Protective role of crocin against sepsis-induced injury in the liver, kidney and lungs via inhibition of p38 MAPK/NF-κB and Bax/Bcl-2 signalling pathways.

CONTEXT: Crocin has been reported to have multiple bioactivities. However, the effect of crocin administration on caecal ligation and puncture (CLP)-induced sepsis remains unknown. OBJECTIVE: We investigated the effects of crocin on CLP-induced sepsis in mice and the underlying mechanism of action. MATERIALS AND METHODS: Five experimental groups (n = 10) of BALB/c mice were used: control, CLP (normal saline) and CLP + crocin (50, 100 and 250 mg/kg, 30 min prior to CLP). Mice were sacrificed 24 h after CLP. Liver, kidney and lung histopathology, indicator levels, apoptotic status, pro-inflammatory cytokines and relative protein levels were evaluated. RESULTS: Compared to the CLP group, crocin treatment significantly increased the survival rate (70%, 80%, 90% vs. 30%). Crocin groups exhibited protection against liver, kidney and lung damage with mild-to-moderate morphological changes and lower indicator levels: liver (2.80 ± 0.45, 2.60 ± 0.55, 1.60 ± 0.55 vs. 5.60 ± 0.55), kidney (3.00 ± 0.71, 2.60 ± 0.55, 1.40 ± 0.55 vs. 6.20 ± 0.84) and lungs (8.00 ± 1.59, 6.80 ± 1.64, 2.80 ± 0.84 vs. 14.80 ± 1.79). The proinflammatory cytokines (IL-1ß, TNF-α, IL-6 and IL-10 levels in the crocin groups) were distinctly lower and the apoptotic index showed a significant decrease. Crocin administration significantly suppressed p38 MAPK phosphorylation and inhibited NF-κB/IκBα and Bcl-2/Bax activation. DISCUSSION AND CONCLUSIONS: Pre-treatment with crocin confers protective effects against CLP-induced liver, kidney and lung injury, implying it to be a potential therapeutic agent.

The selective breeding and mutagenesis mechanism of high-yielding surfactin Bacillus subtilis strains with atmospheric and room temperature plasma.

BACKGROUND: Surfactin, a good biological surfactant, is derived from the metabolites of microorganisms. However, the ability of natural strains to produce surfactin is low, and so the presented study aimed to use a novel mutagenesis technology to increase their yields. RESULTS: Atmospheric and room temperature plasma (ARTP) was used to conduct mutation breeding of Bacillus subtilis CICC 10721, and a mutant strain M45 with a higher surfactin yield of 34.2% and a stable subculture was screened out. From the fermentation kinetics study, it was found that the maximum cell dry weight, maximum growth rate and surfactin synthesis parameters of the mutant strain M45 were all greater than that of the original strain. Scanning electron microscope and laser scanning confocal microscope observations showed that the spore morphology changed after ARTP treating, and the intracellular Ca2+ concentration of the mutant increased. Genome resequencing analysis showed that 66 single nucleotide poymorphism non-synonymous mutation sites occurred in M45, and the identification results of the fermentation broth extract from M45 showed that it is composed of C12 -C16 surfactin. CONCLUSION: ARTP mutagenesis was found to change the morphology of bacteria, membrane permeability and genes related to the synthesis and secretion of surfactin. The present study provides a basis for industrial production of surfactin and an understanding of the mutagenesis mechanism. © 2021 Society of Chemical Industry.

Stimulation of low intensity ultrasound on fermentation of skim milk medium for yield of yoghurt peptides by Lactobacillus paracasei.

Herein the effect of low intensity ultrasound on the fermentation of skim milk medium by Lactobacillus paracasei were investigated to obtain optimum ultrasonic conditions for the highest yield of yoghurt peptides. The results showed that the fermented skim milk medium treated with ultrasound with its seed culture without ultrasonic treatment was an optimum scheme. In this scheme with the ultrasonic conditions of 28 kHz, ultrasonic pulsed model of on-time 100 s and off-time 10 s, 100 W/L for the treatment time of 30 min after the fermentation time of 9 h, the peptide content in the fermented skim milk media increased by 49.5% and the viable cells in the same media increased by 43.5% compared with those in the untreated samples. By response surface methodology (RSM) analysis and its verification experiments, a reasonably accurate empirical model was established for investigating and predicting the relationship between skim milk concentration, ultrasonic treatment time, power and the yield of yoghurt peptides. The former two parameters 12.6% w/v and 35 min were taken in the verification experiments in which the peptide content of the fermented media reached 5.9 mg/mL with an increase by 64.23% and the peptide yield was 14.2%, similar to its theoretical value of 14.6% according to the empirical model. The comparison of extracellular enzyme activities in the fermented skim milk media between with and without ultrasonic treatment under the conditions in the optimum scheme indicated that the mechanism of the ultrasound-activated peptide content increment might be the extracellular enzyme activities immediately activated by the ultrasound, effect of which would disappear in the progress of fermentation after the ultrasound was removed.

Effects of ultrasound on microbial growth and enzyme activity.

Nowadays, ultrasound is widely used in many aspects. In the last few years, many papers have concentrated on the applications of ultrasound in engineering, chemistry, medicine, physics and biology, but few in biological effects such as the acceleration effects on proliferation of microbial cells, the inactivation effects on microorganisms and the influences on the activities of enzyme. Thus, the objective of this review is to investigate the biological effects of ultrasound on these aspects.

Ultrasonic irradiation of low intensity with a mode of sweeping frequency enhances the membrane permeability and cell growth rate of Candida tropicalis.

Here we report the enhancement of both cellular permeability and cell growth rate of Candida tropicalis after treatment with the ultrasonic irradiation of low intensity using a mode of sweeping frequency (UILS) generated by a self-developed ultrasonic device in our lab. After the ultrasonic treatment, remarkable biomass enhancement of the yeast was observed; the hyphae became significantly longer; the seeped cellular protein and nucleic acid from the yeast increased and the cellular Ca2+ content became lower. Illumina transcriptome sequencing showed that the ultrasonic treatment affected the expression of genes involved in diverse cellular components, biological processes and molecular functions. RT-PCR and Western blotting further confirmed the up-/down-regulation of genes in the ultrasound-treated yeasts. The optimal conditions of the ultrasonic treatment for the maximum biomass addition were determined as follows: the yeast was treated for 1h at the mid logarithmic phase, the frequency was 28±2kHz and the power density was 120W/L. Under these conditions, the Candida tropicalis biomass increased by 142.5% compared with the untreated yeast.