solubilization buffer and investigate their effect on solubilization of interferon ß-1b inclusion bodies. Results: Our findings showed the acidic pH in the range of 2-3 is more suitable than alkaline pH >12 for solubilization and achieving higher content of interferon ß-1beta and pure recombinant protein. We have also demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon ß-1b at pH of 2-3. The interferon concentration was 2.35 mg per 100 mg IB when we used 40% (v/v) 1-propanol and 20% (v/v) 2-butanol into the buffer solution as well. Conclusion: The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area.

Effects of synbiotic supplementation on gut microbiome, serum level of TNF-α, and expression of microRNA-126 and microRNA-146a in patients with type 2 diabetes mellitus: study protocol for a double-blind controlled randomized clinical trial.

BACKGROUND: The dramatic increase in the prevalence of type 2 diabetes mellitus (T2DM) is a global major challenge to health. Circulating microRNAs have been suggested as promising biomarkers for different disorders such as diabetes. Imbalances in the gut microbiome have been revealed to contribute to the progression of multiple diseases including T2DM. Recently, the consumption of probiotics and synbiotics in the treatment of various diseases has shown a substantial growth. The anti-diabetes and anti-inflammatory effects of synbiotics have been indicated, which may be due to their beneficial effects on the gut microbiome. However, further research is needed to assess the effects of synbiotics on the microbiota and their impacts on expression of microRNAs relating to T2DM. Thus, we will aim to assess the effects of synbiotics on microbiota, serum level of tumor necrosis factor-α (TNF-α), and expression of microRNA-126 and microRNA-146a in patients with T2DM. METHODS: Seventy-two patients with T2DM will be recruited in this double-blind randomized parallel placebo-controlled clinical trial. After block matching based on age and sex, participants will be randomly assigned to receive 1000 mg/day synbiotic (Familact) or placebo for 12 weeks. The microRNA-126 and microRNA-146a expression levels will be measured by real-time polymerase chain reaction and serum TNF-α level will be assessed by enzyme-linked immunosorbent assay kit at the beginning and at the end of the study. Determination of the gut microbiota will be done by quantitative polymerase chain reaction methods at baseline and at the end of the trial. Biochemical assessments (glycemic and lipid profiles) will also be conducted at onset and end of the study. DISCUSSION: This is the first randomized controlled trial that will determine the effect of synbiotic supplementation on the gut microbiota and its probable impacts on serum levels of TNF-α and expression of related microRNAs in patients with T2DM. TRIAL REGISTRATION: Iranian Registry of Clinical Trials: IRCT20180624040228N2. Registered on 27 March 2019. http://www.irct.ir/trial/38371.

Production of Vitamin D3 Enriched Biomass of Saccharomyces Cerevisiae as A Potential Food Supplement: Evaluation and Optimization of Culture Conditions Using Plackett-Burman and Response Surface Methodological Approaches.

Vitamin D deficiency causes osteoporosis, osteopenia, fractures, rickets, and more recently is linked with some chronic illnesses such as cancer. Because of the safety and probiotic properties of the yeast Saccharomyces cerevisiae, we hypothesized that yeast cells enriched with cholecalciferol (vitamin D3) could represent a solution for prevention or treatment of vitamin D deficiency. In this study S. cerevisiae was used as a vitamin D3 accumulator for the first time and the optimal conditions for enrichment of S. cerevisiae were determined. The Plackett-Burman screening studies were used for selection of the most important factors affecting cholecalciferol entrapment. Response surface methodology was employed for optimization of cholecalciferol accumulation in S. cerevisiae cells by using Box-Behnken design. A modified quadratic polynomial model fit the data appropriately. The optimal points of variables to maximize the response were cholecalciferol initial concentration of 358021.16 IU/mL, tryptone concentration of 1.82 g/L, sucrose concentration of 7.13 % (w/v), and shaking speed of 140.46 rpm. The maximum amount of cholecalciferol in dry cell weight of S. cerevisiae was 4428.11 IU/g. The cholecalciferol entrapment in yeast biomass increased about two-folds in optimized condition which indicates efficiency of optimization.

Down-regulatory effects of green coffee extract on las I and las R virulence-associated genes in Pseudomonas aeruginosa.

BACKGROUND: Antibiotic resistant strains of Pseudomonas aeruginosa are the cause of Gram negative nosocomial infections especially among the immunosuppressed patients. The bacteria contains las I and las R genes that play very important roles in the pathogenesis and mechanisms of aggression. These genes can be influenced by the quorum sensing (QS) system and such mechanism is becoming clinically important worldwide. This study aimed to investigate the preventive effects of green coffee extract (GCE) on the expression of pathogenesis-related genes, las I and las R in P. aeruginosa. METHODS: A total of fifty four P. aeruginosa strains were isolated out of 100 clinical samples collected from the infectious wards in different hospitals (Tehran province) using conventional microscopic and biochemical methods. Susceptibility of the isolates to different antibiotics, GCE and chlorogenic acid were elucidated. Multiplex polymerase chain reaction (PCR) and real-time PCR were performed to detect and quantify the expression levels of las I and las R genes. The presence of chlorogenic acid in GCE was confirmed by HPLC. RESULTS: Antibiotic susceptibility tests revealed multidrug resistance among the clinical isolates of those 40 strains were resistant to ciprofloxacin (74.07%), 43 to ceftazidime (79.26%), 29 to amikacin (53.7%), 42 to ampicillin (77.77%), 17 to colistin (31.48%), 40 to gentamicin (74.77%), and 50 to piperacillin (92.59%). PCR outcomes exhibited that the frequency of las I and las R genes were 100% in resistant and sensitive strains isolated from clinical and standard strains of P. aeruginosa (ATCC 15449). Real-time PCR analyses revealed that GCE significantly prevented the expression of las I and las R genes in P. aeruginosa. GCE at concentration level as low as 2.5 mg/mL could prevent the expression of lasI and lasR genes in P. aeruginosa clinical isolates. CONCLUSION: The presence and expression levels of las I and las R genes in P. aeruginosa isolates were investigated when the bacteria was exposed to GCE. Our results tend to suggest that genes involved in pathogenesis of:Pseudomonas aeruginosa are down regulated by quorum sensing effect of chlorogenic acid and therefore GCE could be useful as an adjuvant in combating multidrug resistance strains of Pseudomonas aeruginosa.

Determination of Biological Activity of Recombinant Reteplase Using Clot Lysis Time and Activated Partial Thromboplastin Time (APTT) Lysis Methods: A Comparative Study.

Recombinant plasminogen activator (reteplase) is a third generation thrombolytic agent which has been used on coronary artery thrombosis and acute myocardial infarction. Clot lysis assay is usually considered as a unique method to evaluate biological activity of reteplase. In this study biological activity of reteplase was determined by APTT (activated partial thromboplastin time) lysis method. Validity of this method was evaluated in comparison with reference method, clot lysis time assay. Results of APTT lysis test showed good reproducibility (relative standard deviation (RSD) 3-5% for within day analysis and 4-7% for between day analysis), and accuracy (101.3-102.7%). APTT lysis responses were linear in range of 0.001-0.1 mg/mL reteplase. Therefore, APTT lysis method is applicable for biological activity determination of reteplase. Although more comprehensive studies are required to approve this test as a reference method, APTT lysis method seems to be valuable to receive more attention due to advantages of technical simplicity, sensitivity, applicability, and cost efficiency.

Effects of a Multispecies Probiotic Supplement on Bone Health in Osteopenic Postmenopausal Women: A Randomized, Double-blind, Controlled Trial.

OBJECTIVE: The development of alternative approaches to prevent and/or treat osteoporosis, as a chronic progressive bone disease, is being considered currently. Among dietary supplements, probiotics may have favorable effects on bone metabolism. Therefore, the aim of this study was to evaluate the effects of a multispecies probiotic supplementation on bone biomarkers and bone density in osteopenic postmenopausal women. METHODS: This randomized double-blind placebo-controlled clinical trial was performed on 50 patients with osteopenia aged 50-72 years. Participants were randomly assigned to take either a multispecies probiotic supplement (GeriLact; n = 25) or placebo (n = 25) for 6 months. GeriLact contains 7 probiotic bacteria species. Participants received 500 mg Ca plus 200 IU vitamin D daily. Bone mineral density (BMD) of lumbar spine and total hip and blood biomarkers including bone-specific alkaline phosphatase (BALP), osteocalcin (OC), collagen type 1 cross-linked C-telopeptide (CTX), deoxypyridinoline (DPD), parathyroid hormone (PTH), 25-OH vitamin D, and serum pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-1ß) were assessed at baseline and at the end of the study. RESULTS: The multispecies probiotic significantly decreased BALP (p = 0.03) and CTX (p = 0.04) levels in comparison with the control group but had no effect on BMD of the spine and total hip. Moreover, there was a statistically significant decrease in serum PTH (p = 0.01) and TNF-α (p = 0.02) in the intervention group compared to the placebo group. CONCLUSIONS: These results may suggest the favorable effects of the multispecies probiotic supplementation for 6 months on bone health in postmenopausal women due to slowing down the rate of bone turnover.

Effect of Nonionic Surfactants (Dodecyl Maltoside and Polysorbate 20) on Prevention of Aggregation and Conformational Changes of Recombinant Human IFNß_1b Induced by Light.

Liquid protein formulations are prone to form aggregates. The effect of nonionic surfactants such as Polysorbate 20 (PS 20) and n-Dodecyl ß-D-maltoside (DDM) on the prevention of aggregation and conformational changes of recombinant human IFNß-1b (rhIFN ß_1b) was explored. Polysorbate has been used in formulations of protein pharmaceuticals. There have been concerns about using PS 20 due to its residual peroxide content which may negatively affect protein efficacy. n-Dodecyl ß-D-maltoside has been of interest and shown to be highly effective in prevention of aggregation. Fresh bulk of rhIFN ß_1b was formulated using DDM or different concentrations of PS 20. Formulations were exposed to light stress condition according to the ICH guideline of Q1b. The overall conformational integrity of individual samples was characterized by a combination of Circular dichroism (CD), Fluorescence spectroscopy and RP_HPLC techniques. The CD spectrum depicting the conformational integrity of rhIFN ß_1b showed 31.9% and 31.2% decreases in α-helix content of protein samples with 0.2% or 0.02% of PS20 compared to only18.2% of that containing 0.2% DDM. The RP-HPLC analysis also showed that the oxidized impurity in formulation containing DDM is less than those contain PS 20. Complementary analysis of the liquid formulations using IFR and UV methods also was in compliance with the data obtained by CD. Compared to PS 20, the sample of rhIFN ß_1b formulation with DDM was more resistant to the destruction effect of light. Results were in accordance with previous studies and could suggest DDM as a reliable anti-aggregation surfactant in biopharmaceutical formulations.

Optimization of Culture Conditions for Enrichment of Saccharomyces cerevisiae with Dl-α-Tocopherol by Response Surface Methodology.

Designing enriched probiotic supplements may have some advantages including protection of probiotic microorganism from oxidative destruction, improving enzyme activity of the gastrointestinal tract, and probably increasing half-life of micronutrient. In this study Saccharomyces cerevisiae enriched with dl-α-tocopherol was produced as an accumulator and transporter of a lipid soluble vitamin for the first time. By using one variable at the time screening studies, three independent variables were selected. Optimization of the level of dl-α-tocopherol entrapment in S. cerevisiae cells was performed by using Box-Behnken design via design expert software. A modified quadratic polynomial model appropriately fit the data. The convex shape of three-dimensional plots reveal that we could calculate the optimal point of the response in the range of parameters. The optimum points of independent parameters to maximize the response were dl-α-tocopherol initial concentration of 7625.82 µg/mL, sucrose concentration of 6.86 % w/v, and shaking speed of 137.70 rpm. Under these conditions, the maximum level of dl-α-tocopherol in dry cell weight of S. cerevisiae was 5.74 µg/g. The resemblance between the R-squared and adjusted R-squared and acceptable value of C.V% revealed acceptability and accuracy of the model.