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Broccoli extract mainly contains polyphenols and glucosinolates (GSLs). GSLs can be hydrolyzed by gut microorganisms into isothiocyanates (ITCs) and other active substances. These substances have anticancer, anti-inflammatory, antimicrobial, and atherosclerosis-reducing functions. In this study, a high concentration (2000 µmol/L GSLs and 24 µmol/L polyphenols) and a low concentration (83 µmol/L GSLs and 1 µmol/L polyphenols) of broccoli extract were prepared. Gut microorganisms from fresh human feces were cultured to simulate the gut environment in vitro. The GSL content decreased and the types and content of ITCs increased with broccoli extract hydrolysis through cyclic condensation and gas chromatography-mass spectrometry (GC-MS) analyses. Broccoli extract significantly increased probiotics and inhibited harmful bacteria through 16S rDNA sequencing. Based on phylum level analysis, Firmicutes and Lachnospiraceae increased significantly (P < 0.05). At the genus level, both high- and low-concentration groups significantly inhibited Escherichia and increased Bilophila and Alistipes (P < 0.05). The high-concentration group significantly increased Bifidobacterium (P < 0.05). The broccoli extract improved the richness of gut microorganisms and regulated their structure. The GSL hydrolysis was significantly correlated with Bilophila, Lachnospiraceae, Alistipes, Bifidobacterium, Escherichia, and Streptococcus (P < 0.05). These study findings provide a theoretical foundation for further exploring a probiotic mechanism of broccoli extract in the intestine.

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The milk-clotting enzyme (MCE) is a crucial ingredient in cheese manufacture. Due to the limits of traditional MCE, finding viable substitute is a pressing issue. This study aims to isolate and identify a wild strain with high milk-clotting activity (MCA) and low proteolytic activity (PA) and optimize the fermentation conditions for MCE production. A strain of Bacillus velezensis DB219 with high MCA/PA value (9.2) was isolated from dairy soil (Wuchang, Heilongjiang, China) and identified through 16S rRNA from 40 strains. The optimal wheat bran, carbon, nitrogen, inoculum size, volume and initial pH were 60 g/L, soluble starch 12.5 g/L, corn steep liquor 3 g/L, 5%, 40 mL and 6.15, respectively for improving DB219 MCE production through single factor experiment. The wheat bran concentration, corn steep liquor concentration and volume were the most critical factor and their changed range was determined through Plackett-Burman design and the steepest ascent/descent experiments. The response surface analysis experiment of three factors and three levels was conducted by Box-Behnken design. The theoretical optimal fermentation conditions for DB219 MCE were as follows: wheat bran concentration 60.14 g/L, soluble starch 12.5 g/L, corn steep liquor 3 g/L, inoculum size 5%, volume 40.08 mL and initial pH 6.15. DB219 MCE achieved the maximal MCA (3164.84 SU/mL) that was 101.9% of the predicted value (3104.49 SU/mL) and 4.3-fold higher than the control.

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Brassica vegetables, especially broccoli, have health benefits such as anticancer activity, which are attributed to isothiocyanate (ITC), products of glucosinolate hydrolysis. This study aimed to explore the effect of cooking time and addition of exogenous myrosinase (MYR) from moringa seeds on the yield of ITCs. The results showed that raw broccoli produced a significantly high amount of ITCs, which decreased by almost 40% after microwaving the broccoli for 1 min. Introducing exogenous MYR by adding ground moringa seeds to cooked broccoli caused a notable increase in ITC of 38%. At pH 4.0-6.0, MYR showed optimal activity, and the thermal stability of MYR from moringa seeds was better than that from broccoli. The kinetic parameters indicated that MYR from moringa seeds had a higher affinity to sinigrin than that from broccoli seeds. This study was novel in reporting that adding ground moringa seeds to cooked broccoli enhanced ITC formation.

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Glucosinolate (GSL) is an important active substance in broccoli and can be hydrolyzed to isothiocyanates (ITCs) by endogenous myrosinase. The ITCs are well-known chemopreventive agents that have received significant attention across the nutrition and pharmaceutical industries due to their anticancer properties. Myrosinase activity decreases during the cooking of broccoli, thus it is essential to study the microbiota involved in GSL hydrolysis to maximize their health benefits. In this study, two strains (Enterococcus gallinarum HG001 and Escherichia coli HG002) isolated from the gut microbiota of C57BL/6 mice were identified through 16 S rRNA gene sequence and characteristic analyses. The maximum GSL hydrolysis activity of 12 strains was observed using the cyclocondensation method. Their growth curves, GSL-hydrolysis curves, ITC generation curves and GSL-hydrolysis products were analyzed. The En. gallisepticum HG001 hydrolyzed GSL to a greater level than the E. coli HG002. It was observed that they could hydrolyze GSL to produce erucin nitrile and 4-methylsulfanylbutyro nitrile through gas chromatography-mass spectrometer analysis. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01006-z.

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BACKGROUND AND AIM: There is evidence of a relationship between infection (and the associated antibiotic exposure) and the risk of celiac disease (CD). This study performed a meta-analysis to investigate this relationship. METHODS: To identify relevant studies, we conducted systematic searches of the PubMed, Embase, and Cochrane databases for articles published up to April 2019. Random effects models were used to determine overall pooled estimates and 95% confidence intervals (CIs). RESULTS: The meta-analysis included 19 observational studies (15 on infection and six on antibiotic exposure). Our results showed that any infection was associated with an increased risk of CD later in life (odds ratio, 1.37; 95% CI: 1.2-1.56; P < 0.001). The I2 was 94% (high heterogeneity among studies). Subgroup analyses suggested that the risk of CD is not affected by the type of infectious agent, timing of exposure, and site of infection. Exposure to antibiotics was also associated with new-onset CD (odds ratio, 1.2; 95% CI: 1.04-1.39; P < 0.001). CONCLUSION: Exposure to early infection or antibiotic appears to increase the odds of developing CD, suggesting that intestinal immune or microbiota dysbiosis may play a role in the pathogenesis of CD. These findings may influence clinical management and primary prevention of CD. However, noncausal explanations for these positive associations cannot be excluded.

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AIM: Epidemiological studies on associations between Caesarean sections (C-sections) and attention-deficit hyperactivity disorder (ADHD) have been inconsistent, and we performed a meta-analysis. METHODS: We systematically searched PubMed and Embase to December 2018 and included nine hospital-based and population registry studies published in 2011-2018. These covered a total study cohort of more than 2.5 million people in eight countries: Australia, Brazil, Denmark, Finland, Germany, Sweden, Turkey and the UK. The analysis provided summary odds ratios (ORs) and 95% confidence intervals (CI) while taking heterogeneity into account. RESULTS: We found that that C-sections were associated with a small increase in the risk of ADHD (OR 1.14, 95% CI 1.11, 1.17, I2 0%) in offspring. In subgroup analyses, the association remained for both infants born after elective C-sections (OR, 1.15, 1.11, 1.19, I2 0%) and emergency C-sections (OR, 1.13, 1.1, 1.17, I2 45.4%). However, these were only marginally significant when we pooled data from siblings from other pregnancies (OR, 1.06, 1.00-1.13, I2 0%), implying that the association was due to confounding. CONCLUSION: The statistically significant association between C-sections and ADHD in children can be partially explained by unmeasured confounding. Further research controlling for important confounders is required before firm conclusions can be drawn.

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