Effects of Lactobacillus Rhamnosus GG on Activated Microglia and Tau Phosphorylation in the Hippocampus of Aged Mice Induced by Anesthesia and Surgery / 中山大学学报(医学科学版)

ObjectiveTo investigate the effects of Lactobacillus rhamnosus GG （LGG）on microglia and Tau phosphorylation in the hippocampus of aged mice induced by anesthesia and surgery. MethodsA total of thirty 18-month-old C57BL/6J mice were randomly divided into three groups： control group， anesthesia surgery group， and anesthesia surgery + LGG group （10 mice/group）. The aged mice were oral administered by NS or LGG 109 CFU 150 μL once a day for 20 days. Then anesthesia surgery group and anesthesia surgery +LGG group received anesthesia with isoflurane and exploratory laparotomy. The activation status of microglia in the hippocampus was detected by immunofluorescence staining 12 hours after surgery. IL-6 concentration changes was detected by ELISA. The expression changes of Tau protein phosphorylation site （Tau-pS202/pT205） and total Tau protein was detected by western blot. ResultsThe microglia in the hippocampus of the control group were in a resting state， and the concentration of inflammatory factor IL-6 was （82.08 ± 12.07） pg/mL in control group. Compared to the control group， the anesthesia surgery group showed microglial cell Microglia were activated， the concentration of inflammatory factors IL-6 increased significantly to （123.7±5.72） pg/mL （P=0.000）， and the expression of phosphorylated Tau-pS202/pT205 increased the hippocampus （P=0.002）. Compared to the anesthesia surgery group， the activated microglia were inhibited， the concentration of IL-6 decreased to （96.68±9.59） pg/mL （P=0.008）， and the expression of phosphorylated Tau-pS202/pT205 reduced significantly in the AS+LGG group （P=0.002）. While there were no significant changes in total Tau protein among 3 groups. ConclusionPreoperative administration of probiotic LGG can alleviate the activation of microglia， increased secretion of inflammatory factors， and increased Tau protein phosphorylation levels in the hippocampus of elderly mice caused by anesthesia surgery.

Effectiveness of Lactobacillus rhamnosusGG as an Adjunct in the Treatment of Enteric Fever in Children: A Double‑Blinded Randomized Controlled Trial in Southern India

Introduction:Aprobiotic used as an adjunct in Salmonella typhi infection along with antibiotic is postulated to interfere with the virulence and growth of Salmonella. To determine the effectiveness of Lactobacillus rhamnosusGG (LGG), as an adjunct with intravenous ceftriaxone, compared with a placebo in defervescence and toxemia resolution in children with enteric fever. Settings and Design: This hospital‑based randomized double‑blinded controlled trial was conducted among 56 study participants who were children below the age of 12 years, admitted as inpatients with fever and whose blood culture grew S. typhi. Materials and Methods: Study participants were equally allocated into intervention or control group by simple randomization. The intervention group received injection ceftriaxone and oral LGG (probiotic) for 7 days while the control group received an injection ceftriaxone and oral placebo for 7 days. Statistical Analysis: Kaplan–Meier curves and mantel cox log‑rank test were used to compare the duration for defervescence and toxemia resolution after treatment initiation. Results: Mean duration for defervescence in the intervention and control groups was 3.87 (1.57) days and 3.35 (1.19) days, respectively. The mean time taken for the resolution of toxemia was 3.00 (1.15) days in the intervention group and 2.64 (0.87) days in the control group. Conclusions:The addition of oral LGG at a dose of 3 × 109 colony‑forming units for 7 days to the standard antibiotic therapy for enteric fever did not show a significant reduction in the time taken for defervescence (P = 0.099) or resolution of toxemia (P = 0.148)

Abordaje de los probióticos en pediatría: el rol de Lactobacillus rhamnosus GG / Approach to probiotics in pediatrics: the role of Lactobacillus rhamnosus GG

El parto prematuro, las cesáreas, los antibióticos y la lactancia materna limitada contribuyen al aumento de enfermedades crónicas no transmisibles. El objetivo fue realizar una revisión descriptiva del uso de probióticos en pediatría, con foco en la cepa Lactobacillus rhamnosus GG. Ciertos probióticos han demostrado ser eficaces en la diarrea aguda y en la diarrea asociada a antibióticos. L. rhamnosus GG y Saccharomyces boulardii pueden acortar la duración y los síntomas. L. reuteri DSM 17938 y L. rhamnosus GG fueron efectivos para el abordaje del cólico del lactante. El uso de esta cepa en fórmulas infantiles para alergia a las proteínas de leche de vaca promovería la adquisición más temprana de tolerancia. En la prevención de dermatitis atópica, la administración de L. rhamnosus GG durante el embarazo redujo su manifestación en el bebé. El empleo de probióticos como coadyuvantes es una posibilidad para considerar en la práctica pediátrica actual.

Preterm birth, C-sections, antibiotics, and limited breastfeeding contribute to the increase in noncommunicable diseases. Our objective was to perform a descriptive review of probiotic use in pediatrics, focused on Lactobacillus rhamnosus GG. Certain probiotics have demonstrated to be effective in acute diarrhea and antibiotic-associated diarrhea. L. rhamnosus GG and Saccharomyces boulardii may shorten their duration and symptoms. L. reuteri DSM 17938 and L. rhamnosus GG were effective to manage infant colic. The use of this strain in infant formulas for cow's milk protein allergy may promote an earlier tolerance acquisition. In relation to the prevention of atopic dermatitis, the administration of L. rhamnosus GG during pregnancy reduced its development in the infant. The use of probiotics as adjuvants is a possibility to consider in current pediatric practice.

Advances in Lactobacillus rhamnosus GG and pediatric nonalcoholic fatty liver disease / 中华实用儿科临床杂志

Nonalcoholic fatty liver disease (NAFLD)has become one of the most common chronic liver diseases in children, which has a wide spectrum including the nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis and hepatocellular carcinoma (HCC)with its clinical consequences.At least 50% of obese children present NAFLD.Increased levels of free fatty acids and insulin resistance play major roles in NAFLD progression, resulting in excessive accumulation of triglycerides in hepatocytes.Lactobacillus rhamnoides is one of the most thoroughly studied intestinal probiotics, and its role in NAFLD progress has been growingly concerned.Lactobacillus rhamnosus GG alleviates NAFLD in children by improving intestinal microbiota dysbiosis, increasing intestinal barrier integrity, promoting fatty acid β-oxidation, and reducing hepatic triglyceride accumulation via regulating bile acid metabolism.Recent studies have shown that microRNAs are involved in multiple pathways to alleviate NAFLD-associated lipid metabolism disorders, which are functional as noninvasive diagnostic marker of fatty liver disease in children.This review mainly summarizes that Lactobacillus rhamnosus GG improves NAFLD in children from multiple metabolic pathways, aiming to provide a new strategy and target for the treatment and early diagnosis of NAFLD in children.

Encapsulation of probiotic strain Lactobacillus rhamnosus GG with black bean extract in alginate-pectin microcapsules

Aims@#Probiotics are claimed to confer many health effects upon consumption. However, the survivability of probiotic under the harsh conditions in the gastrointestinal tract has been a challenge. This study aimed to improve the survivability of Lactobacillus rhamnosus GG under gastrointestinal condition through co-extrusion microencapsulation and the addition of black bean extract. @*Methodology and results@#Optimization was carried out on wall material formulation, types of pectin (low and high methoxyl pectin) and alginate: pectin ratio (2:1 and 3:1), and black bean extract concentration (0 to 1% w/v) to produce capsules with desired properties. The effect of L. rhamnosus GG microencapsulation with and without black bean extract on its survivability under simulated gastrointestinal conditions was also investigated. The optimal formulation that gives the highest microencapsulation efficiency (86.17%) was low methoxyl pectin, alginate: pectin ratio at 3:1, and 0.5% (w/v) of black bean extract. The inclusion of black bean extract into L. rhamnosus GG microencapsulation showed no significant effect (p >0.05) on the capsule diameter, with a mean diameter of 715.44 µm and a high microencapsulation efficiency of 97.4%. The viability of encapsulated L. rhamnosus GG increased with black bean extract after 6 h of sequential digestion with the final viable cell count of 12.47 log10 CFU/mL, which meet the minimum requirement of 10

Advances in Research on Lactobacillus rhamnosus GG in Colorectal Cancer / 胃肠病学

Colorectal cancer is a kind of malignant tumor that occurs in colorectal mucosal epithelium. Its morbidity and mortality are of the leading rank in the world. Probiotics are active microorganisms beneficial to the host, which can reduce the risk of malignant tumor by competing with pathogens, destroying potential carcinogens, reducing microbial genotoxicity, enhancing intestinal barrier and host innate immunity, regulating cell proliferation and apoptosis and other mechanisms. Lactobacillus rhamnosus GG (LGG) is one of the earliest and most widely studied probiotics, which has shown great potential in the tertiary prevention of cancer by regulating intestinal flora, anti-inflammatory, anti-proliferative and pro-apoptotic mechanisms. This article reviewed the advances in research on LGG in colorectal cancer.

Risk and Protective Factors for Gastrointestinal Symptoms associated with Antibiotic Treatment in Children: A Population Study / 대한소아소화기영양학회지

6 years (RR=4.15), with previous abdominal pain (RR=7.2) or constipation (RR=4.06). Constipation was recorded in 23/289 (8.0%), with increased risk in children having surgery (RR=2.56) or previous constipation (RR=7.38). Probiotic supplementation significantly reduced AAD (RR=0.30) and abdominal pain (RR=0.36). Lactobacillus rhamnosus GG (LGG) and L. reuteri significantly reduced AAD (RR=0.37 and 0.35) and abdominal pain (RR=0.37 and 0.24).CONCLUSION: AAD occurred in 20.4% of children, with increased risk at younger age, lower respiratory and urinary tract infections, intravenous treatment and previous AAD. LGG and L. reuteri reduced both AAD and associated abdominal pain.

Protection effect of Lactobacillus rhamnosus GG supplied in early life on intestinal barrier in offspring / 中华临床营养杂志

Objective To investigate the effects of Lactobacillus rhamnosus GG (LGG) colonization in early life on intestinal barrier and intestinal development in offspring mice and its possible mechanism.Methods Six C57BL/6 pregnant mice with the same conception time of 6 weeks were selected and randomly divided into experiment group given 108 cfu/ml LGG live bacteria and control group given LGG inactivated bacteria by gavage from the 18th day of pregnancy until natural birth.The progeny mice in the two groups were continued to be gavaged with 107 cfu/ml of LGG live bacteria or LGG inactivated bacteria on days 1-5 of birth.The body weight changes of 3 week'progeny mice were recorded.The colonization of LGG bacteria in offspring mice was detected at 2nd and 3rd weeks.The mRNA of intestinal proinflammatory cytokines and tight junction molecules were evaluated by real-time PCR method.HE,immunohistochemistry,immunofluorescence staining and enzyme-linked immunosorbent assay were used to evaluate the intestinal barrier of 3-week old off spring mice.Results Compared with the control group,the progeny mice of the experiment group showed no significant difference in body weight at the first week,and the body weight increased at the second week and the third week [2ndweek:(3.790±0.240) g vs.(4.326±0.140) g,t=3.707,P=0.006;3rd week:(7.295±0.326) g vs.(8.040±0.370) g,t=3.130,P=0.011].LGG colonization can be detected only in the feces of progeny mice in the experiment group.Intestinal colonization can promote the growth of small intestine villi and colon crypt depth [jejunum:(320.000±22.514) μm vs.(265.100±15.611) μm,t=8.258,P＜0.001;ileum:(150.500±13.099) μm vs.(111.000±11.308) μm,t=9.958,P＜0.001;colon:(295.000±15.209) μm vs.(233.100±6.678) μm,t=9.129,P＜0.001].Compared with the control group,the number of goblet cells in the colonic crypt of the experiment group increased (11.62 ± 0.780 vs.35.24 ±1.370,t=15.000,P＜0.001),and the relative mRNA expression levels of pro-inflammatory factors as IFN-γ (1.280±0.232 vs.0.512±0.206,t=4.970,P=0.001),IL-6 (1.364±0.271 vs.0.941±0.215,t=2.452,P=0.040),IL-10 (1.341±0.320vs.0.744±0.294,t=2.762,P=0.025)andTNF-α (3.702±0.150 vs.2.581±0.500,t=2.553,P=0.034) in the experiment group decreased;the expression levels of the intimate tight junction molecules (Claudin3) (1.283±0.152 vs.1.881±0.172,t=4.932,P=0.001) and the atresia protein molecule (Occludin) (1.164±0.342 vs.0.812±0.224,t=3.67,P=0.016) significantly increased.Conclusion Early life LGG colonization protects the intestinal barrier by inhibiting lowgrade intestinal inflammation.This study will lay the experimental foundation for the supplementation of probiotics in early life so as to prevent intestinal diseases.

Construction of Lactobacillus rhamnosus GG particles surface display system / 生物工程学报

To describe a novel particles surface display system which is consisted of gram-positive enhancer matrix (GEM) particles and anchor proteins for bacteria-like particles vaccines, we treated Lactobacillus rhamnosus GG bacteria with 10% heated-TCA for preparing GEM particles, and then identified the harvested GEM particles by electron microscopy, RT-PCR and SDS-PAGE. Meanwhile, Escherichia coli was induced to express hybrid proteins PA3-EGFP and P60-EGFP, and GEM particles were incubated with them. Then binding of anchor proteins were determined by Western blotting, transmission electron microscopy, fluorescence microscopy and spectrofluorometry. GEM particles preserved original size and shape, and proteins and DNA contents of GEM particles were released substantially. The two anchor proteins both had efficiently immobilized on the surface of GEM. GEM particles that were bounded by anchor proteins were brushy. The fluorescence of GEM particles anchoring PA3 was slightly brighter than P60, but the difference was not significant (P>0.05). GEM particles prepared from L. rhamnosus GG have a good binding efficiency with anchor proteins PA3-EGFP and P60-EGFP. Therefore, this novel foreign protein surface display system could be used for bacteria-like particle vaccines.

Las posibilidades del empleo de lactobacilos en la gastroenterología y la nutrición humana / Therapeutic potential of the use of lactobacilli in gastroenterology and human nutrition

Lactobacilli are normal inhabitants of the human intestine. They are also present in maternal milk, where Lactobacillus reuteri was first described. This confirms that maternal milk transfers components of the maternal microbiota to the offspring. L. reuteri ATCC 55730 decreases the intensity and duration of infantile colic defined as episodes of strident crying for at least 3 hours, 3 times a week during 3 months. Its etiology is unknown but recent evidence shows that affected infants harbor less Iactobacilli and more Bacteroides in their feces. In 2008 it was shown that this strain carries plasmidia that could transfer resistance to antibiotics. For this reason L. reuteri it was cured of these factors and this resulted in L. reuteri DSM 17938 which is devoid of them; tests in vitro, in laboratory animals and in humans demonstrated that both strains are equivalent. The effect of L. reuteri in infantile colic may be explained by its blocking at the level of the dorsal ganglia of the spinal chord of the transmission to the central nervous system of afferent pain stimuli. L. reuteri has effects on gastroesophageal reflux probably through the acceleration of gastric emptying and it decreases the incidence and severity of necrotizing enterocolitis. Other positive effects are related to chronic constipation and acute diarrhea. L. reuteri synthesizes a molecule with intense bactericidal activity, β-hidroxy propenal o reuterin from glycerol, which may explain some of its effects. Lactobacillus rhamnosus GG ATCC 53103 is probably the probiotic that has been studied the most and for a longer period. It shortens the duration and it decreases the number of evacuations in acute diarrhea as well as the duration and frequency of vomiting. It also induces improvements of antibiotic associated diarrhea and of upper respiratory infections. The positive effects of Lactobacillus GG may be due to the stimulation of innate and adaptive immunity and to the activation of macrophages and of NK lymphocytes. One of its effects refers the response its administration induces in infants with atopy/eccema. Some studies have demonstrated that the severity and extent of the lesions is decreased after Lactobacillus GG administration, that may persist for some years after its administration has been ended. Lactobacillus GG also exerts positive effects on conditions such as the irritable bowel syndrome through mechanisms that are not known with certainty.

Los lactobacilos son habitantes normales de las mucosas de los seres humanos; también están presentes en la leche materna. Lactobacillus reuteri fue descrito en la leche de una madre peruana, lo que confirma que estas bacterias son parte de la microbiota que las madres transfieren a sus hijos. El cólico infantil son episodios de llanto excesivo, estridente, por a lo menos 3 horas 3 veces por semana y por a lo menos 3 meses. L. reuteri ATCC 55730 demostró disminuir su intensidad y duración. La etiología del cólico se desconoce pero en las heces de los lactantes afectados se han demostrado disminuciones de los lactobacilos y aumentos de Bacteroides. En 2008 se demostró que L. reuteri ATCC 55730 era portador de plasmidios capaces de resistencia a antibióticos por lo que fue "curado" de estos factores y resultó una nueva cepa, denominada DSM 17938; pruebas in vitro y en animales mostraron su inocuidad y ensayos en humanos revelaron que ejerce las mismas propiedades benéficas que la cepa original. El efecto de L. reuteri en el cólico infantil puede estar relacionado con su capacidad de bloquear en los ganglios dorsales de la medula espinal la transmisión de estímulos a los centros cerebrales superiores. El efecto antirreflujo de L. reuteri se debería a la aceleración del vaciamiento gástrico. Ambas cepas de esta bacteria disminuyen la progresión, morbilidad y mortalidad de la enterocolitis necrosante. Otros efectos en la constipación crónica y la duración de la diarrea aguda de distintas etiologías. L. reuteri sintetiza una molécula, el β-hidroxipropenal o reuterina, con actividad antibacteriana. Lactobacillus rhamnosus GG ATCC 53103 es probablemente el probiótico más estudiado; acorta la duración de la diarrea aguda, disminuye el número de evacuaciones y la duración de los vómitos e induce mejorías de la diarrea asociada con los tratamientos antibióticos; y disminuiría la intensidad de las infecciones respiratorias altas. Sus efectos se deberían a la estimulación de las inmunidades innata y adquirida y de la activación de los macrófagos y los linfocitos NK. Un efecto notable y discutido de L. ramnosus GG se refiere a sus efectos en la atopia cutánea, ya que disminuiría la extensión e intensidad de sus síntomas, incluso más allá de su administración. También ejercería efectos sobre cuadros funcionales gastrointestinales como el síndrome del intestino irritable, por mecanismos aun no elucidados.

Prophylactic Effect of Lactobacillus GG in Animal Colitis and Its Effect on Cytokine Secretion and Mucin Gene Expressions / 대한소화기학회지

BACKGROUND/AIMS: Lactobacillus rhamnosus GG (LGG) has been used in acute colitis treatment. However, it is unclear whether the LGG prevents chronic colitis. The aim of this study was to examine the prophylactic effect of LGG on animal colitis, cytokine secretion, and mucin gene expression. METHODS: BALB/c mice (n=64) were exposed to 5% dextran sulfate sodium (DSS) for 7 days followed by 10 days recovery period and repeatedly exposed for 4 days. Then, the mice were devided into three group; group of oral LGG adminstration throughout the recovery and repeated colitis period; PBS group of PBS administration; control group. Colon length, histologic score, tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10) levels, mucin gene expressions were determined at each period. RESULTS: In acute colitis period, the LGG group showed higher levels of disease activity index (DAI), histologic score, TNF-alpha, IL-10, but shorter colon length, lower levels of mucin gene expressions than the control group. However, in repeated colitis period, the LGG group showed markedly lower levels of DAI and IL-10 but significantly longer colon length than PBS group (p<0.05). There was no difference in the mucin gene expression. CONCLUSIONS: These results suggest that LGG prevents chronic murine colitis. It may be associated with cytokine modulation and competitive inhibition of pathogenic bacteria. However, it may not be related with gene expression.