Immune System Study

It is probably one of the most complex tasks to understand the mutual relationship between members of the microbial community in the gut and their interaction with the host. Although many animals including humans can live without gut flora, it is beneficial for the growth and protection of the host against pathogens. On the other hand, clinical and experimental intestinal inflammatory reactions have been attributed to immune recognition of the intestinal microflora. This review discusses recent and old findings on function and immunological relevance of the endogenous microflora.

1. Steinhoff U (June 2005). “Who controls the crowd? New findings and old questions about the intestinal microflora”. Immunol. Lett. 99 (1): 12–6.

Connection Between Neuropsychiatry and Gastroenterology Study

Discussion

The idea that implanting the intestines with Lactobacillus strains may improve quality of life and mental health is not a new one. Dr. George Porter Phillips first reported in 1910 that although Lactobacillus tablets and powder were ineffective, a gelatin-whey formula with live lactic acid bacteria improved depressive symptoms in adults with melancholia [15]. In a series of case reports, separate researchers concluded in 1923 that ‘the administration of acidophilus milk is recommended in the treatment of psychoses as a means to physical betterment’ [16]. In this pilot study we found that the oral administration of Lactobacillus casei strain Shirota (LcS, Yakult Honsha, Tokyo, Japan) caused a significant rise in fecal Bifidobacteria spp. and Lactobacillus spp. The rise in Lactobacilli was an expected finding, although the concomitant rise in Bifidobacteria suggests that there may be far reaching effects of oral probiotics on other microbial residents of the gastrointestinal tract. This finding supports previous research showing that the oral administration of Lactobacillus plantarum 299 V caused a significant rise in fecal Bifidobacteria levels [17]. In this case the elevation of Bifidobacteria levels should be considered a positive finding, particularly when considering that Bifidobacteria levels may be low in CFS. Also of relevance is a recent experimental study which has shown that a specific strain of Bifidobacteria can boost plasma tryptophan levels and alter serotonin and dopamine turnover in areas of the brain associated with depression and anxiety [12]. We also found a significant reduction in anxiety scores among those CFS patients consuming the LcS bacteria. The group differences in anxiety are noteworthy since anxiety is a frequent mental health symptom reported by CFS patients.

In recent years the interface between neuropsychiatry and gastroenterology has converged into a new discipline referred to as enteric neuroscience. Emerging studies have shown that intestinal bacteria may directly communicate with the central nervous system by way of the vagal sensory nerve fibers and the peripheral immune system. Indeed, experimental studies have shown that even minute doses of microbes within the gastrointestinal tract, levels that do not trigger an immune response, are capable of influencing neurotransmission in the paraventricular hypothalamus, the central nucleus of the amygdala, and the bed nucleus of the stria terminalis [8]. All three of these regions are involved in the processing of emotions related to anxiety and mood. It is also true that quantitative alterations in the make-up of gastrointestinal microbes are a consequence of states of stress and fear, and alterations in the gut microflora have recently been associated with impaired glucose control and obesity [18, 19].

18. [Bailey MT, Engler H, Sheridan JF: Stress induces the translocation of cutaneous and gastrointestinal microflora to secondary lymphoid organs of C57BL/6 mice. J Neuroimmunol. 2006, 171: 29-37. 10.1016/j.jneuroim.2005.09.008.]
19. [Cani PD, Delzenne NM: Gut microflora as a target for energy and metabolic homeostasis. Curr Opin Clin Nutr Metab Care. 2007, 10: 729-34. 10.1097/MCO.0b013e3282efdebb.]

Mental Health Study

More hints of a connection between intestinal microflora and brain function come from studies in the autistic spectrum. Research has shown marked alterations of the gastrointestinal microflora in autism, with specific elevations in various Clostridium spp. [20]. Some researchers speculate that low-grade chronic intestinal inflammation induced by elevations in bacteria such as potentially pathogenic Clostridium spp may be directly influencing brain centers. Experimental studies show that indeed chronic gut inflammation leads to activation of areas of the brain associated with mental health and behavioral disorders, including the hypothalamus, amygdala and cortical centers [21].

21. [Welch MG, Welch-Horan TB, Anwar M, Anwar N, Ludwig RJ, Ruggiero DA: Brain effects of chronic IBD in areas abnormal in autism and treatment by single neuropeptides secretin and oxytocin. J Mol Neurosci. 2005, 25: 259-74. 10.1385/JMN:25:3:259.]

Anxiety and Aggression Study

While we did not look specifically at Clostridium spp. in this pilot investigation, it has been noted that Lactobacillus can competitively displace Clostridium and other potentially pathogenic gut bacteria [22]. Propionic acid is a short chain fatty acid produced primarily by Clostridium and Bacteroides spp.; emerging research suggests that this acid may be involved in anxiety. Elevated production of propionic acid in the gut has been shown to increase behaviors associated with anxiety and aggression in animals [23]. It has also been shown recently that when propionic acid gains access to the brain it can impair the social behavior of animals. Changes to the animal behavior include decreased playful behavior, increasing social isolation, and an increase in repetitive behaviors that may indicate anxiety [24]. While human data is lacking, a study in animals did show that the LcS as used in our study can lower cecal propionate levels [25].

23. [Hanstock TL, Clayton EH, Li KM, Mallet PE: Anxiety and aggression associated with the fermentation of carbohydrates in the hindguts of rats. Physiol Behav. 2004, 82: 357-68. 10.1016/j.physbeh.2004.04.002.]
24. [Shultz SR, MacFabe DF, Ossenkopp KP, Scratch S, Whelan J, Taylor R, Cain DP: Intercerebroventricular injection of propionic acid, an enteric bacterial metabolic end-product, impairs social behavior in the rat: implications for the animal model of autism. Neuropharmacology. 2008, 54: 901-11. 10.1016/j.neuropharm.2008.01.013.]
25. [Ohashi Y, Tokunaga M, Ushida K: The effect of Lactobacillus casei strain Shirota on the cecal fermentation pattern depends on the individual cecal microflora in pigs. J Nutr Sci Vitaminol (Tokyo). 2004, 50 (6): 399-403.]

Mood and Depression Study

Some researchers have stated that the so-called ‘hygiene hypothesis’ extends into the realm of mental health disorders as well. The hygiene hypothesis is the proposition that the documented rise in chronic inflammatory disorders (allergies, autoimmunity, and inflammatory bowel disease) within developed countries is driven by a changing microbial environment, an absence of beneficial bacteria that has in turn altered the immuno-regulatory circuits which normally keep inflammatory responses in check [26]. Many mental health conditions, and so-called functional somatic disorders such as Chronic Fatigue Syndrome (CFS), have been well-documented to have elevations in inflammatory cytokines, and these inflammatory cytokines at even low levels can produce symptoms of anxiety and depression in otherwise healthy adults [26]. Therefore, since orally administered probiotics can decrease inflammatory cytokines in humans, it has been postulated that bacteria may be used to positively influence mood in patient populations where both emotional symptoms and inflammatory immune chemicals are elevated [10]. It is becoming increasingly clear that anxiety and stress itself may lower levels of fecal lactic acid bacteria, and this, in turn, may compromise various aspects of health [27].

10. [Logan A, Katzman M: Major depressive disorder: probiotics may be an adjuvant therapy. Med Hypotheses. 2005, 64: 533-8. 10.1016/j.mehy.2004.08.019.]
11. [Knowles SR, Nelson EA, Palombo EA: Investigating the role of perceived stress on bacterial flora and salivary cortisol secretion: a possible mechanism underlying susceptibility to illness. Biol Psychol. 2008, 77: 132-37. 10.1016/j.biopsycho.2007.09.010.]

Mood and Cognition Study

Overall the results suggest that specific strains of probiotic bacteria may have a role to play in mediating some of the emotional symptoms of CFS and other related conditions. However, it is important to note that this is a small pilot study and broad conclusions cannot be drawn at this time. Since we did not evaluate bowel function during the study, it is entirely possible that the decreased anxiety was a consequence of improved bowel function. In an unexplained medical condition such as CFS, where over 70% of patients meet the criteria for IBS, it is possible that regulation of bowel movements made a difference in mental state. Indeed LcS has been shown to regulate bowel function and decrease constipation in a controlled trial [28]. It is also true that LcS has been shown to reduce small intestinal bacterial overgrowth and the subjective reporting of the passage of gas in patients with IBS [29]. This is of significance because SIBO and intestinal permeability often overlap, and patients with chronic fatigue syndrome are known to have both increased intestinal permeability and SIBO. Indeed, correction of SIBO and intestinal permeability has been shown to improve symptoms in CFS and depressive disorders [30, 31]. Therefore, it is entirely possible that our results are an artifact of improved gut structure and function via the LcS restoration of a healthy intestinal biofilm. However, a recent study using the same LcS strain in healthy adults suggests that there may be a more direct microbial influence on emotional state. In healthy adults who were reported to be more depressed/less elated in daily functioning at baseline, there was significant improvement in mood scores after taking the probiotic. In that controlled trial the improvements in mood were not related to changes in bowel function [11].

11. [Benton D, Williams C, Brown A: Impact of consuming a milk drink containing a probiotic on mood and cognition. Eur J Clin Nutr. 2007, 61: 355-61. 10.1038/sj.ejcn.1602546.]

Anxiety and Depression Study

This preliminary research raises many questions regarding possible mechanisms whereby probiotics might influence anxiety and depression. The results of the present study should be viewed simply as a stimulus for further research. Follow-up studies with probiotics should further examine specific gut microbes, intestinal structure and function as well as physiological markers associated with anxiety and depression. These may include inflammatory cytokines and other immune chemicals, blood tryptophan levels and urinary metabolites of neurotransmitters.

[3. A Venket Rao, Alison C Bested, Tracey M Beaulne, Martin A Katzman, Christina Iorio5, John M Berardi6 and Alan C Logan. A randomized, double-blind, placebo-controlled pilot study of a probiotic in emotional symptoms of chronic fatigue syndrome; Gut Pathogens 2009, 1:6]

Hypertension Study

Conclusion

This review has illustrated the potential of probiotics in mediating hypertension via positive modulation of several different physiological systems, apart from its conventional benefits for gastrointestinal health. Probiotics have exhibited antihypertensive potential via the improvement of lipid profiles, insulin resistance, modulation of renin and the bioconversion of bioactive isoflavones. These positive findings suggested the potential use of dietary alternatives such as probiotics, to alleviate the occurrence of metabolic diseases via a less radical approach compared to drugs or hormone therapy, with milder, if not none, known side effects. Probiotics could also serve as a complementary supplement to enhance the well-being for those already suffering the diseases and taking drugs or hormonal therapy to medicate the condition. Further revelation on the potential of probiotics in future research could lead to a boost in probiotic-fermented food industries–dairy and non–dairy. Nevertheless, more studies are needed to better understand the exact mechanisms, in vivo target sites, stability and safety, prior to using probiotics as an antihypertensive alternative treatment.

4. [Huey-Shi Lye, Chiu-Yin Kuan, Joo-Ann Ewe, Wai-Yee Fung and Min-Tze Liong. The Improvement of Hypertension by Probiotics: Effects on Cholesterol, Diabetes, Renin, and Phytoestrogens; Int. J. Mol. Sci. 2009, 10, 3755-3775.]

Fever Coughing and Rhinorrhea in Young Children Study

In summary, this paper provides evidence that these probiotic preparations, administered twice daily, at levels of 1010/day, may help reduce the occurrence of symptoms of fever, coughing and rhinorrhea in young children. Whether or not it is practical to administer a dried probiotic to children of this age twice daily over the course of 6 months is another question that must be considered in application of this research, but clearly the effects were demonstrated.

5. [Gregory J. Leyer, PhDa, Shuguang Li, MSb, Mohamed E. Mubasher, PhDc, Cheryl Reifer, PhDd, Arthur C. Ouwehand, PhDe. Probiotic Effects on Cold and Influenza-Like Symptom Incidence and Duration in Children; Pediatrics 2009;124;e172.]

It appears that specific clinical applications of probiotics are safe, effective, and can clearly be recommended. However, the importance of probiotic food items in the “maintenance of health” in healthy individuals as marketed by food industries remains questionable. To date, no generalization can be made from health effects of one probiotic strain to another one and this remains a serious problem within the probiotic research field and its applications. Multi-dimensional research approaches, studying the microflora composition, its metabolic profile, and the impact on host metabolism appear a promising way forward to further describe and explore these complex relationships within the microflora-host “superorganism”.

6. [Silvia Wilson Gratz, Hannu Mykkanen, and Hani S El-Nezami. Probiotics and gut health: A special focus on liver diseases; World J Gastroenterol. 2010 January 28; 16(4): 403–410.]

Food Allergy Study

Conclusion

These results suggest that probiotic bacteria may promote endogenous barrier mechanisms in patients with atopic dermatitis and food allergy, and by alleviating intestinal inflammation, may act as a useful tool in the treatment of food allergy.

9. [Majamaa H, Isolauri E. Probiotics: a novel approach in the management of food allergy; J Allergy Clin Immunol. 1997 Feb;99(2):179-85.]

Here we discuss 1 degrees the specific modulation of the gut microbiota composition by using prebiotics and 2 degrees the novel findings that may explain how gut microbiota can be involved in the development or in the control of obesity and associated low-grade inflammation.

10. [Cani PD, Delzenne NM. Involvement of the gut microbiota in the development of low grade inflammation associated with obesity: focus on this neglected partner; Acta Gastroenterol Belg. 2010 Apr-Jun;73(2):267-9.]

Obesity Study

Obesity results from alterations in the body’s regulation of energy intake, expenditure, and storage. Recent evidence, primarily from investigations in animal models, suggests that the gut microbiota affects nutrient acquisition and energy regulation. Its composition has also been shown to differ in lean vs obese animals and humans. In this article, we review the published evidence supporting the potential role of the gut microbiota in the development of obesity and explore the role that modifying the gut microbiota may play in its future treatment. Evidence suggests that the metabolic activities of the gut microbiota facilitate the extraction of calories from ingested dietary substances and help to store these calories in host adipose tissue for later use. Furthermore, the gut bacterial flora of obese mice and humans include fewer Bacteroidetes and correspondingly more Firmicutes than that of their lean counterparts, suggesting that differences in caloric extraction of ingested food substances may be due to the composition of the gut microbiota. Bacterial lipopolysaccharide derived from the intestinal microbiota may act as a triggering factor linking inflammation to high-fat diet-induced metabolic syndrome. Interactions among microorganisms in the gut appear to have an important role in host energy homeostasis, with hydrogen-oxidizing methanogens enhancing the metabolism of fermentative bacteria. Existing evidence warrants further investigation of the microbial ecology of the human gut and points to modification of the gut microbiota as one means to treat people who are over-weight or obese.

11. [DiBaise JK, Zhang H, Crowell MD, Krajmalnik-Brown R, Decker GA, Rittmann BE. Gut microbiota and its possible relationship with obesity. Mayo Clin Proc. 2008 Apr;83(4):460-9.]

Metabolic Disorders Study

The probiotic LG2055 showed lowering effects on abdominal adiposity, body weight and other measures, suggesting its beneficial influence on metabolic disorders.

12. [Kadooka Y, Sato M, Imaizumi K, Ogawa A, Ikuyama K, Akai Y, Okano M, Kagoshima M, Tsuchida T. Regulation of abdominal adiposity by probiotics (Lactobacillus gasseri SBT2055) in adults with obese tendencies in a randomized controlled trial; Eur J Clin Nutr. 2010 Jun;64(6):636-43.]

Intestinal microflora has metabolic, trophic and protective functions, and can be modified in pathological conditions and by the exogenous administration of probiotics. Probiotics are defined as living microorganisms which resist gastric, bile, and pancreatic secretions, attach to epithelial cells and colonize the human intestine. In the last twenty years research has been focused on the identification of the role of planktonic flora and adhesive bacteria in health and disease, and on the requisite of bacterial strains to become probiotic product which can be marketed. Probiotics can be commercialized either as nutritional supplements, pharmaceuticals or foods, but the marketing as a pharmaceutical product requires significant time, complex and costly research, and the demonstration of a well-defined therapeutic target. This review examines the sequential steps of research which, from the identification of a possible probiotic strain, lead to its production and marketing, summarizing the whole process existing behind its development, through its growth in laboratory, the studies performed to test its resistance to human secretions and stability, microencapsulation technologies, and safety tests.

14. [Del Piano M, Morelli L, Strozzi GP, Allesina S, Barba M, Deidda F, Lorenzini P, Ballaré M, Montino F, Orsello M, Sartori M, Garello E, Carmagnola S, Pagliarulo M, Capurso L. Probiotics: from research to consumer; Dig Liver Dis. 2006 Dec;38 Suppl 2:S248-55.]

Live bacterial cells have been paid considerable attention for treating several diseases including kidney failure uremia, cancer, inflammatory bowel disease, cholesteremia, and others [10- 11, 35, 58]. Probiotic live bacteria are recognized as good or friendly bacteria and thought to reduce potentially harmful bacteria from the intestine [27]. Therefore, these live bacterial microorganisms can improve microbial balances in intestine and exert positive health effects on the host [24]. Lactobacillus and Bifidobacteria are the two most common types of microbes used as probiotics and are extensively investigated for their beneficial importance [72].

Lactobacillus or lactic acid bacteria, a heterogeneous group of Gram-positive, microaerobic, or anaerobic species, are found to be the most beneficial bacteria in the digestive tract.

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Irritable Bowel Syndrome Study

In the past, the relationship between motility and the microbiota was viewed as unidirectional with motility maintaining the sterility of the upper gastrointestinal tract and dysmotility predisposing to small intestinal bacterial overgrowth. This narrow concept has been cast aside with the recognition of the ability of the microbiota to influence the development of normal motor patterns and the potential for disturbances in the microbiota to elicit motor responses or, if sustained, to lead to sensori-motor dysfunction and motility and, perhaps, functional disorders such as IBS. Accordingly, therapeutic approaches which modify the microbiota, including prebiotics, probiotics and antibiotics are being explored in IBS.

20. [Quigley EMM. Microflora Modulation of Motility. J Neurogastroenterol Motil. 2011;17(2):140-147. doi:10.5056/jnm.2011.17.2.140.]

Immune System Study

treatment with IL-33 or transfer of ILC2s ameliorated intestinal disease severity in an AREG-dependent manner. Collectively, these data reveal a critical feedback loop in which cytokine cues from damaged epithelia activate innate immune cells to express growth factors essential for ILC-dependent restoration of epithelial barrier function and maintenance of tissue homeostasis.

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