Rhamnosus GR-1 Research Articles

Editorial: Advances in the Prevention and Treatment of Inflammation-Associated Preterm Birth.

Editorial: Advances in the Prevention and Treatment of Inflammation-Associated Preterm Birth.

Lactobacillus rhamnosus GR-1 Limits Escherichia coli-Induced Inflammatory Responses via Attenuating MyD88-Dependent and MyD88-Independent Pathway Activation in Bovine Endometrial Epithelial Cells.

Intrauterine Escherichia coli infection after calving reduces fertility and causes major economic losses in the dairy industry. We investigated the protective effect of the probiotic Lactobacillus rhamnosus GR-1 on E. coli-induced cell damage and inflammation in primary bovine endometrial epithelial cells (BEECs). L. rhamnosus GR-1 reduced ultrastructure alterations and the percentage of BEECs apoptosis after E. coli challenge. Increased messenger RNA (mRNA) expression of immune response indicators, including pattern recognition receptors (toll-like receptor [TLR]2, TLR4, nucleotide-binding oligomerization domain [NOD]1, and NOD2), inflammasome proteins (NOD-like receptor family member pyrin domain-containing protein 3, apoptosis-associated speck-like protein, and caspase-1), TLR4 downstream adaptor molecules (myeloid differentiation antigen 88 [MyD88], toll-like receptor adaptor molecule 2 [TICAM2]), nuclear transcription factor kB (NF-kB), and the inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-8, IL-10, IL-18, and interferon (IFN)-β, was observed following E. coli challenge. However, these increases were attenuated by L. rhamnosus GR-1 pretreatment. Our data indicate that L. rhamnosus GR-1 ameliorates the E. coli-induced disruption of cellular ultrastructure, subsequently reducing the percentage of BEECs apoptosis and limiting inflammatory responses, partly via attenuation of MyD88-dependent and MyD88-independent pathway activation. Certain probiotics could potentially prevent postpartum uterine diseases in dairy cows, ultimately reducing the use of antibiotics.

Probiotics [LGG-BB12 or RC14-GR1] versus placebo as prophylaxis for urinary tract infection in persons with spinal cord injury [ProSCIUTTU]: a study protocol for a randomised controlled trial.

BackgroundUrinary tract infections [UTIs] are very common in people with Spinal Cord Injury [SCI]. UTIs are increasingly difficult and expensive to treat as the organisms that cause them become more antibiotic resistant. Among the SCI population, there is a high rate of multi-resistant organism [MRO] colonisation. Non-antibiotic prevention strategies are needed to prevent UTI without increasing resistance. Probiotics have been reported to be beneficial in preventing UTIs in post-menopausal women in several in vivo and in vitro studies. The main aim of this study is to determine whether probiotic therapy with combinations of Lactobacillus reuteri RC-14 + Lactobacillus rhamnosus GR-1 [RC14-GR1] and/or Lactobacillus rhamnosus GG + Bifidobacterium BB-12 [LGG-BB12] are effective in preventing UTI in people with SCI compared to placebo.MethodThis is a multi-site randomised double-blind double-dummy placebo-controlled factorial design study conducted in New South Wales, Australia. All participants have a neurogenic bladder as a result of spinal injury. Recruitment started in April 2011.Participants are randomised to one of four arms, designed for factorial analysis of LGG-BB12 and/or RC14-GR1 v Placebo. This involves 24 weeks of daily oral treatment with RC14-GR1 + LGG-BB12, RC14-GR1 + placebo, LGG-BB12 + placebo or two placebo capsules. Randomisation is stratified by bladder management type and inpatient status. Participants are assessed at baseline, three months and six months for Short Form Health Survey [SF-36], microbiological swabs of rectum, nose and groin; urine culture and urinary catheters for subjects with indwelling catheters. A bowel questionnaire is administered at baseline and three months to assess effect of probiotics on bowel function.The primary outcome is time from randomisation to occurrence of symptomatic UTI. The secondary outcomes are change of MRO status and bowel function, quality of life and cost-effectiveness of probiotics in persons with SCI. The primary outcome will be analysed using survival analysis of factorial groups, with Cox regression modelling to test the effect of each treatment while allowing for the other, assuming no interaction effect. Hazard ratios and Kaplan-Meier survival curves will be used to summarise results.DiscussionIf these probiotics are shown to be effective in preventing UTI and MRO colonisation, they would be a very attractive alternative for UTI prophylaxis and for combating the increasing rate of antibiotic resistance after SCI.Trial registrationAustralian New Zealand Clinical Trials Registry [ACTRN 12610000512022]. Date of registration: 21 June 2010.Electronic supplementary materialThe online version of this article (doi:10.1186/s12894-016-0136-8) contains supplementary material, which is available to authorized users.

Non-Antibiotic Prophylaxis for Urinary Tract Infections.

Increasing antimicrobial resistance has stimulated interest in non-antibiotic prophylaxis of recurrent urinary tract infections (UTIs). Well-known steps in the pathogenesis of UTIs are urogenital colonization and adherence of uropathogens to uroepithelial cell receptors. To prevent colonization in postmenopausal women, vaginal, but not oral, estrogens have been shown to restore the vagina lactobacilli flora, reduce vaginal colonization with Enterobacteriaceae, and reduce the number of UTIs compared to placebo. Different lactobacilli strains show different results in the prevention of recurrent UTIs. Intravaginal suppositories with Lactobacillus crispatus in premenopausal women and oral capsules with Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 in postmenopausal women are promising. Ascorbic acid (vitamin C) cannot be recommended for the prevention of UTIs. Cranberries are thought to contain proanthocyanidins that can inhibit adherence of P-fimbriated E. coli to the uroepithelial cell receptors. Cranberry products decreased UTI recurrences about 30%–40% in premenopausal women with recurrent UTIs, but are less effective than low-dose antimicrobial prophylaxis. However, the optimal dose of cranberry product has still to be determined. Initially OM-89, a vaccine with 18 heat-killed E. coli extracts, seemed promising, but this was not confirmed in a recently randomized trial.

Outcome of a public consultation on the draft guidance of the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) on the scientific requirements for health claims related to the immune system, the gastrointestinal tract and defence against pathogenic microorganisms

Outcome of a public consultation on the draft guidance of the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) on the scientific requirements for health claims related to the immune system, the gastrointestinal tract and defence against pathogenic microorganisms

Preferential production of G-CSF by a protein-like Lactobacillus rhamnosus GR-1 secretory factor through activating TLR2-dependent signaling events without activation of JNKs.

BackgroundDifferent species and strains of probiotic bacteria confer distinct immunological responses on immune cells. Lactobacillus rhamnosus GR-1 (GR-1) is a probiotic bacterial strain found in both the intestinal and urogenital tracts, and has immunomodulatory effects on several cell types including macrophages. However, detailed immunological responses and the signaling mechanism involved in the response are largely unknown.ResultsWe examined the production of GR-1-induced cytokines/chemokines and signaling events in macrophages. Among 84 cytokines and chemokines examined, GR-1 discretely induced granulocyte colony-stimulating factor (G-CSF) mRNA at highest levels (>60-fold) without inducing other cytokines such as IL-1α, IL-1β, IL-6 and TNF-α (<5-fold). The toll-like receptor (TLR) 2/6-agonist PAM2CSK4, TLR2/1-agonist PAM3CSK4 and TLR4-agonist lipopolysaccharide induced all of these inflammatory cytokines at high levels (>50-fold). The TLR2 ligand lipoteichoic acid activated all mitogen-activated kinases, Akt and NF-κB; whereas, GR-1 selectively activated extracellular regulated kinases and p38, NF-κB and Akt, but not c-Jun N-terminal kinases (JNKs) in a TLR2-dependent manner. Using specific inhibitors, we demonstrated that lack of JNKs activation by GR-1 caused inefficient production of pro-inflammatory cytokines but not G-CSF production. A secreted heat-labile protein-like molecule, 30–100 kDa in size, induced the preferential production of G-CSF.ConclusionThis study elucidated unique signaling events triggered by GR-1, resulting in selective production of the immunomodulatory cytokine G-CSF in macrophages.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0578-2) contains supplementary material, which is available to authorized users.

Changes in vaginal microbiota following antimicrobial and probiotic therapy.

BackgroundThe composition of the vaginal microbiota is known to be important for health. When infections occur, antimicrobial therapy is often poorly efficacious.Objective and designWe used 16S rRNA gene sequencing to characterize changes in the bacterial microbiota following oral antimicrobial and probiotic interventions.ResultsWhile the bacterial vaginal profiles of women with vulvovaginal candidiasis were dominated by lactobacilli as in healthy women, and unchanged by therapy, Gardnerella vaginalis, Prevotella, Atopobium, Sneathia, and Megasphaera dominated the vagina of women with bacterial vaginosis (BV), and treatment with tinidazole plus Lactobacillus reuteri RC-14+L. rhamnosus GR-1 resulted in an increased relative abundance of indigenous L. iners or L. crispatus.ConclusionsThe ability to restore homeostasis provides a rationale for conjoint use of probiotics with antibiotic treatment of BV.

In vitro modulation of probiotic bacteria on the biofilm of Candida glabrata

A conspicuous new concept of pathogens living as the microbial societies in the human host rather than free planktonic cells has raised considerable concerns among scientists and clinicians. Fungal biofilms are communities of cells that possess distinct characteristic such as increased resistance to the immune defence and antimycotic agents in comparison to their planktonic cells counterpart. Therefore, inhibition of the biofilm may represent a new paradigm for antifungal development. In this study, we aim to evaluate the in vitro modulation of vulvovaginal candidiasis (VVC)-causing Candida glabrata biofilms using probiotic lactobacilli strains. Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 were shown to have completely inhibited C. glabrata biofilms and the results were corroborated by scanning electron microscopy (SEM), which revealed scanty structures of the mixed biofilms of C. glabrata and probiotic lactobacilli strains. In addition, biofilm-related C. glabrata genes EPA6 and YAK1 were downregulated in response to the probiotic lactobacilli challenges. The present study suggested that probiotic L. rhamnosus GR-1 and L. reuteri RC-14 strains inhibited C. glabrata biofilm by partially impeding the adherence of yeast cells and the effect might be contributed by the secretory compounds produced by these probiotic lactobacilli strains. Further investigations are required to examine and identify the biofilm inhibitory compounds and the mechanism of probiotic actions of these lactobacilli strains.

Use of a Fluorometric Microplate Assay to Assess the Activity of Prebiotics and Probiotics against Uropathogenic E. coli Adherence to Human Uroepithelial Cells

Urinary tract infections impact ~40-50% of women in the US. Probiotics have gained interest as an alternative treatment as they may compete with pathogens for adherence and colonization of mucosal surfaces. We used a fluorometric microplate assay to determine the ability of prebiotics and probiotics to inhibit E. coli adherence to human uroepithelial cells (UEC). Probiotic bacteria and p-fimbriated E. coli were labeled for 30 min with 400 μM BacLight Red or Green; E. coli adherence to UEC was measured after pre- (exclusion), co- (competition), and post-incubation (displacement) of probiotics with UEC. Probiotics inhibited E. coli adherence in a dose dependent manner. B. infantis reduced E. coli adherence up to 80, 77, and 27% for exclusion, competition, and displacement. In addition, L. plantarum reduced E. coli adherence up to 62, 70, and 41% and L. rhamnosus GR-1 inhibited E. coli adherence up to 45, 61, and 25% for exclusion, competition, and displacement, respectively. Incubation of probiotics with fructooligosaccharides, inulin, and pectin further reduced E. coli adherence; however, these prebiotics did not increase probiotic adherence to UEC. Probiotics inhibited the adherence of uropathogenic E. coli to human UEC; the addition of prebiotics further promoted this anti-E. coli adhesion activity, but did not increase probiotic adherence to UEC. Therefore, probiotics may produce metabolites that are inhibitory to urinary tract pathogens. Supported by the ARC, Washington State University.

Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 exhibit strong antifungal effects against vulvovaginal candidiasis-causing Candida glabrata isolates.

AimsThis study investigates the antagonistic effects of the probiotic strains Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 against vulvovaginal candidiasis (VVC)-causing Candida glabrata.Methods and ResultsGrowth inhibitory activities of Lact. rhamnosus GR-1 and Lact. reuteri RC-14 strains against C. glabrata were demonstrated using a spot overlay assay and a plate-based microtitre assay. In addition, these probiotic lactobacilli strains also exhibited potent candidacidal activity against C. glabrata, as demonstrated by a LIVE/DEAD yeast viability assay performed using confocal laser scanning microscopy. The metabolic activities of all C. glabrata strains were completely shut down in response to the challenges by the probiotic lactobacilli strains. In addition, both probiotic lactobacilli strains exhibited strong autoaggregation and coaggregation phenotypes in the presence of C. glabrata, which indicate that these lactobacilli strains may exert their probiotic effects through the formation of aggregates and, thus the consequent prevention of colonization by C. glabrata.ConclusionsProbiotic Lact. rhamnosus GR-1 and Lact. reuteri RC-14 strains exhibited potent antagonistic activities against all of the tested C. glabrata strains. These lactobacilli exhibited antifungal effects, including those attributed to their aggregation abilities, and their presence caused the cessation of growth and eventual cell death of C. glabrata.Significance and Impact of the StudyThis is the first study to report on the antagonistic effects of these probiotic lactobacilli strains against the non-Candida albicans Candida (NCAC) species C. glabrata.

The evaluation of recognize of clinical factors and microscopyexamination vaginal discharge in vaginal candidias diagnostic among the women who reffered to kohasht health –medical center in 2011

Abstract AbstIntroduction: The reproductive system infections are most common womens reasons for repetitive refers to gynecological clinic and the second problem is vulvovaginal candidiasis. since, distribution of patients and clinical signs may cause the wrong diagnose and treatment , in this study clinical signs and examination microscopy and vaginal discharge were compared. Method: In this descriptive study that has done among 18-44 married women years with complain from vaginal itching or irritation (july-may 2011). After diagnose the vaginite associate of patients complains and rigest the clinical signs, three sampling were done from every women. Two of them for wet mount with saline /KOH 10% and the other sample for sending to lab to culture purpose(golden standard test). data analysis was performed by spss 17 soft ware. Result: Among the factors that evaluate the most sensitivity belong to vulval erythema (89/1%) and the less sensitivity belong to cottage discharge(74/3%) and in the microscopic examination, the most specificity(100%) and the less specificity belong to themselves in vaginal candidiases diagnose between studied factors. Conclusion: Using the lab method (culture) and microscopic study on vaginite diagnose. Keyword: candida vaginitis, microscopy examination, candidiasis REFERENCES Ali B Sh,Tohidi A(2000)” Prevalence of candida vaginitis among symptomatic patiets in kerman”. The Journal of Qazvin University of Medical Science. (13). 42-48 Bansal KM, Singh K and Bhatnagar S (2001)” Prevalence of lower RTI among married female in the reproductive age group(15-45). Health and population. 24(3)157-163. Berek JS & Novak E. (2007). Berek & Novak’s Gynecology. 14th ed. Philadelphia, Lippincott Williams & Wilkins. 542-547. Buyukbayrak EE, Kars B, Karageyim karsidag AY, Ilkay karadenis B, Kaymas O, Geneer S(2010)”Diagnosis of vulvovaginitis: comparison of clinical and microbiological diagnosis”. Archive of Gynecology and Obstetrics. 282(5): 515-519. Ehrstrom S, Kornfeld D, Rylander E (2007)” Perceived stress in women with recurrent vulvovaginal candidiasis”. Jurnal of Psychosomatic Obstetrics and Gynecology. 28(3)169-176 Kariman N, Shafai Z, Afrakhte M, Valai N, Ahmadi M(2002)”Comparative study of traputic effects of fluconazole and clotrimazole on candida vaginitis”. Jornal Behbood. 6(13): 9-16 Martinez RCR, Franceschini SA, Patta MC, Quintana SM, Candido RC, Frreira ECP and etal(2009)”Improved treatment of vulvovaginal candidiasis with fluconazole plus probiotic lactobacillus rhamnosus GR-1 and lactobacillus reuteri RC-14”. Letters in Applied Microbiology. 48(3): 269-274. Moreira D, Paula CR(2006)” Vulvovaginal candidiasis”. International Journal Gynecology and Obestetrics. 92: 266-267. Ryan KJ, Berkovitz R, Robert L, Ryan BJ. (1999). Kistner’s gynecology. 5th ed. Medical Publisher Inc. 476-480 Sobel DJ(2007)” Vulvovaginal candidosis”. Lancet. 396(9577): 1961-1971.

Lipopolysaccharide-Induced Profiles of Cytokine, Chemokine, and Growth Factors Produced by Human Decidual Cells Are Altered by Lactobacillus rhamnosus GR-1 Supernatant

The aim of this study was to assess the effects of bacterial lipopolysaccharide (LPS) and Lactobacillus rhamnosus GR-1 supernatant (GR-1SN) on secretion profiles of cytokines, chemokines, and growth factors from primary cultures of human decidual cells. Lipopolysaccharide significantly increased the output of proinflammatory cytokines (interleukin [IL]-1B, IL-2, IL-6, IL-12p70, IL-15, IL-17A, interferon gamma [IFN-γ], and tumor necrosis factor [TNF]); anti-inflammatory cytokines (IL-1RN, IL-4, IL-9, and IL-10); chemokines (IL-8, eotaxin, IFN-inducible protein 10 [IP-10], monocyte chemoattractant protein 1 [MCP-1], macrophage inflammatory protein-1α [MIP-1α], macrophage inflammatory protein-1β [MIP-1β], and regulated on activation normal T cell expressed and secreted [RANTES]); and growth factors (granulocyte colony-stimulating factor [CSF] 3, CSF-2, and vascular endothelial growth factor A [VEGFA]). Lactobacillus rhamnosus GR-1SN alone significantly increased CSF-3, MIP-1α MIP-1β, and RANTES but decreased IL-15 and IP-10 output. The GR-1SN also significantly or partially reduced LPS-induced proinflammatory cytokines TNF, IFN-γ, IL-1β, IL-2 IL-6, IL-12p70, IL-15, IL-17, and IP-10; partially reduced LPS-induced anti-inflammatory cytokines IL-1RN, IL-4 and IL-10, and LPS-induced VEGFA output but did not affect CSF-3, MIP-1α, MIP-1β, MCP-1, IL-8, and IL-9. Our results demonstrate that GR-1SN attenuates the inflammatory responses to LPS by human decidual cells, suggesting its potential role in ameliorating intrauterine infection.

Probiotic Lactobacillus rhamnosus GR-1 supernatant prevents lipopolysaccharide-induced preterm birth and reduces inflammation in pregnant CD-1 mice

The objective of this study was to determine the effect of probiotic Lactobacillus rhamnosus GR-1 supernatant (GR-1 SN) on lipopolysaccharide-induced preterm birth (PTB) and outputs of cytokines, chemokines, and progesterone in pregnant CD-1 mice. We compared PTB rates after intrauterine injection of lipopolysaccharide with and without previous GR-1 SN treatment. Cytokines and chemokines in the maternal plasma, myometrium, placenta, and amniotic fluid were examined with multiplex assay, and circulating maternal progesterone was measured with enzyme-linked immunoassay. Statistical significance was assessed with 2-tailed 1-way analysis of variance or analysis of variance on ranks. Fetal sex ratios in mice that delivered preterm were compared with those that delivered at term after lipopolysaccharide and GR-1 SN treatments. GR-1 SN reduced lipopolysaccharide-induced PTB by 43%. GR-1 SN significantly decreased the lipopolysaccharide-induced production of interleukin (IL)-1β, -6, and -12p40, tumor necrosis factor-α, CCL4, and CCL5 in maternal plasma; IL-6, -12p70, -17, and -13 and tumor necrosis factor-α in myometrium; IL-6, -12p70, and -17 in placenta; and IL-6, tumor necrosis factor-α, CCL3, and CCL4 in amniotic fluid. Maternal plasma progesterone was reduced significantly after lipopolysaccharide injection with and without GR-1 SN pretreatment. There was no difference in fetal sex ratios between mice that delivered preterm and those that did not after lipopolysaccharide and GR-1 SN treatments. The supernatant of probiotic L rhamnosus GR-1 attenuated lipopolysaccharide-induced inflammation and PTB in vivo. GR-1 SN may confer therapeutic benefits in the prevention of infection-associated PTB by controlling systemic and intrauterine inflammation.

Effects of Probiotics on the Recurrence of Bacterial Vaginosis

Bacterial vaginosis (BV) is a common cause of genital discomfort in women in reproductive ages, which causes many complications. Bacterial vaginosis is usually treated by metronidazole and clindamycin. However, this protocol does not prevent its recurrence, which is a main complaint of the patients. The number of lactobacilli in the vagina of women with BV is significantly lower than that in healthy women. Hence, efforts have been made to normalize vaginal flora by oral or vaginal administration of lactobacilli. The objective of the present study was to review clinical evidences available regarding the efficacy of probiotics in the prevention and treatment of BV. Published randomized controlled trials were searched in PubMed, Science Direct, and the Cochrane Database between 1990 and 2011. Search terms included bacterial vaginosis, urinary tract infection, lactobacillus, and probiotics. Orally consumed probiotics are believed to ascend to the vaginal tract after they are excreted from the rectum; vaginal administration allows for direct replacement of the probiotics for unhealthy vaginal microbiota and occupation of specific adhesion sites at the epithelial surface of the urinary tract, which consequently results in maintenance of a low pH and production of antimicrobial substances like acids and hydrogen peroxide. Receiving Lactobacillus acidophilus, Lactobacillus rhamnosus GR-1, and Lactobacillus fermentum RC-14 at a dose of at least 10 CFU/day for 2 months has been shown to present the patients with better results. Although the results of different studies are controversial, most studies have been in favor of the probiotics in the prevention or treatment of BV, and no adverse effects have been reported. Therefore, it may be helpful to recommend daily consumption of probiotic products to improve public health among women.

Microbial Properties of Probiotic Fermented Milk Supplemented with Ginseng Extracts

The therapeutic properties of Lactobacillus rhamnosus GR-1 and ginseng extracts have been well documented. The objective of this study was to measure growth and survival of L. rhamnosus GR-1 in milk fortified with ginseng extracts during storage period of 28 days. Five formulations of milk (1% fat) were prepared. Two samples were fortified with alcoholic ginseng extract (AGE: 150 μg/mL, AGE: 500 μg/mL) and the other two samples were fortified with aqueous ginseng extract (QGE: 150 μg/mL, QGE: 500 μg/ml) and one with no ginseng extract (C). The mixtures were autoclaved for 15 min, cooled to 37?C and inoculated with 1% starter culture. The samples were then incubated anaerobically at 37℃ overnight. Selective MRS agar containing 0.015 g/L fusidic acid was used to enumerate L. rhamnosus GR-1 after 1, 14, and 28 d storage at 4℃. All ginseng-fortified samples supported the growth and survival of L. rhamnosus GR-1, as viable counts were detected at each measurement point throughout the 28 d storage period. After 1 d of storage, total colony counts for L. rhamnosus GR-1 in AGE: 150 μg/mL and AGE: 500 μg/mL were 1.3 × 109 and 1.4 × 109 CFU/mL, respectively; probiotic counts in QGE 150 μg/mL and QGE: 500 μg/mL were 1.5 × 109 and 1.4 × 109 CFU/mL, respectively. Additionally, L. rhamnosus GR-1 viability remained stable throughout the 28 days storage period, as no significant decrease (p L. rhamnosus GR-1 can remain viable in presence of ginseng extracts during storage period and there is potential for incorporating ginseng extracts in other probiotic dairy products.

Fortification of Probiotic Yogurt with Ginseng (Panax quinquefolius) Extract

Fortification of Probiotic Yogurt with Ginseng (Panax quinquefolius) Extract Research suggests that the bioactive components of ginseng can decrease oxidative stress, which is a mechanism associated with prevention of chronic disease development. Since the efficacy of ginseng is dependent on gut microbiota, combining ginseng with a probiotic yogurt may improve microflora health and enhance the health benefits associated with consumption of this herb.To identify if yogurt is a suitable medium for North American Ginseng and probiotic bacteria Lactobacillus rhamnosus GR-1, ginseng fortified probiotic yogurt was developed by inoculating milk with probiotic mother culture, starter cultures, and various concentrations of aqueous ginseng extract. L. rhamnosus GR-1 viability and ginsenoside stability were measured at 1, 14, and 28 days of refrigerated storage through microbial analysis and high performance liquid chromatography, respectively.

In vitro interactions between probiotic bacteria and milk proteins probed by atomic force microscopy

Interactions between microbial cells and milk proteins are important for cell location into dairy matrices. In this study, interactions between two probiotic strains, Lactobacillus rhamnosus GG and Lactobacillus rhamnosus GR-1, and milk proteins (micellar casein, native and denatured whey proteins) were studied. The bacterial surface characterization was realized with X-ray photoelectron spectroscopy (XPS) to evaluate surface composition (in terms of proteins, polysaccharides and lipid-like compounds) and electrophoretic mobility that provide information on surface charge of both bacteria and proteins along the 3–7 pH range. In addition, atomic force microscopy (AFM) enabled the identification of specific interactions between bacteria and whey proteins, in contrast to the observed nonspecific interactions with micellar casein. These specific events appeared to be more important for the GG strain than for the GR-1 strain, showing that matrix interaction is strain-specific. Furthermore, our study highlighted that in addition to the nature of the strains, many other factors influence the bacterial interaction with dairy matrix including the nature of the proteins and the pH of the media.

Substances released from probioticLactobacillus rhamnosusGR-1 potentiate NF-κB activity inEscherichia coli-stimulated urinary bladder cells

Lactobacillus rhamnosus GR-1 is a probiotic bacterium used to maintain urogenital health. The putative mechanism for its probiotic effect is by modulating the host immunity. Urinary tract infections (UTI) are often caused by uropathogenic Escherichia coli that frequently evade or suppress immune responses in the bladder and can target pathways, including nuclear factor-kappaB (NF-κB). We evaluated the role of L.rhamnosus GR-1 on NF-κB activation in E.coli-stimulated bladder cells. Viable L.rhamnosus GR-1 was found to potentiate NF-κB activity in E.coli-stimulated T24 bladder cells, whereas heat-killed lactobacilli demonstrated a marginal increase in NF-κB activity. Surface components released by trypsin- or LiCl treatment, or the resultant heat-killed shaved lactobacilli, had no effect on NF-κB activity. Isolation of released products from L.rhamnosus GR-1 demonstrated that the induction of NF-κB activity was owing to released product(s) with a relatively large native size. Several putative immunomodulatory proteins were identified, namely GroEL, elongation factor Tu and NLP/P60. GroEL and elongation factor Tu have previously been shown to elicit immune responses from human cells. Isolating and using immune-augmenting substances produced by lactobacilli is a novel strategy for the prevention or treatment of UTI caused by immune-evading E.coli.

Micronutrient supplemented probiotic yogurt for HIV-infected adults taking HAART in London, Canada

The Human Immunodeficiency Virus has devastating effects worldwide. The burden is less pronounced, but still present in Canada where approximately 64,000 men and women are HIV positive. The virus and the life-saving antiretroviral therapies often contribute to diarrhea and other gastrointestinal disturbances. Certain probiotic organisms, such as Lactobacillus rhamnosus GR-1, have been shown to alleviate diarrhea as well as delay the decline of CD4 lymphocytes in some subjects. In addition, micronutrient formulae have been used extensively among HIV positive persons as a cost-effective method for improving quality of life and immune function. The objective of the present study was to combine probiotics and micronutrients into an affordable and highly palatable nutritional supplement and assess outcomes in 21 HIV-positive participants receiving highly active antiretroviral therapy in London, Ontario, Canada. The design was a randomized, double blind, three-period, cross-over controlled trial with three different formulations of supplemented yogurt; micronutrient and probiotic (A), micronutrient alone (B) and probiotic alone (C). The period of intake for each of the types was 30 days with a 14 day wash-out period between the intervention types. The mean increase in CD4 was greatest with B (41 cells/µL, SD 221). Supplement A showed a mean change of +19 cells/µL (SD 142) and supplement C a mean change of – 7 cells/µL (SD 154). All yogurt types caused an increase in subjective energy and ability to perform daily activity scores. According to the safety measures taken to assess the tolerance of the yogurt, there were no adverse events and the yogurt was well-tolerated. These preliminary findings suggest that micronutrient supplemented probiotic yogurt may support immune function among people living with HIV.

A review of probiotics studies in HIV research suggests improved immunological presentation and preservation of viral host restrictive factors of TH17 in HIV patients

Background Recovery of gut mucosal immune system is slow and incomplete during HAART therapy, leading to elevated inflammation rates, increased mitochondrial damage and the pathogenesis of replicatively competent escape mutations. A recent study of Ganeden BC30, Bacillus coagulans GBI-30, demonstrated safety, increased CD4+ counts in patients on HARRT, and a significant increase in CD69 and maturation of dendritic cells in vitro. Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 demonstrate efficacy in treating naive HIV patients against gastrointestinal and urogenital infections . Restoration of viral host restrictive factors of Th17 cell lines in GALT may prevent the evolving viral diversity, provoking increased CD4 presentation and regulation of inflammatory cytokine levels responsible for increased viral replication.