Lactobacillus casei CRL 431 improves endothelial and platelet functionality in a pneumococcal infection model

2019 ◽  
Vol 10 (5) ◽  
pp. 533-541
Author(s):  
C. Haro ◽  
M. Medina
Keyword(s):  
Cell Death ◽  
Lactobacillus Casei ◽  
Pneumococcal Infection ◽  
Probiotic Strain ◽  
Endothelial Activation ◽  
Control Group ◽  
Infection Model ◽  
Soluble Adhesion Molecules ◽  
Induce Platelet Aggregation ◽  
Platelet Counts

Streptococcus pneumoniae is able to activate coagulation and induce platelet aggregation, both of which are typical responses to systemic inflammation. The interactions between inflammation and coagulation and between soluble adhesion molecules and endothelial cells are important in the pathogenesis of an unbalanced haemostatic system. Therefore, an exaggerated and/or insufficiently controlled haemostatic activity may appreciably contribute to the severity of the disease. The aim of the present study was to evaluate the effect of the oral administration of Lactobacillus casei CRL 431 on platelet and endothelial activation mechanisms in a respiratory pneumococcal infection model in mice. S. pneumoniae induced an increase in platelet counts and enhanced the expression of P-selectin in control group, with higher endothelial activation in lung shown by the increase in von Willebrand factor (vWF) and vascular cell adhesion molecule 1 (VCAM-1) expression. Also, infection induced a decrease in CXCR-4 leukocytes, increased expression in annexinV and cell death at the pulmonary level and decreased antithrombin levels in bronchoalveolar lavage. In contrast, L. casei mice restored platelet counts, favoured faster P-selectin expression, lower vWF levels and VCAM-1 expression than control group. Also, L. casei induced higher levels of annexinV expression and lower cell death in the lung. Moreover, it was able to modulate antithrombin levels within the normal range, which would indicate lower coagulation activation and a protective effect locally exerted by L. casei. In this work, the ability of L. casei to favourably modulate platelet and endothelial functionality during a pulmonary infection with S. pneumoniae was demonstrated. Our findings offer a promising perspective for the use of this probiotic strain in the prevention of thrombotic complications associated with pneumococcal pneumonia, especially in at-risk patients. In addition, the use of L. casei would provide novel alternatives for the prevention and treatment of thrombosis associated with various diseases.

scholarly journals Can phenotypic data complement our understanding of antimycobacterial effects for drug combinations?

2019 ◽  
Vol 74 (12) ◽  
pp. 3530-3536 ◽  
Author(s):  
Frank Kloprogge ◽  
Robert Hammond ◽  
Andrew Copas ◽  
Stephen H Gillespie ◽  
Oscar Della Pasqua
Keyword(s):  
Cell Death ◽  
Cell Viability ◽  
Cell Injury ◽  
Multinomial Logistic Regression ◽  
Onset Time ◽  
Control Group ◽  
Infection Model ◽  
Pharmacodynamic Interaction ◽  
Phenotypic Data ◽  
Bacterial Killing

Abstract Objectives To demonstrate how phenotypic cell viability data can provide insight into antimycobacterial effects for the isoniazid/rifampicin treatment backbone. Methods Data from a Mycobacterium komossense hollow-fibre infection model comprising a growth control group, rifampicin at three different exposures (Cmax = 0.14, 0.4 and 1.47 mg/L with t½ = 1.57 h and τ = 8 h) and rifampicin plus isoniazid (Cmax rifampicin = 0.4 mg/L and Cmax isoniazid = 1.2 mg/L with t½ = 1.57 h and τ = 8 h) were used for this investigation. A non-linear mixed-effects modelling approach was used to fit conventional cfu data, quantified using solid-agar plating. Phenotypic proportions of respiring (alive), respiring but with damaged cell membrane (injured) and ‘not respiring’ (dead) cells data were quantified using flow cytometry and Sytox Green™ (Sigma–Aldrich, UK) and resazurin sodium salt staining and fitted using a multinomial logistic regression model. Results Isoniazid/rifampicin combination therapy displayed a decreasing overall antimicrobial effect with time (θTime1/2 = 438 h) on cfu data, in contrast to rifampicin monotherapy where this trend was absent. In the presence of isoniazid a phenotype associated with cell injury was displayed, whereas with rifampicin monotherapy a pattern of phenotypic cell death was observed. Bacterial killing onset time on cfu data correlated negatively (θTime50 = 28.9 h, θLAGRIF50 = 0.132 mg/L) with rifampicin concentration up to 0.165 mg/L and this coincided with a positive relationship between rifampicin concentration and the probability of phenotypic cell death. Conclusions Cell viability data provide structured information on the pharmacodynamic interaction between isoniazid and rifampicin that complements the understanding of the antibacillary effects of this mycobacterial treatment backbone.

scholarly journals MicroRNA-513b-5p targets COL1A1 and COL1A2 associated with the formation and rupture of intracranial aneurysm

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Zheng Zheng ◽  
Yan Chen ◽  
Yinzhou Wang ◽  
Yongkun Li ◽  
Qiong Cheng
Keyword(s):  
Cell Death ◽  
Intracranial Aneurysm ◽  
Luciferase Reporter Assay ◽  
Luciferase Reporter ◽  
Control Group ◽  
Type I ◽  
Mrna Levels ◽  
Reporter Assay ◽  
Over Expression ◽  
Tnf Α

AbstractCollagen-type I alpha 1 chain (COL1A1) and COL1A2 are abnormally expressed in intracranial aneurysm (IA), but their mechanism of action remains unclear. This study was performed to investigate the mechanism of COL1A1 and COL1A2 affecting the occurrence and rupture of IA. Quantitative real-time polymerase chain reaction was used to measure the expression of hsa-miR-513b-5p, COL1A1, COL1A2, TNF-α, IL-6, MMP2, MMP3, MMP9 and TIMP4 in patients with ruptured IA (RA) (n = 100), patients with un-ruptured IA (UA) (n = 100), and controls (n = 100). Then, human vascular smooth muscle cells (HASMCs) were cultured, and dual luciferase reporter assay was performed to analyse the targeting relationship between miR-513b-5p and COL1A1 or COL1A2. The effects of the miR-513b-5p mimic and inhibitor on the proliferation, apoptosis, and death of HASMC and the RIP1-RIP3-MLKL and matrix metalloproteinase pathways were also explored. The effect of silencing and over-expression of COL1A1 and COL1A2 on the role of miR-513b-5p were also evaluated. Finally, the effects of TNF-α on miR-513b-5p targeting COL1A1 and COL1A2 were tested. Compared with those in the control group, the serum mRNA levels of miR-513b-5p, IL-6 and TIMP4 were significantly decreased in the RA and UA groups, but COL1A1, COL1A2, TNF-α, IL-1β, MMP2, MMP3 and MMP9 were significantly increased (p < 0.05). Compared with those in the UA group, the expression of COL1A1, COL1A2, TNF-α, IL-1β and MMP9 was significantly up-regulated in the RA group (p < 0.05). Results from the luciferase reporter assay showed that COL1A1 and COL1A were the direct targets of miR-513b-5p. Further studies demonstrated that miR-513b-5p targeted COL1A1/2 to regulate the RIP1-RIP3-MLKL and MMP pathways, thereby enhancing cell death and apoptosis. Over-expression of COL1A1 or COL1A2, rather than silencing COL1A1/2, could improve the inhibitory effect of miR-513b-5p on cell activity by regulating the RIP1-RIP3-MLKL and MMP pathways. Furthermore, over-expression of miR-513b-5p and/or silencing COL1A1/2 inhibited the TNF-α-induced cell proliferation and enhanced the TNF-α-induced cell death and apoptosis. The mechanism may be related to the inhibition of collagen I and TIMP4 expression and promotion of the expression of RIP1, p-RIP1, p-RIP3, p-MLKL, MMP2 and MMP9. MiR-513b-5p targeted the inhibition of COL1A1/2 expression and affected HASMC viability and extracellular mechanism remodelling by regulating the RIP1-RIP3-MLKL and MMP pathways. This process might be involved in the formation and rupture of IA.

Developing probiotic pumpkin juice by fermentation with commercial probiotic strain Lactobacillus casei 431

2021 ◽  
Author(s):  
Darko Dimitrovski ◽  
Maja Dimitrovska‐Vetadjoka ◽  
Hristo Hristov ◽  
Donka Doneva‐Shapceska
Keyword(s):  
Lactobacillus Casei ◽  
Probiotic Strain

scholarly journals Hippocampal distribution of IL-1β and IL-1RI following lithium-pilocarpine-induced status epilepticus in the developing rat

2016 ◽  
Vol 88 (suppl 1) ◽  
pp. 653-663 ◽  
Author(s):  
Dulce-Mariely Álvarez-Croda ◽  
Juan Santiago-García ◽  
Jesús S. Medel-Matus ◽  
Joel Martínez-Quiroz ◽  
Angel A. Puig-Lagunes ◽  
...  
Keyword(s):  
Cell Death ◽  
Status Epilepticus ◽  
Mrna Expression ◽  
Dorsal Hippocampus ◽  
Neuronal Cell Death ◽  
Neuronal Cell ◽  
Control Group ◽  
Rat Pups ◽  
Fluoro Jade B ◽  
Developing Rat

The contribution of Interleukin-1β (IL-1β) to neuronal injury induced by status epilepticus (SE) in the immature brain remains unclear. The goal of this study was to determine the hippocampal expression of IL-1β and its type 1 receptor (IL-1RI) following SE induced by the lithium-pilocarpine model in fourteen-days-old rat pups; control animals were given an equal volume of saline instead of the convulsant. IL-1β and IL-1RI mRNA hippocampal levels were assessed by qRT-PCR 6 and 24 h after SE or control conditions. IL-1β and IL-1RI expression was detected in the dorsal hippocampus by immunohistochemical procedures; Fluoro-Jade B staining was carried out in parallel sections in order to detect neuronal cell death. IL-1β mRNA expression was increased 6 h following SE, but not at 24 h; however IL-1RI mRNA expression was unaffected when comparing with the control group. IL-1β and IL-1RI immunoreactivity was not detected in control animals. IL-1β and IL-1RI were expressed in the CA1 pyramidal layer, the dentate gyrus granular layer and the hilus 6 h after SE, whereas injured cells were detected 24 h following seizures. Early expression of IL-1β and IL-1RI in the hippocampus could be associated with SE-induced neuronal cell death mechanisms in the developing rat.

scholarly journals Probiotic Potential of a Novel Vitamin B2-Overproducing Lactobacillus plantarum Strain, HY7715, Isolated from Kimchi

2021 ◽  
Vol 11 (13) ◽  
pp. 5765
Author(s):  
Joo-Yun Kim ◽  
Eun-Jung Choi ◽  
Jae-Ho Lee ◽  
Myeong-Seok Yoo ◽  
Keon Heo ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Probiotic Strain ◽  
Optimal Growth ◽  
Control Group ◽  
High Survival ◽  
Vitamin B2 ◽  
Riboflavin Deficiency

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.

Combined administration of bacteriocin- -producing, probiotic strain Enterococcus faecium CCM7420 with Eleutherococcus senticosus and their effect in rabbits

2013 ◽  
Vol 16 (4) ◽  
pp. 619-627 ◽  
Author(s):  
M. Pogány Simonová ◽  
A. Lauková ◽  
L. Chrastinová ◽  
I. Plachá ◽  
V. Strompfová ◽  
...  
Keyword(s):  
Growth Performance ◽  
Enterococcus Faecium ◽  
Average Daily Gain ◽  
Probiotic Strain ◽  
Cholesterol Concentration ◽  
Control Group ◽  
Feed Conversion ◽  
Eleutherococcus Senticosus ◽  
Commercial Diet ◽  
And Control

Abstract The effect of Enterococcus faecium CCM7420 (EF) - enterocin-producing and probiotic strain of rabbit origin, Eleutherococcus senticosus extract (ES) and their combination (ES+EF) was determined on selected bacteria in faeces and caecum content, leukocytes phagocytosis, blood biochemistry and growth performance. Ninety-six weaned rabbits were divided into 3 experimental (ES, EF, ES+EF) and control group (CG). The rabbits in the groups ES and EF+ES were fed commercial diet enriched with E. senticosus extract (30 g/100 kg feed), rabbits in groups EF and CG were fed untreated diet. The rabbits in the EF and ES+EF groups were administered with an overnight culture of E. faecium CCM7420 strain (500 μl/animal/day into water, 109 CFU/ml). The treatment period lasted 21 days. The microbiological examinations in faecal samples confirmed the presence of E. faecium CCM7420 strain. In groups EF and ES+EF, the reduction of faecal coliforms, Pseudomonas-like sp., Clostridium-like sp. and S. aureus was recorded. Leucocyte phagocytosis significantly increased in all experimental groups (P<0.0001) compared to CG. The lowest GPx values were measured in the ES+EF group. Higher total protein, triglycerides and calcium concentrations were detected in experimental groups compared to CG. The cholesterol concentration decreased in the ES group. The highest average daily gain was recorded in EF group; in ES+EF the better feed conversion ratio and no mortality was recorded. These results indicated that the dietary supplementation with the E. faecium CCM7420 and E. senticosus extract stimulate the leukocytes phagocytosis and reduces the potential pathogens in rabbits digestive tract without oxidative stress and improve the growth performance.

scholarly journals PSVIII-8 Effect of Lactobacillus Casei Zhang on E. coli induced injury of blood and milk barrier in dairy cow

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 259-259
Author(s):  
Yuhui Zheng ◽  
Shengli Li
Keyword(s):  
Bacterial Infection ◽  
Lactobacillus Casei ◽  
Bacterial Resistance ◽  
Bovine Mastitis ◽  
Production Performance ◽  
Economic Losses ◽  
Control Group ◽  
Bovine Mammary Epithelial Cells ◽  
E Coli ◽  
Lactobacillus Casei Zhang

Abstract Bovine mastitis is one of the major diseases which directly affects the milk production performance and it causes huge economic losses in the dairy industry. Bacterial infection is the main risk factor of bovine mastitis and the antibiotic therapy is the primary choice to control the disease. However, persistence use of antibiotic increases the incidence of bacterial resistance and traces of antibiotic residues in animal products. Lactobacillus casei Zhang is one of the probiotics with multiple biological functions, which has certain bacteriostatic effect on pathogenic microorganism. The purpose of this study was to explore the effect of Lactobacillus casei Zhang (L. casei Zhang) on the prevention of E. coli-induced milk-blood barrier damage. Bovine mammary epithelial cells (BMECs) were used to establish a milk-blood model and Control group (PBS), E. coli group, and L. casei Zhang pretreatment plus E. coli group were set up respectively. The results showed that: L. casei Zhang could significantly reduce the increase of LDH release caused by E. coli treatment (P&lt; 0.05). And it can also significantly reduce the decrease of transmembrane resistance of monolayer cells caused by E. coli treatment (P&lt; 0.05). In addition, L. casei Zhang could significantly reduce the expression of tight junction proteins ZO-1, Claudin-1, Claudin-4 and Occludin (P &lt; 0.05). In conclusion, L. casei Zhang could effectively improve the damage of the blood-milk barrier caused by E. coli and could protect BMECs during bacterial infection.

Age-associated features of the expression level of apoptosis markers in cardiomyocytes of patients with dilated cardiomyopathy

2022 ◽  
pp. 885-890
Author(s):  
К.П. Кравченко ◽  
К. Л. Козлов ◽  
А.О. Дробинцева ◽  
Д.С. Медведев ◽  
В.О. Полякова
Keyword(s):  
Cell Death ◽  
Dilated Cardiomyopathy ◽  
Molecular Mechanisms ◽  
Molecular Level ◽  
Confocal Laser ◽  
Control Group ◽  
Cellular Mechanisms ◽  
Apoptosis Pathway ◽  
Bax Protein

Для понимания патогенеза дилатационной кардиомиопатии (ДКМП) необходимо установить молекулярно-клеточные механизмы старения миокарда, в том числе связанные с программируемой клеточной гибелью, молекулярные механизмы которого практически не изучены. Цель работы - изучение маркеров апоптоза в кардиомиоцитах у пациентов с ДКМП in vitro. В работе использовали метод первичных диссоциированных клеточных культур и метод иммунофлюоресцентной конфокальной лазерной микроскопии. Для моделирования клеточного старения использовали клетки 3-го и 14-го пассажей, соответствующие «молодым» и «старым» культурам. На молекулярном уровне старение клеток кардиомиоцитов сопровождалось повышением экспрессии р16 в 2 раза по сравнению с «молодыми культурами» как в контрольной, так и в группе с ДКМП. Также установлено, что экспрессия р16 в культурах, взятых от пациентов с патологией, была в 2 раза выше, чем в аналогичных культурах от здоровых пациентов. Экспрессия р21 была повышена в группе с ДКМП по сравнению с контрольной группой, однако при старении культуры экспрессия p21 не изменялась, оставаясь на высоком уровне. Наиболее значимые различия были получены при сравнении экспрессии Bax в культуре клеток кардиомиоцитов из группы с ДКМП в «молодой» культуре с нормой - в 3,2 раза. Старение клеток миокарда на молекулярном уровне проявлялось в повышении экспрессии белка Baх, именно он является запускающим механизмом митохондриального пути апоптоза. Возможно, этот путь клеточной гибели является превалирующем при ДКМП. To understand the pathogenesis of dilated cardiomyopathy (DCMP), it is necessary to establish the molecular-cellular mechanisms of myocardial aging, including those associated with programmed cell death, the molecular mechanisms of which have not been practically studied. The aim of this work is to study markers of apoptosis in cardiomyocytes of patients with DCMP in vitro. We used the method of primary dissociated cell cultures and the method of immunofluorescence confocal laser microscopy. Cells of the 3 and 14 passages, corresponding to «young» and «old» cultures, were used to simulate cellular senescence. Results. At the molecular level, aging of cardiomyocyte cells was accompanied by a twofold increase in the expression of p16 compared to «young cultures» both in the control group and in the group with DCMP. It was also found that the expression of p16 in cultures taken from patients with pathology was 2 times higher than in similar cultures from healthy patients. The expression of p21 was increased in the group with DCMP compared to the control; however, with aging of the culture, the expression of p21 did not change, remaining at a significant level. The most significant differences were obtained when comparing the expression of Bax in the cell culture of cardiomyocytes from the group with DCMP in a «young» culture compared with the norm, 3,2 times. Aging of myocardial cells at the molecular level was manifested in an increase in the expression of the Bax protein, which is the triggering mechanism of the mitochondrial apoptosis pathway. It is possible that this pathway of cell death is prevalent in DCMP.

MO739EX VIVO THROMBOCYTE FUNCTION IN HEMODIALYSIS PATIENTS

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Vera Bonell ◽  
Georg Lorenz ◽  
Thorsten Kessler ◽  
Uwe Heemann ◽  
Christoph Schmaderer ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Clinical Outcome ◽  
Platelet Function ◽  
Primary Endpoint ◽  
Hemodialysis Patients ◽  
Healthy Controls ◽  
Induce Platelet Aggregation ◽  
Platelet Counts ◽  
Hospitalization Rates ◽  
Multiplate Analyzer

Abstract Background and Aims Coagulation disorders with both risk for bleeding and thrombotic events are common in hemodialysis (HD) patients. Altered thrombocyte counts and function may account for that. Here, we sought to better characterize thrombocyte function in hemodialysis patients. Method Platelet function was investigated using the Multiplate analyzer (Roche) based on impedance aggregometry. Adenosine diphosphate (ADP) was used to induce platelet aggregation and area under the curve (AUC) was used as primary endpoint. Platelet counts and C-reactive protein (CRP) levels were measured. Hospitalization was the primary clinical outcome. Pearson regression was used to test for associations of thrombocyte function and the primary endpoint. Results In total 60 chronic HD patients undergoing dialysis 3 times per week, and 67 healthy controls were included. In general, HD patients presented with significantly lower thrombocyte numbers compared to healthy controls (Median: 221 vs. 245 G/l, p=0.029). Further, thrombocyte function as determined by AUC was significantly altered in HD patients versus healthy controls (Median: 455 vs. 677 AU*min, p&lt;0.001; figure 1) with a significant correlation for platelet count and platelet function (r=0.42, p=0.001). Platelet function also correlated with the inflammatory state as seen by systemic CRP levels (r=0.28, p=0.033). Regarding the clinical outcome, platelet function correlated with hospitalization rates for infectious disease (r=0.27; p=0.040) and cardiovascular events (r=0.30; p=0.022). In case of hospitalization rates for infectious disease this correlation remained stable irrespective of adjustment for thrombocyte counts (r=0.27, p=0.036). Conclusion Lower platelet counts and altered function in HD patients was associated with risk of hospitalization and markers of inflammation in this cohort. The Multiplate analyzer appeared to be a valid and easily accessible method to assess thrombocyte function. Further studies are needed to determine whether assessment of thrombocyte function in clinical routine should be used to stratify risk in the vulnerable population of HD patients.

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