scholarly journals Assessment of the Immunomodulatory Properties of the Probiotic Strain Lactobacillus paracasei K5 In Vitro and In Vivo

2020 ◽  
Vol 8 (5) ◽  
pp. 709 ◽  
Author(s):  
Pelagia Chondrou ◽  
Athanasios Karapetsas ◽  
Despoina Eugenia Kiousi ◽  
Stavros Vasileiadis ◽  
Petros Ypsilantis ◽  
...  

Lactobacillus paracasei K5 is a lactic acid bacteria (LAB) strain that has been isolated from dairy products. Previous studies have established its probiotic potential in a series of in vitro tests, including molecular characterization, safety profiling, and tolerability of the gastrointestinal tract conditions. To characterize its beneficial actions on the host, we have shown previously that L. paracasei K5 adheres to Caco-2 cells and exerts anti-proliferative effects through the induction of apoptosis. In the present study, we focused on the immunomodulatory potential of this strain. We employed the dorsal-air-pouch mouse model of inflammation and recorded an eight-fold increase in the recruitment of immune cells in mice treated with the probiotic strain, compared to the control group. Analysis of the exudates revealed significant changes in the expression of pro-inflammatory mediators on site. Treatment of Caco-2 cells with L. paracasei K5 induced significant upregulation of cytokines interleukin-1α (IL-1α), ΙL-1β, IL-6, tumor necrosis factor-alpha (TNF-α), the chemokine C-X-C motif ligand 2 (CXCL2), and the inflammation markers soluble intercellular adhesion molecule (sICAM) and metallopeptidase inhibitor-1 (TIMP-1). Transient induction of the Toll-like receptors (TLRs) 2, 4, 6, and 9 expression levels was recorded by real-time PCR analysis. These results highlight the immunomodulatory potential of this strain and further support its probiotic character.

2018 ◽  
Vol 6 (4) ◽  
pp. 121 ◽  
Author(s):  
Ioanna Mantzourani ◽  
Antonia Terpou ◽  
Athanasios Alexopoulos ◽  
Pelagia Chondrou ◽  
Alex Galanis ◽  
...  

In the present study 38 lactic acid bacteria strains were isolated from kefir grains and were monitored regarding probiotic properties in a series of established in vitro tests, including resistance to low pH, resistance to pepsin and pancreatin, and tolerance to bile salts, as well as susceptibility against common antibiotics. Among them, the strain SP3 displayed potential probiotic properties. Multiplex PCR analysis indicated that the novel strain belongs to the paracasei species. Likewise, the novel strain (Lactobacillus paracasei SP3) was applied as a starter culture for Feta-type cheese production. Feta-type cheese production resulted in significantly higher acidity; lower pH; reduced counts of coliforms, yeasts and fungi; and improved quality characteristics compared with cheese samples produced with no starter culture. Finally, it is highlighted that the application of the novel strain led to Feta-type cheese production with improved overall quality and sensory characteristics.


2021 ◽  
Vol 11 (13) ◽  
pp. 5765
Author(s):  
Joo-Yun Kim ◽  
Eun-Jung Choi ◽  
Jae-Ho Lee ◽  
Myeong-Seok Yoo ◽  
Keon Heo ◽  
...  

Vitamin B2, also known as riboflavin, is essential for maintaining human health. The purpose of this study was to isolate novel lactic acid bacteria that overproduce vitamin B2 and to validate their potential as probiotics. In this study, Lactobacillus plantarum HY7715 (HY7715) was selected among lactic acid bacteria isolated from Kimchi. HY7715 showed a very high riboflavin-producing ability compared to the control strain due to the high expression of ribA, ribB, ribC, ribH, and ribG genes. HY7715 produced 34.5 ± 2.41 mg/L of riboflavin for 24 h without consuming riboflavin in the medium under optimal growth conditions. It was able to produce riboflavin in an in vitro model of the intestinal environment. In addition, when riboflavin deficiency was induced in mice through nutritional restriction, higher levels of riboflavin were detected in plasma and urine in the HY7715 administration group than in the control group. HY7715 showed high survival rate in simulated gastrointestinal conditions and had antibiotic resistance below the cutoff MIC value suggested by the European Food Safety Authority; moreover, it did not cause hemolysis. In conclusion, HY7715 could be considered a beneficial probiotic strain for human and animal applications, suggesting that it could be a new alternative to address riboflavin deficiency.


2019 ◽  
Vol 18 (1) ◽  
pp. 50-54
Author(s):  
S. E. Gostischeva ◽  
N. V. Abzaeva ◽  
E. L. Rakitina ◽  
D. G. Ponomarenko ◽  
M. V. Kostuchenko ◽  
...  

Research objective–studying of a possibility of application antigen – stimulated cellular in vitro tests and technology of the cytometric analysis for control of immunogene activity of batches of vaccine plague live.Materials and methods.As biomodels used white laboratory mice, immunized commercial medicine of vaccine of the plague NIIEG line, live from a strain of Yersinia pestis EV, in doses – 8 х 102, 4 х 103, 2 х 104 and 1 х 105 of living microbic cells. Blood for a research was taken from intact mice and on 7, 14 and 21 days after immunization. The intensity of an antigenreaktivnost of lymphocytes was defined in cellular in vitro tests, analyzing a marker of early activation (CD45+CD3+CD25+) of lymphocytes with use of the monoclonal antibodies conjugated from fluorokhroma. As specific antigen used a complex of water-soluble antigens of a plague microbe.Results.As a result of a research it is shown that at the animals vaccinated by doses 4 х 103 – 1 х 105 living microbic cells, the highest level of an expression activation marker lymphocytes at anti-gene stimulation of in vitro is registered on 14 days after immunization, at the same time the quantity of CD25 – positive lymphocytes are on average 6.8 times higher, than in control group. High degree of direct link (coefficient of correlation of r = 1,000) quantities of the survived animals with increase in level of lymphocytes, expressiruyushchy markers of early activation – CD25 is established.Conclusions.The offered technique can be used as the additional test when studying degree of immunogenicity of new (kandidatny) vaccines against plague.


2020 ◽  
Vol 2020 ◽  
pp. 1-22
Author(s):  
Abulaihaiti Maitiseyiti ◽  
Hongbo Ci ◽  
Qingbo Fang ◽  
Sheng Guan ◽  
Alimujiang Shawuti ◽  
...  

Objective. Long noncoding RNAs (lncRNAs) have emerged as critical molecular regulators in various diseases. However, the potential regulatory role of lncRNAs in the pathogenesis of abdominal aortic aneurysm (AAA) remains elusive. The aim of this study was to identify crucial lncRNAs associated with human AAA by comparing the lncRNA and mRNA expression profiles of patients with AAA with those of control individuals. Materials and Methods. The expression profiles of lncRNAs and mRNAs were analyzed in five dilated aortic samples from AAA patients and three normal aortic samples from control individuals using microarray technology. Functional annotation of the screened lncRNAs based on the differentially expressed genes was performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Results. Microarray results revealed 2046 lncRNAs and 1363 mRNAs. Functional enrichment analysis showed that the mRNAs significantly associated with AAA were enriched in the NOD-like receptor (NLR) and nuclear factor kappa-B (NF-κB) signaling pathways and in cell adhesion molecules (CAMs), which are closely associated with pathophysiological changes in AAA. The lncRNAs identified using microarray analysis were further validated using quantitative real-time polymerase chain reaction (qRT-PCR) analysis with 12 versus 11 aortic samples. Finally, three key lncRNAs (ENST00000566954, ENST00000580897, and T181556) were confirmed using strict validation. A coding-noncoding coexpression (CNC) network and a competing endogenous RNA (ceRNA) network were constructed to determine the interaction among the lncRNAs, microRNAs, and mRNAs based on the confirmed lncRNAs. Conclusions. Our microarray profiling analysis and validation of significantly expressed lncRNAs between patients with AAA and control group individuals may provide new diagnostic biomarkers for AAA. The underlying regulatory mechanisms of the confirmed lncRNAs in AAA pathogenesis need to be determined using in vitro and in vivo experiments.


2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Weitao Ji ◽  
Hongyun Shi ◽  
Hailin Shen ◽  
Jing Kong ◽  
Jiayi Song ◽  
...  

Krüppel-like factor 4 (KLF4) is a key transcription factor that regulates genes involved in the proliferation or differentiation in different tissues. Apelin plays roles in cardiovascular functions, metabolic disease, and homeostatic disorder. However, the biological function of apelin in liver disease is still ongoing. In this study, we investigated the mechanism of KLF4-mediated protection against acute liver injury via the inhibition of the apelin signaling pathway. Mice were intraperitoneally injected with carbon tetrachloride (CCl4; 0.2 mL dissolved in 100 mL olive oil, 10 mL/kg) to establish an acute liver injury model. A KLF4 expression plasmid was injected through the tail vein 48 h before CCl4 treatment. In cultured LX-2 cells, pAd-KLF4 or siRNA KLF4 was overexpressed or knockdown, and the mRNA and protein levels of apelin were determined. The results showed that the apelin serum level in the CCl4-injected group was higher than that of control group, and the expression of apelin in the liver tissues was elevated while KLF4 expression was decreased in the CCl4-injected group compared to the KLF4-plasmid-injected group. HE staining revealed serious hepatocellular steatosis in the CCl4-injected mice, and KLF4 alleviated this steatosis in the mice injected with KLF4 plasmid. In vitro experiments showed that tumor necrosis factor-alpha (TNF-α) could downregulate the transcription and translation levels of apelin in LX-2 cells and also upregulate KLF4 mRNA and protein expression. RT-PCR and Western blotting showed that the overexpression of KLF4 markedly decreased basal apelin expression, but knockdown of KLF4 restored apelin expression in TNF-α-treated LX-2 cells. These in vivo and in vitro experiments suggest that KLF4 plays a key role in inhibiting hepatocellular steatosis in acute liver injury, and that its mechanism might be the inhibition of the apelin signaling pathway.


Author(s):  
Anastasia Drakou ◽  
Despoina Mavrogianni ◽  
Konstantinos Ntzeros ◽  
Athanasios Protopapas ◽  
Petros Drakakis ◽  
...  

Abstract Background Tumor necrosis factor-alpha (TNF-α) is a pro-inflammatory cytokine which plays an important role in the pathogenesis of many diseases. Endometriosis is one of the most common gynecological diseases. The purpose of this study was to investigate the association of TNF-α-1031T/C polymorphism with the genetic susceptibility of endometriosis in a European population. Materials and methods In this case-control study, 51 endometriosis patients and 67 healthy control women participated. We used endometrial tissue from the patients and peripheral blood from the healthy women to extract DNA. Polymerase chain reaction (PCR) analysis and the restriction enzyme Bbs I were used to analyze the -1031 T/C polymorphism in the TNF-α gene promoter region. Statistical analysis was performed using Fisher’s exact test. We also calculated the odds ratios. Results In the group of patients, 66.7% of women were detected with the TT genotype, 33.3% with the TC genotype and 0% with the CC genotype while in the control group, 46.3% had the TT genotype, 47.8% had the TC genotype and 6% had the CC genotype. There was a significant association between the TT genotype with endometriosis (p = 0.03). There was no significant deviation from the Hardy-Weinberg equilibrium. Conclusions The TC and CC genotypes appeared more often in the healthy women than the endometriosis patients and this shows that the C allele might have a protective role in endometriosis in the Greek population. Further studies are needed to specify the role of this polymorphism in pathogenesis of endometriosis and the mechanisms that protect the patients from the disease.


2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Hua Guan ◽  
Yan Lin ◽  
Liang Bai ◽  
Yingfeng An ◽  
Jianan Shang ◽  
...  

Cocoa powder is rich in flavonoids, which have many beneficial effects on human health, including antioxidative and anti-inflammatory effects. The aim of our study was to investigate whether the intake of cocoa powder has any influence on hyperlipidemia and atherosclerosis and examine the underlying molecular mechanisms. We fed apoE knockout mice a Western diet supplemented with either 0.2% (low group) or 2% (high group) cocoa powder for 12 weeks. The groups fed dietary cocoa powder showed a significant reduction in both plasma cholesterol levels and aortic atherosclerosis compared to the control group. Analysis of mRNA profiling of aortic atherosclerotic lesions revealed that the expression of several genes related to apoptosis, lipid metabolism, and inflammation was significantly reduced, while the antiapoptotic gene Bcl2 was significantly increased in the cocoa powder group compared to the control. RT-PCR analysis along with Western blotting revealed that a diet containing cocoa powder inhibited the expression of hepatic endoplasmic reticulum stress. These data suggest that cocoa powder intake improves hyperlipidemia and atherosclerosis, and such beneficial effects are possibly mediated through the suppression of hepatic endoplasmic reticulum stress.


2021 ◽  
Vol 9 (D) ◽  
pp. 1-7
Author(s):  
Filip Koneski ◽  
David Tipton ◽  
Jegdish Babu ◽  
Danica Popovic-Monevska ◽  
Vladimir Popovski ◽  
...  

BACKGROUND: Medication-related osteonecrosis of the jaw (MRONJ) is of considerable concern among clinicians and researchers, with no clear pathology mechanism, preventive, or treatment protocols. AIM: This study aimed to assess the effects of geranylgeraniol (GGOH) on the toxicity induced by clinical doses of zoledronic acid (ZOL) on gingival epithelial cells and gingival fibroblasts in vitro. METHODS: Human gingival fibroblasts and gingival epithelial cells were treated with 5, 25, or 50 μM ZOL ± 50 μM GGOH for 3 days. Viability of the cells was determined using the 3-[4, 5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay. Calculation of percentage of the control group, analysis of variance and Tukey post-hoc comparisons were performed to test the significance between groups, which was set at p = 0.05. Cell morphology was evaluated using light microscopy. RESULTS: ZOL significantly reduced the viability of both epithelial cells and fibroblasts at all concentrations (p < 0.05), with the exception of fibroblasts at concentration of 5 μM (p = 0.44). GGOH had positive effects on the viability of the cells treated with ZOL at all concentrations. However, statistically significant improvement was obtained only in epithelial cells at 5 and 25 μM ZOL. The cell morphology of both types of cells was improved after addition of GGOH. CONCLUSION: GGOH reverses the toxic effects of clinical doses of ZOL on gingival epithelial cells and has slightly positive, but not significant effects on gingival fibroblasts. This study suggests that GGOH may be effective in the prevention and treatment of MRONJ.


1997 ◽  
Vol 272 (3) ◽  
pp. G408-G416 ◽  
Author(s):  
A. R. Nagendra ◽  
J. K. Mickelson ◽  
C. W. Smith

We investigated the hypothesis that CD54 (intercellular adhesion molecule-1) expressed on hepatocytes will support beta2-integrin (CD18)-dependent adhesion of neutrophils. An in vitro model using C3A cells (a human hepatoblastoma cell line exhibiting many characteristics of normal hepatocytes) and human neutrophils was utilized. C3A cells were stimulated with interleukin-1beta (IL-1beta), tumor necrosis factor-alpha, or interferon-gamma (IFN-gamma) for 24 h to induce expression of CD54, and adhesion of neutrophils was found to be markedly increased. Detailed studies with IFN-gamma-stimulated (100 U/ml) C3A cells revealed that this adhesion involved CD11a/CD18 [lymphocyte function-associated antigen-1 (LFA-1)] and CD54 and was dependent on low levels of IL-8 produced by the stimulated hepatocytes. Addition of higher concentrations of chemotactic factor (e.g., IL-8) further augmented adhesion and recruited contributions of CD11b/CD18 (Mac-1). In contrast to LFA-1, Mac-1 appeared to recognize a CD54-independent ligand constitutively expressed on the hepatocytes. Such close apposition of neutrophils to hepatocytes may increase the potential for parenchymal cell injury by providing a short distance through which cytotoxic factors such as reactive oxygen or proteolytic enzymes could act.


Author(s):  
Debanga Das ◽  
Jashabir Chakraborty ◽  
Suvakanta Dash

Objective: Bioequivalence studies are the commonly accepted methods displaying therapeutic equivalence between two products. This study was conducted to evaluate the bioequivalence study of anti-diabetic activity between two formulations of metformin tablets which were marketed in India.Methods: In in vitro study five essential in vitro tests including disintegration, weight variation, hardness, friability and a comparative in vitro dissolution study were performed.Results: For in vivo study adult male rabbits were divided into three groups of two each. The first group is regarded as control group received 3 ml of normal saline daily by using the gastric tube for 15 d and the second and third group received (0.35 mg/Kg B.W. single dosage) of dexamethasone tablets which were powdered, dissolved in 3 ml of normal saline daily for 15 d. After 15 d the blood glucose of second and third group was estimated and after that received formulation X and formulation Y, dissolved in 3 ml of normal saline daily for 15 d at the dose of 0.5 gm/kg body weight orally. After 15 d again blood glucose of second and third group was estimated and compare the results of both the group. This shows the favourable response of metformin against glucocorticoid-induced renal damage and hyperglycemia.Conclusion: Results of this study showed that the extent, rate of absorption and anti-diabetic activity of two different formulations of metformin tablets are bioequivalent to each other.


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