Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk

Abstract Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 g.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

Resumo O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 g.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

Partial purification of linoleic acid isomerase enzyme from Lactobacillus paracasei bacteria isolated from milk / Purificação parcial da enzima isomerase do ácido linoleico da bactéria Lactobacillus paracasei isolada do leite

Conjugated Linoleic Acid (CLA) has attracted the attention of many researchers, especially that of microbial origin due to its biological importance to the consumer. The current study aims to extract LA Isomerase enzyme from Lactobacillus paracasei bacteria from milk and to use the enzyme in the production of CLA. Selective media, including MRS and MRS-Dagatose, were used in isolating local strains. The selected bacterial isolates were tested for their ability to produce LA-Isomerase enzyme. The isolate with high enzymatic activity was selected. After extraction and partial purification of the enzyme, the optimal conditions for the production of conjugated fatty acid were studied, and the reaction products were diagnosed using GC-MS technology. It was found that 11 isolates have the ability to produce CLA at different concentrations, H1 isolate showed the highest production of conjugated fatty acid at a concentration of 120.45 g.ml-1, this isolate was selected as the source for enzyme extraction. The enzymatic activity of the crude extract and partially purified with ammonium sulfate was estimated using color methods at wavelength of 233 nm. The effect of the optimum conditions (pH, temperature, linoleic acid concentration and enzyme concentration) on the CLA product was studied using the partially purified LA Isomerase enzyme, the optimum conditions for production were 6.5, 45 °C, 100 µg.ml-1 and 0.7 ml, respectively. The GC-MS technique showed the presence of a number of reaction products that are isomers of conjugated linoleic acid (C9T11, T9T12, T10C12) with different concentrations.

O Ácido Linoleico Conjugado (CLA) tem chamado a atenção de diversos pesquisadores, principalmente aquele de origem microbiana, devido à sua importância biológica para o consumidor. O presente estudo visa extrair a enzima LA Isomerase da bactéria Lactobacillus paracasei do leite e usar essa enzima na produção de CLA. Meios seletivos, incluindo MRS e MRS-Dagatose, foram usados no isolamento de cepas locais. Os isolados bacterianos selecionados foram testados quanto à sua capacidade de produzir a enzima LA-Isomerase. Foi selecionado o isolado com alta atividade enzimática. Após a extração e purificação parcial da enzima, as condições ideais para a produção de ácido graxo conjugado foram estudadas e os produtos da reação foram identificados usando a tecnologia GC-MS. Verificou-se que 11 isolados possuem capacidade de produzir CLA em diferentes concentrações. O isolado H1 apresentou a maior produção de ácido graxo conjugado, na concentração de 120,45 g.ml-1, e este isolado foi selecionado como fonte para extração enzimática. A atividade enzimática do extrato bruto e parcialmente purificado com sulfato de amônio foi estimada por métodos de coloração em comprimento de onda de 233 nm. O efeito das condições ótimas (pH, temperatura, concentração de ácido linoleico e concentração de enzima) no produto CLA foi estudado usando a enzima LA Isomerase parcialmente purificada e as condições ótimas para produção foram 6,5, 45 °C, 100 µg.ml-1 e 0,7 mL, respectivamente. A técnica de GC-MS mostrou a presença de uma série de produtos de reação que são isômeros do ácido linoleico conjugado (C9T11, T9T12, T10C12) com diferentes concentrações.

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.

Effect of Lactobacillus crispatus on Female Reproductive Tract Health and Its Antibacterial and Anti-inflammatory Mechanisms / 中山大学学报(医学科学版)

Lactobacilli are important colonizing bacteria in female reproductive tract， among which Lactobacillus crispatus is closely associated with reproductive tract health and plays a crucial role in maintaining the vaginal microbiota balance. A decrease in the abundance of Lactobacillus crispatus may be correlated with various female reproductive tract diseases， such as bacterial vaginosis and vulvovaginal candidiasis， and it can even lead to adverse pregnancy outcomes such as infertility and embryo arrest. This article provides an overview of the basic characteristics of Lactobacillus crispatus， relationship between the decrease of its quantity and reproductive tract diseases， its ability to inhibit pathogenic bacteria such as Candida albicans and Chlamydia trachomatis， and the anti-inflammatory effects of Lactobacillus crispatus. The aim is to provide references for the use of Lactobacillus crispatus in vaginal therapeutics.

Isolation and identification of Lactobacillus paracasei and Bacillus subtilis in goose feces / 中国生物制品学杂志

@#Objective To isolate and identify Lactobacillus paracasei and Bacillus subtilis from feces of northern white geese.Methods Lactobacillus paracasei and Bacillus subtilis,numbered S00834-8 and IS00439-2 respectively,were isolated from feces of 30 52-week-old northern white geese and cultured in specific medium,and identified by 16S DNA sequencing. The two strains were cultured for different time duration(0,2,4,6,8,10,12,14,16,18,20,22,24,36 and 48 h),at different temperature(34,36,37,38 and 40 ℃)and pH(5. 5,6. 0,6. 5,7. 0,7. 5 and 8. 0). The growth characteristics of the strains were evaluated according to the A600values measured by UV-Vis spectrophotometer. At the same time,the acid and bile salt tolerance(pH 2. 0,2. 5,3. 0,3. 5 and 7. 0,bile salt concentration 0,0. 3%,0. 5%,1. 0% and 1. 5%)and the antibiotic tolerance were tested. Results The strains S00834-8 and IS00439-2 were Lactobacillus paracasei subspecies tolerance H and Bacillus subtilis KA9,respectively. The strain S00834-8 cultured under the optimum condition of the culture medium with pH 6. 0,at 37 ℃,for 12 h,had good bile resistance and gastric acid resistance,which was moderately susceptible to piperacillin,norfloxacin,amikacin,gentamicin and tetracycline,and susceptible to levofloxacin,streptomycin and chloramphenicol. The strain IS00439-2 best cultured in the culture medium with pH 7. 5,at 36 ℃,for 8 h,showed good bile resistance and gastric acid resistance,which was moderately sensitive to levofloxacin and tetracycline,while sensitive to piperacillin,norfloxacin,streptomycin and chloramphenicol. Conclusion Goose-derived Lactobacillus paracasei subspecies tolerance H and Bacillus subtilis KA9 have good acid resistance and bile salt resistance,which are expected to be candidate strains for goose-derived microecological preparations.

Analysis of Bifidobacterium animalis subsp. lactis BB-12 ® and Lactobacillus rhamnosus GG on underweight and malabsorption in premature infants

SUMMARY OBJECTIVE: This study aimed to explore and analyze the therapeutic effect of the combination of Bifidobacterium animalis subsp. lactis BB-12® and Lactobacillus rhamnosus GG on underweight and malabsorption in premature infants. METHODS: This is a retrospective study. The clinical data of 68 premature infants admitted to Beijing United Family Hospital (Private Secondary Comprehensive Hospital, Chaoyang District, Beijing, China) from January 2016 to January 2022 were analyzed retrospectively. Preterm infants less than 37 weeks of gestational age admitted to the neonatal intensive care unit were included in the study. Patients with intestinal malformations, necrotizing enterocolitis, etc., who require long-term fasting were excluded. A telephone follow-up was performed 3-6 months after discharge. They were classified as treatment groups A and B according to the treatment plan. The treatment group A included parenteral nutrition, enteral nutrition, etc. In treatment group B, based on treatment group A, the premature infants were treated with Bifidobacterium animalis subsp. lactis BB-12® and Lactobacillus rhamnosus GG. The time to regain birthweight and the weight on day 30 were compared between the two groups, as was the duration of transition from parenteral nutrition to total enteral nutrition. RESULTS: The time of weight regain birthweight in group B was shorter than that in group A (t=-2.560; t=-4.287; p<0.05). The increase of weight on day 30 in group B was significantly higher than that in group A (t=2.591; t=2.651; p<0.05). The time from parenteral nutrition to total enteral nutrition in group B was shorter than that in group A (z=-2.145; z=-2.236; p<0.05). CONCLUSION: In the treatment of premature infants, the combination of Bifidobacterium animalis subsp. lactis BB-12® and Lactobacillus rhamnosus GG can have a better therapeutic effect on the underweight and malabsorption of premature infants, and this treatment method can be popularized in clinics.

Vaginosis citolítica: actualización diagnóstica y terapéutica / Cytolytic vaginosis: diagnostic and therapeutic update

El objetivo es conocer el abordaje diagnóstico y terapéutico de la vaginosis citolítica. Para ello se hizo una búsqueda sistemática de la literatura médica mediante las bases de datos: PubMed, Central, etc. Se limitó a ensayos clínicos aleatorizados, metaanálisis y revisiones bibliográficas, con disponibilidad del texto completo para evaluarlos en su totalidad e incluirlos en la revisión; publicados entre 1980 y 2021. Se incluyeron 27 publicaciones. La vaginosis citolítica es un trastorno infradiagnosticado. En mujeres con síntomas y signos de candidiasis vulvovaginal, que no responden a la terapia, se debe considerar la probabilidad de estar en presencia de una vaginosis citolítica. El tratamiento obliga a elevar el pH vaginal a valores básicos y a disminuir el número excesivo de Lactobacillus, resultando de utilidad las duchas vaginales con bicarbonato de sodio y/o un tratamiento con antibióticos derivados de la penicilina junto a un inhibidor de ß-lactamasas o doxiciclina en las pacientes alérgicas a la penicilina. Finalmente, se concluye que la vaginosis citolítica es una afección común, frecuentemente se diagnostica de forma errónea porque se confunde con la candidiasis vulvovaginal; se caracteriza por producir un cuadro clínico similar. El tratamiento se enfoca en disminuir el número de Lactobacillus y la elevación del pH vaginal.

The objective is to know the diagnostic and therapeutic approach of cytolytic vaginosis. A systematic search of the medical literature was carried out using the following databases: Medline via PubMed, Central, and Cochrane Database of Systematic Reviews, among others. The search was limited to randomized clinical trials, meta-analyses, and literature reviews that had the full text available for full evaluation and inclusion in the review; published between 1980 and 2021. Twenty-seven publications were included. Cytolytic vaginosis is a frequently underdiagnosed disorder, which mimics Candida vaginitis. In women with symptoms and signs of vulvovaginal candidiasis who do not respond to antifungal therapy, the possibility of cytolytic vaginosis should be considered. The treatment of this condition requires raising the vaginal pH to basic values and reducing the excessive number of Lactobacillus, resulting in useful vaginal douches with sodium bicarbonate and/or treatment with antibiotics derived from penicillin together with a ß-lactamases inhibitor or doxycycline in patients allergic to penicillin. Finally, we conclude that cytolytic vaginosis is a common condition, frequently misdiagnosed because it is confused with vulvovaginal candidiasis, since it is characterized by producing a similar clinical picture. Treatment focuses on reducing the number of Lactobacillus and raising vaginal pH.

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)

The Efficacy and Safety of a Combination of Amino Acids, Vitamins and Probiotics in PROM and PPROM

The prevalence and severity of prelabor rupture of the membranes (PROM)/preterm PROM (PPROM) are a worldwide public health concern. PROM is the result of a cascade of events involving matrix metalloproteinase (MMP)-9, tissue inhibitor of metalloproteinases 1 (TIMP1), cytokines and proapoptotic genes, which is initiated by several factors such as infection, genotoxic agents or some unknown etiology. In PROM, there is an increased expression and activation of MMP-2, MMP-3 and MMP-9 and a reduction of TIMP1. p53 and tumor necrosis factor (TNF)-? mediate the major apoptotic pathway of PROM. p53 can transactivate some MMP genes, resulting in the overexpression of MMPs. This leads to apoptosis. MMP-2 and MMP-9 degrades type-IV collagen, which is the major structural component of chorioamnion. Understanding the fundamental pathology at the molecular level, it appears necessary to adjust the biologically protective mechanism to prevent spontaneous preterm labor. Our findings show that the novel combination of arginine, ascorbic acid, folic acid, glutamine, glutathione, thiamine, lactic acid bacillus spores, vitamin E acetate and pyridoxine is safe and effectively prevents PROM and PPROM (97% patients) and prolongs pregnancy term.

Heat-killed Lactobacillus murinus confers neuropro-tection against dopamine neuronal loss by targeting NLRP3 inflammasome / 中国药理学与毒理学杂志

OBJECTIVE In this study,the effects of live Lactobacillus murinus(L.m)and heat-killed L.muri-nus(H-k L.m)on DA neuronal damage in rats and the underlying mechanisms were investigated.METHODS Male SD rats were randomly divided into five groups:vehicle group,L.m/H-k L.m(1×109 cfu)group,6-OHDH group,6-OHDH + L.m/H-k L.m(1×107 cfu)group,and 6-OHDH + L.m/H-k L.m(1×109 cfu)group.Wild-type and NLRP3 knockout mice were divided into three groups:sham(vehicle),6-OHDH,and 6-OHDH + H-k L.m(1×109 cfu).The model was established after five weeks of pre-administration.Motor ability of experimental mice was assessed by rotarod,mine,and stepping experiments;the expression of dopaminergic neuron markers—tyro-sine hydroxylase(TH),microglial cell markers—ionized calnexin 1(IBA-1),and NOD-like receptor family protein 3(NLRP3)in the substantia nigra was detected by immunohistochemistry and immunofluorescence experi-ments.The expression changes of TH,IBA-1,NLRP3,apoptosis-associated microparticle protein(ASC),cas-pase 1,and inflammatory factors such as interleukin-1β(IL-1β),IL-18,and tumor necrosis factor-α(TNF-α)were detected by immunoblotting experiments.RESULTS H-k L.m ameliorated 6-OHDH-induced motor dysfunctions and loss of substantia nigra DA neurons,while no protec-tion was shown in live L.m treatment.At the same time,H-k L.m reduced the activation of NLRP3 inflammasome in microglia and the secretion of pro-inflammatory factors,thus inhibiting the development of neuroinflammation.Fur-thermore,H-k L.m failed to display its original neuropro-tective properties in NLRP3 inflammasome knockout mice.CONCLUSION H-k L.m conferred neuroprotec-tion against DA neuronal loss via the inhibition of microglial NLRP3 inflammasome activation,these findings provide a promising potential for future applications of L.m,and also beneficial strategy for PD treatment.

Effect of Lactobacillus plantarum WCFS1 on pancreatic and ileal injury in mice with acute necrotizing pancreatitis / 中华胰腺病杂志

Objective:To explore the effect of probiotics Lactiplantibacillus plantarum(LP) WCFS1 by gavage on acute necrotizing pancreatitis (ANP) and associated ileum injury in mice. Methods:Twenty-four healthy male mice were gavaged with broad-spectrum antibiotics for 3 weeks to establish microbiota-depleted mice, and then randomly divided into control group (CON), ANP model group (ANP), LP gavage group (LP) and LP gavage and ANP induced group (LP+ ANP) , with 6 mice in each group. Mice in LP and LP+ ANP group were treated by gavage of LP (1×10 9 CFU/ml, 0.2 ml/day per mouse) for 1 week, while CON and ANP were gavaged with sterile phosphate buffered saline for 1 week instead. The ANP model was induced by intraperitoneal injection with caerulein (100 μg/kg) for 10 times with 1-hour interval between two injections and the 10th injection with lipopolysaccharide(LPS) 5 mg/kg intraperitoneally, and the mice were sacrificed 2 h later. Levels of LP in stool and ileal mucosa were detected by real-time PCR; the pancreas and ileum were collected for pathological examination to observe the extent of tissue inflammation and to score the pathology. Serum amylase activities were determined by enzymatic kinetic chemistry; serum inflammators levels and intestinal permeability were detected by ELISA; levels of inflammators in pancreatic and ileal tissues were detected by real-time PCR; ileal tight-junction proteins (occludin, claudin-1 and ZO-1) were measured by immunofluorescence staining. Results:LP levels in the stool and ileal mucosa of mice were significantly increased after LP gavage, and the differences were statistically significant (913.30±39.12 vs 2.39±1.39, 23.11±0.50 vs 1.38±0.28, all P value <0.05). The pathological scores of pancreatic tissue of CON, LP, ANP and LP+ ANP group were (0.26±0.41), (0.17±0.26), (8.55±0.46) and (6.30±0.45); the serum amylase activities were (219.70±19.73), (217.60±11.30), (2896.24±98.32) and (1837.13±131.60)U/L, IL-1β were (0.87±0.28), (1.4±0.85), (67.41±6.45) and (36.33±5.65)pg/ml, IL-6 were (0.74±0.27), (0.16±0.16), (280586.12±39163.92) and (107912.75±31283.47)pg/ml, IL-10 were (35.52±5.27), (50.99±15.34), (2008.45±184.83) and (3070.35±403.71)pg/ml; the expression level of pancreatic IL-1β mRNA was 1.42±0.39, 0.95±25, 20.53±0.50 and 10.69±1.01, IL-6 mRNA was 1.31±0.44, 0.93±0.023, 21.97±1.71 and 11.54±1.75, IL-10 mRNA was 0.93±0.14, 0.75±0.15, 0.99±0.21 and 1.76±0.19; there was no significant difference between LP and CON group, and pancreatic pathological scores, serum amylase、IL-1β and IL-6 levels, and the expression level of pancreatic IL-1β and IL-6 mRNA were significantly decreased in LP+ ANP group compared with those in ANP group, while serum IL-10 levels and the expression level of pancreatic IL-10 mRNA were significantly increased compared with ANP group, and all the differences were statistically significant (all P values <0.05). The pathological scores of ileal tissue of CON, LP, ANP and LP+ ANP group were 0, 0, (3.17±0.41) and (1.67±0.52); the levels of serum DAO of CON, LP, ANP and LP+ ANP group were (0.03±0.03), (0.02±0.02), (0.50±0.05) and (0.49±0.06)ng/ml; LPS levels were (2.75±0.35), (3.74±0.28), (7.19±0.92) and (5.88±0.38)ng/ml; the expression level of ileal IL-1β mRNA was 1.21±0.20, 1.17±0.09, 1.81±0.25 and 1.63±0.21; IL-6 mRNA was 1.01±0.29, 2.83±0.42, 54.45±8.50 and 16.87±4.42; IL-10 mRNA was 1.12±0.41, 6.09±2.51, 11.65±1.47 and 29.86±2.93. There was no significant difference between LP and CON group, except that the ileal IL-10 mRNA expression was significantly higher than that of CON group. Ileal pathological scores, serum LPS levels and the expression level of ileal IL-6 mRNA were significantly lower in LP+ ANP group than those in ANP group, while the expression level of ileal IL-10 mRNA was significantly higher than that of ANP group; the expression of ileal tight junction proteins (ocludin, claudin-1, ZO-1) was significantly higher than those in ANP group, and all the differences were statistically significant (all P values <0.05). Conclusions:LP WCFS1 gavage could ameliorate the injury of pancreatic and ileal barrier in caerulein-induced ANP mice.

Effect of Anchang Decoction on intestinal flora and fecal calcitonin in rats with severe ulcerative colitis / 国际中医中药杂志

Objective:To investigate the effects of Anchang Decoction on intestinal microflora and faecal calprotectin (FC) in rats with severe ulcerative colitis based on probiotics.Methods:Totally 50 rats were taken to prepare a model of severe ulcerative colitis. The rats who successfully modeled were divided into model group, Anchang Decoction low-, medium-, high-dosage groups, Lizhu Changle group, and mesalazine group, with 5 rats in each group. Another 6 rats were set and the blank group. Lizhu Changle group received Lizhu Changle suspension (containing 50 million live bacteria/ml) for gavage; Anchang Decoction low-, medium-, high-dosage groups received Anchang Decoction 1, 5, 10 ml/(kg·d) for gavage; Mesalazine group received Mesalazine suspension (10.5 mg/100 g) for gavage. Rats in the blank group and model group were gavaged with the same volume of normal saline for 7 days. Body mass and disease activity index (DAI) before and after administration were measured. 16s rDNA of lactobacillus and bifidobacterium in feces of rats was detected. FC content of rats was detected by ELISA.Results:Compared with the model group, the weight of rats in each administration group increased ( P<0.05), DAI score decreased ( P<0.05), and the level of calprotectin in feces decreased ( P<0.05), and there was no significant difference in the levels of lactobacillus and bifidobacterium in each administration group ( P>0.05). Conclusion:Anchang Decoction can improve the intestinal micro-ecology by regulating the level of calprotectin in rats with severe ulcerative colitis, thereby playing a therapeutic role.

Effect of Liquiritigenin on Intestinal Flora in Ovariectomized APP/PS1 Transgenic Mice / 中山大学学报(医学科学版)

ObjectiveTo investigate the effect of liquiritigenin （LG） on intestinal flora in menopausal APP/PS1 mice. MethodsA total of forty 3-month-old female APP/PS1 mice were randomly divided into sham surgery group （n=20） and ovariectomy group （n=20）. Seven days after surgery， the ovariectomy group was randomly divided into ovariectomy control group （OVX， n=10）， ovariectomy + liquiritigenin treatment group （OVX + LG， n=10）， and the sham surgery group was randomly divided into liquiritigenin treatment group （LG， n=10） and reagent control group （Sham， n=10）， and ten C57BL/6J mice were taken as WT group. The dose of LG group and OVX + LG group was 30 mg•kg-1•d-1. After 90 days of drug treatment， fecal samples were gathered， genomes were extracted， and intestinal flora were analyzed by 16S rDNA Amplicon Sequencing. Morris water maze was performed to evaluate learning and memory abilities of mice. Immunofluorescence was used to observe the deposition of senile plaques （SP） in the brain of mice. ResultsThe results of water maze showed that LG significantly improved the learning memory ability of APP/PS1 mice with/without OVX （P<0.05）， and reduced the number of SPs in the brain of APP/PS1 mice with/without OVX， and the differences were statistically significant （P<0.000 1）. 16s rDNA sequencing analysis of the relative abundance of gut microbiota proved that LG treatment significantly increased the relative abundance of Firmicutes and Lactobacillus （P<0.05） and reduced the relative abundance of harmful bacteria belong to Bacteroidetes （P<0.05） in APP/PS1 mice intestines with/without menopause. After LG treatment， the relative abundance of Allobaculun elevated in the intestines of APP/PS1 mice， while declined in the intestines of menopausal APP/PS1 mice， but the difference was not statistically significant. LEfSe analysis revealed the bacteria with the most differential abundance of the gut microbiota of WT mice were Firmicutes， Bacillus， and Lactobacillales （P<0.05）； Lactobacillus reuteri had a greater influence on the LG group （P<0.05）； Bacteroidia， Bacteroidales and Bacteroides gathered in the intestines of mice in the Sham group （P<0.05）. Firmicutes and Allobaculum were the dominant in the WT group （P<0.05）； Bacteroides， Bacteroidia and Bacteroidales were more abundant in the Sham group（P<0.05）； Bacterroidaceae and Bacteroides had the most differential abundances in the OVX group （P<0.05）； Lactobacillaceae and Lactobacillus were more abundant in the intestines in the OVX + LG group （P<0.05）. ConclusionLG could improve the ratio of beneficial and harmful bacteria in the intestines of APP/PS1 mice before and after menopause. Liquiritigenin treatment showed consistent variations in intestinal flora in APP/PS1 mice with or without ovariectomy. It is presumed that menopausal APP/PS1 mice have lipid metabolism disorders which requires further study.

Lactobacillus gasseri LA39 promotes hepatic primary bile acid biosynthesis and intestinal secondary bile acid biotransformation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

A growing body of evidence has linked the gut microbiota to liver metabolism. The manipulation of intestinal microflora has been considered as a promising avenue to promote liver health. However, the effects of Lactobacillus gasseri LA39, a potential probiotic, on liver metabolism remain unclear. Accumulating studies have investigated the proteomic profile for mining the host biological events affected by microbes, and used the germ-free (GF) mouse model to evaluate host-microbe interaction. Here, we explored the effects of L. gasseri LA39 gavage on the protein expression profiles of the liver of GF mice. Our results showed that a total of 128 proteins were upregulated, whereas a total of 123 proteins were downregulated by treatment with L. gasseri LA39. Further bioinformatics analyses suggested that the primary bile acid (BA) biosynthesis pathway in the liver was activated by L. gasseri LA39. Three differentially expressed proteins (cytochrome P450 family 27 subfamily A member 1 (CYP27A1), cytochrome P450 family 7 subfamily B member 1 (CYP7B1), and cytochrome P450 family 8 subfamily B member 1 (CYP8B1)) involved in the primary BA biosynthesis pathway were further validated by western blot assay. In addition, targeted metabolomic analyses demonstrated that serum and fecal β‍-muricholic acid (a primary BA), dehydrolithocholic acid (a secondary BA), and glycolithocholic acid-3-sulfate (a secondary BA) were significantly increased by L. gasseri LA39. Thus, our data revealed that L. gasseri LA39 activates the hepatic primary BA biosynthesis and promotes the intestinal secondary BA biotransformation. Based on these findings, we suggest that L. gasseri LA39 confers an important function in the gut‒liver axis through regulating BA metabolism.

Changes in intestinal microbiota and application of probiotics in patients with acute pancreatitis / 临床肝胆病杂志

Intestinal microbiota is the most complex and important microecosystem in the human body， and gut microbiota dysbiosis is closely associated with the development and progression of acute pancreatitis. Targeted regulation of intestinal microecology in assisting the treatment of acute pancreatitis has attracted more attention in recent years. This article describes the changes in intestinal microbiota and related mechanisms in patients with acute pancreatitis， summarizes the current research status of the use of probiotics， points out the research direction of probiotics as the adjuvant treatment regime， and proposes a new method for predicting the dominant flora in patients with acute pancreatitis， in order to bring new ideas for the treatment of acute pancreatitis.

Distribution and function of cariogenic bacteria in supragingival plaque in patients with type 2 diabetes mellitus / 口腔疾病防治

Objective@#To investigate the distribution characteristics and functional genes of cariogenic bacteria in oral microorganisms of patients with type 2 diabetes mellitus and to improve the understanding of the relationship between type 2 diabetes mellitus and dental caries.@*Methods@#The experimental group included 10 patients with type 2 diabetes treated in the Department of Endocrinology, the First Affiliated Hospital of Air Force Medical University. The normal control group included healthy oral subjects without type 2 diabetes in the community population (10 cases). Samples of supragingival plaque from patients with type 2 diabetes mellitus and normal controls were collected and sequenced. Bioinformatics and statistical analysis of cariogenic bacteria such as Streptococcus mutans, Lactobacillus, Actinomyces viscosus and Candida albicans were carried out.@*Results@#There were slightly fewer cariogenic bacteria such as Streptococcus mutans, Lactobacillus, Actinomyces viscosus and Candida albicans in supragingival plaque samples of type 2 diabetic patients than in normal controls, but the difference was not statistically significant (P>0.05). The results of KEGG pathway functional metabolic differences showed that the metabolic pathways of D-arginine and D-ornithine metabolism, biofilm formation-Escherichia coli, carolactam degradation and arginine biosynthesis were more abundant in the T2DM group than in the normal control group, while metabolic pathways such as tyrosine metabolism, selenocompound metabolism and pyruvate metabolism showed the opposite trend. @*Conclusion @#There was no significant difference in the content of cariogenic microorganisms between type 2 diabetic patients and normal control group. The differential metabolic pathways of the functional genes indicated that an increase in the arginine metabolic pathway was beneficial to the maintenance of acid-base balance in the oral microecological environment.

Herbal formula BaWeiBaiDuSan alleviates polymicrobial sepsis-induced liver injury via increasing the gut microbiota Lactobacillus johnsonii and regulating macrophage anti-inflammatory activity in mice

Sepsis-induced liver injury (SILI) is an important cause of septicemia deaths. BaWeiBaiDuSan (BWBDS) was extracted from a formula of Panax ginseng C. A. Meyer, Lilium brownie F. E. Brown ex Miellez var. viridulum Baker, Polygonatum sibiricum Delar. ex Redoute, Lonicera japonica Thunb., Hippophae rhamnoides Linn., Amygdalus Communis Vas, Platycodon grandiflorus (Jacq.) A. DC., and Cortex Phelloderdri. Herein, we investigated whether the BWBDS treatment could reverse SILI by the mechanism of modulating gut microbiota. BWBDS protected mice against SILI, which was associated with promoting macrophage anti-inflammatory activity and enhancing intestinal integrity. BWBDS selectively promoted the growth of Lactobacillus johnsonii (L. johnsonii) in cecal ligation and puncture treated mice. Fecal microbiota transplantation treatment indicated that gut bacteria correlated with sepsis and was required for BWBDS anti-sepsis effects. Notably, L. johnsonii significantly reduced SILI by promoting macrophage anti-inflammatory activity, increasing interleukin-10+ M2 macrophage production and enhancing intestinal integrity. Furthermore, heat inactivation L. johnsonii (HI-L. johnsonii) treatment promoted macrophage anti-inflammatory activity and alleviated SILI. Our findings revealed BWBDS and gut microbiota L. johnsonii as novel prebiotic and probiotic that may be used to treat SILI. The potential underlying mechanism was at least in part, via L. johnsonii-dependent immune regulation and interleukin-10+ M2 macrophage production.

Immobilization of Lactobacillus bulgaricus with gellan gum and its application in continuous fermentation of D-lactic acid from corn straw / 生物工程学报

Biorefinery of chemicals from straw is an effective approach to alleviate the environmental pollution caused by straw burning. In this paper, we prepared gellan gum immobilized Lactobacillus bulgaricus T15 gel beads (LA-GAGR-T15 gel beads), characterized their properties, and established a continuous cell recycle fermentation process for D-lactate (D-LA) production using the LA-GAGR-T15 gel beads. The fracture stress of LA-GAGR-T15 gel beads was (91.68±0.11) kPa, which was 125.12% higher than that of the calcium alginate immobilized T15 gel beads (calcium alginate-T15 gel beads). This indicated that the strength of LA-GAGR-T15 gel beads was stronger, and the strain was less likely to leak out. The average D-LA production was (72.90±2.79) g/L after fermentation for ten recycles (720 h) using LA-GAGR-T15 gel beads as the starting strain and glucose as the substrate, which was 33.85% higher than that of calcium alginate-T15 gel beads and 37.70% higher than that of free T15. Subsequently, glucose was replaced by enzymatically hydrolyzed corn straw and fermented for ten recycles (240 h) using LA-GAGR-T15 gel beads. The yield of D-LA reached (1.74±0.79) g/(L·h), which was much higher than that of using free bacteria. The wear rate of gel beads was less than 5% after ten recycles, which indicated that LA-GAGR is a good carrier for cell immobilization and can be widely used in industrial fermentation. This study provides basic data for the industrial production of D-LA using cell-recycled fermentation, and provides a new way for the biorefinery of D-LA from corn straw.

Administração de Lactobacillus para casei 28.4 como terapia adjuvante no tratamento da periodontite: desenvolvimento in vitro de formulação probiótica usando gellan gum e ensaio clínico randomizado placebo controlado / Administration of Lactobacillus paracasei 28.4 as adjuvant therapy in the treatment of periodontitis: in vitro development of the probiotic formulation using gellan gum and placebo-controlled randomized clinical trial

A periodontite é uma doença inflamatória do periodonto associada ao acúmulo de biofilme dental, com consequente disbiose da microbiota oral e alteração da resposta imuno-inflamatória. O controle da periodontite realizado por meio do tratamento periodontal não-cirúrgico altera o ambiente subgengival e pode ser associado a terapias adjuntas. O uso de probióticos como terapia adjuvante a esse tratamento parece favorecer a modificação do biofilme bacteriano e resposta imuno-inflamatória. Entretanto, os probióticos empregados até o momento não são próprios da cavidade oral e, por isso, a busca por cepas originadas da microbiota oral tem ganhado espaço na tentativa de favorecer a aderência e colonização permanente dessas bactérias. Recentemente, a cepa de Lactobacillus paracasei 28.4 foi isolada da cavidade oral, exibindo atividades antimicrobianas promissoras sobre o controle de patógenos orais. O presente estudo teve como objetivos o desenvolvimento de uma formulação probiótica de L. paracasei 28.4 incorporado em gellan gum para uso humano, bem como a avaliação dos efeitos clínicos da administração desta formulação como adjuvante no tratamento periodontal. Para tal, o trabalho foi dividido em 2 etapas, contemplando uma etapa laboratorial (parte A), para desenvolvimento da formulação probiótica; e um ensaio clínico randomizado (parte B, n=40), envolvendo um protocolo de tratamento periodontal não cirúrgico associado à terapia adjunta probiótica ou placebo. Os resultados da parte A permitiram encontrar a melhor forma de apresentação e armazenamento da formulação de gellan gum. Na parte B, aos 3 e 6 meses de acompanhamento, os dois grupos apresentaram melhoras significativas dos parâmetros periodontais (índice de sangramento, índice gengival, profundidade de sondagem e ganho de nível de inserção) em relação ao baseline, sem diferença estatística na comparação intergrupo. Como conclusão, uma formulação probiótica segura e possível de ser aplicada na prática clínica foi obtida; no entanto, sua administração não promoveu efeitos clínicos adicionais ao tratamento de pacientes com periodontite generalizada estágios III/IV e graus B/C (AU)

Periodontitis is an inflammatory disease of the periodontium associated with the accumulation of dental biofilm, with consequent dysbiosis of the oral microbiota and alteration of the immune-inflammatory responseThe control of periodontitis carried out through non-surgical periodontal treatment alters the subgingival environment and can be associated with adjunctive therapies.The use of probiotics as an adjuvant therapy in the periodontal treatment seems to favor the modification of the bacterial biofilm and modulation of the immuneinflammatory response. However, the probiotics used so far are not specific to the oral cavity and, therefore, the search for strains originating from the oral microbiota has gained space to favor the adherence and permanent colonization of these bacteria. Recently, strain of Lactobacillus paracasei 28.4 was isolated from the oral cavity itself, showing promising antimicrobial activities in the control of oral pathogens. The present study aimed to develop and characterize a probiotic formulation of L. paracasei 28.4 incorporated into gellan gum for human use, as well as to evaluate the clinical effects of administering this formulation as an adjuvant in the treatment of periodontitis. To this end, the study was divided into 2 stages, comprising a laboratory stage (part A), for the development of the probiotic, and a randomized clinical trial (part B, n=40) contemplating a non-surgical periodontal treatment protocol associated with adjunctive probiotic therapy or placebo. The results from part A made it possible to find the best way to present and store the gellan gum formulation. In part B, at 3 and 6 months of follow-up, both groups showed significant improvements in periodontal parameters (bleeding index, gingival index, probing depth and attachment level gain) in relation to baseline, with no statistical difference in the intergroup comparison. In conclusion, a safe probiotic formulation that can be applied in clinical practice was obtained; however, its administration did not promote additional clinical effects in the treatment of patients with stage III/IV and grades B/C generalized periodontitis (AU)

Is the use of Lactobacillus reuteri probiotic efficient as adjunctive therapy in the treatment of periodontitis? A systematic review / O uso do probiótico Lactobacillus reuteri como terapia adjuvante no tratamento da periodontite é eficaz? Uma revisão sistemática

Objective: This systematic review had the purpose to validate the probiotic Lactobacillus reuteri as adjuvant therapy in the periodontal treatment of periodontitis, by the analysis of randomized controlled trial, controlled clinical trial, and observational studies. Material and Methods: Search keys related to the subject were defined, and the following databases were used as search strategies: MEDLINE via PubMed, Scopus, Web of Science, Cochrane Central Controlled Trials Registry, and EMBASE. The data selection and study were performed by two independent evaluators: first, they selected the article by title and abstract and subsequently qualified according to the bias risk analysis. A narrative synthesis has been performed based on the data obtained from the best-quality articles. After data extraction, their heterogeneity was analyzed, and a meta-analysis was performed. Results: In general, the results of the meta-analysis were positive for the use of probiotics as an adjuvant treatment. Considering the limitations of the comparisons between the analyzed articles studied, the conclusion was that probiotics may provide supplementary benefits to the treatment of periodontitis, with improvement in bleeding on probing rates and probing depth (AU)

Objetivo: Esta revisão sistemática teve como objetivo validar o uso do probiótico Lactobacillus reuteri como terapia adjuvante no tratamento periodontal da periodontite, por meio da análise de ensaios clínicos randomizados, ensaios clínicos controlados e estudos observacionais. Material e Métodos: Chaves de busca relacionadas ao assunto foram definidas, e as seguintes bases de dados foram utilizadas como estratégias de busca: MEDLINE via PubMed, Scopus, Web of Science, Cochrane Central Controlled Trials Registry e EMBASE. A seleção dos dados e o estudo foram realizados por dois avaliadores independentes: primeiro, selecionaram o artigo pelo título e resumo e posteriormente qualificaram de acordo com a análise de risco de viés. Uma síntese narrativa foi realizada com base nos dados obtidos dos artigos de melhor qualidade. Após a extração dos dados, sua heterogeneidade foi analisada e uma meta-análise foi realizada. Resultados: Em geral, os resultados da meta-análise foram positivos para o uso de probióticos como tratamento adjuvante. Considerando as limitações das comparações entre os artigos analisados estudados, concluiu-se que os probióticos podem trazer benefícios complementares ao tratamento da periodontite, com melhora nas taxas de sangramento à sondagem e na profundidade de sondagem (AU)