植物乳杆菌ATCC8014对寄生曲霉NRRL 2999孢子形态及生长与产毒影响的研究

为探讨植物乳杆菌ATCC 80 1 4对寄生曲霉NRRL 2 999生长与产毒的影响 ,将植物乳杆菌与寄生曲霉同时接种到MRS肉汤和预先厌氧培养植物乳杆菌后再接种寄生曲霉孢子 ,2 8℃培养 1 5天。在培养的第3、6、9、1 2和 1 5天测定培养液中的pH值、黄曲霉毒素B1 和菌丝体重量。结果显示 :植物乳杆菌可显著抑制寄生曲霉生长 (P <0 0 1 ) ,抑制的机制在于孢子灭活 ,使其不能发育成菌丝 ,故也不能产生黄曲霉毒素B1 ,在电子显微镜下可观察到寄生曲霉孢子充水肿胀死亡 ,而植物乳杆菌的细胞形态也由长杆状变为椭圆形 ;当植物乳杆菌ATCC 80 1 4接种到已经培养了 3天的寄生曲霉NRRL 2 999培养液中时 ,则植物乳杆菌对其生长与产毒无显著影响。在以乳酸调整的pH4 5MRS肉汤中 ,乳酸对寄生曲霉生长有显著的抑制作用 (P <0 0 1 ) ,但并不影响黄曲霉毒素B1 的合成     

Fermentation of Vaccinium floribundum Berries with Lactiplantibacillus plantarum Reduces Oxidative Stress in Endothelial Cells and Modulates Macrophages Function

Accumulating evidence suggests that high consumption of natural antioxidants promotes health by reducing oxidative stress and, thus, the risk of developing cardiovascular diseases. Similarly, fermentation of natural compounds with lactic acid bacteria (LAB), such as Lactiplantibacillus plantarum, enhances their beneficial properties as regulators of the immune, digestive, and cardiovascular system. We investigated the effects of fermentation with Lactiplantibacillus plantarum on the antioxidant and immunomodulatory effects of Pushgay berries (Vaccinium floribundum, Ericaceae family) in human umbilical vein endothelial cells (HUVECs) and macrophage cell line RAW264.7. Polyphenol content was assayed by Folin–Ciocalteu and HPLC-MS/MS analysis. The effects of berries solutions on cell viability or proliferation were assessed by WST8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt and Lactate dehydrogenase (LDH) release, Trypan blue exclusion test, and Alamar blue assay. Antioxidant activity was evaluated by a cell-based chemiluminescent probe for the detection of intracellular H₂O₂ production in HUVECs. Heme oxygenase-1 (HO-1) expression levels were investigated by RT-qPCR. Glutathione reductase (GR), glutathione peroxidase (Gpx), superoxide dismutase (SOD), and catalase (CAT) activities, as markers of intracellular antioxidant defense, were evaluated by spectrophotometric analysis. The immunomodulatory activity was examined in RAW 264.7 by quantification of inducible nitric oxide synthase (iNOS) and Tumor Necrosis Factor—alpha (TNFα) by RT-qPCR. Data showed that fermentation of Pushgay berries (i) enhances the content of quercetin aglycone, and (ii) increases their intracellular antioxidant activity, as indicated by the reduction in H₂O₂-induced cell death and the decrease in H₂O₂-induced HO-1 gene expression in HUVECs treated for 24 h with fermented berries solution (10 µg/mL). Moreover, treatment with Pushgay berries for 72 h (10 µg/mL) promotes cells growth in RAW 264.7, and only fermented Pushgay berries increase the expression of iNOS in the same cell line. Taken together, our results show that LAB fermentation of Pushgay berries enhances their antioxidant and immunomodulatory properties.     

In Vitro and In Vivo Antihypertensive Effect of Milk Fermented with Different Strains of Common Starter Lactic Acid Bacteria

Currently, functional dairy products pave a promising way for the prophylaxis of essential hypertension, and the search for new strains capable of producing such products is a constant challenge for scientists around the world. In this study, the antihypertensive properties of milk fermented with several strains of traditional yogurt starters (Lactobacillus delbrueckii strains Lb100 and Lb200; Lactococcus lactis strains dlA, AM1 and MA1; Streptococcus thermophilus strains 159 and 16t) and one strain of non-conventional probiotic starter (Lacticaseibacillus paracasei ABK) were assessed. The in vitro assessment using angiotensin-converting enzyme inhibition assay was performed for all fermentation products, and the best performed products were tested in vivo using Spontaneously Hypertensive Rat (SHR) animal model. In addition, for the best performed products the fatty acid (FA) composition and FA-related nutritional indices were determined. As a result, the milk fermented with two strains (Lb. delbrueckii LB100 and Lc. lactis AM1) demonstrated significant antihypertensive effect during both in vitro and in vivo experiments. Moreover, the milk fermented with Lb. delbrueckii Lb100 demonstrated significantly better FA-related nutritional indexes and lowered total cholesterol in SHRs upon regular consumption. The obtained results can be used in the future to develop new starter cultures producing effective functional antihypertensive dairy products.     

Cytoplasmic expression of a model antigen with M Cell-Targeting moiety in lactic acid bacteria and implication of the mechanism as a mucosal vaccine via oral route

Lactic acid bacteria (LAB) have been widely studied as mucosal vaccine delivery carriers against many infectious diseases for heterologous expression of protein antigens. There are three antigen expression strategies for LAB: cytoplasmic expression (CE), cell surface display (SD), and extracellular secretion (ES). Despite the generally higher protein expression level and many observations of antigen-specific immunogenicity in CE, its application as a mucosal vaccine has been overlooked relative to SD and ES because of the antigens enclosed by the LAB cell wall. We hypothesized that the antigens in CE could be released from the LAB into the intestinal lumen before host bacterial access to gut-associated lymphoid tissue (GALT), which could contribute to antigen-specific immune responses after oral administration. To elucidate this hypothesis, three recombinant Lactobacillus plantarum (LP) strains were constructed to produce a model antigen, BmpB, with or without an M cell-targeting moiety, and their immunogenicities were analyzed comparatively as oral vaccines in mouse model. The data indicated that the recombinant LPs producing BmpBs with different conformations could induce mucosal immunity differentially. This suggests that the cytoplasmic antigens in LAB could be released into the intestinal lumen, subsequently translocated through M cells, and stimulate the GALT to generate antigen-specific immune responses. Therefore, the CE strategy has great potential, especially in the application of oral LAB vaccines as well as SD and ES strategies. This research provides a better understanding of the mechanism for recombinant oral LAB vaccines and gives insight to the future design of LAB vaccines and oral delivery applications for useful therapeutic proteins.     

Effects of Lactofermented Beetroot Juice Alone or with N-nitroso-N-methylurea on Selected Metabolic Parameters,Composition of the Microbiota Adhering to the Gut Epithelium and Antioxidant Status of Rats

An objective of this work was to assess the biological activity of beetroot juice (Chrobry variety, Beta vulgaris L. ssp. vulgaris), which was lactofermented by probiotic bacteria Lactobacillus brevis 0944 and Lactobacillus paracasei 0920. The oxidative status of blood serum, kidneys, and liver of rats consuming the fermented beetroot juice were determined. The experimental rats were divided into four groups on diet type: Basal diet, basal diet supplemented with fermented beetroot juice, basal diet and N-nitroso-N-methylurea treatment, and basal diet supplemented with fermented beetroot juice and N-nitroso-N-methylurea treatment. Mutagen N-nitroso-N-methylurea, which was added to diet in order to induce aberrant oxidative and biochemical processes and disadvantageous changes in the count and metabolic activity of the gut epithelium microbiota. The nutritional in vivo study showed that supplementing the diet of the rats with the lactofermented beetroot juice reduced the level of ammonia by 17% in the group treated with N-nitroso-N-methylurea. Furthermore, the positive modulation of the gut microflora and its metabolic activity was observed in groups of rats fed with the diet supplemented with the fermented beetroot juice. A concomitant decrease in the β-glucuronidase activity was a consequence of the gut epithelium microbiota modulation. The antioxidant capacity of blood serum aqueous fraction was increased by about 69% in the group of rats treated N-nitroso-N-methylurea mixed with the fermented beetroot juice and N-nitroso-N-methylurea versus to the N-nitroso-N-methylurea treatment, whereas the antioxidant parameters of the blood serum lipid fraction, kidneys, and liver remained unchanged.     

An observational study of 127 preschool children at their homes and daycare centers in Ohio: environmental pathways to cis- and trans-permethrin exposure

The potential exposures of 127 preschool children to the pyrethroid insecticides, cis- and trans-permethrin, in their everyday environments were examined. Participants were recruited randomly from 127 homes and 16 daycare centers in six Ohio (OH) counties. Monitoring was performed over a 48-h period at the children's homes and/or daycare centers. Samples collected included soil, carpet dust, indoor air, outdoor air, diet, hand wipes, surface wipes, transferable residues, and urine. The environmental samples were analyzed for the cis and trans isomers of permethrin, and the urine samples were analyzed for the pyrethroid urinary metabolite, 3-phenoxybenzoic acid (3-PBA), by gas chromatography/mass spectrometry. The isomers were detected most often in the dust (100%) and hand wipe (>78%) samples collected at both homes and daycare centers. The median levels of cis-permethrin (470 and 1010 ng/g) were higher than the median levels of trans-permethrin (344 and 544 ng/g) in the dust samples at both the children's homes and daycare centers, respectively. In the children's hand wipe samples, the median levels of cis- and trans-permethrin were similar, ranging from 0.03 to 0.04 ng/cm(2), at both locations. The urinary metabolite 3-PBA was detected in 67% of the children's urine samples. The median urinary 3-PBA concentration for the children was 0.3 ng/mL, and the maximum value for one child was 33.8 ng/mL. The primary route of the children's exposure to the combined isomers was through dietary ingestion, followed by indirect ingestion. In addition, our calculated aggregate absorbed doses of permethrin accounted for about 60% of the excreted amounts of 3-PBA found in the children's urine. In conclusion, these children were potentially exposed to low levels of permethrin from several sources, and through several pathways and routes.     

Changes in hearing,cardiovascular functions,haemodynamics, upright body sway,urinary catecholamines and their correlates after prolonged successive exposures to complex environmental conditions

Summary This study deals with changes in temporary threshold shifts of hearing (TTS₂), cardiovascular functions (HR, SBP, DBP, PP, RWA), haemodynamic activity (HDI), upright body sway (VUBSA), ratios of urinary catecholamines (10A/DA, 10NA/DA, A/NA, NA/A) and correlations between these changes in complex exposure situations. The study was carried out in a special exposure chamber on 60 healthy male students. It was based on a factorial experimental design with a total of 12 exposure combinations. Each individual experiment took 6 h with a pause of 1 h at noon. There were 12 sequential exposure periods lasting 16 min each. A pause of 4 min followed each exposure period. The subjects were exposed to noise and whole body vibration at two different dry bulb temperatures. Noise classes were: (1) no noise, and (2) stable broad-band noise of 90 dBA. Vibration classes were: (1) no vibration, (2) a sinusoidal 5 Hz vibration, and (3) a stochastic vibration with a frequency range of 2.8 to 11.2 Hz along the Z-axis and with an acceleration of 2.12 m/s². Temperature classes were: (1) 20°C and (2) 35°C. Changes in body functions were registered during the pauses. Urine samples were gathered for the morning and afternoon sessions and for the preceding and the following night of the test. The changes were dependent on the combinations of noise, vibration and temperature to which the subjects were exposed. The TTS₂ values at 4 kHz were associated with the HDI-values when subjects were exposed simultaneously to noise and stochastic vibration at 35°C. The TTS₂ values at 6 kHz were associated most strongly with the HDI values after exposure to a combination of noise, stochastic or sinusoidal vibration and a temperature of 20°C. The TTS₂ values at 4 and 6 kHz correlated positively with the NA/A ratio when subjects were exposed to noise at 35°C. The association between TTS₂ values and the 10A/NA ratio and especially the A/NA ratio was very strong when subjects had been simultaneously exposed to noise and sinusoidal or stochastic vibration at 35°C. Furthermore, the highest positive correlation coefficients were found between TTS₂ values at 4 kHz and VUBSA values in the X-direction when subjects had been exposed to noise or simultaneously to noise and sinusoidal vibration at 20°C. With vibration exposure at 35°C, noise led to an increase in the 10NA/DA and NA/A ratios, but with vibration exposure at 20°C, noise decreased the 10NA/DA and NA/A ratios. The exposure was reflected as a rise in the NA/A ratio during the night following the experiment. The variance analytical model as a whole explained 58 to 62% in the morning and 55 to 60% in the afternoon (TTS₂ values at 4 kHz), 50 to 62% and 45 to 50% (TTS₂ values at 6 kHz), 9 to 12% and 9 to 19% (HR values), 16 to 31% and 23 to 36% (SBP values), 3 to 7% and 2 to 6% (DBP values), 7 to 12% and 9 to 13% (PP values), 5 to 8% and 1 to 3% (RWA values), 10 to 18% and 12 to 23% (HDI values), 6 to 7% and 7% (VUBSA values in X direction), 1% and 1 to 8% (VUBSA values in the Y direction), 18% and 20% (10A/DA), 17% and 20% (10NA/DA), 28% and 24% (A/NA), and 27% and 21% (NA/A) of the variation in the values of the variables involved. Some results were presented in Kanazawa, Japan, at the Second International Conference on the Combined Effects of Environmental Factors, September 28–October 1, 1986