Lactobacillus rhamnosus GG and Bifidobacterium bifidum TMC3115 Can Affect Development of Hippocampal Neurons Cultured In Vitro in a Strain-Dependent Manner

Abstract Objectives This study aimed to examine whether probiotics could morphologically or physiologically influence hippocampal neuron development. Methods Hippocampal neurons cultured in vitro were exposed to live or heat-inactivated Lactobacillus rhamnosus GG (LGG), or live or heat-inactivated Bifidobacterium bifidum TMC3115 (TMC3115), for either 6 or 24 h. Neuron viability was then tested using the methyl thiazolyl tetrazolium assay. Neuronal morphological changes and drebrin (DRB) and synaptophysin (SYP) protein levels were monitored using immunofluorescence. And the levels of DRB, SYP, and brain-derived neurotrophic factor (BDNF), and cAMP-response element binding protein (CREB) mRNA were detected using RT-PCR. The BDNF, CREB and phosphorylated-CREB (P-CREB) protein levels were detected by ELISA or Western blot assays. Results We found exposure to probiotics could enhance neuron viability, although no significant differences were found in neuronal morphology among the groups following exposure to the test bacteria. However, the synapse development-related proteins, DRB and SYP, as well as BDNF and P-CREB protein levels, were significantly altered in this specific culture system. Conclusions These results demonstrated that LGG and TMC3115 exposure can affect neuronal viability, along with synaptic and brain function development, in a strain-dependent manner, which may also be closely associated with the physiological and cultural conditions of each strain, The up-regulated P-CREB protein level may be one of the underlying mechanisms by which the tested bacteria, especially live TMC3115 following exposure for 24 h, are able to regulate neuronal BDNF protein production. Further studies are needed to explore other possible effects probiotic exposure may have on hippocampal neurons, as well as the corresponding mechanisms that underlie them. Funding Sources This work was funded by the National Natural Science Foundation of China (Grant number 81872606).

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An aniline-substituted bile salt analog protects both mice and hamsters from multiple Clostridioides difficile strains

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01435-21 ◽

2021 ◽

Author(s):

Jacqueline R. Phan ◽

Dung M. Do ◽

Minh Chau Truong ◽

Connie Ngo ◽

Julian H. Phan ◽

...

Keyword(s):

Bile Salt ◽

Spore Germination ◽

Dependent Manner ◽

Germination Inhibitors ◽

New Methods ◽

Clostridioides Difficile ◽

Antibiotic Associated Diarrhea ◽

Careful Screening ◽

Strain Dependent

Clostridioides difficile infection (CDI) is the major identifiable cause of antibiotic-associated diarrhea. The emergence of hypervirulent C. difficile strains has led to increases in both hospital- and community-acquired CDI. Furthermore, CDI relapse from hypervirulent strains can reach up to 25%. Thus, standard treatments are rendered less effective, making new methods of prevention and treatment more critical. Previously, the bile salt analog CamSA was shown to inhibit spore germination in vitro and protect mice and hamsters from C. difficile strain 630. Here, we show that CamSA was less active at preventing spore germination of other C. difficile ribotypes, including the hypervirulent strain R20291. Strain-specific in vitro germination activity of CamSA correlated with its ability to prevent CDI in mice. Additional bile salt analogs were screened for in vitro germination inhibition activity against strain R20291, and the most active compounds were tested against other strains. An aniline-substituted bile salt analog, (CaPA), was found to be a better anti-germinant than CamSA against eight different C. difficile strains. In addition, CaPA was capable of reducing, delaying, or preventing murine CDI signs in all strains tested. CaPA-treated mice showed no obvious toxicity and showed minor effects on their gut microbiome. CaPA’s efficacy was further confirmed by its ability to prevent CDI in hamsters infected with strain 630. These data suggest that C. difficile spores respond to germination inhibitors in a strain-dependent manner. However, careful screening can identify anti-germinants with broad CDI prophylaxis activity.

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In Vivo, In Vitro and Transplacental Immune Effects of a Probiotic Bacterium Lactobacillus rhamnosus GG in Humans

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2008.12.761 ◽

2009 ◽

Vol 123 (2) ◽

pp. S200-S200

Author(s):

R.J. Boyle ◽

L. Mah ◽

S. Kivivuori ◽

A. Chen ◽

S.J. Lahtinen ◽

...

Keyword(s):

Lactobacillus Rhamnosus ◽

Probiotic Bacterium ◽

Lactobacillus Rhamnosus Gg

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Lactobacillus rhamnosus GG in Experimental Oral Biofilms Exposed to Different Carbohydrate Sources

Caries Research ◽

10.1159/000479380 ◽

2018 ◽

Vol 52 (3) ◽

pp. 220-229 ◽

Cited By ~ 5

Author(s):

Qingru Jiang ◽

Veera Kainulainen ◽

Iva Stamatova ◽

Riitta Korpela ◽

Jukka H. Meurman

Keyword(s):

Lactobacillus Rhamnosus ◽

In Vitro Study ◽

Viable Cell ◽

Lactobacillus Rhamnosus Gg ◽

Carbohydrate Source ◽

Ph Values ◽

Dental Hard Tissues ◽

Oral Biofilms ◽

Spent Media

Probiotic administration may favour caries prevention, as recent research has shown. This in vitro study aimed to investigate the growth of Lactobacillus rhamnosus GG (LGG) in experimental biofilms exposed to various carbohydrates, and also to assess its cariogenic potential. Multispecies experimental oral biofilms with or without LGG were grown with a sole-carbohydrate source (fructose/glucose/lactose/sorbitol/sucrose). The viable cells of LGG and structure of the biofilms were examined after 64.5 h of incubation, and pH values of spent media were measured at 16.5, 40.5, and 64.5 h. Fermentation profiles of LGG in biofilm media were assessed with study carbohydrate as the sole energy source. Our results showed that LGG reached higher viable cell numbers with glucose and sucrose in 64.5-h multispecies experimental oral biofilms compared to other carbohydrates. When LGG was incorporated in biofilms, no distinct pH changes at any time points were observed under any of the carbohydrates used; the pH values of spent media at each time point were lower when lactose was used, compared to other carbohydrates. The fermentation profiles of LGG in biofilm media were similar to its growth in MRS (no obvious growth with lactose or sucrose). In conclusion, LGG in our in vitro multispecies experimental oral biofilms was capable of surviving and growing well in each carbohydrate source. LGG might not have harmful effects on dental hard tissues. Another finding from our study was that the lowest pH values were observed in the presence of lactose, and the thickest biofilms were in sucrose.

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Blanching effect on the bioactive compounds and on the viability of Lactobacillus rhamnosus GG before and after in vitro simulation of the digestive system in jabuticaba juice

Semina Ciências Agrárias ◽

10.5433/1679-0359.2017v38n3p1277 ◽

2017 ◽

Vol 38 (3) ◽

pp. 1277 ◽

Cited By ~ 1

Author(s):

Daniane Campos de Oliveira ◽

Eliane Maurício Furtado Martins ◽

Maurilio Lopes Martins ◽

Giovanna Bretas Martins ◽

Mirella Lima Binoti ◽

...

Keyword(s):

Ascorbic Acid ◽

Phenolic Compounds ◽

Antioxidant Capacity ◽

Lactobacillus Rhamnosus ◽

Total Phenolic ◽

Total Phenolic Compounds ◽

Lactobacillus Rhamnosus Gg ◽

In Vitro Simulation ◽

Hedonic Scale

The viability of Lactobacillus rhamnosus GG (LGG) in jabuticaba juices and its survival in the gastrointestinal tract (GIT), simulated in vitro, was studied. Two juices were prepared: A - with non-blanched fruits, and B - with blanched fruits. LGG was then added and the juices maintained at 8 ºC for 28 days. The control treatment consisted of juices without the added probiotic. The following were determined in the juices: the viability and in vitro survival of LGG, fecal coliforms, Salmonella sp., pH, acidity, total soluble solids (TSS), color, antioxidant capacity, total phenolic compounds, anthocyanins and ascorbic acid. The sensory acceptability was also determined using a 9-point hedonic scale. Blanching interfered (p < 0.05) with the viability of LGG, juice A showing the greatest viability as compared to juice B. After in vitro simulation, the probiotic bacterial count was < 1.0 log CFU mL-1, which demonstrates the low resistance of the strain to the simulated GIT conditions. The juices were conformed to the microbiological standards established by law. The pH, acidity and TSS were influenced by blanching (p < 0.05), with values of 5.03, 0.46% and 15.38 °Brix for juice A and 5.12, 0.66% and 16.05 °Brix for juice B, respectively. The addition of LGG did not influence these characteristics. Only the pH value was influenced by the storage time (p < 0.05), increasing throughout storage. Juice B showed lower luminosity (L*) and a greater value for a* as compared to juice A, indicating that the former became darker and redder due to the blanching process. Both juices showed positive values for the b* coordinate. The juice was found to be a good source of polyphenols. Neither the time nor the addition of LGG affected the antioxidant capacity, total phenolic compounds or anthocyanin contents. However, blanching contributed (p < 0.05) to an increase in the contents of these compounds in the juices. Values for antioxidant capacity of 186.20 and 2552.59 uM Trolox g-1, for total phenolic compounds of 275.06 and 1163.18 mg GAE 100 g-1-wwb, and for anthocyanins as cyanidin 3-glucoside of 12.71 and 90.99 mg 100 g-1 were found for juices A and B, respectively. The juices contained 72.87 mg 100 mL-1 of ascorbic acid. Scores of above 6.0 (liked slightly) were awarded on the hedonic scale for the attributes evaluated. The addition of probiotics in jabuticaba juices needs to be further studied to ensure the viability of the cultures during storage and their survival in the gastrointestinal tract.

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Protective Effects of Scolopendra Water Extract on Trimethyltin-Induced Hippocampal Neurodegeneration and Seizures in Mice

Brain Sciences ◽

10.3390/brainsci9120369 ◽

2019 ◽

Vol 9 (12) ◽

pp. 369

Author(s):

Yun-Soo Seo ◽

Mary Jasmin Ang ◽

Byeong Cheol Moon ◽

Hyo Seon Kim ◽

Goya Choi ◽

...

Keyword(s):

Hippocampal Neurons ◽

Primary Cultures ◽

Water Extract ◽

Model Systems ◽

Inflammatory Factor ◽

Dependent Manner ◽

Protective Effects ◽

Cell Degeneration

Trimethyltin (TMT) is an organotin compound with potent neurotoxic action characterized by neuronal degeneration in the hippocampus. This study evaluated the protective effects of a Scolopendra water extract (SWE) against TMT intoxication in hippocampal neurons, using both in vitro and in vivo model systems. Specifically, we examined the actions of SWE on TMT- (5 mM) induced cytotoxicity in primary cultures of mouse hippocampal neurons (7 days in vitro) and the effects of SWE on hippocampal degeneration in adult TMT- (2.6 mg/kg, intraperitoneal) treated C57BL/6 mice. We found that SWE pretreatment (0–100 μg/mL) significantly reduced TMT-induced cytotoxicity in cultured hippocampal neurons in a dose-dependent manner, as determined by lactate dehydrogenase and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assays. Additionally, this study showed that perioral administration of SWE (5 mg/kg), from −6 to 0 days before TMT injection, significantly attenuated hippocampal cell degeneration and seizures in adult mice. Furthermore, quantitative analysis of Iba-1 (Allograft inflammatory factor 1)- and GFAP (Glial fibrillary acidic protein)-immunostained cells revealed a significant reduction in the levels of Iba-1- and GFAP-positive cell bodies in the dentate gyrus (DG) of mice treated with SWE prior to TMT injection. These data indicated that SWE pretreatment significantly protected the hippocampus against the massive activation of microglia and astrocytes elicited by TMT. In addition, our data showed that the SWE-induced reduction of immune cell activation was linked to a significant reduction in cell death and a significant improvement in TMT-induced seizure behavior. Thus, we conclude that SWE ameliorated the detrimental effects of TMT toxicity on hippocampal neurons, both in vivo and in vitro. Altogether, our findings hint at a promising pharmacotherapeutic use of SWE in hippocampal degeneration and dysfunction.

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Intestinal Mucus Alters the Ability of Probiotic Bacteria To Bind Aflatoxin B1 In Vitro

Applied and Environmental Microbiology ◽

10.1128/aem.70.10.6306-6308.2004 ◽

2004 ◽

Vol 70 (10) ◽

pp. 6306-6308 ◽

Cited By ~ 50

Author(s):

S. Gratz ◽

H. Mykkänen ◽

A. C. Ouwehand ◽

R. Juvonen ◽

S. Salminen ◽

...

Keyword(s):

Aflatoxin B1 ◽

Probiotic Bacteria ◽

Dependent Manner ◽

Surface Binding ◽

Intestinal Mucus ◽

Aflatoxin B ◽

Strain Dependent

ABSTRACT Several probiotics are known to bind aflatoxin B1 (AFB1) to their surfaces and to adhere to intestinal mucus. In this study, preincubation of two probiotic preparations with either AFB1 or mucus reduced the subsequent surface binding of mucus and AFB1, respectively, in a strain-dependent manner.

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Effect of probiotics on the viability of porcine and human neutrophils in vitro

Veterinární Medicína ◽

10.17221/72/2017-vetmed ◽

2017 ◽

Vol 62 (No. 12) ◽

pp. 637-646

Author(s):

T. Sustrova ◽

P. Ondrackova ◽

L. Leva ◽

M. Kolarova ◽

P. Kulich ◽

...

Keyword(s):

Tissue Regeneration ◽

Lactobacillus Rhamnosus ◽

Bifidobacterium Bifidum ◽

Human Neutrophils ◽

Neutrophil Apoptosis ◽

Large White ◽

Healthy Human ◽

Lactate Dehydrogenase Activity ◽

Late Apoptosis

Lactobacillus, Bifidobacterium and Enterococcus cultures are increasingly used as probiotics for humans and pigs. The aim of this study was to investigate if co-cultivation of porcine and human neutrophils with probiotics can lead to increased apoptosis in vitro. Ten adult Large white pigs and 10 healthy human donors were used in this study. Neutrophils were isolated by dextran sedimentation and cultivated with and without the lactic acid bacteria Bifidobacterium bifidum, Lactobacillus rhamnosus, and Enterococcus faecium for 2, 4, 24 and 48 h. Early and late apoptosis was measured using flow cytometry, and cell lysis was detected based on lactate dehydrogenase activity (LDH). A significant (P < 0.05; P < 0.01) increase in apoptotic neutrophils and LDH was observed at 24 h and 48 h in vitro. All probiotics exerted their greatest effects on the early apoptosis of porcine neutrophils, while the effects of L. rhamnosus were most pronounced on late apoptosis and those of B. bifidum on LDH release of human neutrophils. The increased neutrophil apoptosis caused by probiotic bacteria can be beneficial for more efficient efferocytosis and faster resolution of inflammation and tissue regeneration. In conclusion, we have demonstrated that the interaction of B. bifidum, L. rhamnosus, and E. faecium with human and porcine neutrophils leads to their apoptosis.

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