PROBIOTICS WATCH

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					A SCIENTIFIC TOOL SUPPORTED BY DANONE
                                                                                                                               Issue N°2
                                                                                                                               July 2010

PROBIOTICS WATCH
  Contents :
                           This probiotics watch is designed as a time-saving tool for scientists and clinicians interested in
  EDITO                    probiotic research. In an interactive format, the quarterly report provides timely, quasi-exhaustive
                           lists of the scientific publications of the previous three months. It sorts them by topic and highlights
  THE MAIN POINTS OF       some of the most relevant results. Readers can also check out upcoming scientific events and
  THE QUARTER              regular bibliometric analyses.
 1. CHARACTERISATION OF    Objectivity is a strong commitment, that’s why the articles are selected by an editorial committee,
    PROBIOTICS AND         composed of renowned scientists in the field. Editorial committee members also comment on
    PROBIOTIC FOODS
                           what they believe are the quarter’s most relevant publications.
 2. PHYSIOLOGICAL
                           Last but not least each issue features an editorial by a probiotic expert, which offers special
    INTERACTION WITH THE   insight into this fascinating field of science.
    HOST

 3. IN VIVO OUTCOMES OF
    PROBIOTIC INTAKE
                                   EDITO
  REFERENCES

  KEY DATES                                                CAN WE TRUST META-ANALYSES ON PROBIOTICS?
                                                           by Pr. Hania Szajewska, M.D.
                                                           The Medical University of Warsaw, Poland

                                                           «While systematic reviews, particularly compared to traditional
                                                           review articles (narrative reviews), are generally well accepted,
                                                           a meta-analytical approach has been the subject of many
                                                           controversies...»
                                                                                                                            > Read more




                                    NOTEWORTHY PUBLICATIONS

                                       Metabolic regulations induces in a probiotic during GABA production

                                       Beneficial impact of a probiotic on human intestinal barrier function

                                       Anti-inflammatory effect of DNA from commensal intestinal bacteria – Now does it work?

                                       B. Lactis does not prevent nosocomial infections in very-low-birth-weight infants

                                       Lactobacillus GG prevents nosocomial diseases in hospitalized children

                                       Lactobacillus GG failed to treat diarrhea in aboriginal children

                                       An aditional benefit provided by a probiotic against allergic colitis in infants

                                       A probiotic mixture ameliorates IBS symptoms in children


                                       Perinatal probiotic intake may limit excess weight gain during the first years of life



                           Method
                           > Monitoring period : 18/02/2009 to 18/05/2010 > Database : Medline > Result : 339 publications
                           > Keywords : yogurt / yoghurt / probiotic / lactic acid bacteria / streptococcus thermophilus / lactobacill / fermented
                           milk / bifidobacteria

                                                                                                                                                     1
A SCIENTIFIC TOOL SUPPORTED BY DANONE
                                                                                                                               Issue N°2
                                                                                                                               July 2010

PROBIOTICS WATCH
  Contents :
  EDITO

  THE MAIN POINTS OF
  THE QUARTER
                                                  CAN WE TRUST META-ANALYSES ON PROBIOTICS?
 1. CHARACTERISATION OF
    PROBIOTICS AND                                                       Nowadays, systematic reviews with or without a meta-analysis
    PROBIOTIC FOODS                                                      are a well-established means of reviewing existing evidence and
                                                                         integrating findings from various studies, including those related
 2. PHYSIOLOGICAL                                                        to probiotics. Some such meta-analyses1, 2 are summarized in
    INTERACTION WITH THE                                                 this issue of the Probiotics Watch.
    HOST
                                                                       While systematic reviews, particularly compared to traditional
 3. IN VIVO OUTCOMES OF                                                review articles (narrative reviews), are generally well accepted,
    PROBIOTIC INTAKE                                                   a meta-analytical approach has been the subject of many
                                                                       controversies. Despite some criticisms, the number of meta-
  REFERENCES                                                           analyses is increasing rapidly and they are unlikely to disappear.
                                             Pr. Hania Szajewska, M.D. On the contrary, they continue to gain popularity. Considering
  KEY DATES                                  The Medical University of this, it is important to be aware of the problems and limitations of
                                             Warsaw, Poland            meta-analyses, particularly with regard to probiotics.
                                                                      The analysis of the results of published meta-analyses has
                                                                      revealed that, with few exceptions, probiotics administered for
                                                                      the treatment of a specific disease or condition are all evaluated
   together. One of the most relevant questions is whether it is appropriate to pool data on different probiotic microorganisms. Can we
   trust the results of such meta-analyses?
   There are arguments that support the pooling of data on different probiotics. For example, pooled data on different probiotics allows
   one to establish whether there is evidence of an effect, to determine the direction of the effect, and to determine the size of the
   effect (and the 95% CI around the effect). In addition, pooled data allows one to assess the consistency of the effect across studies.
   If many trials exist involving the administration of different probiotics to different participants, with similar results seen consistently
   in the various trials, the effect of the probiotic(s) has some applicability to a wider population. In addition, pooled data on different
   probiotics is important for demonstrating whether further research on these probiotics is required. If so, this pooled data may
   potentially help to identify the most promising microorganisms as well as the research questions to be addressed in future studies.
   However, there are a number of arguments against the pooling of data. Firstly, there is evidence that the beneficial effects of
   probiotics, particularly the immunomodulatory effects of individual probiotics observed in the host, differ greatly and are strain
   specific.3, 4 Secondly, probiotics vary by organism. In addition to the most commonly used lactic acid bacteria (e.g., lactobacilli,
   bifidobacteria), the yeast Saccharomyces boulardii is often used. All of these microorganisms have different properties and
   antipathogenic mechanisms. Consequently, their efficacy, as well as maybe their safety, may vary.5 Thirdly, the dose of probiotics may
   be important, as documented in a number of studies.6, 7, 8, 9 And finally, the results of some studies do suggest a different response
   to probiotics in different populations.10, 11, 12 Collectively, this data suggests that it is difficult to consider probiotic supplementation
   as a homogeneous intervention. Pooling data from different genera, species, strains, and doses of probiotics obtained in different
   settings and/or populations, presumably with variations in their native intestinal microbiota, may result in misleading conclusions.
   The risk is that the results could be erroneously extrapolated to other probiotics. This concern is shared by many experts in the field
   of probiotics.13
   In the context of the considerations discussed above, an updated meta-analysis of the effects of probiotics in the prevention of
   necrotizing enterocolitis (NEC) in preterm infants published in Pediatrics by Deshpande et al.1 is of major interest.
   NEC is one of the most serious, life-threatening gastrointestinal diseases, and it is characterized by varying degrees of mucosal or
   transmural necrosis of the intestine. The highest incidence is reported in infants with birth weights below 1000 g, and the incidence
   decreases with increasing birth weights.14 The exact cause of NEC remains unclear. However, in addition to prematurity, factors
   such as formula feeding rather than breastfeeding, intestinal hypoxia-ischemia, and colonization with pathogenic microbiota are
   considered to play a role in the pathogenesis of NEC.15 It also has been suggested that the enteral administration of probiotics to
   preterm newborns could prevent infections, prevent NEC, and reduce the use of antibiotics.16
   The updated meta-analysis by Deshpande et al. (search date: March 2009) identified 11 RCTs (N=2176), including four recent trials,
   and involved 2176 preterm infants. The authors concluded that probiotic supplements provide significant benefits in reducing the
   risk of NEC (RR 0.35, 95% CI 0.23 to 0.55) and all-cause mortality (RR 0.42, 95% CI 0.29 to 0.62) in preterm neonates. According to
   the authors, ‘the dramatic effect sizes, tight confidence intervals, extremely low P values, and overall evidence indicate that additional
   placebo-controlled trials are unnecessary if a suitable probiotic product is available’.
   Concurrently with the updated meta-analysis, in the same issue of Pediatrics, two accompanying editorials were published. Their
   titles speak for themselves. Tarnow-Mordi et al.17 stated that when it comes to probiotics and NEC, ‘it is time to change practice.’
   In contrast, Soll asked ‘are we ready for routine use?’18 Also, some rapid responses to the updated meta-analysis suggest that the
   issue remains controversial.19

                                                                                                                                                  2
A SCIENTIFIC TOOL SUPPORTED BY DANONE
                                                                                                                                                                   Issue N°2
                                                                                                                                                                   July 2010

PROBIOTICS WATCH
  Contents :
  EDITO
                                                          From a methodological point of view, the meta-analysis by Deshpande et al. is very well
  THE MAIN POINTS OF                                      done. In my opinion, however, caution must be exercised to avoid over-interpreting the
  THE QUARTER                                             results. The major concern with regard to this meta-analysis, as with many other meta-
                                                          analyses in the area of probiotics, is whether it is appropriate to pool data on different
 1. CHARACTERISATION OF                                   microorganisms. Arguments for and against such an approach were discussed above. All
    PROBIOTICS AND                                        probiotics are not equally effective in preventing NEC. The strong statement in favor of the
    PROBIOTIC FOODS                                       routine use of probiotics for preventing NEC without clearly stating which probiotic(s), at what
                                                          dose, for how long, etc. may be considered by some as beyond the evidence. It is also in
 2. PHYSIOLOGICAL
                                                          contrast to the recent recommendation made by the Committee on Nutrition of the European
    INTERACTION WITH THE
                                                          Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN).20 The
    HOST
                                                          Committee concluded that the data currently available does not allow the recommendation
 3. IN VIVO OUTCOMES OF                                   of the routine use of prebiotics or probiotics as food supplements in preterm infants; the
    PROBIOTIC INTAKE                                      Committee also recommended that each probiotic strain and potential combinations need
                                                          to be characterized separately for each product. The ESPGHAN Committee on Nutrition
  REFERENCES                                              considers it premature to support the routine use of probiotics to prevent NEC. However, it
                                                          also stated that in settings where the incidence of NEC is high, one may consider the use of
  KEY DATES
                                                          probiotics – single or in combination – that are the best studied, with the greatest effect size,
                                                          and the best safety profile.21

                                            So can we trust the results of meta-analyses on probiotics?
                                            Systematic reviews with or without a meta-analysis are now a well-established means of
                                            reviewing existing evidence and integrating findings from various studies. An understanding
   of the strengths and limitations of the meta-analytical approach is needed by everyone involved in decision-making regarding the
   use of probiotics. Meta-analyses on probiotics do provide valid information. However, caution should be exercised so one does
   not over-interpret the results of a meta-analysis when all probiotics have been evaluated together. Given these concerns, the best
   approach to trusting the results of a meta-analysis would be to perform a meta-analysis that evaluates the effect of administering a
   clearly-defined, single-organism, probiotic preparation or an equally well-defined combination of probiotic microorganisms for the
   treatment of a specific disease or condition. Another approach could be, and often is, to perform a review of all probiotics and then
   to perform subgroup analyses based on factors considered to potentially influence the magnitude of the treatment response (e.g.
   the type of probiotic administered, the study population).
   1
      Deshpande G, Rao S, Patole S, Bulsara M. Updated meta-analysis of probiotics for preventing necrotizing enterocolitis in preterm neonates. Pediatrics 2010;125:921-30.
   2
      Moayyedi P, Ford AC, Talley NJ, et al. The efficacy of probiotics in the treatment of irritable bowel syndrome: a systematic review. Gut. 2010 Mar;59(3):325-32
   3
      O’Mahony L, McCarthy J, Kelly P, et al. Lactobacillus and bifidobacterium in irritable bowel syndrome: symptom responses and relationship to cytokine profiles. Gastroenterology
   2005;128:541-51.
   4
      Wickens K, Black PN, Stanley TV, Mitchell E, Fitzharris P, Tannock GW, Purdie G, Crane J; Probiotic Study Group. A differential effect of 2 probiotics in the prevention of eczema
   and atopy: a double-blind, randomized, placebo-controlled trial. J Allergy Clin Immunol 2008;122:788-94.
   5
      Canani RB, Cirillo P, Terrin G, Cesarano L, Spagnuolo MI, De Vincenzo A, Albano F, Passariello A, De Marco G, Manguso F, Guarino A. Probiotics for treatment of acute diarrhoea in
   children: randomised clinical trial of five different preparations. BMJ 2007;335:340.
   6
      Whorwell PJ, Altringer L, Morel J, et al. Efficacy of an encapsulated probiotic Bifidobacterium infantis 35624 in women with irritable bowel syndrome. Am J Gastroenterol
   2006;101:1581-90.
   7
      Basu S, Chatterjee M, Ganguly S, Chandra PK. Efficacy of Lactobacillus rhamnosus GG in acute watery diarrhoea of Indian children: a randomized controlled trial. J Paediatr Child
   Health 2007;43:837-42.
   8
      Basu S, Paul DK, Ganguly S, Chatterjee M, Chandra PK. Efficacy of high-dose Lactobacillus rhamnosus GG in controlling acute watery diarrhea in Indian children: a randomized
   controlled trial. J Clin Gastroenterol 2009;43:208-13.
   9
      Misra S, Sabui TK, Pal NK. A randomized controlled trial to evaluate the efficacy of lactobacillus GG in infantile diarrhea. J Pediatr 2009;155:129-32.
   10
       Vanderhoof JA, Whitney DB, Antonson DL, Hanner TL, Lupo JV, Young RJ. Lactobacillus GG in the prevention of antibiotic-associated diarrhea in children. J Pediatr
   1999;135:564-8.
   11
      Arvola T, Laiho K, Torkkeli S, et al. Prophylactic Lactobacillus GG reduces antibiotic-associated diarrhea in children with respiratory infections: a randomized study. Pediatrics
   1999;104:e64.
   12
      Thomas MR, Litin SC, Osmon DR, Corr AP, Weaver AL, Lohse CM. Lack of effect of Lactobacillus GG on antibiotic-associated diarrhea: a randomized, placebo controlled trial.
   Mayo Clinic Proc 2001;76:883-9.
   13
       Rijkers GT, Bengmark S, Enck P, Haller D, Herz U, Kalliomaki M, Kudo S, Lenoir-Wijnkoop I, Mercenier A, Myllyluoma E, Rabot S, Rafter J, Szajewska H, Watzl B, Wells J, Wolvers
   D, Antoine JM. Guidance for substantiating the evidence for beneficial effects of probiotics: current status and recommendations for future research. J Nutr 2010;140:671S-6S.
   14
      Llanos AR, Moss ME, Pinzòn MC, Dye T, Sinkin RA, Kendig JW. Epidemiology of neonatal necrotizing enterocolitis: a population-based study. Paediatr Perinat Epidemiol
   2002;16:342-9.
   15
       Hsueh W, Caplan MS, Qu XW, Tan XD, De Plaen IG, Gonzalez-Crussi F. Neonatal necrotizing enterocolitis: clinical considerations and pathogenetic concepts. Pediatr Dev Pathol
   2003;6:6–23.
   16
      Caplan MS, Jilling T. Neonatal necrotizing enterocolitis: possible role of probiotic supplementation. J Pediatr Gastroenterol Nutr 2000;30:S18-S22.
   17
       Tarnow-Mordi WO, Wilkinson D, Trivedi A, Brok J. Probiotics reduce all-cause mortality and necrotizing enterocolitis: it is time to change practice. Pediatrics 2010;125:1068-70.
   18
       Soll RF. Probiotics: are we ready for routine use? Pediatrics 2010;125:1071-2.
   19
       Szajewska H, Shamir R. Is a meta-analytical approach appropriate to assess the efficacy of probiotics? http://pediatrics.aappublications.org/cgi/eletters/125/5/921. Accessed on
   24 June 2010.
   20
       ESPGHAN Committee on Nutrition: Agostoni C, Buonocore G, Carnielli VP, De Curtis M, Darmaun D, Decsi T,Domellöf M, Embleton ND, Fusch C, Genzel-Boroviczeny O, Goulet
   O, Kalhan SC, Kolacek S, Koletzko B, Lapillonne A, Mihatsch W, Moreno L, Neu J, Poindexter B, Puntis J, Putet G, Rigo J, Riskin A, Salle B, Sauer P, Shamir R, Szajewska H, Thureen
   P, Turck D, van Goudoever JB, Ziegler EE; ESPGHAN Committee on Nutrition. Enteral nutrient supply for preterm infants: commentary from the European Society of Paediatric
   Gastroenterology, Hepatology and Nutrition Committee on Nutrition. J Pediatr Gastroenterol Nutr 2010;50:85-91.
   21
       van Goudoever JB, Koletzko B, Rigo J, Mihatsch W, Szajewska H, Shamir, on behalf of the ESPGHAN Committee on Nutrition. J Pediatr Gastroenterol Nutr 2010;50:694; author
   reply 694-5. Re: ESPGHAN commentary and education that probiotics substantially reduce all-cause mortality and necrotizing enterocolitis in preterm infants. Tarnow-Mordi W,
   Wilkinson D, Trivedi A, Sinn J, Dutta S, Parikh T, Lin HC. Comment on: J Pediatr Gastroenterol Nutr 2010;50:85-91.

                                                                                                                                                                                           3
 PROBIOTICS
 WATCH
 ISSUE N°2                              THE MAIN POINTS OF THE QUARTER



 Contents :                             Europe : Leader of research on probiotics
 EDITO

 THE MAIN POINTS OF
 THE QUARTER

1. CHARACTERISATION OF
   PROBIOTICS AND
   PROBIOTIC FOODS

2. PHYSIOLOGICAL
   INTERACTION WITH THE
   HOST

3. IN VIVO OUTCOMES OF
   PROBIOTIC INTAKE

 REFERENCES

 KEY DATES
                                        With 158 publications, Europe is still the leading region for scientific production in the field of probiotics.
                                        Within Europe, the top 3 most productive countries are Italy (n=25), Spain (n=21), and France (n=19). The
                                        USA (n=23) is the leader in the Americas. Japan and China (n=24), which are joint leaders in Asia, are
                                        closely followed by South Korea (n=19).



                                        A quarter with many in vitro studies


                                                Meta-analysis 1,1%



                                                Clinical
                                                16,2%

                                                                  In vitro
                                             Animal               64,0%
                                             18,7%                                 During the second quarter of 2010, 339 publications were
                                                                                   retrieved from Medline, including 18% of reviews. This survey
                                                                                   period, like the preceding ones, is characterised by the
                                                                                   publication of a majority of in vitro studies.




                                        Nearly 50% of the articles are related to probiotics characterisation

                                            46,8%                                  Most of the 278 articles retrieved from Medline were related to
                                                                                   probiotic characterisation (strain identification, characterisation,
                                                                                   metabolism and physiology; and technology of probiotic and
                                                                                   probiotic foods). The papers dealing with in vivo and clinical
                                                                                   effects (infections, allergy & atopy, IBS & functional GI disorders,
                                                                                   IBD and other inflammatory diseases, nutrition, etc.) represent
         Characterisation of
         probiotics & probiotic foods
                                                                                   24.5% of the total number of published articles. A third type
                                                      31,7%                        of study assesses (31.7%) the physiological interaction of
         Physiological interaction                                                 probiotics with the host (survival & physical interactions of
         with the host                                          24,5%
                                                                                   probiotics with mucosa, interaction with pathogens, mucosa
         In vivo outcomes of                                                       response to probiotics, effects on microbiota and immunity,
         probiotic intake
                                                                                   etc.)


                                                                                                                                                    4
   PROBIOTICS
   WATCH
   ISSUE N°2                          1. CHARACTERISATION OF PROBIOTICS AND PROBIOTIC FOODS



   Contents :
                                      STRAIN METABOLISM AND PHYSIOLOGY
   EDITO
                                                 Metabolic regulations induced in a probiotic during GABA production
   THE MAIN POINTS OF
   THE QUARTER

 1. CHARACTERISATION OF               OBJECTIVES
    PROBIOTICS AND                    Biogenic amines can be found in a great number of different foods, above all those involving a microbial
    PROBIOTIC FOODS                   fermentation process. High amounts of these compounds, in particular histamine and tyramine, are toxic
                                      for consumers. Different from the other biogenic amines, -aminobutyric acid (GABA) is known to have
 2. PHYSIOLOGICAL                     positive effects on human health. GABA is of increasing nutraceutical interest due to its modulatory
    INTERACTION WITH THE
    HOST                              activity on the central nervous system and smooth muscle relaxation. Probiotic bacteria can produce it
                                      by glutamate decarboxylation; the aim of the present research was to study the metabolic modifications
 3. IN VIVO OUTCOMES OF               occurring at microbial level during GABA production.
    PROBIOTIC INTAKE
                                      METHODS & MEASURES
   REFERENCES                         A GABA-producing Lactococcus lactis NCDO2118 strain grown in a medium supplemented with or
                                      without glutamate was studied using a combined transcriptome/proteome analysis.
   KEY DATES
                                      RESULTS
                                      A tenfold increase in GABA production in the glutamate medium was observed only during the stationary
                                      phase and at low pH. About 30 genes and/or proteins were shown to be differentially expressed in
                                      glutamate-stimulated conditions as compared to control conditions, and the modulation exerted by
                                      glutamate on entire metabolic pathways was highlighted by the complementary nature of transcriptomics
                                      and proteomics. Most glutamate-induced responses consisted of the under-expression of metabolic
                                      pathways, with the exception of glycolysis where either over- or under-expression of specific genes was
                                      observed. The energy-producing arginine deiminase pathway, the ATPase, and also some stress proteins
                                      were down-regulated, suggesting that glutamate is not only an alternative means to get energy, but also
                                      a protective agent against stress for the strain studied.

                                      CONCLUSION
                                      The present investigation has shed light on some metabolic modulations occurring in L. lactis NCDO
                                      2118 during GABA production, revealing that no significant stimulation of pathways generating toxic
                                      metabolites takes place. On the contrary it seems that the ammonia-generating arginine deiminase route
                                      is lightly negatively regulated, thus creating the possibility of using this strain for probiotic applications.
                                      Mazzoli R, Pessione E, Dufour M, Laroute V, Giuffrida MG, Giunta C, Cocaign-Bousquet M, Loubière P. Glutamate-induced metabolic changes in
                                      Lactococcus lactis NCDO 2118 during GABA production: combined transcriptomic and proteomic analysis. Amino Acids. 2010 Feb 21.


                                                 COMMENTARY

                                          from James Versalovic, Baylor College of Medicine, Houston, US
                                          «This manuscript describes the changes in gene expression that occur in Lactococcus lactis in the presence of
                                          the amino acid, glutamate. Glutamate serves as a precursor to the biogenic amine and neurotransmitter, gamma-
> The studies performed in humans
                                          aminobutyric acid (GABA). The study describes the pattern of GABA production by lactic acid bacteria (LAB),
are highlighted using the following
                                          in this case Lactococcus lactis, in order to understand how probiotics may function in the presence of amino
symbols :
                                          acids and biogenic amines in vivo. GABA was only produced in the late stationary phase and at low pH (around
                                          4.4). Despite the presence of glutamate in the medium, the LAB only produced significant amounts of GABA at
       for the GENERAL POPULATION         increased cell densities and acidic pH. GABA production clearly depended on the presence of glutamate in the
                                          media, as GABA quantities in the media were markedly enhanced in glutamate-fortified media.
                                          The comprehensive assessment of L. lactis by combined transcriptomics and proteomics analyses has
       for CHILDREN                       augmented our understanding of metabolic pathways and the physiology of this industrially important LAB.
                                          One key benefit of this study is that new proteins of L. lactis were identified, although most of them were not
                                          differentially produced in glutamate-fortified media. Interestingly, although this bacterium contains more than
       for the ELDERLY                    2200 genes, only 27 genes and 30 proteins were significantly altered as evaluated respectively by microarray
                                          and proteomic studies. Glutamate decarboxylase genes and enzymes were not significantly affected, despite
                                          the fact that this enzyme is responsible for the conversion of glutamate to GABA. It appears that catalytic
In VITRO/ANIMALS                          activation of this enzyme results in the increased synthesis of GABA during stationary phase. Cytosolic and
studies are highlighted                   membrane proteins were studied by proteomics approaches using two-dimensional gels. Most glutamate-
with the symbol :                         induced responses consisted of the under-expression of genes and proteins in metabolic pathways including
                                          arginine deiminase, pyrimidine biosynthesis and ATPase. Glycolysis was differentially regulated, either induced
                                          or suppressed, at discrete steps in the pathway. Interestingly, only six genes demonstrated to be consistently
                                          up- or downregulated by transcriptomics and proteomics. In summary, production of neurotransmitters and
                                          biogenic amines by LAB can be enhanced by the presence of precursors such as dietary amino acids.»



                                                                                                                                                                            5
 PROBIOTICS
 WATCH
 ISSUE N°2                2. PHYSIOLOGICAL INTERACTION WITH THE HOST



 Contents :
                          MUCOSA RESPONSE TO PROBIOTICS (TROPHICITY & STRUCTURE)
 EDITO
                                      Beneficial impact of a probiotic on human intestinal barrier function
 THE MAIN POINTS OF
 THE QUARTER
                          OBJECTIVES
1. CHARACTERISATION OF    Increased permeability of the intestinal epithelium or “leaky gut” is now recognized as having a role in
   PROBIOTICS AND         the pathophysiology of a variety of gastrointestinal disorders. Consequently, the modulation of epithelial
   PROBIOTIC FOODS
                          permeability is a highly relevant target for novel therapeutic or prophylactic treatments against a range
2. PHYSIOLOGICAL          of diseases. The objective of this study was to assess the effect of Lactobacillus plantarum WCFS1 on
   INTERACTION WITH THE   human barrier function.
   HOST
                          METHODS & MEASURES
3. IN VIVO OUTCOMES OF    Seven healthy nonsmoking subjects participated in a double-blind randomized crossover study. After
   PROBIOTIC INTAKE       an overnight fast, an intraduodenal feeding catheter was placed orogastrically for continuous probiotic
                          injection for 6 hours. Food or beverage consumption was not allowed during the experiment. After this
 REFERENCES
                          6-hour period, tissue samples were obtained from the horizontal part of the duodenum.
 KEY DATES                RESULTS
                          After administration of L. plantarum, the scaffold protein zonula occludens (ZO)-1 and transmembrane
                          protein occludin were found to be significantly increased in the vicinity of the tight-junction structures,
                          which form the paracellular seal between cells of the epithelium. In an in vitro model of the human
                          epithelium (Caco-2 cells), L. plantarum induced translocation of ZO-1 to the TJ region; however, the
                          effects on occludin were minor compared with those seen in vivo. L. plantarum was shown to activate Toll-
                          like receptor 2 (TLR2) signaling, and treatment of Caco-2 monolayers with the TLR2 agonist Pam3-Cys-
                          SK4 (PCSK) significantly increased fluorescent staining of occludin in the tight-junctions. Pretreatment
                          of Caco-2 monolayers with L. plantarum or PCSK significantly attenuated the effects of phorbol ester-
                          induced dislocation of ZO-1 and occludin and the associated increase in epithelial permeability.

                          CONCLUSION
                          This study demonstrates that, in healthy subjects, the administration of L. plantarum WCFS1 induced
                          changes in the epithelial tight junctions of the intestine, resulting in increased staining of the scaffold
                          protein ZO-1 and the transmembrane protein occludin. Probiotic-induced translocation of ZO-1 to
                          the tight-junction region was also seen in an in vitro model, and this significantly protected against
                          chemically induced disruption of the tight junction and the associated increase in epithelial permeability.
                          The mechanism was shown to be dependent on TLR2 signaling and highlights the homeostatic role of
                          innate signaling pathways in maintaining human intestinal epithelial barrier functions.
                          Karczewski J, Troost FJ, Konings I, Dekker J, Kleerebezem M, Brummer RJ, Wells JM. Regulation of human epithelial tight junction proteins
                          by Lactobacillus plantarum in vivo and protective effects on the epithelial barrier. Am J Physiol Gastrointest Liver Physiol. 2010 Mar 11;298:
                          G851-G859.




                                     COMMENTARY

                              from Bruno Pot, Institut Pasteur de Lille, France
                              «The manuscript by Karczewski and his colleagues is interesting for several reasons. First of all, it shows that
                              probiotic administration (in casu of L. plantarum WCFS1) to healthy persons can actually induce measurable
                              beneficial effects: changes in the epithelial tight junctions (TJ) resulted in increased staining of the proteins
                              ZO-1 and occludin in the duodenum of healthy subjects. Secondly, it illustrates that, at least in this case,
                              it is possible to use in vitro models to select strains with an in vivo activity, as the same probiotic strain, in
                              an in vitro model of human epithelial cells, was shown to induce TJ modifications which protected against
                              chemically-induced disruption of the paracellular seal between epithelial cells and thereby decreased epithelial
                              permeability. And finally, the study suggested that the underlying mechanism for this improved barrier, although
                              not completely clarified at this stage, may involve TLR2 receptors of the gut epithelium. The latter finding is
                              interesting as some immunological effects of probiotics also involve TLR2 signaling on immune cells, raising
                              some new questions on the possible mechanistic link between both probiotic functionalities and the
                              role of TLR2 in the control of intestinal homeostasis.»




                                                                                                                                                                    6
 PROBIOTICS
 WATCH
 ISSUE N°2                2. PHYSIOLOGICAL INTERACTION WITH THE HOST



 Contents :
                                     Anti-inflammatory effect of DNA from commensal intestinal bacteria
 EDITO                               – How does it work?
 THE MAIN POINTS OF       OBJECTIVES
 THE QUARTER              The aim of this study was to test the effect of natural commensal-origin DNA on the toll-like receptors
                          TLR9 signaling cascade and the barrier integrity of polarized intestinal epithelial cells (IECs).
1. CHARACTERISATION OF
   PROBIOTICS AND         METHODS & MEASURES
   PROBIOTIC FOODS        Polarized HT-29 and T84 cells were treated with TNF-alpha in the presence or absence of DNA from
                          Lactobacillus rhamnosus GG (LGG) and Bifidobacterium longum. TLR9 and IL-8 mRNA expression was
2. PHYSIOLOGICAL
   INTERACTION WITH THE
                          assessed by semiquantitative and TaqMan real-time RT-PCR. Expression of TLR9 protein, degradation
   HOST                   of inhibitor of kappa B alpha (IkappaBalpha), and p38 mitogen-activated protein kinase (p38 MAP)
                          phosphorylation were assessed by Western blotting. To further reveal the role of TLR9 signaling, the TLR9
3. IN VIVO OUTCOMES OF    gene was silenced by siRNA. IL-8 secretion was measured by ELISA. Nuclear factor-kappa B (NF-kappaB)
   PROBIOTIC INTAKE       activity was assessed by the electrophoretic mobility shift assay and NF-kappaB-dependent luciferase
                          reporter gene assays. As an indicator of tight junction formation and monolayer integrity of epithelial
 REFERENCES
                          cell monolayers, transepithelial electrical resistance (TER) was repetitively monitored. Transmonolayer
                          movement of natural commensal-origin DNA across monolayers was monitored using qRT-PCR and
 KEY DATES
                          nested PCR based on bacterial 16S rRNA genes. Experiments for gene silencing and TLR9-associated
                          intracellular signalling cascade, focused on LGG DNA although preliminary results showed that both
                          strains of probiotics affected TLR9 expression and IL-8 secretion.

                          RESULTS
                          In response to apically applied natural commensal-origin DNA, HT-29 and T84 cells enhanced expression
                          of TLR9 in a specific manner, which was subsequently associated with the attenuation of TNF-alpha-
                          induced NF-kappaB activation and NF-kappaB-mediated IL-8 expression. TLR9 silencing abolished
                          this inhibitory effect. Apically applied LGG DNA attenuated TNF-alpha-enhanced NF-kappaB activity
                          by reducing IkappaBalpha degradation and p38 phosphorylation. LGG DNA did not decrease the TER
                          but rather diminished the TNF-alpha-induced TER reduction. Translocation of natural commensal-origin
                          DNA into basolateral compartments did not occur under tested conditions.

                          CONCLUSION
                          These results indicate that TLR9 signaling mediates, at least in part, the anti-inflammatory effects of
                          natural commensal-origin DNA on the gut because TLR9 silencing abolished the inhibitory effect of
                          natural commensal-origin DNA on TNF-alpha-induced IL-8 secretion in polarized IECs. The nature of the
                          TLR9 agonist, the polarity of cells, and the tight junction integrity of IECs has to be taken into account in
                          order to predict the outcome of TLR9 signaling.
                          Ghadimi D, Vrese M, Heller KJ, Schrezenmeir J. Effect of natural commensal-origin DNA on toll-like receptor 9 (TLR9) signaling cascade,
                          chemokine IL-8 expression, and barrier integritiy of polarized intestinal epithelial cells. Inflamm Bowel Dis. 2010 Mar;16(3):410-27.




                                    COMMENTARY
                             from Bruno Pot, Institut Pasteur de Lille, France
                             «Already in 2004, Daniel Rachmilewitz and his colleagues suggested a role for methylated and unmethylated
                             genomic DNA, isolated from probiotics, in lowering inflammation in mice (Gastroenterology 126, p 520-528).
                             Since then, the topic has been controversial, especially in understanding the overall impact on the immune
                             system of the DNA of 1014 commensal microorganisms from our microbiome. In an attempt to help understand
                             the mechanisms, this study used an interesting approach to evaluate the effect of DNA from natural commensal-
                             origin on the TLR9 signaling cascade and the barrier integrity of polarized IECs. The (partial) role of TLR9
                             signaling in anti-inflammatory effects of natural commensal-origin DNA was confirmed, as TLR9 silencing
                             abolished the inhibitory effect of the DNA on TNF-alpha-induced IL-8 secretion in polarized IECs. The authors
                             furthermore show that the nature of the TLR9 agonist, the polarity of cells, and the tight junction integrity of
                             IECs had to be taken into account in order to predict the outcome of TLR9 signaling.»




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                          3.1. INFECTIONS (GI, RESPIRATORY INCL. FLU, GENITO-URINARY, OTHER)
 EDITO
                                     B. Lactis does not prevent nosocomial infections in very-low-birth-
 THE MAIN POINTS OF                  weight infants
 THE QUARTER

1. CHARACTERISATION OF
                          OBJECTIVES/BACKGROUND
   PROBIOTICS AND         Nosocomial infections are a significant cause of morbidity and mortality in infants with very low birth
   PROBIOTIC FOODS        weight (VLBW). Several pieces of data support the hypothesis that probiotics prevent nosocomial
                          diseases. The aim of this clinical trial was to investigate whether Bifidobacterium lactis reduces the
2. PHYSIOLOGICAL          incidence of nosocomial infections in infants with very low birth weights (<1,500 g).
   INTERACTION WITH THE
   HOST                   DESIGN
3. IN VIVO OUTCOMES OF
                          Randomized, placebo-controlled trial.
   PROBIOTIC INTAKE
                          SETTINGS & PARTICIPANTS
 REFERENCES               183 VLBW infants were stratified according to gestational age (23-26 and 27-29 weeks) and early
                          antibiotic therapy (days 1-3, yes or no) and randomly assigned to have their milk feedings supplemented
 KEY DATES                with B. lactis (n=93) or placebo (n=90) for the first 6 weeks of life.

                          MAIN OUTCOME
                          The main outcome was the incidence of nosocomial diseases; the main secondary outcome was the
                          incidence of necrotizing enterocolitis (NEC; ≥stage 2).

                          RESULTS
                          There was no significant difference between the two groups with regard to the incidence density of
                          nosocomial infections (0.021 vs. 0.016; P=0.9). There were two cases of NEC in the B. lactis group and
                          four in the placebo group. None of the blood cultures grew B. lactis.

                          CONCLUSION
                          In the present setting, B. lactis at a dosage of 12 billion CFU/kg/day did not reduce the incidence density
                          of nosocomial infections in VLBW infants. No adverse effect of the probiotic was observed.

                          Mihatsch WA, Vossbeck S, Eikmanns B, Hoegel J, Pohlandt F. Effect of Bifidobacterium lactis on the Incidence of Nosocomial Infections in Very-
                          Low-Birth-Weight Infants: A Randomized Controlled Trial. Neonatology. 2010 Mar 16;98(2):156-163.




                                     COMMENTARY

                              from Hania Szajewska, The Medical University of Warsaw, Poland
                              «The strengths of the study by Mihatsch et al. include adequate sequence generation, blinding, intention-to-
                              treat analysis, and the selection of a clinically-relevant primary outcome. One potential limitation is the unclear
                              allocation concealment.
                              The study by Mihatsch et al. is the largest of those aimed at evaluating the efficacy and safety of B. lactis
                              (currently known as Bifidobacterium animalis ssp. lactis CNCMI-3446i ) supplementation in preterm infants.
                              With regard to clinically-relevant outcomes, a recent systematic reviewii of these studies found that, compared
                              with controls, B. lactis supplementation had no effect on the risk of sepsis (2 randomized controlled trials
                              [RCTs], 397 cultures, RR 0.6, 95% CI 0.07 to 5.2), the use of antibiotics (2 RCTs, n=255, RR 0.67, 95% CI 0.28
                              to 1.62), and the risk of necrotizing enterocolitis (NEC) stage ≥2 (3 RCTs, n=293, RR 0.53, 95% CI 0.16 to 1.83).
                              However, the power of these studies does not allow for a definitive statement regarding a reduced risk of NEC.
                              Given the positive results with other probiotics, as documented in a recent meta-analysis by Desphande et al. iii,
                              a large RCT to investigate the efficacy of B. lactis supplementation in the prevention of severe-stage NEC is
                              worthwhile. Importantly, no adverse events associated with B. lactis supplementation in preterm infants have
                              been reported.»
                             i
                                 Masco L, Ventura M, Zink R, et al. Polyphasic taxonomic analysis of Bifidobacterium animalis and Bifidobacterium lactis reveals
                             relatedness at the subspecies level: reclassification of Bifidobacterium animalis as Bifidobacterium animalis subsp. animalis subsp. nov.
                             and Bifidobacterium lactis as Bifidobacterium animalis subsp. lactis subsp. nov. Int J Syst Evol Microbiol 2004;54:1137-43.
                             ii
                                 Szajewska H, Guandalini S, Morelli L, Van Goudoever JB, Walker A. Effect of Bifidobacterium animalis subsp lactis supplementation
                             in preterm infants: A systematic review of randomized controlled trials. J Pediatr Gastroenterol Nutr 2010 Jun 9. [Epub ahead of print]
                             PubMed PMID: 20543719.
                             iii
                                 Deshpande G, Rao S, Patole S, Bulsara M. Updated meta-analysis of probiotics for preventing necrotizing enterocolitis
                             in preterm neonates. Pediatrics 2010;125:921-30.




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                                     Lactobacillus GG prevents nosocomial diseases in hospitalized
 EDITO                               children
                          OBJECTIVES / BACKGROUND
 THE MAIN POINTS OF
                          The incidence of nosocomial infections, predominantly gastrointestinal and respiratory, in children in
 THE QUARTER
                          developed countries is high. There is no effective strategy for preventing these infections. The objective
                          of this clinical trial was to investigate the role of Lactobacillus casei GG (LGG) in preventing nosocomial
1. CHARACTERISATION OF
   PROBIOTICS AND
                          gastrointestinal and respiratory tract infections in a pediatric hospital.
   PROBIOTIC FOODS        DESIGN
2. PHYSIOLOGICAL          Randomized, double-blind, placebo-controlled trial.
   INTERACTION WITH THE
                          SETTINGS & PARTICIPANTS
   HOST
                          742 hospitalized children were randomly allocated to receive a fermented milk containing LGG (n=376;
3. IN VIVO OUTCOMES OF    mean age =9.9±5.1 yrs) or placebo that was the same postpasteurized fermented milk product without
   PROBIOTIC INTAKE       LGG (n=366; mean age = 10.6±5.0 yrs). Both study products were administered for the duration of the
                          hospitalization.
 REFERENCES               MAIN OUTCOME
                          The primary end points were: gastrointestinal tract infections and respiratory tract infections, including
 KEY DATES                upper and lower respiratory tract infections. Secondary end points were number of: vomiting
                          episodes; diarrheal episodes; gastrointestinal infections; respiratory tract infections; and duration of
                          hospitalization.
                          RESULTS
                          The LGG group, compared with the placebo group, shows a significantly reduced risk for gastrointestinal
                          and respiratory infections as well as for episodes of vomiting, diarrhea, and gastrointestinal infections.
                          Groups did not differ in hospitalization duration (P=0.1).




                          CONCLUSION
                          The results suggest that Lactobacillus GG administration decreases the risk of nosocomial gastrointestinal
                          and respiratory tract infections in hospitalized children.
                          Hojsak I, Abdovic S, Szajewska H, Milosevic M, Krznaric Z, Kolacek S. Lactobacillus GG in the prevention of nosocomial gastrointestinal and
                          respiratory tract infections. Pediatrics. 2010 May;125(5):e1171-7.




                                    COMMENTARY
                             from James Versalovic, Baylor College of Medicine, Houston, US
                             «This manuscript describes the application of Lactobacillus rhamnosus GG (LGG) for the prevention of
                             nosocomial gastrointestinal and respiratory infections in children older than 12 months of age. In this study,
                             hospitalized children in the Children’s Hospital of Zagreb were selected to receive either an oral dose of LGG
                             in fermented milk (109 cfu per day) or placebo for the duration of the hospitalization. The children were then
                             followed for the incidence of gastrointestinal infection (diarrheal illness), respiratory tract infection (upper
                             or lower respiratory tract), and individual episodes of diarrhea or vomiting. Ingestion of the probiotic LGG
                             prevented both gastrointestinal and respiratory tract infections during hospitalization. Using the chi-squared
                             test, the relative risks were 0.40 and 0.38 for gastrointestinal infections and respiratory infections, respectively,
                             acquired during hospitalization. Using Cox proportional-hazards regression model analysis, the patients who
                             received placebo were almost 3 times more likely to contract a gastrointestinal infection and 3.17 times more
                             likely to contract a respiratory infection.
                             These infections prolonged hospitalization so that probiotic LGG could prevent nosocomial infections and
                             reduce length of stay. The length of stay data was not formally analyzed and could be addressed in follow-up
                             studies. No etiologic agents were identified in most cases, although the disease patterns were consistent with
                             viral infections. Similar findings have been found in other studies, so prophylactic administration of probiotic
                             LGG and other strains could be an effective strategy for the prevention of nosocomial infections. Additional
                             studies should address the issue of nosocomial infections in infants, as this study did not include infants. This
                             study was relatively large in terms of the number of patients (376 and 366 children in treatment and placebo
                             groups, respectively), so the study size was not limiting and this report can be viewed as an authoritative
                             reference for studies on probiotics in children.»

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 Contents :
                                     Lactobacillus GG failed to treat diarrhea in aboriginal children
 EDITO

 THE MAIN POINTS OF       OBJECTIVES / BACKGROUND
 THE QUARTER              The effectiveness of probiotic therapy for acute rotavirus infectious diarrhea in an indigenous setting
                          with bacterial/parasitic diarrhea is unclear. This study assessed the efficacy of probiotics in Australian
1. CHARACTERISATION OF    Aboriginal children admitted to hospital with diarrheal disease.
   PROBIOTICS AND
   PROBIOTIC FOODS        DESIGN
                          Randomised, double-blind, placebo-controlled study.
2. PHYSIOLOGICAL
   INTERACTION WITH THE   SETTINGS & PARTICIPANTS
   HOST                   Aboriginal children (ages 4 months-2 years), admitted to hospital with acute diarrheal disease (>3 loose
                          stools per day) received either capsules of Lactobacillus casei GG (n = 33) or placebo (n =31) for 3
3. IN VIVO OUTCOMES OF    days.
   PROBIOTIC INTAKE
                          MAIN OUTCOME
 REFERENCES               The primary outcome is definied as the short-term change in small intestinal absorption capacity during
                          the study intervention. This was assessed by a noninvasive C-sucrose breath test on days 1 and 4.
 KEY DATES                Secondary endpoints were: duration and frequency of diarrhea, total stool output, and proportion of
                          subjects with diarrhea on days 3 and 4.
                          RESULTS
                          Both groups showed a mean improvement in the sucrose breath test after 4 days; however, there was
                          no difference between probiotic and placebo groups. Probiotics did not change the duration of diarrhea,
                          total diarrhea stools, or diarrhea score compared with placebo. There was a significant (P<0.05) difference
                          in diarrhea frequency on day 2 between probiotics (3.3 [loose stools]; 2.5-4.3) and placebo (4.7; 3.8-5.7)
                          groups.
                          CONCLUSION
                          Lactobacillus casei GG had no significant impact on minimising any of the important diarrhea symptoms
                          in this setting. The strengths of this trial were to include children with nongastrointestinal infectious
                          cormorbidity and malnutrition, commonly seen in the developing world setting; no selective exclusion of
                          participants based on the presence or type of enteric pathogens identified. A limitation of this study was
                          the small number of participants enrolled.
                          Ritchie BK, Brewster DR, Tran CD, Davidson GP, McNeil Y, Butler RN. Efficacy of Lactobacillus GG in Aboriginal Children With Acute Diarrheal
                          Disease: A Randomised Clinical Trial. J Pediatr Gastroenterol Nutr. June 2010;50(6):619–624.



                                     COMMENTARY

                              from Hania Szajewska, The Medical University of Warsaw, Poland
                              «According to recent European evidence-based guidelines, Lactobacillus rhamnosus GG (LGG) has proven
                              efficacy and may be used as an adjuvent to rehydration therapy for the management of acute gastroenteritis
                              in children.iv However, the study by Ritchie et al. found that LGG at the dose 5 x 109 CFU 3 times per day
                              for 3 days, compared with placebo, administered to Aboriginal children with acute diarrhea resulted in no
                              significant differences in the number of outcomes related to diarrhea. Strengths of the study include appropriate
                              randomization and allocation concealment, and intention-to-treat analysis. Perhaps the most important
                              limitation is the small sample size. The study was underpowered for addressing clinical diarrheal outcomes.

                              Still, this negative study is important as it was carried out in children with malnutrition and underlying
                              environmental enteropathy. Almost all positive studies with LGG were conducted in otherwise healthy children,
                              living in developed countries. In addition to the small sample size, this difference in the study population may
                              be one of the factors that could have contributed to the lack of effect. Another factor that may explain lack of
                              effect may be the inadequate dosage of LGG. Recent studies suggest that the minimal dose is needed. Once
                              a therapeutic effect is achieved, there is no need to further increase the dose.v vi vii Finally, the duration of LGG
                              supplementation was shorter (3 days) than in many of the earlier studies (on average, 5-7 days).viii

                              If confirmed, the trial by Ritchie et al., along with the other recent negative studies carried out in less affluent
                              populations,vii highlights the need for careful consideration of the patient population when making decisions
                              regarding the choice of probiotic(s) for the management of children with acute diarrhea. This study adds to
                              our knowledge that the effects of probiotics are not only strain specific, but also population specific, and they
                              cannot be generalized.»
                              iv
                                   Guarino A, Albano F, Ashkenazi S, Gendrel D, Hoekstra JH, Shamir R, Szajewska H; European Society for Paediatric Gastroenterology,
                              Hepatology, and Nutrition; European Society for Paediatric Infectious Diseases. European Society for Paediatric Gastroenterology,
                              Hepatology, and Nutrition/European Society for Paediatric Infectious Diseases evidence-based guidelines for the management of acute
                              gastroenteritis in children in Europe. J Pediatr Gastroenterol Nutr 2008;46 Suppl 2:S81-122.
                              v
                                  Basu S, Chatterjee M, Ganguly S, Chandra PK. Efficacy of Lactobacillus rhamnosus GG in acute watery diarrhea of Indian children: a
                              randomized controlled trial. J Paediatr Child Health 2007;43:837-42.
                              vi
                                  Basu S, Paul DK, Ganguly S, Chatterjee M, Chandra PK. Efficacy of high-dose Lactobacillus rhamnosus GG in controlling acute watery
                              diarrhea in Indian children: a randomized controlled trial. J Clin Gastroenterol 2009;43:208-13.
                              vii
                                   Misra S, Sabui TK, Pal NK. A randomized controlled trial to evaluate the efficacy of lactobacillus GG in infantile diarrhea. J Pediatr
                              2009;155:129-32.
                              viii
                                   Szajewska H, Skórka A, Ruszczynski M, Gieruszczak-Białek D. Meta-analysis: Lactobacillus GG for treating acute diarrhea in
                              children. Aliment Pharmacol Ther 2007;25:871-81.


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                                     A study does not support the value of a probiotic yogurt in reducing
 EDITO                               illness in children
 THE MAIN POINTS OF
 THE QUARTER              OBJECTIVES / BACKGROUND
                          This clinical trial tried to determine if consumption of fermented milk improves health in children aged 1-3
1. CHARACTERISATION OF    years attending daycare/school centres.
   PROBIOTICS AND
   PROBIOTIC FOODS        DESIGN
                          Double-blind, randomized, placebo-controlled trial.
2. PHYSIOLOGICAL
   INTERACTION WITH THE   SETTINGS & PARTICIPANTS
   HOST                   182 healthy children (age of 1-3 yrs) attending daycare/school at least 3 days a week were administered
                          active or control drink for 90 days (4 ounces/d). Active and control drinks combined a blend of
3. IN VIVO OUTCOMES OF    Streptococcus thermophilus and Lactobacillus bulgaricus; both drinks were identical except that prior to
   PROBIOTIC INTAKE       packaging the active one was supplemented with the probiotic Bifidobacterium lactis BB-12.
 REFERENCES               MAIN OUTCOME
                          Primary outcome was to determine if consumption of the probiotic decreases absences due to illnesses
 KEY DATES                from daycare. Secondary was to determine if the probiotic improves overall parental satisfaction due to
                          decreased absences from work and an overall healthier child.

                          RESULTS
                          There were no significant differences in the number of school days missed per group, with 51.9% in
                          the active group and 47.1% in the placebo group missing at least 1 day of school throughout the study.
                          Additionally, there were no differences in any secondary outcomes among the groups.

                          CONCLUSION
                          Consumption of a yogurt-based drink delivering B. lactis BB-12 did not decrease the number of school
                          days missed due to illness.
                          Merenstein DJ, Smith KH, Scriven M, Roberts RF, Sanders ME, Petterson S. The study to investigate the potential benefits of probiotics
                          in yogurt, a patient-oriented, double-blind, cluster-randomised, placebo-controlled, clinical trial. Eur J Clin Nutr. 2010 Mar 10.




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                          3.2. ALLERGY & ATOPY
 EDITO                           An aditional benefit provided by a probiotic against allergic colitis in
 THE MAIN POINTS OF              infants
 THE QUARTER              OBJECTIVES / BACKGROUND
                          Cow’s milk allergic colitis (CMAC), a benign condition that affects term infants and children younger
1. CHARACTERISATION OF
                          than 3 years of age, is suspected in otherwise healthy infants with blood in the stools, mucus, with
   PROBIOTICS AND
   PROBIOTIC FOODS
                          or without diarrhea. The condition occurs in breast- and formula-fed infants. The aims of this clinical
                          trial, conducted in infants, were to determine the benefits of Lactobacillus rhamnosus GG (LGG) in an
2. PHYSIOLOGICAL          extensively hydrolyzed casein formula (EHCF) in improving hematochezia and fecal calprotectin over
   INTERACTION WITH THE   EHCF alone.
   HOST
                          DESIGN
3. IN VIVO OUTCOMES OF    For formula-fed infants, the trial was randomized, double-blind, and placebo-controlled.
   PROBIOTIC INTAKE
                          SETTINGS & PARTICIPANTS
                          Infants from 0 to 12 months of age, fed a casein-based routine formula or breast milk, with presumptive
 REFERENCES
                          diagnosis of CMAC were enrolled in group A (n=30); group B (comparison group; n=32) included healthy
                          infants of the same sex and age, matched for the type of feeding (formula or breast milk), recruited
 KEY DATES                from the general population of a local paediatric practice. Fecal calprotectin was compared in 30
                          infants with hematochezia and 4 weeks after milk elimination with that of a healthy group. Calprotectin
                          and hematochezia were also compared on 26 formula-fed infants randomly assigned to extensively
                          hydrolyzed casein formula with LGG (EHCF+LGG) or without (EHCF-LGG) and on 4 breastfed infants
                          whose mothers eliminated dairy.
                          MAIN OUTCOME
                          The main outcome was to determine the benefits of LGG over EHCF alone in CMAC infants. The main
                          secondary outcomes were to determine whether fecal calprotectin can be used as a marker of intestinal
                          inflammation in infants suspected of CMAC; and whether fecal calprotectin can be used to monitor
                          clinical improvement in CMAC.
                          RESULTS
                          Fecal calprotectin in those with hematochezia was significantly higher than in comparisons (P<0.0001).
                          At 4 weeks, fecal calprotectin decreased to 50% of baseline but was still significantly higher than in
                          comparisons (P=0.03). The mean fecal calprotectin decrease was significantly higher among EHCF+LGG
                          compared with EHCF-LGG (P=0.02). At 4 weeks, none of the EHCF+LGG had blood in stools, and 5/14
                          on EHCF-LGG did (P=0.002).
                          CONCLUSION
                          Fecal calprotectin is high in infants with hematochezia and possible allergic colitis. Extensively hydrolyzed
                          casein formula with L. rhamnosus GG resulted in significant improvement of hematochezia and fecal
                          calprotectin compared with the same milk without the probiotic.
                          Baldassarre ME, Laforgia N, Fanelli M, Laneve A, Grosso R, Lifschitz C. Lactobacillus GG improves recovery in infants with blood in the stools and
                          presumptive allergic colitis compared with extensively hydrolyzed formula alone. J Pediatr. 2010 Mar;156(3):397-401.


                                     COMMENTARY
                              from James Versalovic, Baylor College of Medicine, Houston, US
                              «This report describes the laboratory evaluation of cow’s milk allergic colitis and effects of the probiotic strain
                              Lactobacillus rhamnosus GG in infants with hematochezia due to this form of allergic colitis. This study was
                              conducted in the outpatient setting with infants 1-10 months of age in Bari, Italy and included 62 infants in
                              two groups. A minimum dose of 1.46 x 107 organisms per 100 ml of formula was administered daily to the
                              infants in an extensively hydrolyzed casein formula (EHCF). Infants with evidence of hematochezia due to
                              cow’s milk allergic colitis had significantly elevated calprotectin levels (as evaluated by stool ELISA), when
                              compared to controls. Fecal calprotectin is a protein found in high concentrations in neutrophils, and it serves
                              as a reliable biomarker for intestinal inflammation. Calprotectin appears to be a useful marker for allergic colitis
                              in infants, based on this study. During a period of antigen elimination from the diet, infants receiving EHCF
                              plus LGG demonstrated a more complete recovery, based on greater reduction of fecal calprotectin levels
                              when compared to infants not receiving LGG. The addition of LGG to the diet of these infants also ameliorated
                              hematochezia in these infants, as evaluated by fecal occult blood testing. Significantly fewer infants consuming
                              LGG had persistence of occult blood in stools one month after dietary antigen eradication.
                              This study provided evidence that probiotics can reduce hematochezia and intestinal inflammation due to
                              exposure to cow’s milk antigens during infancy. Fecal calprotectin may be a useful biomarker to study responses
                              of infants to dietary modification and probiotics in the context of allergic colitis. However, the presence of
                              colitis was not established by rectal mucosal biopsy so the diagnoses were unconfirmed. Prior studies have
                              indicated that these cases may be misdiagnosed without a biopsy, resulting in some degree of uncertainty
                              regarding the pathophysiology in the infants under study. Additionally the breastfed infant group was small as
                              a control group, and it is not clear if probiotics truly resolve the allergic colitis or disease phenotype.»

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                          3.3.        IBS & FUNCTIONAL GI DISORDERS
 EDITO
                                      A probiotic mixture ameliorates IBS symptoms in children
 THE MAIN POINTS OF
 THE QUARTER              OBJECTIVES / BACKGROUND
                          Irritable bowel syndrome (IBS) is a common problem in pediatrics, for which no safe and effective
1. CHARACTERISATION OF    treatment is available. Probiotics have shown some promising results in adult studies, but no positive
   PROBIOTICS AND         study has been published on pediatric age. This study investigated the efficacy of VSL#3 in a population
   PROBIOTIC FOODS        of children and teenagers affected by IBS.
2. PHYSIOLOGICAL          DESIGN
   INTERACTION WITH THE   Randomized, double-blind, placebo-controlled, crossover study conducted in 7 pediatric gastroenterology
   HOST
                          divisions.
3. IN VIVO OUTCOMES OF    SETTINGS & PARTICIPANTS
   PROBIOTIC INTAKE
                          Children 4 to 18 years of age, meeting eligibility criteria, were enrolled. The patients were assessed by
                          a questionnaire for a 2- week baseline period. They were then randomized to receive either VSL#3 or a
 REFERENCES
                          placebo for 6 weeks, with controls every 2 weeks. At the end, after a ‘‘wash-out’’ period of 2 weeks, each
                          patient was switched to the other group and followed for a further 6 weeks.
 KEY DATES
                          MAIN OUTCOME
                          The primary endpoint was improvement in the subject’s overall assessment of relief (SGARC). The
                          secondary endpoints were improvements in abdominal pain/discomfort, stool pattern, bloating/
                          gassiness, and family assessment of the impact of their child’s IBS on the family’s life.

                          RESULTS
                          A total of 59 children completed the study. Although placebo was effective in some of the parameters
                          and in as many as half of the patients, VSL#3 was significantly superior to it (P<0.05) in the primary
                          endpoint, the subjective assessment of relief of symptoms; as well as in 3 of 4 secondary endpoints:
                          abdominal pain/discomfort (P<0.05), abdominal bloating/gassiness (P<0.05), and family assessment of
                          life disruption (P<0.01). No significant difference was found (P=0.06) in the stool pattern. No adverse
                          effects were recorded in any of the patients.

                          CONCLUSION
                          VSL#3 is safe and was more effective than placebo in ameliorating symptoms and improving the quality
                          of life in children affected by IBS.
                          Guandalini S, Magazzù G, Chiaro A, La Balestra V, Di Nardo G, Gopalan S, Sibal A, Romano C, Canani RB, Lionetti P, Setty M. VSL#3 Improves
                          Symptoms in Children With Irritable Bowel Syndrome: A Multicenter, Randomized, Placebo-Controlled, Double-Blind, Crossover Study. J Pediatr
                          Gastroenterol Nutr. 2010 May 4.



                                     COMMENTARY

                             from Hania Szajewska, The Medical University of Warsaw, Poland
                             «The results of the crossover randomized controlled trial (RCT) by Guandalini et al. suggest that VSL#3 is safe
                             and effective in ameliorating symptoms and improving the quality of life in children affected by irritable bowel
                             syndrome (IBS). This study is the first one to evaluate VSL#3 in children with IBS. The findings are in line with the
                             evidence obtained in adults. Given the lack of effective therapy for IBS, this new data is welcome. Nonetheless,
                             even if the results are encouraging, repeat studies, with longer follow-up periods, are needed. Perhaps studies
                             that include such an important outcome measure as the percentage of patients who are asymptomatic after
                             the intervention are needed before VSL#3 could be routinely recommended for the treatment of pediatric IBS.

                             With regard to other probiotics evaluated in children with IBS, only Lactobacillus rhamnosus GG (LGG) has
                             been studied in two double-blind RCTs, with conflicting results. The first RCT (n=50) concluded that LGG at
                             the dose of 1010 CFU twice daily administered for 6 weeks was not superior to placebo in the treatment of
                             abdominal pain in children with IBS but may be of help in relieving such symptoms as perceived abdominal
                             distention (bloating). The second RCT (n=37) found that those in the LGG group (3 x 109 CFU twice daily, for 4
                             weeks) were more likely to respond to treatment than those in the placebo group (33% vs. 5%, relative benefit
                             6.3, 95% CI 1.2 – 38).x More participants in the LGG group reported a reduced frequency of pain but not
                             decreased pain severity. With regard to adults with IBS, more probiotic preparations have been investigated,
                             and the American College of Gastroenterology recently concluded that ‘in single organism studies, lactobacilli
                             do not appear effective for patients with IBS; bifidobacteria and certain combinations of probiotics demonstrate
                             some efficacy.’xi

                             Still, both in adults and children, more data is needed. While awaiting new studies, it seems reasonable to
                             discuss current IBS evidence regarding probiotics with patients and let them (or their parents) decide whether
                             the intervention might be beneficial.»
                              ix
                                 Bausserman M, Michail S. The use of Lactobacillus GG in irritable bowel syndrome in children: a double-blind randomized control trial.
                              J Pediatr 2005;147:197-201.
                              x
                                 Gawronska A, Dziechciarz P, Horvath A, Szajewska H. A randomized double-blind placebo-controlled trial of Lactobacillus GG for
                              abdominal pain disorders in children. Aliment Pharmacol Ther 2007;25:177-84.
                              xi
                                 American College of Gastroenterology Task Force on Irritable Bowel Syndrome, Brandt LJ, Chey WD, Foxx-Orenstein AE, Schiller
                              LR, Schoenfeld PS, Spiegel BM, Talley NJ, Quigley EM. An evidence-based position statement on the management of irritable bowel
                              syndrome. Am J Gastroenterol 2009;104 Suppl 1:S1-35.


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 Contents :
                                     Probiotics prevent the incidence of pelvic radiation-induced
 EDITO                               diarrhea in women
                          OBJECTIVES/BACKGROUND
 THE MAIN POINTS OF
                          Radiation-induced diarrhea is frequently observed during pelvic radiotherapy. This study was performed
 THE QUARTER
                          to determine the ability of Lactobacillus acidophilus and Bifidobacterium bifidum to reduce the incidence
                          of radiation-induced diarrhea in locally-advanced cervical cancer patients.
1. CHARACTERISATION OF
   PROBIOTICS AND         DESIGN
   PROBIOTIC FOODS
                          Randomized, double-blind, placebo-controlled study.
2. PHYSIOLOGICAL
   INTERACTION WITH THE
                          SETTINGS & PARTICIPANTS
   HOST                   A total of 63 patients (18 yrs≤age≤65 yrs) who were undergoing pelvic radiotherapy (against cervical
                          cancer) concurrent with weekly cisplatin were randomly assigned to receive capsules of L. acidophilus
3. IN VIVO OUTCOMES OF    plus B. bifidum (n=32) twice-daily, beginning 7 days before starting radiotherapy and continuing every
   PROBIOTIC INTAKE       day during radiotherapy. In the control group (n=30), placebo capsules were administered using the
                          same schedule. Diarrhea was graded weekly according the Common Toxicity Criteria (CTC) system.
 REFERENCES               Stool consistency and white and red blood cell count in stool were also assessed.
                          MAIN OUTCOME
 KEY DATES
                          The primary endpoint was the reduction of diarrhea incidence, defined by a CTC grade 2 or more, and
                          the need for anti-diarrheal medication.

                          RESULTS
                          Grade 2 -3 diarrhea was observed in 45% of the placebo group and 9% of the study drug group (P=0.002).
                          Anti-diarrheal medication use was significantly reduced in the placebo group (P=0.03). The patients in
                          the study drug group had a significantly improved stool consistency (P<0.001).

                          CONCLUSION
                          Administration of live L. acidophilus plus B. bifidum to patients undergoing pelvic radiotherapy reduced
                          the incidence of radiation-induced diarrhea and the need for anti-diarrheal medication; it also led to a
                          significant benefit in stool consistency.
                          Chitapanarux I, Chitapanarux T, Traisathit P, Kudumpee S, Tharavichitkul E, Lorvidhaya V. Randomized controlled trial of live lactobacillus
                          acidophilus plus bifidobacterium bifidum in prophylaxis of diarrhea during radiotherapy in cervical cancer patients. Radiat Oncol. 2010 May
                          5;5(1):31.


                                     A systematic review concludes that probiotics are efficacious in IBS

                          OBJECTIVES
                          Probiotics may benefit irritable bowel syndrome (IBS) symptoms, but randomised controlled trials (RCTs)
                          have been conflicting; therefore a systematic review was conducted.

                          DESIGN
                          Systematic review of published data.

                          RETRIEVAL STRATEGY
                          Information was retrieved from MEDLINE (1966 to May 2008), EMBASE (1988 to May 2008) and the
                          Cochrane Controlled Trials Register (2008) electronic databases were searched, as were abstracts from
                          Digestive Diseases Week and United European Gastroenterology Week, and authors were contacted for
                          extra information. Only parallel group RCTs with at least one week of treatment comparing probiotics
                          with placebo or no treatment in adults with IBS according to any acceptable definition were included.

                          MAIN OUTCOME
                          The main outcome is the improvement in abdominal pain or overall IBS symptoms.

                          RESULTS
                          19 RCTs (18 papers) in 1650 patients with IBS were identified. Trial quality was generally good, with nice
                          reporting adequate methods of randomisation and six a method of allocation concealment. There were 10
                          RCTs involving 918 patients providing outcomes as a dichotomous variable. Probiotics were statistically
                          significantly better than placebo (RR of IBS not improving=0.71; 95% CI 0.57-0.88) with a number needed
                          to treat NNT=4 (95% CI 3-12.5). There was significant heterogeneity (X2=28.3, P=0.001, I2=68%) and
                          possible funnel plot asymmetry. Fifteen trials assessing 1351 patients reported an improvement in IBS
                          score as a continuous outcome (SMD=-0.34; 95% CI -0.60 to-0.07). There was statistically significant
                          heterogeneity (X2=67.04, P<0.001, I2=79%), but this was explained by one outlying trial.

                          CONCLUSION
                          These results support the efficacy of probiotics in IBS, which is strengthened by the fact that this
                          systematic review retrieved a large number of RCTs.The main limitation is that there were a variety of
                          species, strains and doses of probiotics used, and therefore it was difficult to come to any conclusion
                          about the magnitude of benefit and optimum probiotic species and strain to use in IBS.
                          Moayyedi P, Ford AC, Talley NJ, Cremonini F, Foxx-Orenstein AE, Brandt LJ, Quigley EM. The efficacy of probiotics in the treatment      14
                          of irritable bowel syndrome: a systematic review. Gut. 2010 Mar;59(3):325-32.
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 Contents :
                          3.4.       IBD AND OTHER INFLAMMATORY DISEASES
 EDITO
                                     A meta-analysis highlights the benefits of probiotics against
 THE MAIN POINTS OF                  ulcerative colitis
 THE QUARTER
                          OBJECTIVES / BACKGROUND
1. CHARACTERISATION OF    Many studies have discussed the positive effects of probiotics for treating gastro-intestinal diseases,
   PROBIOTICS AND         including ulcerative colitis. However, the sample size was relatively small, so there is no definitive
   PROBIOTIC FOODS        evidence as to whether probiotics are helpful. This paper systematically evaluates probiotics’ curative
2. PHYSIOLOGICAL
                          effects for treating ulcerative colitis based on existing random control trials.
   INTERACTION WITH THE
                          DESIGN
   HOST
                          Meta-analysis.
3. IN VIVO OUTCOMES OF
   PROBIOTIC INTAKE       RETRIEVAL STRATEGY
                          Information was retrieved from MEDLINE (1966 to August 2009), EMBASE (1980 to August 2009), and
 REFERENCES               the Cochrane Controlled Trials Register (1995 to August 2009). All the random control experiments that
                          compared probiotics with ulcerative colitis treatment medicine or placebos were collected, including
 KEY DATES
                          abstracts and full texts. The selection criteria were as follows: (1) adult and pediatric studies were included;
                          (2) the experiments compared the curative effect of probiotics with standard therapy for ulcerative colitis
                          or placebo; (3) all were random control tests; (4) abstracts and meeting presentations were selected; and
                          (5) patients had a definite diagnosis of ulcerative colitis using definite diagnosis standards. Reviews and
                          case reports were excluded.

                          MAIN OUTCOME
                          The induction of remission and promotion of maintenance were compared between probiotics treatment
                          and non-probiotics treatment in ulcerative colitis.

                          RESULTS
                          A total of 286 papers were retrieved, 13 randomized controlled studies met the selection criteria. Seven
                          studies evaluated the remission rate, and eight studies estimated the recurrence rate; two studies
                          evaluated both remission and recurrence rates. Compared with the non-probiotics group, the remission
                          rate for ulcerative colitis patients who received probiotics was 1.35 (95% CI: 0.98-1.85). Compared with
                          the placebo group, the remission rate of ulcerative colitis patients who received probiotics was 2.00
                          (95% CI: 1.35-2.96). During the course of treatment, in patients who received probiotics for less than
                          12 months compared with the group treated by non-probiotics, the remission rate of ulcerative colitis
                          was 1.36 (95% CI: 1.07-1.73). Compared with the non-probiotics group, the recurrence rate of ulcerative
                          colitis patients who received probiotics was 0.69 (95% CI: 2.47-1.01). In the mild to moderate group who
                          received probiotics, compared to the group who did not receive probiotics, the recurrence rate was 0.25
                          (95% CI: 0.12-0.51).

                          CONCLUSION
                          It seems that probiotic treatment was more effective than placebo in maintaining remission in ulcerative
                          colitis. However, in this meta-analysis, the evaluation criteria of the results were not totally consistent. To
                          a certain extent, this condition added to the heterogeneity of the systemic review. Thus, further clinical
                          trials are still needed to determine whether the use of probiotics is better than non-probiotics therapy
                          during the remission induction stage and maintenance stage of ulcerative colitis, including variations of
                          bacterial species, applied dosage, treatment timing, and the course of treatment.
                          Sang LX, Chang B, Zhang WL, Wu XM, Li XH, Jiang M. Remission induction and maintenance effect of probiotics on ulcerative colitis: a meta-
                          analysis. World J Gastroenterol. 2010 Apr 21;16(15):1908-15.




                                     A mathematical model as a preliminary tool for exploring the effects
                                     of probiotic treatment for NEC
                          OBJECTIVES
                          The objective of this study is to use a mathematical model to predict the conditions under which
                          probiotics may be successful in promoting the health of infants suffering from necrotizing enterocolitis
                          (NEC). Based on experimental and clinical studies, the developed tool is a simplified mathematical model
                          of the host-pathogen interaction that occurs in the setting of NEC and it is used to analyze the impact of
                          probiotic administration on the ensuing dynamics. The predictions derived from this computational study
                          may help to explain the diverse outcomes that may arise in this setting and may be useful for guiding
                          future experimental and clinical studies.




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 Contents :
                          METHOD & MEASURES
                          An ordinary differential equation model was developed, which tracks the populations of pathogenic
 EDITO                    and probiotic bacteria in the intestinal lumen and in the blood/tissue region. The permeability of the
                          intestinal epithelial layer is treated as a variable, and the role of the inflammatory response is included.
 THE MAIN POINTS OF
                          To investigate various features of probiotic treatment for NEC, equations in the absence of probiotics
 THE QUARTER
                          for varying levels of initial pathogenic attack were first considered. Next, the effects of probiotics on
                          the growth of pathogenic bacteria in the lumen, on the permeability of the epithelial wall, and on the
1. CHARACTERISATION OF
   PROBIOTICS AND         activation of the inflammatory response are analyzed.
   PROBIOTIC FOODS
                          RESULTS
2. PHYSIOLOGICAL          The model predicts that in the presence of probiotics, health is restored in many cases that would have
   INTERACTION WITH THE   otherwise been pathogenic. The timing of probiotic administration is also shown to determine whether
   HOST                   or not health is restored. Finally, the model predicts that probiotics may be harmful to the NEC patient
                          under very specific conditions, perhaps explaining the detrimental effects of probiotics observed in some
3. IN VIVO OUTCOMES OF    clinical studies.
   PROBIOTIC INTAKE
                          CONCLUSION
 REFERENCES               The reduced, experimentally motivated mathematical model suggests how a certain general set of
                          characteristics of probiotics can lead to beneficial or detrimental outcomes for infants suffering from
 KEY DATES                NEC, depending on the influences of probiotics on defined features of the inflammatory response.
                          Arciero JC, Ermentrout GB, Upperman JS, Vodovotz Y, Rubin JE. Using a mathematical model to analyze the role of probiotics and inflammation
                          in necrotizing enterocolitis. PLoS One. 2010 Apr 19;5(4):e10066.




                                      A meta-analysis confirms that probiotics prevent NEC in preterm
                                      neonates
                          OBJECTIVES / BACKGROUND
                          Systematic reviews of randomized, controlled trials (RCTs) indicate lower mortality and necrotizing
                          enterocolitis (NEC) and shorter time to full feeds after probiotic supplementation in preterm (<34 weeks’
                          gestation) very low birth weight (VLBW; birth weight <1500 g) neonates. The objective of this study
                          was to update the 2007 systematic review of RCTs of probiotic supplementation for preventing NEC in
                          preterm VLBW neonates.
                          DESIGN
                          Meta-analysis.

                          RETRIEVAL STRATEGY
                          Data was collected from the Cochrane Central register; Medline, Embase, and Cinahl databases; and
                          proceedings of the Pediatric Academic Society meetings and gastroenterology conferences. Cochrane
                          Neonatal Review Group search strategy was followed. Selection criteria were RCTs of any enteral
                          probiotic supplementation that started within first 10 days and continued for ≥7 days in preterm VLBW
                          neonates and reported on stage 2 NEC or higher (Modified Bell Staging).
                          MAIN OUTCOME
                          The primary outcome was efficacy of probiotic supplement in preventing stage 2 NEC or higher, safety
                          in terms of blood culture-positive sepsis including that by the organism(s) in the probiotic supplement,
                          and any other adverse events reported by the authors. The secondary outcomes included the time to
                          reach full feeds and duration of hospital stay. An intervention effect of 30% for the prevention of NEC
                          was regarded as clinically significant.
                          RESULTS
                          A total of 11 (n=2176), including 4 new (n=783), trials were eligible for inclusion in the meta-analysis by
                          using a fixed-effects model. The risk for NEC and death was significantly lower. Risk for sepsis did not
                          differ significantly. No significant adverse effects were reported. Trial sequential analysis showed 30%
                          reduction in the incidence of NEC (alpha = 0.05 and 0.01; power: 80%).
                          CONCLUSION
                          The dramatic effect sizes, tight confidence intervals, extremely low P values, and overall evidence confirm
                          the significant benefits of probiotic supplements in reducing death and disease in preterm neonates.
                          Deshpande G, Rao S, Patole S, Bulsara M. Updated meta-analysis of probiotics for preventing necrotizing enterocolitis in preterm
                          neonates. Pediatrics. 2010 May;125(5):921-30.




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 Contents :
                          3.5.        NUTRITION: OBESITY, CHOLESTEROL, ETC.
 EDITO
                                      Perinatal probiotic intake may limit excess weight gain during the
 THE MAIN POINTS OF                   first years of life
 THE QUARTER
                          OBJECTIVES / BACKGROUND
1. CHARACTERISATION OF    The recently discovered role of the gut microbiota in host metabolism may offer a new target in the
   PROBIOTICS AND         prevention and management of obesity. The composition of gut microbiota may thus be taken to
   PROBIOTIC FOODS        represent a new contributor to obesity above the traditional and well-known risk factors, excessive
                          energy intake and sedentary behaviour. On this basis, it may be hypothesized that early modification
2. PHYSIOLOGICAL
   INTERACTION WITH THE   of the gut microbial and immunological environment with probiotics will reduce the risk of becoming
   HOST                   overweight and, thus, beneficially influence the child’s early growth pattern. This study aimed to evaluate
                          the impact of perinatal probiotic intervention on childhood growth patterns and excess weight gain
3. IN VIVO OUTCOMES OF    during a 10-year follow-up period.
   PROBIOTIC INTAKE
                          DESIGN
 REFERENCES               The trial with the mothers was randomized, double-blind, and placebo-controlled; these mothers’
                          newborns were followed-up for 10 years.
 KEY DATES
                          SETTINGS & PARTICIPANTS
                          Altogether, 159 women were randomized to receive Lactobacillus rhamnosus GG or placebo four weeks
                          before expected delivery; the intervention extending for six months postnatally. After delivery, the probiotic
                          was given either to the mothers, if they were breastfeeding, or otherwise to the children mixed in water.

                          MAIN OUTCOME
                          Anthropometric measurements of the children were taken at the ages of 3, 6, 12 and 24 months and at
                          4, 7 and 10 years in 113 (72%) children.

                          RESULTS
                          Excess weight gain was observed to occur in two stages; the initial phase of excess weight gain occurs
                          during the foetal period and continues until 24-48 months of age and a second phase of excess weight
                          gain starts after the age of 24-48 months. The perinatal probiotic intervention appeared to moderate the
                          initial phase of excessive weight gain, especially among children who later became overweight, but not
                          the second phase of excessive weight gain, the impact being most pronounced at the age of 4 years
                          (P=0.063, analysis of variance for repeated measures). The effect of intervention was also shown as a
                          tendency to reduce the birth-weight-adjusted mean body mass index at the age of 4 years (P=0.080,
                          analysis of covariance).

                          CONCLUSION
                          Early gut microbiota modulation with probiotics may modify the growth pattern of the child by restraining
                          excessive weight gain during the first years of life. This new observation calls for further epidemiological
                          and clinical trials, with precise data on early growth patterns and on confounding factors influencing
                          weight development.

                          Luoto R, Kalliomäki M, Laitinen K, Isolauri E. The impact of perinatal probiotic intervention on the development of overweight and obesity: follow-
                          up study from birth to 10 years. Int J Obes (Lond). 2010 Mar 16.




                                     COMMENTARY

                              from Bruno Pot, Institut Pasteur de Lille, France
                              «For many years it has been accepted that probiotic intervention during the first months of life offers possibilities
                              that are no longer possible later in life. Furthermore, gut microbiota composition has been shown to be
                              important in host energy harvest and nutrient metabolism through intestinal glucose absorption, lipogenesis
                              and fat deposition regulation. The article by Luoto et al. illustrates that perinatal probiotic intervention will
                              not only consolidate immunological responsiveness, but may have an impact on childhood growth patterns
                              and excess weight gain during a 10-year follow-up period. It was shown that excess weight gain was seen
                              to occur in two stages: an initial phase during the foetal period and continuing until 24–48 months after birth
                              and a second phase starting after that age. The perinatal intervention with LGG moderated the initial phase
                              of excess weight gain, but not the second phase. The results of the study confirmed the potential of probiotic
                              interventions to fight obesity and possibly other Western lifestyle diseases, but also point out the limitations in
                              terms of the susceptibility period for such an intervention.»




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                          Lactococcus lactis by Matrix-assisted Laser Desorption Ionization -        2010 May;27(3):381-9.
                          Time-of-Flight Mass Spectrometry. Appl Environ Microbiol. 2010 Apr
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                          for the detection and enumerization of Lactobacillus acidophilus and       lactic acid bacteria and effect of PlnA on bacterial and Caco-2 cells.
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                          Wenning M, Büchl NR, Scherer S. Species and strain identification           Lactobacillus plantarum strains under in vitro acidic conditions using
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                          Aredes Fernández PA, Farías ME, de Nadra MC. Interaction between           Galle S, Schwab C, Arendt E, Gänzle M. Exopolysaccharide-forming
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                          Audy J, Labrie S, Roy D, Lapointe G. Sugar source modulates                and antiadhesive properties of a biosurfactant isolated from
                          exopolysaccharide biosynthesis in Bifidobacterium longum subsp.             Lactobacillus paracasei ssp. paracasei A20. Lett Appl Microbiol. 2010
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                          Synergysm between surface charge density and chain packing.                   via incorporation and conversion to coprostanol. J Dairy Sci. 2010
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 THE QUARTER
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                          domain from Lactobacillus fermentum bacteriophage endolysin and               linoleate isomerase in Lactobacillus acidophilus. Appl Microbiol
1. CHARACTERISATION OF    its use as an anchor to display heterologous proteins on the surfaces         Biotechnol. 2010 May 12.
   PROBIOTICS AND         of lactic acid bacteria. Appl Environ Microbiol. 2010 Apr;76(8):2410-8.
   PROBIOTIC FOODS                                                                                      Maischberger T, Mierau I, Peterbauer CK, Hugenholtz J, Haltrich
                          Hynönen U, Avall-Jääskeläinen S, Palva A. Characterization and                D. High-level expression of Lactobacillus beta-galactosidases in
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2. PHYSIOLOGICAL          S-layer protein gene. Appl Microbiol Biotechnol. 2010 Mar 13.                 system NICE. J Agric Food Chem. 2010 Feb 24;58(4):2279-87.
   INTERACTION WITH THE
                          Iqbal S, Nguyen TH, Nguyen TT, Maischberger T, Haltrich D. beta-              Mazzoli R, Pessione E, Dufour M, Laroute V, Giuffrida MG, Giunta
   HOST                   Galactosidase from Lactobacillus plantarum WCFS1: biochemical                 C, Cocaign-Bousquet M, Loubière P. Glutamate-induced metabolic
                          characterization and formation of prebiotic galacto-oligosaccharides.         changes in Lactococcus lactis NCDO 2118 during GABA production:
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                          Khaleghi M, Kermanshahi RK, Yaghoobi MM, Zarkesh-Esfahani SH,                 a coupled reaction catalysed by Lactobacillus acidophilus NCFM
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                          Li H, Qiu T, Gao D, Cao Y. Medium optimization for production of
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                          Li Z, Lu J, Zhao L, Xiao K, Tan T. Improvement of L: -Lactic Acid
                          Production under Glucose Feedback Controlled Culture by                       Rehaiem A, Martínez B, Manai M, Rodríguez A. Production of enterocin
                          Lactobacillus rhamnosus. Appl Biochem Biotechnol. 2010 Apr 15.                A by Enterococcus faecium MMRA isolated from ‘Rayeb’, a traditional
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                          Liu S, Bischoff KM, Qureshi N, Hughes SR, Rich JO. Functional
                          expression of the thiolase gene thl from Clostridium beijerinckii P260        Rhimi M, Boisson A, Dejob M, Boudebouze S, Maguin E, Haser
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 ISSUE N°2                REFERENCES



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   PROBIOTIC FOODS        Shimosato T, Fujimoto M, Tohno M, Sato T, Tateo M, Otani H, Kitazawa         Pategrás cheese as a suitable carrier for six probiotic cultures. J Dairy
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                                                                                                                                                                             20
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 ISSUE N°2                REFERENCES



 Contents :                     Tholpady SS, Sifri CD, Sawyer RG, Hazen KC, Pruett TL, Bonatti
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                               Vidal M, Forestier C, Charbonnel N, Henard S, Rabaud C, Lesens       Staphylococcus hominis MBBL 2-9 as anti-Staphylococcus aureus
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1. CHARACTERISATION OF       SURVIVAL & PHYSICAL INTERACTIONS OF PROBIOTICS WITH                    Stimulate in Vitro Bacteriocin Production by Lactobacillus Spp. and
   PROBIOTICS AND            MUCOSA                                                                 Enterococcus Spp. Strains. J Agric Food Chem. 2010 Apr 29.
   PROBIOTIC FOODS        Buriti FC, Castro IA, Saad SM. Viability of Lactobacillus acidophilus     Trejo FM, Pérez PF, De Antoni GL. Co-culture with potentially probiotic
                          in synbiotic guava mousses and its survival under in vitro simulated      microorganisms antagonises virulence factors of Clostridium difficile in
2. PHYSIOLOGICAL          gastrointestinal conditions. Int J Food Microbiol. 2010 Feb               vitro. Antonie Van Leeuwenhoek. 2010 Jun;98(1):19-29.
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                                                                                                    Yang EJ, Chang HC. Purification of a new antifungal compound
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                          Yiangou M. Evaluation of adhesion capacity, cell surface traits and       Food Microbiol. 2010 Apr 30;139(1-2):56-63.
3. IN VIVO OUTCOMES OF    immunomodulatory activity of presumptive probiotic Lactobacillus
                          strains. Int J Food Microbiol. 2010 Apr 10.                                   MUCOSA RESPONSE             TO    PROBIOTICS       (TROPHICITY      &
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                          effects of Bifidobacterium lactis HN019 on intestinal epithelial cells
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                                                                                                    Maragkoudakis PA, Mountzouris KC, Rosu C, Zoumpopoulou G,
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                          Generoso SV, Viana M, Santos R, Martins FS, Machado JA, Arantes
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                                                                                                                                                                          21
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1. CHARACTERISATION OF    Th1 autophagy-promoting cytokine (IFN-gamma) and nitric oxide
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                          Yazdi MH, Soltan Dallal MM, Hassan ZM, Holakuyee M, Agha Amiri
                                                                                                                                                                          22
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 ISSUE N°2                REFERENCES



 Contents :                     Montalto M, Gallo A, Curigliano V, D’Onofrio F, Santoro L,
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 THE QUARTER                    supplementation with Lactobacillus reuteri or tilactase in lactose           Saudi Med J. 2010 Mar;31(3):280-3.
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                          porcine erythrocytes. Appl Environ Microbiol. 2010 May 14.                   Elderly. Biosci Biotechnol Biochem. 2010 May 7.

                          Yeo SK, Liong MT. Angiotensin I-converting enzyme inhibitory activity        Pascual L, Ruiz F, Giordano W, Barberis IL. Vaginal colonization and
                          and bioconversion of isoflavones by probiotics in soymilk supplemented        activity of the probiotic bacterium Lactobacillus fermentum L23
                          with prebiotics. Int J Food Sci Nutr. 2010 Mar;61(2):161-81.                 in a murine model of vaginal tract infection. J Med Microbiol. 2010
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                                                                                                       experimentally infected with Toxoplasma gondii. Comp Immunol
                                Arroyo R, Martín V, Maldonado A, Jiménez E, Fernández L,               Microbiol Infect Dis. 2010 Apr 19.
                                Rodríguez JM. Treatment of Infectious Mastitis during Lactation:
                                Antibiotics versus Oral Administration of Lactobacilli Isolated        Salva S, Villena J, Alvarez S. Immunomodulatory activity of Lactobacillus
                                from Breast Milk. Clin Infect Dis. 2010 May 10.                        rhamnosus strains isolated from goat milk: Impact on intestinal and
                                                                                                       respiratory infections. Int J Food Microbiol. 2010 Mar 18.
                          Bisson JF, Hidalgo S, Rozan P, Messaoudi M. Preventive effects
                          of different probiotic formulations on travelers’ diarrhea model in          Salva S, Villena J, Alvarez S. Immunomodulatory activity of Lactobacillus
                          wistar rats : preventive effects of probiotics on TD. Dig Dis Sci. 2010      rhamnosus strains isolated from goat milk: Impact on intestinal and
                          Apr;55(4):911-9.                                                             respiratory infections. Int J Food Microbiol. 2010 Mar 18.

                                Chitapanarux I, Chitapanarux T, Traisathit P, Kudumpee S,              Santini C, Baffoni L, Gaggia F, Granata M, Gasbarri R, Di Gioia D,
                                Tharavichitkul E, Lorvidhaya V. Randomized controlled trial of         Biavati B. Characterization of probiotic strains: An application as feed
                                live lactobacillus acidophilus plus bifidobacterium bifidum in           additives in poultry against Campylobacter jejuni. Int J Food Microbiol.
                                prophylaxis of diarrhea during radiotherapy in cervical cancer         2010 Apr 8.
                                patients. Radiat Oncol. 2010 May 5;5(1):31.
                                                                                                       Zhang L, Xu YQ, Liu HY, Lai T, Ma JL, Wang JF, Zhu YH. Evaluation of
                                 Chou IC, Kuo HT, Chang JS, Wu SF, Chiu HY, Su BH, Lin HC. Lack        Lactobacillus rhamnosus GG using an Escherichia coli K88 model of
                                 of effects of oral probiotics on growth and neurodevelopmental        piglet diarrhoea: Effects on diarrhoea incidence, faecal microflora and
                                 outcomes in preterm very low birth weight infants. J Pediatr.         immune responses. Vet Microbiol. 2010 Feb 24;141(1-2):142-8.
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                          Deshpande G, Rao S, Patole S, Bulsara M. Updated meta-analysis of
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                          Ehrström S, Daroczy K, Rylander E, Samuelsson C, Johannesson U,              by the mitochondrial pathway. Int J Food Microbiol. 2010
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                          Outcome after Probiotic Supplementation in Conventionally Treated            Baldwin C, Millette M, Oth D, Ruiz MT, Luquet FM, Lacroix M. Probiotic
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                                 Frohmader TJ, Chaboyer WP, Robertson IK, Gowardman J.
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 ISSUE N°2                REFERENCES



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 THE QUARTER                    Bifidobacterium lactis, Lactobacillus acidophilus) in the primary
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1. CHARACTERISATION OF          2):e386-93.                                                          G, Langella P, Bermúdez-Humarán LG. Intragastric administration of
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2. PHYSIOLOGICAL                23;153(2):201-206.                                                       NUTRITION: OBESITY, CHOLESTEROL…
   INTERACTION WITH THE
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3. IN VIVO OUTCOMES OF    placebo-controlled study. Aliment Pharmacol Ther. 2010 Apr 7.
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                                                                                                     Malik R, Bandla S. Effect of source and dose of probiotics and
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                          Arciero JC, Ermentrout GB, Upperman JS, Vodovotz Y, Rubin JE.                    de Regt MJ, Willems RJ, Hené RJ, Siersema PD, Verhaar HJ,
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                          Kobayashi T, Kato I, Nanno M, Shida K, Shibuya K, Matsuoka Y, Onoue        Karen M, Yuksel O, Akyürek N, Ofluo lu E, Ca lar K, Sahin TT, Pa ao lu H,
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                                                                                                                                                                             24
 PROBIOTICS
 WATCH
 ISSUE N°2                REFERENCES



 Contents :               Maragkoudakis PA, Mountzouris KC, Rosu C, Zoumpopoulou G,
                          Papadimitriou K, Dalaka E, Hadjipetrou A, Theofanous G, Strozzi
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                          Lactobacillusplantarum PCA 236 modulates gut microbiota and milk              feeding for safe food production. Int J Food Microbiol. 2010 Mar 10.
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1. CHARACTERISATION OF
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                                leukocytes from patients with chronic infected leg ulcers treated
                                with Lactobacillus plantarum. Clin Microbiol Infect. 2010               Gómez-Llorente C, Muñoz S, Gil A. Session 5: Early programming of
2. PHYSIOLOGICAL                Mar;16(3):281-6.                                                        the immune system and the role of nutrition Role of Toll-like receptors
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                                                                                                        pancreatitis. Langenbecks Arch Surg. 2010 Apr;395(4):309-16.
                          Flichy-Fernández AJ, Peñarrocha-Diago M. Probiotic treatment in the
                          oral cavity: an update. Med Oral Patol Oral Cir Bucal. 2010 Feb 21.           Ozdemir O. Any benefits of probiotics in allergic disorders? Allergy
                                                                                                        Asthma Proc. 2010 Mar;31(2):103-11.                             25
                          Ford AC. Probiotics in irritable bowel syndrome: underachievers
                          or underpowered? Aliment Pharmacol Ther. 2010 Apr;31(8):922-3;
 PROBIOTICS
 WATCH
 ISSUE N°2                REFERENCES



 Contents :               Ozdemir O. Various effects of different probiotic strains in allergic
                          disorders: an update from laboratory and clinical data. Clin Exp
                          Immunol. 2010 Mar 16.

 EDITO                    Packey CD, Ciorba MA. Microbial influences on the small intestinal
                          response to radiation injury. Curr Opin Gastroenterol. 2010
                          Mar;26(2):88-94.
 THE MAIN POINTS OF
 THE QUARTER              Parkes GC, Sanderson JD, Whelan K. Treating irritable bowel syndrome
                          with probiotics: the evidence. Proc Nutr Soc. 2010 May;69(2):187-94.
1. CHARACTERISATION OF    Petrovsky N. Immunomodulation with microbial vaccines to prevent
   PROBIOTICS AND         type 1 diabetes mellitus. Nat Rev Endocrinol. 2010 Mar;6(3):131-8.
   PROBIOTIC FOODS
                          Salminen S, Nybom S, Meriluoto J, Collado MC, Vesterlund S, El-
                          Nezami H. Interaction of probiotics and pathogens--benefits to human
2. PHYSIOLOGICAL          health? Curr Opin Biotechnol. 2010 Apr;21(2):157-67.
   INTERACTION WITH THE
                          Samaraee AA, McCallum IJ, Coyne PE, Seymour K. Nutritional
   HOST                   strategies in severe acute pancreatitis: a systematic review of the
                          evidence. Surgeon. 2010 Apr;8(2):105-10.
3. IN VIVO OUTCOMES OF
                          Sans M. Probiotics for inflammatory bowel disease: a critical appraisal.
   PROBIOTIC INTAKE       Dig Dis. 2009;27 Suppl 1:111-4.

                          Shanahan F. 99th Dahlem conference on infection, inflammation and
 REFERENCES               chronic inflammatory disorders: host-microbe interactions in the
                          gut: target for drug therapy, opportunity for drug discovery. Clin Exp
 KEY DATES                Immunol. 2010 Apr;160(1):92-7.

                          Shaukat A, Levitt MD, Taylor BC, Macdonald R, Shamliyan TA, Kane
                          RL, Wilt TJ. Systematic Review: Effective Management Strategies for
                          Lactose Intolerance. Ann Intern Med. 2010 Apr 19.

                          Singh M, Ranjan Das R. Probiotics for allergic respiratory diseases-
                          -putting it into perspective. Pediatr Allergy Immunol. 2010 Mar;21(2
                          Pt 2):e368-76.

                          Solís G, de Los Reyes-Gavilan CG, Fernández N, Margolles A,
                          Gueimonde M. Establishment and development of lactic acid bacteria
                          and bifidobacteria microbiota in breast-milk and the infant gut.
                          Anaerobe. 2010 Feb 20.

                          Soll RF. Probiotics: are we ready for routine use? Pediatrics. 2010
                          May;125(5):1071-2.

                          Tarnow-Mordi WO, Wilkinson D, Trivedi A, Brok J. Probiotics reduce
                          all-cause mortality and necrotizing enterocolitis: it is time to change
                          practice. Pediatrics. 2010 May;125(5):1068-70.

                          Thulkar J, Kriplani A, Agarwal N. Probiotic and metronidazole
                          treatment for recurrent bacterial vaginosis. Int J Gynaecol Obstet.
                          2010 Mar;108(3):251-2.

                          Tiihonen K, Ouwehand AC, Rautonen N. Human intestinal microbiota
                          and healthy ageing. Ageing Res Rev. 2010 Apr;9(2):107-16.

                          van der Aa LB, Heymans HS, van Aalderen WM, Sprikkelman AB.
                          Probiotics and prebiotics in atopic dermatitis: review of the theoretical
                          background and clinical evidence. Pediatr Allergy Immunol. 2010
                          Mar;21(2 Pt 2):e355-67.

                          Weichselbaum E. Potential benefits of probiotics--main findings of an
                          in-depth review. Br J Community Nurs. 2010 Mar;15(3):110, 112, 114.

                          Williams MD, Ha CY, Ciorba MA. Probiotics as Therapy
                          in Gastroenterology: A Study of Physician Opinions and
                          Recommendations. J Clin Gastroenterol. 2010 Mar 8.

                          Williams NT. Probiotics. Am J Health Syst Pharm. 2010 Mar
                          15;67(6):449-58.

                          Yan F, Polk DB. Probiotics: progress toward novel therapies for
                          intestinal diseases. Curr Opin Gastroenterol. 2010 Mar;26(2):95-101.




                                                                                                      26
PROBIOTICS
WATCH
ISSUE N°2                                                                                                                       HEALTH AFFAIRS

                                                                                                           probioticswatch@danone.com




  KEY DATES
                                                                                        Kobe, Japan
                                                      August 22 - 27                    http://www.ici2010.org/

                                                                                        Boston, USA
                                                      August 26 - 29                    http://www.motilitysociety.org/

                                                                                        St Louis, USA
                                                      Aug 31 - Sept 2                   http://www.hmpdacc.org/

                                                                                        Florence, Italy
                                                      September 13 - 15                 http://www.polosci.unifi.it/phenotype/

                                                                                        Malinska, Croatia
                                                      September 22 - 25
                                                                                        http://www.hmd-cms.hr/power2010/

                                                                                        Barcelona, Spain
                                                      October 23 - 27                   http://uegw10.uegf.org/

                                                      October 25 - 29                   Miami, USA
                                                                                        http://www.asm.org/

                                                      October 28 - 29                   Montreal, Canada
                                                                                        http://www.initia.org/




      METHOD
       > Monitoring period : 18/02/2009 to 18/05/2010            > Keywords : yogurt / yoghurt / probiotic / lactic
       > Database : Medline                                      acid bacteria / streptococcus thermophilus / lac-
       > Result : 339 publications                               tobacill / fermented milk / bifidobacteria


      This probiotics watch is designed as a time-saving tool for scientists and clinicians interested in probiotic research.
      In an interactive format, the quarterly report provides timely, quasi-exhaustive lists of the scientific publications of
      the previous three months. It sorts them by topic and highlights some of the most relevant results. Readers can
      also check out upcoming scientific events and regular bibliometric analyses.

      Objectivity is a strong commitment, that’s why the articles are selected by an editorial committee, composed of
      renowned scientists in the field. Editorial committee members also comment on what they believe are the quarter’s
                                                                                                                                           Realized with iNTUiTiO and Strat & Gérie, 2010




      most relevant publications.
      Last but not least each issue features an editorial by a probiotic expert, which offers special insight into this
      fascinating field of science.

       THE EDITORIAL COMMITTE
       The members :                                                    Its missions :
           Bruno Pot,                                                        Providing international and multidisciplinary
           Institut Pasteur de Lille, FRANCE.                                vision.
           Hania Szajewska,                                                  Selecting the articles of interest in the
           The Medical University of Warsaw, Warsaw, POLAND.                 probiotics field.
           James Versalovic,                                                 Commenting the most relevant publications.
           Baylor College of Medicine, Houston, Texas, US.

				
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