scholarly journals Real-Time Analysis of Antiproliferative Effects of Mouthwashes Containing Alcohol, Sodium Fluoride, Cetylpyridinium Chloride, and Chlorhexidine In Vitro

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Mustafa Ülker ◽  
A. C. Tutku Çelik ◽  
Emine Yavuz ◽  
Firdevs Kahvecioğlu ◽  
H. Esra Ülker
Keyword(s):  
Real Time ◽  
Sodium Fluoride ◽  
Mtt Assay ◽  
Cetylpyridinium Chloride ◽  
The Other ◽  
Control Group ◽  
Label Free ◽  
L929 Cells ◽  
Antiproliferative Effects

Objectives. In this study, the cytotoxic responses of six different over-the-counter mouthwashes on L929 cells were analyzed by two different techniques: the traditional colorimetric tetrazolium-based reduction assay (MTT) and the modern impedance-based real-time cell analysis (RTCA) system to investigate their biocompatibility in vitro. Thus, the investigation of the antiproliferative effects of the specified materials via different techniques is vital to reach this goal. Materials and Methods. First, L929 mouse fibroblasts were exposed to the dilutions of mouthwashes for 2 minutes. After incubation, the tetrazolium reduction method was used to assess the metabolic viability of cells measured by colorimetric MTT assay and morphological inspection of cells was performed via phase-contrast microscopy. Furthermore, the effect of each mouthwash on the proliferation, morphology, and adhesion of L929 cells was monitored continuously by a noninvasive and label-free RTCA system for 140 h. Results. Our data showed that all of the mouthwashes had varying cytotoxic effects on fibroblasts compared to the control group in MTT assay. In addition to that, RTCA technology has provided the growth kinetic profiles that can be used to analyze if the treatment is causing antimitotic or DNA-damaging effect on cells. Thus, analysis via this system can tell us the mechanism of toxicity behind the cell growth inhibition in vitro. Here, we found that only mouthwash 1 moderately maintained the viability of the L929 cells, yet displaying antimitotic effects and the other mouthwashes (mouthwash 2-mouthwash 6) showed toxicity via DNA-damaging effects. Conclusions. Of the six types of mouthwash tested, the most biocompatible result was obtained from a mouthwash containing alcohol (i.e., mouthwash 1). On the other hand, sodium fluoride- (NaF-) and cetylpyridinium chloride- (CPC-) containing mouthwash (i.e., mouthwash 2) showed the most cytotoxic effect.

scholarly journals Quantifying single-cell secretion in real time using resonant hyperspectral imaging

2018 ◽  
Vol 115 (52) ◽  
pp. 13204-13209 ◽  
Author(s):  
José Juan-Colás ◽  
Ian S. Hitchcock ◽  
Mark Coles ◽  
Steven Johnson ◽  
Thomas F. Krauss
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Protein Secretion ◽  
Cell Communication ◽  
Label Free ◽  
Cell Secretion ◽  
Physiological Disorder ◽  
Real Time Analysis ◽  
Technology Heterogeneity

Cell communication is primarily regulated by secreted proteins, whose inhomogeneous secretion often indicates physiological disorder. Parallel monitoring of innate protein-secretion kinetics from individual cells is thus crucial to unravel systemic malfunctions. Here, we report a label-free, high-throughput method for parallel, in vitro, and real-time analysis of specific single-cell signaling using hyperspectral photonic crystal resonant technology. Heterogeneity in physiological thrombopoietin expression from individual HepG2 liver cells in response to platelet desialylation was quantified demonstrating how mapping real-time protein secretion can provide a simple, yet powerful approach for studying complex physiological systems regulating protein production at single-cell resolution.

scholarly journals Transcriptional, Mitochondrial Activity, and Viability of Egyptian Buffalo’s Granulosa Cells In Vitro Cultured under Heat Elevation

2021 ◽  
Vol 11 (2) ◽  
pp. 193-201
Author(s):  
Nasser Ghanem ◽  
Marwa Said Faheem ◽  
Romysa Samy ◽  
Ashraf Hesham Barkawi
Keyword(s):  
Heat Stress ◽  
Granulosa Cells ◽  
Transcriptional Activity ◽  
The Other ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Fluorescent Intensity ◽  
Stressed Group ◽  
Other Hand

It is documented that heat stress caused impairment on the reproductive performance of dairy animals. However, there are few reports that have focused on the molecular and intracellular responses of in vitro cultured buffalo granulosa cells during heat elevation. The present study was conducted to investigate the effect of heat elevation during in vitro culture of buffalo granulosa cells on their viability, quality, mitochondrial activity, and transcriptional activity. Granulosa cells were harvested after aspiration of cumulus-oocytes complexes that were collected from abattoir ovaries. The granulosa cells were cultured in vitro either at a normal physiological temperature suitable for oocyte maturation and embryo development (38.5°C) or exposed to the elevated temperature of 40.5°C on day 3 of culture (the first two days were for confluence) for two hours of culture then continued at 38.5°C up to day 7 of culture. The viability of granulosa cells was measured using trypan blue and quality was estimated by measuring the level of intracellular reactive oxygen species (ROS) on day 7. Moreover, metabolic activity was performed by measuring the fluorescent intensity of mitochondria. Moreover, transcriptional activity was done by profiling four selected candidate genes using quantitative real-time PCR. The results indicated that the granulosa cells viability rate significantly decreased in the heat stress group (25.1 ± 3.7), compared to the control group (36.6 ± 5.3) on confluence day (day 3). In addition, the viability rate on the last day of culture (day 7) decreased in heat stress, compared to control (83.7 ± 4.5 and 97.4 ± 0.4, respectively). On the other hand, there was a nonsignificant difference in ROS profile between the control (21.7*104 ± 1.3) and the heat-stressed group (15.7 ± 0.7) on day 7 of culture. However, the mitochondrial fluorescent intensity was higher in the control (21.9 ± 1.9) than in the heat-stressed group (15.4 ± 0.8) on day 7 of culture. The expression of cellular defense (HSF1) and apoptosis-inducing gene (P53) were significantly up-regulated in granulosa cells exposed to heat elevation, compared to the control group. On the other hand, the steroidogenesis-regulating gene (StAR) was down-regulated in granulosa cells cultured under heat shock, compared to the control group. In conclusion, heat stress reduced the viability of granulosa cells by inducing the expression of an apoptosis-related gene (P53) and compromised expression of genes regulating the steroid biosynthesis, which resulted in up-regulation of cell defense gene (HSF1) in an attempt to ameliorate the deleterious effect of heat stress on the biological activity of the granulosa cells.

scholarly journals Jianpi Yangwei decoction promotes apoptosis and suppresses proliferation of 5-fluorouracil resistant gastric cancer cells in vitro and in vivo

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Huijuan Tang ◽  
Wenjie Huang ◽  
Qiang Yang ◽  
Ying Lin ◽  
Yihui Chen ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Drug Resistance ◽  
Cell Line ◽  
Real Time ◽  
Mtt Assay ◽  
Xenograft Tumor ◽  
Rt Pcr ◽  
Induced Apoptosis

Abstract Background The exploration of new therapeutic agents targeting 5-Fu resistance may open a new opportunity to gastric cancer treatment. The objective is to establish a 5-Fu resistant gastric cancer cell line and observe the effect of Jianpi Yangwei decoction (JPYW) on its apoptosis and drug-resistance related proteins. Methods MTT assay was used to measure the effect of JPYW on the BGC823 cells proliferation, and the apoptosis was observed by flow cytometry and Hoechst fluorescence staining. The BGC823 xenograft tumor nude mice models were established, the apoptosis was detected by Tunel method. BGC-823/5-Fu was established by repeated low-dose 5-Fu shocks, the drug resistance index and proliferation were detected by the MTT assay; MDR1 mRNA was detected by real-time RT-PCR; Western blot was used to detect the ratio of p-AKT to AKT; The BGC823/5-Fu xenograft tumor nude mice models were established and apoptosis was measured. The expressions of MRP1, MDR1, ABCG2, AKT, p-AKT, caspase-3 and bcl-2 were detected by immunohistochemistry and the AKT mRNA expression was detected by real-time RT-PCR. Results JPYW induced apoptosis in BGC823 cells; Drug-resistant cell line BGC-823/5-Fu was sucessfully established; JPYW induced apoptosis of BGC823/5-Fu cells, down-regulated the expression of MRP1, MDR1 and ABCG2 in vitro and in vivo, and further decreased MDR1 expression when combined with pathway inhibitor LY294002 (P < 0.05); JPYW down-regulated the ratio of p-AKT to AKT in vitro in a dose-dependent manner, the same as after the combination with LY294002 (P < 0.05). Conclusion JPYW can induce apoptosis of BGC823 and BGC823/5-Fu cells, and down-regulate the expression of MDR1, MRP1, ABCG2 in vitro and in vivo. Its in vitro effect is related to the PI3K/AKT signaling pathway.

scholarly journals In VitroAdherence of Oral Bacteria to Different Types of Tongue Piercings

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Lucas Pereira Borges ◽  
Julio Cesar Campos Ferreira-Filho ◽  
Julia Medeiros Martins ◽  
Caroline Vieira Alves ◽  
Bianca Marques Santiago ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Bacterial Adhesion ◽  
Oral Bacteria ◽  
The Other ◽  
Control Group ◽  
Colony Forming Units ◽  
Different Types ◽  
Scanning Electron

The purpose of this work was to verifyin vitroadherence ofE. corrodensandS. oralisto the surface of tongue piercings made of surgical steel, titanium, Bioplast, and Teflon. For this, 160 piercings were used for the count of Colony Forming Units (CFU) and 32 piercings for analysis under scanning electron microscopy. Of these, 96 (24 of each type) were individually incubated in 5 mL of BHI broth and 50 μL of inoculum at 37°C/24 h. The other 96 piercings formed the control group and were individually incubated in 5 mL of BHI broth at 37°C/24 h. Plates were incubated at 37°C/48 h for counting of CFU/mL and data were submitted to statistical analysis (pvalue<0.05). ForE. corrodens, difference among types of material was observed (p<0.001) and titanium and surgical steel showed lower bacterial adherence. The adherence ofS. oralisdiffered among piercings, showing lower colonization (p<0.007) in titanium and surgical steel piercings. The four types of piercings were susceptible to colonization byE. corrodensandS. oralis, and bacterial adhesion was more significant in those made of Bioplast and Teflon. The piercings presented bacterial colonies on their surface, being higher in plastic piercings probably due to their uneven and rough surface.

404 DNA TAKE-UP BY SPERM AND IN VITRO EMBRYO DEVELOPMENT BY SPERM-MEDIATED GENE TRANSFER IN THE PIG

2007 ◽  
Vol 19 (1) ◽  
pp. 317
Author(s):  
T. S. Kim ◽  
Y. Cao ◽  
H. T. Cheong ◽  
B. K. Yang ◽  
C. K. Park
Keyword(s):  
Gene Transfer ◽  
Transfection Efficiency ◽  
The Other ◽  
Control Group ◽  
Dna Uptake ◽  
Alkaline Lysis ◽  
Exogenous Dna ◽  
Other Hand ◽  
Dna Complex

Sperm mediated gene transfer (SMGT) is based on the ability of spermatozoa to bind and internalize exogenous DNA and transfer it into the oocytes at fertilization. The purpose of this study was to assess introducing exogenous DNA into boar spermatozoa by DNA solution or DNA/liposome complex under different conditions (period of incubation, exogenous DNA, liposome, and concentration of spermatozoa). Genomic DNA of sperm loaded with DNA by treatment was isolated by alkaline lysis. Quantitation of exogenous DNA amplified by PCR was analyzed by agarose electrophoresis densitometry. The quality of treated spermatozoa under the best conditions or no treatment (control) was evaluated during incubation (0, 2, 4, and 6 h) for viability (SYBR-14/PI), motility (Makler counting chamber), morphology (rose bengal staining), and acrosomal status (Coomassie staining). Sperm loaded with DNA also were used for in vitro fertilization. Immature oocytes incubated in TCM-199 medium for 44 h were fertilized in mTBM medium for 6 h and cultured in PZM-3. Cleavage and development of embryos were assessed on Days 2 and 7 of culture, respectively. Transfection rates at the blastocyst stage were assessed by PCR analysis. Data were evaluated by Duncan&apos;s multiple-range test using the GLM procedure. In the preliminary experiment, DNA uptake of spermatozoa by DNA solution and liposome/DNA complex was completed within 90-120 min. Transfection efficiency of spermatozoa was significantly (P &lt; 0.05) higher in the 105 spermatozoa group than in the other groups (104, 106, and 107 spermatozoa). The transfection efficiency was gradually increased by increasing the concentration of exogenous DNA. On the other hand, viability of transfected spermatozoa by all treatments (control, DNA solution, and DNA/liposome) at 0 h (72.3 � 0.2, 70.8 � 1.8, and 68.0 � 2.2%, respectively) of storage was significantly (P &lt; 0.05) lower than for fresh spermatozoa (83.3 � 1.7%). Survival and motility of all treatments after 4 h of storage were significantly (P &lt; 0.05) lower than at 0 and 2 h. Both abnormality and acrosome reaction of spermatozoa were gradually increased with prolonged storage periods. On the other hand, the cleavage rate of embryos by DNA/liposome complex (56.3 � 2.3%) was significantly (P &lt; 0.05) lower compared to both DNA solution (64.0 � 1.1%) and control (67.8 � 2.3%). The developmental rates of blastocysts were significantly (P &lt; 0.05) lower in the liposome/DNA complex and DNA solution groups (9.1 � 1.3 and 11.3 � 0.8%) than in the control group (22.2 � 0.6%). The transfection rates of blastocysts were higher in the liposome/DNA group (54.3 � 12.0%) than in the DNA solution group (38.7 � 6.6%). These results show that the SMGT method under the control conditions efficiently transfers exogenous DNA into the porcine oocytes. This work was supported by the Research on the Production of Bio-organs (No. 2005 03020302) Ministry of Agriculture and Forestry, Republic of Korea

4 MYOSTATIN GENE KNOCKDOWN THROUGH LENTIVIRAL VECTOR-MEDIATED DELIVERY OF shRNA FOR IN VITRO PRODUCTION OF TRANSGENIC BOVINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 120 ◽  
Author(s):  
M. P. Milazzotto ◽  
W. B. Feitosa ◽  
B. E. Strauss ◽  
M. Bajgelman ◽  
C. M. Mendes ◽  
...  
Keyword(s):  
Real Time ◽  
Cell Morphology ◽  
Real Time Pcr ◽  
Lentiviral Vector ◽  
Gene Knockdown ◽  
Control Group ◽  
Bovine Embryos ◽  
Myostatin Gene ◽  
Transgenic Embryo

The main goal of husbandry and beef cattle production is to enhance performance rates, for example, weight gain. Myostatin is referred to as a negative regulator of skeletal muscle growth. Genetic engineering of this character in order to produce double muscling animals that can transmit to future progeny will enhance its usefulness. The present research aimed to analyze myostatin inhibition through lentiviral-mediated delivery of shRNA in mouse myoblast culture and the feasibility of the lentiviral-mediated delivery of shRNA into in vitro-produced transgenic bovine embryos. In order to achieve knockdown of myostatin in cell and embryo culture, a lentiviral vector was constructed with ubiquitin C promoter-driven GFP gene (green fluorescent protein) and shRNA to suppress myostatin gene expression driven by the U6 promoter. Vector efficiency was verified through in vitro murine myoblast (C2C12) cell morphology after inductive differentiation and by means of real-time PCR of myostatin and GAPDH genes. Later, bovine oocytes were in vitro-matured and the lentiviral vector was microinjected into the oocyte perivitelline space (2.5 � 106 IU mL-1) after mechanical and chemical cumulus cell removal. Non-microinjected mature oocytes were considered as control. After microinjection, oocytes were fertilized and cultured in vitro. After 4 and 9 days of culture, embryos were evaluated by epifluorescence microscopy. The GFP-positive embryos were green under fluorescence. Cell morphology and embryo development rate data were analyzed by Minitab Release 14 Statistical Software (Minitab, Inc., State College, PA, USA), submitted to ANOVA, and compared by Tukey test (P d 0.05). Real-time PCR data were analyzed by Pair-Wise Fixed Reallocation Randomization Test using REST2005 software. Cell morphology results demonstrated that the vector was able to inhibit myostatin mRNA in C2C12 cells as the transducted group progressed less to myotubes than in the control group. A lower amount of myostatin mRNA after 72 h of differentiation indicated an inhibition tendency by real-time PCR. In relation to the transgenic embryo production, 96.9 � 0.34% (62.65) developed to cleavage, 80.24 � 4.38% (51/65) were GFP-positive, and 50.95 � 3.37% (26/65) achieved blastocyst stage. After hatching, 3.07% (2/65) of GFP-positive embryos maintained fluorescence. In relation to the control group, the cleavage rate was 93.81 � 0.68% (61/65); the blastocyst rate 38.34 � 2.36% (25/65), and none were fluorescent. In conclusion, myostatin gene knockdown was effectively performed by lentiviral vector-mediated delivery of shRNA. Thus, novel studies about the efficiency of this vector on transgenic embryo production can be performed. This work was supported financially by FAPESP 03/0156-9.

scholarly journals A lectin-coupled porous silicon-based biosensor: label-free optical detection of bacteria in a real-time mode

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Mona Yaghoubi ◽  
Fereshteh Rahimi ◽  
Babak Negahdari ◽  
Ali Hossein Rezayan ◽  
Azizollah Shafiekhani
Keyword(s):  
Fourier Transform ◽  
Porous Silicon ◽  
Real Time ◽  
Limit Of Detection ◽  
Principal Component ◽  
The Other ◽  
Detection Methods ◽  
Carbohydrate Binding ◽  
Label Free ◽  
E Coli

Abstract Accuracy and speed of detection, along with technical and instrumental simplicity, are indispensable for the bacterial detection methods. Porous silicon (PSi) has unique optical and chemical properties which makes it a good candidate for biosensing applications. On the other hand, lectins have specific carbohydrate-binding properties and are inexpensive compared to popular antibodies. We propose a lectin-conjugated PSi-based biosensor for label-free and real-time detection of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) by reflectometric interference Fourier transform spectroscopy (RIFTS). We modified meso-PSiO2 (10–40 nm pore diameter) with three lectins of ConA (Concanavalin A), WGA (Wheat Germ Agglutinin), and UEA (Ulex europaeus agglutinin) with various carbohydrate specificities, as bioreceptor. The results showed that ConA and WGA have the highest binding affinity for E. coli and S. aureus respectively and hence can effectively detect them. This was confirmed by 6.8% and 7.8% decrease in peak amplitude of fast Fourier transform (FFT) spectra (at 105 cells mL−1 concentration). A limit of detection (LOD) of about 103 cells mL−1 and a linear response range of 103 to 105 cells mL−1 were observed for both ConA-E. coli and WGA-S. aureus interaction platforms that are comparable to the other reports in the literature. Dissimilar response patterns among lectins can be attributed to the different bacterial cell wall structures. Further assessments were carried out by applying the biosensor for the detection of Klebsiella aerogenes and Bacillus subtilis bacteria. The overall obtained results reinforced the conjecture that the WGA and ConA have a stronger interaction with Gram-positive and Gram-negative bacteria, respectively. Therefore, it seems that specific lectins can be suggested for bacterial Gram-typing or even serotyping. These observations were confirmed by the principal component analysis (PCA) model.

Noninvasive label-free nanoplasmonic optical imaging for real-time monitoring of in vitro amyloid fibrogenesis

Nanoscale ◽  
2014 ◽  
Vol 6 (7) ◽  
pp. 3561-3565 ◽  
Author(s):  
Sung Sik Lee ◽  
Luke P. Lee
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Optical Imaging ◽  
Real Time ◽  
Label Free ◽  
Real Time Monitoring ◽  
Pathological Mechanism

We utilize nanoplasmonic optical imaging as the noninvasive and label-free method in order to monitorin vitroamyloid fibrogenesis in real-time, which is considered as the primary pathological mechanism of Alzheimer's disease.

Cytotoxicity and Hemolytic Properties of Nano-Hydroxyapatite/Polyetheretherketone Biocomposites

2016 ◽  
Vol 848 ◽  
pp. 567-572 ◽  
Author(s):  
Lin Wang ◽  
Shen Hua Song ◽  
Zheng Zhi Wu ◽  
Li Hong Duan ◽  
Chun Bao Wang
Keyword(s):  
Negative Control Group ◽  
Negative Control ◽  
National Standard ◽  
Control Group ◽  
Culture Solution ◽  
Contact Method ◽  
Cytotoxicity Test ◽  
L929 Cells ◽  
Hemolysis Test

In order to evaluate the cytocompatibility and hemolytic properties of n-HA/PEEK biocomposites the nanohydroxyapatite/polyetheretherketone (n-HA/PEEK) biocomposites were successfully prepared. The mechanical properties of the biocomposites were proximal to human bone, at the same time, they had the optimal value with the HA volume content of 5%. The PEEK and n-HA/PEEK biocomposites with different HA content extraction medium was prepared with fresh medium. Simple DMEM culture solution was taken as negative control group. The pure PEEK and 5vol.%, 15vol.%, 30vol.% n-HA/PEEK biocomposites were the testing group. The relative proliferation rate of L929 cells was determined on the 1st, 2nd, 3rd and 6th days with CCK-8 assay. The cytotoxicity of n-HA/PEEK biocomposites were evaluated according to ISO 10993-5: 2009. The L929 cells morphology and growth on the 1st, 2nd, 3rd and 6th days were determined under inverted microscope. The hemolysis test in vitro of n-HA/PEEK biocomposites were evaluated through measuring erythrocyte lysis and ferro-hemoglobin freeing degree with indirect contact method basing on ISO 10993-4:2009. The experimental results showed that the growth and morphology of cells in pure PEEK and n-HA/PEEK biocomposites extraction medium had no difference from negative control group. Cytotoxicity test showed that PEEK and n-HA/PEEK biocomposites did not have obvious toxicity on L929 cells, and the cytotoxicity of these extracts was in grade 0-1. Hemolysis test suggested that PEEK and n-HA/PEEK biocomposites did not have obvious hemolysis reaction, and the hemolysis rate of PEEK and n-HA/PEEK biocomposites were 2.37%, 1.71%, 1.05% and 1.32% respectively, which are less than the national standard (5%). It may be concluded that the n-HA/PEEK biocomposites did not have obvious cytotoxicity and hemolysis reaction, which demonstrated that n-HA/PEEK biocomposites had good cytocompatibility.

scholarly journals Effects of a mixture of organisms,Lactobacillus acidophilusorStreptococcus faecalison cholesterol metabolism in rats fed on a fat- and cholesterol-enriched diet

1996 ◽  
Vol 76 (6) ◽  
pp. 857-867 ◽  
Author(s):  
Michihiro Fukushima ◽  
Masuo Nakano
Keyword(s):  
Rice Bran ◽  
Cholesterol Metabolism ◽  
Micelle Formation ◽  
The Other ◽  
Cholesterol Concentration ◽  
Control Group ◽  
Treatment Groups ◽  
Colony Forming Units ◽  
Organism Group

The effect of a mixture of organisms (a probiotic mixture) comprisingBacillus, Lactobacillus, Streptococcus, Clostridium, SaccharomycesandCandida(107–8colony-forming units/g rice bran of each component) on lipid metabolism was compared with that ofL. acidophilusand that ofS. faecalis. There were four treatment groups: rice bran (control), the mixture of organisms,L. acihphifusorS. faecds(30g/kg) were given to rats in a fat- and cholesterol-enriched diet for 4 weeks. The serum total cholesterol concentration of the group fed on the mixture of organisms was reduced by 15–33% compared with the other groups at the end of the 4week feeding period (P< 0·05). This group also had a lower hepatic cholesterol concentration (36–44%) than the two single-bacteria groups (P< 0·05). 3-Hydmxy-3-methylglutaryl-Co A reductase (NADPH; EC 1.1.1.34) activities of the mixed-organism andL. acidophifusgroups were significantly lower (61–63%) than those of the other groups (P< 0·05); the activity of the S. faecalis group was also signikantly lower (42%) than that of the control group (P< 0·05). The faecal cholesterol and bile acid concentrations of the mixed-organism group increased compared with those of theL. acidophilusandS. faecalisgroups (P< 0·05). The capacity of the mixed- organism cells to bind bile saltin vitrowas significantly higher (approximately 50%) than that of the singlebacteria cells (P< 0·05). On the other hand, cholesterol micelle formation for the mixed-organism cells was significantly (approximately 9%) lower than that of the singlebacteria cells (P< 0·05). These results indicate that the mixture of organisms decreased the synthesis of cholesterol in the liver and increased the loss of steroids from the intestine, in rats. Thus, the mixture of organisms had a hypocholeaterolaemic role

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