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.

Surface functionalization of endotracheal tubes coated with laccase-gadolinium phosphate hybrid nanoparticles for antibiofilm activity and contrasting properties.

Ventilator-associated pneumonia (VAP) is a severe hospital-acquired infection that endangers patients' treatment in intensive care units (ICUs). One of the leading causes of VAP is biofilm formation on the endotracheal tube (ETT) during ventilation. This study reports a combination of laccase-gadolinium phosphate hybrid nanoparticles (laccase@GdPO4·HNPs) and enzyme mediator with an antibiofilm property coated on the surface of the ETT. The hybrid nanostructures were fabricated through a simple, rapid, and facile laccase immobilization method, resulting in efficiency and yield percentages of 82 ± 6% and 83 ± 5%, respectively. The surface of the ETT was then functionalized and coated with the constructed HNP/catechol. The layered ETT was able to reduce the surface adhesion of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus by 82.1%, 84.5%, and 77.1%, respectively. The prepared ETT did not affect the viability of human lung epithelial cells L929 and A549 at concentrations of 1-5 mg mL-1. The layered ETT produced a strong computed tomography (CT) signal in comparison with iobitridol. The HNP/catechol-coated ETT exhibited a Gd3+ release of 0.45 ppm over 72 h, indicating reduced risks of cytotoxicity arising from the metal ions. In this research we develop a biofilm-resistant and contrasting agent-based ETT coated with green synthesized laccase@GdPO4·HNPs.

Cedrus deodara: In Vivo Investigation of Burn Wound Healing Properties.

Objective: Cedrus deodara (Roxb. Ex Lamb.) G. Don possesses various biological activities, which have been documented in modern and traditional medicine. In this study, burn wound healing activity of the methanol extract of C. deodara wood was evaluated via a burn wound model in Wistar rats. Methods: The methanol extract of C. deodara was evaluated for the contents of phenolic compounds, flavonoids, and tannins. Also, its antioxidant activity was determined using the DPPH assay. Then, a topical ointment containing the methanol extract of C. deodara (10%) was used to evaluate the healing effects on a model of second-degree thermal burn in 4 groups of 7 rats within 21 days. In this respect, average wound surface area, wound closure, and various histological features were examined. Results: Our findings revealed that the wounds treated with the methanol extract of C. deodara showed higher wound contraction (33.6, 87.1, and 93.4% on days 7, 14, and 21, respectively) compared with the positive control (27.6, 80.7, and 88.3% on days 7, 14, and 21, respectively) and the negative control (20.1, 77.9, and 80.2% on days 7, 14, and 21, respectively). According to the results from epitheliogenesis score, the number of inflammatory cells, neovascularization, and collagen density, good burn wound healing activity of the methanol extract of C. deodarawas demonstrated. Conclusion: Using the methanol extract of C. deodara in an ointment formulation can be developed to prevent or reduce burn injury progression.

Improved infectious burn wound healing by applying lyophilized particles containing probiotics and prebiotics.

Lactiplantibacillus plantarum cells were encapsulated in a mixture of cationic and anionic polymers, with the final composition stabilized through freeze-drying. A D-optimal design was used to examine the effects of different polymer concentrations as well as adding prebiotics on the probiotic viability and swelling behavior of the formulations. Scanning electron micrographs revealed stacked particles capable of rapidly absorbing significant amounts of water. These images corresponded to initial swelling percentages of around 2000% for the optimal formulation. The optimized formula had a viability percentage of more than 82%, with the stability studies suggesting that the powders should be stored at refrigerated temperatures. The physical characteristics of the optimized formula were examined to ensure compatibility with its application. According to antimicrobial evaluations, the difference in pathogen inhibition between formulated and fresh probiotics was less than a logarithm. The final formula was tested in vivo and showed improved wound healing indicators. The optimized formula resulted in a higher rate of wound closure and infection clearance. Furthermore, the molecular studies for oxidative stress indicated that the formula could modify wound inflammatory responses. In histological investigations, the probiotic-loaded particles functioned exactly as efficaciously as silver sulfadiazine ointment did.

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.

A survey on the stabilizing effect of osmolytes on the ultrasound-irradiated lipase for efficient enzymatic hydrolysis of coconut oil.

The stabilizing effect of some osmolytes including betaine, mannitol, proline, sorbitol, and trehalose (each 0.5 M) was investigated on the ultrasound-irradiated (60 kHz and 138 W, for 240 min) lipase by determination of the enzyme half-life time, evaluation of the enzymatic reaction velocity (Vmax), and hydrolysis of coconut oil for production of lauric acid (the main saturated fatty acid of the oil). The enzyme conformational stability was also assessed by circular dichroism (CD) and fluorescence spectroscopy. The average half-life time of mannitol- and sorbitol-treated lipase under the ultrasound irradiation was 511 ± 3 min and 531 ± 2 min, respectively; 3-fold higher than the unirradiated enzyme. The Vmax value of the ultrasound-treated lipase increased from 100 ± 3 nmol min-1 in the absence of osmolyte to 500 ± 7 nmol min-1 and 500 ± 9 nmol min-1 in the presence of mannitol and sorbitol, respectively. CD and fluorescence spectra indicated that mannitol and sorbitol enhanced the rigidity of the lipase molecular conformational structure, increasing the enzyme stability against the ultrasonic field. The ultrasound-irradiated lipase was then used to hydrolyze coconut oil in the absence or presence of the selected osmolytes, which led to liberate 310 ± 6 mg g-1, 413 ± 7 mg g-1, and 420 ± 4 mg g-1 of lauric acid in the absence or presence of sorbitol and mannitol, respectively. In the absence of an ultrasonic field, the non-osmotically-treated lipase was able to liberate only 211 ± 5 mg g-1 of lauric acid. These promising results indicate that sorbitol and mannitol stabilize the structural conformation of lipase under an ultrasonic field which in turn could improve the enzymatic hydrolysis of coconut oil.

Organic-inorganic hybrid nanoflowers: The known, the unknown, and the future.

Organic-inorganic hybrid nanoflowers (HNFs) are hierarchical flower-shaped microstructures that are assembled by nanoscale petal-like nanosheets composed of both organic and inorganic constituents. Generally, inorganic parts of HNFs are transition metal phosphates and organic components are mostly enzymes and proteins; however, non-protein molecules could be also used as organic phase in some types of newly described HNFs. Recent findings indicate that they are constructed through the coordination between organic and inorganic components. HNFs are mainly used for efficient biocatalysis and highly sensitive biosensing, while they have also some other noteworthy applications such as antimicrobial agents, antigen careers, and delivery platforms for anticancer drugs. It is believed that the high surface-to-volume ratio of HNFs could tackle mass transfer limitations leading to enhance the activity of their organic constituents. The environment-friendly route of synthesis and stabilization of biomolecules upon storage are the advantages of enzyme-based HNFs. In the present review, the focuses are on designs, preparations, formation mechanisms, and remarkable applications of the conventional forms and also magnetic, multi-component, and enzyme-free HNFs. Considering the fact that HNFs are in the early stages of development, the unknown aspects and future directions of research in this field are also discussed.

The antimicrobial and antibiofilm effects of three herbal extracts on Streptococcus mutans compared with Chlorhexidine 0.2% (in vitro study).

There is a special focus on using natural materials and herbal plants to prevent dental caries. Previous studies showed that some herbal plants have antimicrobial effects on oral pathogens. Thus we investigated the antimicrobial effects of three herbal extracts (Carum copticum, Phlomis bruguieri, and Marrubium parviflorum) on the growth of Streptococcus mutans, as the most important bacteria causing dental caries. First, plant methanolic extracts were prepared. Then, to evaluate the antimicrobial activity of the three herbal extracts, the agar well diffusion method and MIC were performed. The biofilm formation was carried out using a broth dilution method with 2% glucose-supplemented BHIS in sterile 96-well microplates. Serial dilutions (50, 25, 12.5, 6.25, 3.12 mg/ml) of extracts were prepared. Next, a 0.5 McFarland Suspension of S. mutans was added to wells. The inhibitory effect on biofilm formation was measured by the ELISA reader apparatus. The assay was repeated three times, and the average was calculated as 3. The results were compared with those of Chlorhexidine 0.2%. Carum copticum showed a better effect in the agar well diffusion method than others. MIC of the extracts of Carum coptimum, Phlomis bruguieri, and Marrubium parviflorum were 3.12, 6.25, and 12.5 mg/ml, respectively. Overall, the highest activity belonged to Carum copticum extract. For the anti-biofilm effect, the OD values of Carum copticum and Marrubium parviflorum were significantly different from that of Phlomis bruguieri. Although all of the methanolic herbal extracts can inhibit S. mutans growth and remove the biofilm, the effect of Carum copticum was better than Phlomis bruguieri and Marrubium parviflorum. Further studies are recommended to indicate how these extracts perform against the bacteria.

In Vitro-In Vivo Correlation for the Antibacterial Effect of Lactiplantibacillus plantarum as a Topical Healer for Infected Burn Wound.

Difficulties in delivering antimicrobial agents to wound areas and emersion of multiple drug resistant organisms (MDROs) have converted managing burn infections into a complicated task in medicine. Probiotics emerged not only as a probable solution for burn infections but also as an accelerator in the healing process. The probability of in vitro-in vivo correlation (IVIVC) in probiotic activity leads to lower costs in finding new therapeutic options. Simulated wound fluid (SWF) was used to evaluate the antibacterial function of Lactiplantibacillus plantarum in wounds. The growth parameters in SWF were evaluated using a logistic model to predict growth behavior in the wound area. In addition, probiotic antimicrobial activity and secretion of antibacterial substances in SWF were also studied. Data were used to select the initial dose and apply frequency for in vivo study. The wound models were infected by two main pathogens (Pseudomonas aeruginosa or Staphylococcus aureus). In vitro results showed less lag time associated with considerable acid production in SWF. In the following, secretion of antimicrobial substances and co-aggregation with pathogens became more important. The susceptibility of pathogens to these factors was different, and culture medium affected the yield of each factor involved in eliminating pathogens. Histological analysis and macroscopic examination of wounds revealed probiotics as effective as positive control or more. There were some differences in the antibacterial functions of probiotics in simulated and real wound environments. The in vitro effect of probiotics on removal of pathogens was not the same as the trend seen in vivo.

High efficiency of osmotically stable laccase for biotransformation and micro-detoxification of levofloxacin in the urea-containing solution: Catalytic performance and mechanism.

Laccase-catalyzed oxidation was applied in the biotransformation of levofloxacin (a potentially environmental antibiotic contamination); however, the enzyme may denature in urea-containing wastewater and lead to the formation of an inactive form followed by decreasing the yield of the bio-removal. In this study, the osmolytes-stabilized laccase was used to eliminate levofloxacin in the urea-containing solution. Sorbitol and proline 100 mM appeared to be the two most efficient laccase protectants against the urea-induced denaturation. In a 1-M urea solution, the maximum velocity (Vmax) of laccase was estimated to be 39.1 µmol min-1 mg-1. This value was improved to 101.7 and 51.8 µmol min-1 mg-1 in the presence of sorbitol and proline, respectively. In optimal conditions for the elimination of levofloxacin, sorbitol- and proline-treated laccase led to 82.9 % and 76.2 % bio-removal of the applied fluoroquinolone in 1 M urea solution, respectively. Biotransformation products of the parent antibiotic were spectroscopically analyzed that assigned to different reaction pathways including demethylation, defluorination, decarboxylation, deamination, and hydroxylation. A micro-toxicity study concerning the growth of some Gram+ and Gram- bacteria exhibited decreasing in inhibition of laccase-treated levofloxacin after a 10-h incubation at 37 °C.

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.

The health-related quality of life in Iranian patients with COVID-19.

BACKGROUND: COVID-19 is a public health emergency with a high mortality rate and it reduces the patient's Health-Related Quality of Life (HRQoL) significantly. This effect is measured in the current study. METHODS: In a cross-sectional study in Iran, 320 randomly selected treated patients from COVID-19 were studied. To collect the required data, we applied a questionnaire that included socio-demographic factors, clinical characteristics, and questions on the patients' HRQoL. Time trade-off (TTO) approach was used to measure the lost HRQoL attributed to COVID-19. Besides, we applied a two-limit Tobit regression model to determine the effects of the socio-demographic factors on patients' health utility and the visual analogue scale approach was used to estimate the perceived total current health status. RESULTS: The overall mean (SE) and median (IQR) of the health utility values were 0.863 (0.01) and 0.909 (0.21) respectively. These values for the traders (those who were willing to lose a part of their remaining time of life to avoid the disease) were estimated at 0.793 (0.01) and 0.848 (0.17), respectively. The lowest amount of utility value belonged to the elderly (mean (SE) = 0.742 (0.04); median (IQR) = 0.765 (0.42)) and those living in rural areas (mean (SE)) = 0.804 (0.03); median (IQR) = 0.877 (0.30)). The univariate analysis showed that age, place of residence, and household size had a statistically significant effect on health utility. Moreover, findings of the regression analysis indicated that the participants' age and hospitalization status were the key determinants of COVID-19 health utility value. CONCLUSION: COVID-19 is associated with a substantial and measurable decrease in HRQoL. This decline in HRQoL can be directly compared with that induced by systemic health states.

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.

Evaluation of Anti-oxidant and Anti-biofilm Activities of Biogenic Surfactants Derived from Bacillus amyloliquefaciens and Pseudomonas aeruginosa.

Biosurfactants, the microbial originated surface active agents, can modify the physicochemical properties of surfaces and reduce the bacterial adhesion via changing bacterial adhesion interactions on surfaces. They were also able to block oxidative chain reactions and might show antioxidant properties. The goal of this study was to evaluate the antioxidant and antibiofilm activities of biosurfactants which were derived from two autochthonous biosurfactant-producing strains, Bacillus amyloliquefaciens NS6 (surfactin), and Pseudomonas aeruginosa MN1 (rhamnolipids). Their antioxidant activities were determined by ferric reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) methods. Ferric thiocyanate (FTC) assay was used for determination of their lipid peroxidation inhibition capacity. Their effect to reduce the adhesion of Streptococcus mutans on polystyrene surfaces and disruption of its pre-formed biofilms were also investigated. Our results indicated that surfactin showed higher antioxidant activity than rhamnolipids and showed relatively similar efficiency to BHA that suggests it as a good alternative for synthetic antioxidants. In other hand, rhamnolipid conditioned surfaces showed higher antiadhesive and antibiofilm activity in comparison with surfactin treated surfaces.

Experiences of nurses within a nurse-led multidisciplinary approach in providing care for patients with diabetic foot ulcer.

BACKGROUND AND AIMS: Diabetic foot ulcers require special attention due to their complex and chronic nature and special care and the Nurse-led approach has emerged in the current situation in response to the increasing need for health care as a way to treat chronic patients and provide them with ongoing care. Providing this specialized duty in nursing can be promoted and enhanced the professional independence of nurses and presents challenges. This study has been conducted to make an understanding of the nurses' experiences about their participation in a nurse-led multidisciplinary team to providing care for patients with Diabetic Foot Ulcer. METHODS: This was a qualitative, phenomenological research, carried out with the participation of 8 nurses of Shariaty Hospital of Tehran. Data collection was done through in-depth and semi structured interview. Dieklemann method was used for data analysis. RESULTS: 2 themes and 4 categories emerged during data analysis: antecedents (background and nursing skills) and manifestation of multidisciplinary function (from despair to tranquility, professional worthiness, promotion of self-confidence, preference of multidisciplinary approach). CONCLUSION: The findings of this study, which were the lived experiences of the participants, indicated that most of them had experienced positive effect of nurse-led multidisciplinary team care, training and development, appropriate skill mix, quality and outcome of care. Therefore, despite working on a multidisciplinary team is complex and demanding, management attributions and communication strategies is well situated to meet the challenge.

Comparative evaluation of hydrogen peroxide sporicidal efficacy by different standard test methods.

BACKGROUND AND OBJECTIVES: There are different sporicidal standard tests with various specifications to deal with products that are claimed for sporicidal activity. The aim of this study was to compare the 7% H2O2 sporicidal efficacy against Bacillus subtilis spores using different standard test methods. MATERIALS AND METHODS: The 7% H2O2 sporicidal efficacy against Bacillus subtilis spores was determined according to the AOAC MB-15-04 standard of carrier test and two standard suspension tests (BS EN 13704, AFNOR NF 72-230) in both clean and dirty conditions and by using different interfering substances including bovine serum albumin, yeast extract and skimmed milk. RESULTS: The results of suspension tests with 3 × 105 and 2 × 107 CFU/ml of B. subtilis spore concentration demonstrated that the higher spore counts lead to lower efficacy of 7% H2O2. Also, the sporicidal activity of 7% H2O2 was reduced in the presence of interfering substances. Bovine serum albumin, yeast, and skimmed milk showed similar interfering effects in suspension test with 3 × 105 CFU/ml. While, in suspension tests with higher initial spore count (2 × 107 CFU/ml) severity of interfering effects were intensified and distinct. Our results indicated that the carrier sporicidal test in comparison with suspension tests required more contact time to kill B. subtilis spores. CONCLUSION: The results of this study showed that it is reasonable to use interfering substances and inoculated carriers in accordance with actual conditions of product usage in a sporicidal test. Interfering substances may reduce the contact surface between H2O2 and test spores; therefore, the sporicidal efficacy of H2O2 was diminished. So applying suspension test in clean condition to verify the claim of sporicidal activity is strongly discouraged.

Effect of nurse-led care on quality of care and level of HbA1C in patients with diabetic foot ulcer: A randomized clinical trial.

Diabetic foot ulcer (DFU) is a serious and costly complication in diabetes which affects different aspects of life and can reduce patient's quality of life. Various views to manage DFU have been introduced. The nurse-led team, as a multidisciplinary team, can be effective due to a holistic approach to some disease management; but in patients with DFU it has not been assessed. Therefore, this study was designed to determine the effect of nurse-led care on quality of care (QOC) and improvement of HbA1C in Patients with DFU. This was a randomized clinical trial study performed on 52 patients with DFU. Patients were randomly assigned into two groups: nurse-led care intervention and standard care. The study was conducted in one of the hospitals affiliated to Tehran University of Medical Sciences. Data were collected using two questionnaires: Demographic characteristics and Quality Patient Care Scale (QUALPACS) and taking a blood sample for HbA1C. The nurse-led Care interventions were conducted in three stages: Integrated, Interdisciplinary, and Comprehensive. Descriptive and analytic statistical methods were used to analyze the data. P < .05 was considered significant. The results demonstrated that according to repeated measures test, (before, 4 and 12 weeks after the intervention), the level of QOC dimensions (Psychosocial, communication and physical aspect) in the nurse-led group had a significant difference with control group (P < .0001). Also, according to the results of Independent t test, there was a significant difference in total QOC scoring and HbA1c between intervention and control groups after the intervention and follow up (P < .0001). Considering the burden of diabetes and DFUs, it seems that the establishment of a nurse-led care approach can be an effective strategy to manage and treat these patients, and eliminate the disruption of care and achieve optimal care quality.

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.