Probiotic mixture of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 attenuates hippocampal apoptosis induced by lipopolysaccharide in rats

In recent years, the beneficial impact of targeted gut microbiota manipulation in various neurological disorders has become more evident. Therefore, probiotics have been considered as a promising approach to modulate brain gene expression and neuronal pathways even in some neurodegenerative diseases. The purpose of this study was to determine the effect of probiotic biotherapy with combination of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 on the expression levels of proteins critical to neuronal apoptosis in hippocampus of lipopolysaccharide (LPS)-exposed rats. Four groups of animals (Control, LPS, Probiotic + LPS, and Probiotic) were treated with maltodextrin (placebo) or probiotic (109 CFU/ml/rat) for 2 weeks by gavage. On the 15th day, a single intraperitoneal dose of saline or LPS (1 mg/kg) was injected and 4 h later, protein assessment was performed by western blotting in hippocampal tissues. LPS significantly increased the Bax, Bax/Bcl-2 ratio, and cleaved caspase-3 expression along with decreased the Bcl-2 and procaspase-3 protein levels. However, probiotic pretreatment (L. helveticus R0052 + B. longum R0175) significantly downregulated the Bax and Bax/Bcl-2 ratio accompanied with upregulated Bcl-2 expression. Prophylactic treatment with these bacteria also attenuated LPS-induced caspase-3 activation by remarkably increasing the expression of procaspase-3 while reducing the level of cleaved caspase-3 in target tissues. Our data indicate that probiotic formulation (L. helveticus R0052 + B. longum R0175) alleviated hippocampal apoptosis induced by LPS in rats via the gut-brain axis and suggest that this probiotic could play a beneficial role in some neurodegenerative conditions

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Probiotic mixture of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 attenuates hippocampal apoptosis induced by lipopolysaccharide in rats.

In recent years, the beneficial impact of targeted gut microbiota manipulation in various neurological disorders has become more evident. Therefore, probiotics have been considered as a promising approach to modulate brain gene expression and neuronal pathways even in some neurodegenerative diseases. The purpose of this study was to determine the effect of probiotic biotherapy with combination of Lactobacillus helveticus R0052 and Bifidobacterium longum R0175 on the expression levels of proteins critical to neuronal apoptosis in hippocampus of lipopolysaccharide (LPS)-exposed rats. Four groups of animals (Control, LPS, Probiotic + LPS, and Probiotic) were treated with maltodextrin (placebo) or probiotic (109 CFU/ml/rat) for 2 weeks by gavage. On the 15th day, a single intraperitoneal dose of saline or LPS (1 mg/kg) was injected and 4 h later, protein assessment was performed by western blotting in hippocampal tissues. LPS significantly increased the Bax, Bax/Bcl-2 ratio, and cleaved caspase-3 expression along with decreased the Bcl-2 and procaspase-3 protein levels. However, probiotic pretreatment (L. helveticus R0052 + B. longum R0175) significantly downregulated the Bax and Bax/Bcl-2 ratio accompanied with upregulated Bcl-2 expression. Prophylactic treatment with these bacteria also attenuated LPS-induced caspase-3 activation by remarkably increasing the expression of procaspase-3 while reducing the level of cleaved caspase-3 in target tissues. Our data indicate that probiotic formulation (L. helveticus R0052 + B. longum R0175) alleviated hippocampal apoptosis induced by LPS in rats via the gut-brain axis and suggest that this probiotic could play a beneficial role in some neurodegenerative conditions.

The Effects of Probiotic Formulation Pretreatment (Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) on a Lipopolysaccharide Rat Model.

OBJECTIVE: The role of gut microbiota in the pathogenesis of several neurodegenerative disorders, including Alzheimer's disease (AD), via the gut-brain axis has recently been demonstrated; hence, modification of the intestinal microbiota composition by probiotic biotherapy could be a therapeutic target for these conditions. The aim of this study was to assess the effects of a probiotic formulation (Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) on inflammatory and memory processes in lipopolysaccharide (LPS)-induced rats, one of the animal models used in peripherally induced neuroinflammation and neurodegeneration. METHODS: Rats were randomly divided into four groups (Control, LPS, Probiotic + LPS, and Probiotic). All experimental groups were orally administrated maltodextrin (placebo) or probiotic (109 CFU/ml/rat) for 14 consecutive days and then were injected with saline or LPS (1 mg/kg, intraperitoneally [i.p.], single dose) 20 hours later. Memory retention ability and systemic and neuroinflammatory markers were assessed 4 hours after the injections. RESULTS: Systemic exposure to LPS resulted in significant elevation of both the circulating and hippocampal levels of proinflammatory cytokines, which decreased remarkably following probiotic pretreatment. Oral bacteriotherapy with a combination of L. helveticus R0052 and B. longum R0175 also attenuated the decremental effect of LPS on memory through brain-derived neurotrophic factor (BDNF) expression at the molecular level; however, this effect was not significant in the passive avoidance test at the behavioral level. CONCLUSIONS: These results suggest that the management of gut microbiota with this probiotic formulation could be a promising intervention to improve neuroinflammation-associated disorders such as AD.

Diagnostic efficacy of lsa63 antigen for human leptospirosis.

BACKGROUND: Timely diagnosis of leptospirosis is essential for early and effective treatment, for there are many differential diagnoses for it.. Leptospiral researchers have an increasing interest in developing new serological methods with recombinant antigens to improve the Leptospirosis diagnosis. Several serological tests have been developed for the proper diagnosis of leptospirosis. OBJECTIVES: To improve the previous works we developed an enzyme linked immunosorbent assay (ELISA) with novel recombinant leptospiral surface adhesion (Lsa63) protein to offer a new test. MATERIALS AND METHODS: In an experimental study, Recombinant Lsa63 (rLsa63) was produced in Escherishia coli (E.coli) BL21 (DE3). By using rLsa63, we generated IgM and IgG ELISA. Performance of these tests was compared to microscopic agglutination golden test (MAT). Two hundred twenty human serum samples were obtained from individuals suspicious of leptospirosis who were referred to Guilan Province Central Leptospira Laboratory for definitive diagnosis. The sensitivity, specificity and other statistical indexes of Lsa63-ELISAs were also determined. RESULTS: Among 220 serum samples, 30% (n = 65) had positive MAT responses, and also 38% (n = 84) and 40.9% (n = 90) showed positive reaction to IgG and IgM rLsa63-ELISA, respectively. The sensitivity, specificity and accuracy were 93.8%, 81.29 % and 85.0 for IgM-Lsa63- ELISA and 83.07, 80, 64 and 81.36 for IgG-Lsa63- ELISA, respectively. CONCLUSIONS: Our results demonstrated that the sensitivity and specificity of Lsa63-ELISAs are promising for the detection of Leptospira serovars.

Evaluation of New ELISA based on rLsa63 - rLipL32 antigens for serodiagnosis of Human Leptospirosis.

BACKGROUND AND OBJECTIVES: Timely diagnosis of leptospirosis is essential for an effective treatment. Large diversity of clinical symptoms has led leptospirosis diagnosis difficult. Researchers have conducted many tests with wide-range of sensitivity and specificity to achieve novel diagnostic procedures which have higher sensitivity and specificity compared with previous tests and which are more reliable and available to public laboratories. This study aimed to introduce Lsa63 and LipL32 proteins-based ELISA tests with more sensitivity, specificity, accuracy and convenience for public laboratories. MATERIALS AND METHODS: Recombinant forms of Lsa63 and LipL32 proteins were first generated. After coating these proteins, IgM and IgG ELISA tests were performed. 220 patients with suspicion of leptospirosis infection were selected for serum collection. The sera tests were carried out using MAT, IgM and IgG ELISA tests. In order to assess the performance of ELISA, the results of this test were compared with MAT. RESULTS: 30% of serum samples (n=65) in MAT were positive for leptospirosis infection, while ELISA tests including rLipL32- rLsa63-IgM and rLipL32-rLsa63-IgG showed 40.45% (n=89) and 38.63% (n=80) positive reaction, respectively. CONCLUSION: Our results demonstrated that new ELISA tests based on mixing LipL32 and Lsa63 proteins, a novel mixture of recombinant antigens, are valuable to detect specific antibodies against pathogenic Leptospira in human serum and could be considered as helpful techniques in leptospirosis diagnosis.

An old man with a mass in the retropharyngeal space.

In this article, we discuss clinical manifestations, laboratory results, and radiological findings in an old man with suppurative tender mass in the neck, accompanied by, fever, weight loss, malaise, fatigue, night sweat, cough, vomiting, and dysphagia. Pharyngeal exam revealed a huge retropharyngeal abscess.