Enhanced production of fructosyltransferase in Aspergillus oryzae by genome shuffling.

OBJECTIVE: To breed Aspergillus oryzae strains with high fructosyltransferase (FTase) activity using intraspecific protoplast fusion via genome-shuffling. RESULTS: A candidate library was developed using UV/LiCl of the conidia of A. oryzae SBB201. By screening for enzyme activity and cell biomass, two mutants (UV-11 and UV-76) were chosen for protoplast fusion and subsequent genome shuffling. After three rounds of genome recombination, a fusion mutant RIII-7 was obtained. Its FTase activity was 180 U g-1, approximately double that of the original strain, and RIII-7 was genetically stable. In fermentation culture, FTase activity of the genome-shuffled strain reached a maximum of 353 U g-1 using substrate-feeding method, and this value was approximately 3.4-times higher than that of the original strain A. oryzae SBB201. CONCLUSIONS: Intraspecific protoplast fusion of A. oryzae significantly enhanced FTase activity and generated a potentially useful strain for industrial production.

Improving the electrochemical characteristics and performance of a neutral all-iron flow battery by using the iron reduction bacteria.

At present, the all-iron redox flow batteries (RFBs) have greater application potential due to high accessibility of electrolytes compared to traditional RFBs. Meanwhile, although electroactive bacteria can accelerate the electrons transfer, their potential to improve the performance of RFBs has been overlooked. Previously, we had confirmed that ferrous-oxidizing bacteria (FeOB) could enhance the performance of an all-iron RFB, therefore we conducted several batch experiments and chronopotentiometry experiments by using the ferric-reducing bacteria (FeRB) or mixed culture (FeOB and FeRB) to demonstrate whether they have the same or stronger effects on Fe3+-DTPA/Na4[Fe(CN)6] RFB. The results showed that the experimental reactors could achieve higher charging current density and initial cathodic potential during constant voltage charging process. The electrochemical impedance spectroscopy data and cyclic voltammetry curves demonstrated that the polarization impedance increased slower and reduction peak potential of experimental groups also emerged a positive shift compared to CK. According to chronopotentiometry experiments results, the microbes could function at maximum 0.3 M, 12 mA/cm2, and also improved the charging specific capacity. Combined the SEM pictures and microbial composition analysis, the main functional electroactive FeRB were Alcaligenes, Corynebacterium and Bacillus, which indicated to have important potential in improving the performance of RFBs.

Using ferrous-oxidizing bacteria to enhance the performance of a pH neutral all-iron flow battery.

Among various redox flow batteries (RFBs), the all-iron RFBs have greater application potential due to high accessibility of electrolytes. However, the potential of microaerobic ferrous-oxidizing bacteria (FeOB) to improve the performance of RFB has been neglected. Here, several experiments were conducted using Fe2+-diethylenetriaminepentaacetic acid (DTPA)/Na3[Fe(CN)6] as a redox couple for investigating the enhanced performance by FeOB in this RFB. Results showed that the maximum current density of experimental reactors could achieve 22.56 A/m2 at 0.1 M, whereas power density could still maintain 3.42 W/m2(16.96 A/m2 and 1.58 W/m2 for control group); meantime, the polarization impedance of anode increased slower and Fe2+-DTPA oxidation peak emerged maximum 494 mV negative shift. With increased electrolyte concentration in chronopotentiometry experiments, the experimental reactor achieved higher discharging specific capacity at 0.3 M, 10 mA/cm2. Microbial composition analysis showed maximum 75% is Brucella, indicating Brucella has ferrous-oxidizing electroactivity.

Effects of nitrogen supply on hydrogen-oxidizing bacterial enrichment to produce microbial protein: Comparing nitrogen fixation and ammonium assimilation.

To investigate the effects of nitrogen source supply on microbial protein (MP) production by hydrogen-oxidizing bacteria (HOB) under continuous feed gas provision, a sequencing batch culture comparison (N2 fixation versus ammonium assimilation) was performed. The results confirmed that even under basic cultivation conditions, N2-fixing HOB (NF-HOB) communities showed higher levels of CO2 and N2 fixation (190.45 mg/L Δ CODt and 11.75 mg/L Δ TNbiomass) than previously known, with the highest biomass yield being 0.153 g CDW/g COD-H2. Rich ammonium stimulated MP synthesis and the biomass accumulation of communities (increased by 7.4 ～ 14.3 times), presumably through the enhancement of H2 and CO2 absorption. The micro mechanism may involve encouraging the enrichment of species like Xanthobacter and Acinetobacter then raising the abundance of nitrogenase and glutamate synthase to facilitate the nitrogen assimilation. This would provide NF-HOB with ideas for optimizing their MP synthesis activity.

Analyzing the defense response mechanism of Atractylodes macrocephala to Fusarium oxysporum through small RNA and degradome sequencing.

Introduction: Fusarium oxysporum is a significant soil-borne fungal pathogen that affects over 100 plant species, including crucial crops like tomatoes, bananas, cotton, cucumbers, and watermelons, leading to wilting, yellowing, growth inhibition, and ultimately plant death. The root rot disease of A. macrocephala, caused by F. oxysporum, is one of the most serious diseases in continuous cropping, which seriously affects its sustainable development. Methods: In this study, we explored the interaction between A. macrocephala and F. oxysporum through integrated small RNA (sRNA) and degradome sequencing to uncover the microRNA (miRNA)-mediated defense mechanisms. Results: We identified colonization of F. oxysporum in A. macrocephala roots on day 6. Nine sRNA samples were sequenced to examine the dynamic changes in miRNA expression in A. macrocephala infected by F. oxysporum at 0, 6, and 12 days after inoculation. Furthermore, we using degradome sequencing and quantitative real-time PCR (qRT-PCR), validated four miRNA/target regulatory units involved in A. macrocephala-F. oxysporum interactions. Discussion: This study provides new insights into the molecular mechanisms underlying A. macrocephala's early defense against F. oxysporum infection, suggesting directions for enhancing resistance against this pathogen.

Specific convulsions and brain damage in children hospitalized for Omicron BA.5 infection: an observational study using two cohorts.

BACKGROUND: SARS-CoV-2 continues to mutate over time, and reports on children infected with Omicron BA.5 are limited. We aimed to analyze the specific symptoms of Omicron-infected children and to improve patient care. METHODS: We selected 315 consecutively hospitalized children with Omicron BA.5 and 16,744 non-Omicron-infected febrile children visiting the fever clinic at our hospital between December 8 and 30, 2022. Specific convulsions and body temperatures were compared between the two cohorts. We analyzed potential associations between convulsions and vaccination, and additionally evaluated the brain damage among severe Omicron-infected children. RESULTS: Convulsion rates (97.5% vs. 4.3%, P < 0.001) and frequencies (median: 2.0 vs. 1.6, P < 0.001) significantly differed between Omicron-infected and non-Omicron-infected febrile children. The body temperatures of Omicron-infected children were significantly higher during convulsions than when they were not convulsing and those of non-Omicron-infected febrile children during convulsions (median: 39.5 vs. 38.2 and 38.6 °C, both P < 0.001). In the three Omicron-subgroups, the temperature during convulsions was proportional to the percentage of patients and significantly differed ( P < 0.001), while not in the three non-Omicron-subgroups ( P = 0.244). The convulsion frequency was lower in the 55 vaccinated children compared to the 260 non-vaccinated children (average: 1.8 vs. 2.1, P < 0.001). The vaccination dose and convulsion frequency in Omicron-infected children were significantly correlated ( P < 0.001). Fifteen of the 112 severe Omicron cases had brain damage. CONCLUSIONS: Omicron-infected children experience higher body temperatures and frequencies during convulsions than those of non-Omicron-infected febrile children. We additionally found evidence of brain damage caused by infection with omicron BA.5. Vaccination and prompt fever reduction may relieve symptoms.

Polysaccharides as antioxidants and prooxidants in managing the double-edged sword of reactive oxygen species.

Polysaccharides, a class of naturally occurring carbohydrates, were widely presented in animals, plants, and microorganisms. Recently, health benefits of polysaccharides have attracted much attention due to their unique characteristics in reactive oxygen species (ROS) management. ROS, by-products of aerobic metabolism linked to food consumption, exhibited a dual role in protecting cells and fostering pathogenesis collectively termed double-edged sword. Some interesting studies reported that polysaccharides could behave as prooxidants under certain conditions, besides antioxidant capacities. Potentiation of the bright side of ROS could contribute to the host defense that was vitally important for the polysaccharides acting as biological response modifiers. Correspondingly, disease prevention of polysaccharides linked to the management of ROS production was systematically described and discussed in this review. Furthermore, major challenges and future prospects were presented, aiming to provide new insight into applying polysaccharides as functional food ingredients and medicine.

White hyacinth bean polysaccharide ameliorates diabetes via microbiota-gut-brain axis in type 2 diabetes mellitus rats.

Gut played a potent role in onset and progression of metabolic disorders, presenting an exciting direction for diabetes prevention. Here, the anti-diabetic effects of White hyacinth bean polysaccharides (WHBP) were observed, including the reduction of blood glucose levels and improvement of intestinal impairment in type 2 diabetes mellitus (T2DM) rats. Further data concerning intestinal protection suggested that WHBP restored intestinal barrier, as evidenced by inhibition of intestinal pathological damage, up-regulation of Zonula occluden-1 expression and manipulation of the redox system in T2DM rats. Moreover, WHBP-mediated anti-diabetic effects were in parallel with the adjustment of changes in gut microbiota composition of T2DM rats. Meanwhile, hypersecretion of corticotropin-releasing hormone, adrenocorticotropic hormone, and corticosterone levels, which were critical coordinators of the hypothalamic-pituitary-adrenal (HPA) axis, were suppressed in T2DM rats exposed to WHBP, indicating that WHBP-mediated health benefits were referring to regulate brain feedback in reduction of HPA axis. Concomitantly, further suggested and expanded on gut-brain communication by data of microbial metabolites short-chain fatty acids, mediators of gut-brain interactions, were remarkably raised in cecum contents of T2DM rats subjected to WHBP. Collectively, WHBP performed anti-diabetic effects were associated with control of microbiota-gut-brain axis implicated in intestinal barrier, HPA axis, gut microbiota and their metabolites.

Hyperlipidemic plasma molecules bind and inhibit adiponectin activity.

INTRODUCTION: Adiponectin is a potent vascular protective molecule. Recent findings have suggested adiponectin resistance during early diabetes. However, the molecular mechanisms responsible remain unidentified. Here, we took an unbiased approach to identify whether hyperlipidemic plasma molecules exist that bind and inhibit adiponectin function, contributing to adiponectin resistance and diabetic vascular injury. METHODS: Adult rats were randomly assigned to receive either a normal or a high-fat diet for 8 weeks. Plasma was co-immunoprecipitated with anti-APN antibody and analyzed by mass spectrometry. The APN binding molecules and their effect upon APN biological activity were determined. RESULTS: As expected, the high-fat-diet increased plasma triglyceride, total cholesterol, and low-density lipoprotein. Importantly, the circulating APN level was significantly increased at this time point. Mass spectrometry identified 18 proteins with increased APN binding in hyperlipidemic plasma, among which four proteins critical in lipid metabolism, including apolipoprotein A1 (APOA1), APOA4, APOC1, and paraoxonase 1, were further investigated. Incubating recombinant APN with APOA1 markedly (P < 0.01), and incubating with APOC1 significantly (P < 0.05), inhibited APN activity as evidenced by the reduced AMPK activation in HUVECs. APOA4 and paraoxonase 1 incubation had no effect upon APN activity. Finally, plasma APOA1 was significantly increased (P < 0.05) in hyperlipidemic plasma compared with the control plasma. CONCLUSIONS: It was demonstrated for the first time that increased APOA1 and APOC1 in hyperlipidemic plasma binds and inhibits APN activity. This result not only identifies a novel molecular mechanism responsible for adiponectin resistance during early stage diabetes, but also provides additional new insight into the diverse/controversial (protective and harmful) functions of high-density lipoprotein.

[6]-Gingerol Ameliorates ISO-Induced Myocardial Fibrosis by Reducing Oxidative Stress, Inflammation, and Apoptosis through Inhibition of TLR4/MAPKs/NF-κB Pathway.

SCOPE: [6]-Gingerol is one of the primary pungent constituents of ginger. While [6]-gingerol has many pharmacological effects, its benefits for myocardial fibrosis, including its exact role and underlying mechanisms, remain largely unexplored. The present study is designed to characterize the cardio-protective effects of [6]-gingerol in myocardial fibrosis mice and possible underlying mechanisms. METHODS AND RESULTS: Mice are subcutaneously injected with isoproterenol (ISO, 10 mg kg-1 ) and gavaged with [6]-gingerol (10, 20 mg kg-1 day-1 ) for 14 days. Pathological alterations, fibrosis, oxidative stress, inflammation response, and apoptosis are examined. In ISO-induced myocardial fibrosis, [6]-gingerol treatment decreases the J-point, heart rate, cardiac weight index, left ventricle weight index, creatine kinase (CK), and lactate dehydrogenase serum levels, calcium concentration, reactive oxygen species, malondialdehyde, and glutathione disulfide (GSSG), and increases levels of superoxide dismutase, catalase, glutathione, and GSH/GSSG. Further, [6]-gingerol improved ISO-induced morphological pathologies, inhibited inflammation and apoptosis, and suppressed the toll-like receptor-4 (TLR4)/mitogen-activated protein kinases (MAPKs)/nuclear factor κB (NF-κB) signaling pathways. CONCLUSION: The protective effect of [6]-gingerol in mice with ISO-induced myocardial fibrosis may be related to the inhibition of oxidative stress, inflammation, and apoptosis, potentially through the TLR4/MAPKs/NF-κB signaling pathway.

Physical activity level and incident type 2 diabetes among Chinese adults.

PURPOSE: The objective is to examine the association between physical activity level (PAL) and incident type 2 diabetes among middle-age and older Chinese men and women in urban China. METHODS: This prospective study included 6348 participants (age 35 to 74 yr) who were free of diabetes and cardiovascular disease at baseline. PAL was estimated on the basis of self-reported overall physical activity on a typical day. According to PAL, participants were classified into four groups: sedentary (PAL, 1.00-1.39), low active (PAL, 1.40-1.59), active (PAL, 1.60-1.89), and very active (PAL, >1.89). The association of PAL with incident diabetes was examined by Cox proportional hazards model. RESULTS: During 7.9 yr of follow-up (50,293 person-years), 478 incident cases of type 2 diabetes were identified. After adjustment for age, sex, geographic region, educational level, smoking, alcohol use, and family history of diabetes, the HR (95% CI) values for type 2 diabetes across increasing categories of PAL were 1.00 (reference), 0.82 (0.62-1.09), 0.63 (0.47-0.83), and 0.47 (0.36-0.61), respectively (P for trend <0.0001). Additional adjustment for baseline body mass index or waist circumference attenuated the magnitude of risk reduction, but it remained significant. The inverse association between PAL and risk of incident diabetes was persistent in subgroup analyses according to age, sex, hypertension, smoking, body mass index, waist circumference, and fasting plasma glucose level. CONCLUSIONS: Higher PAL is associated with substantial reduction in risk of type 2 diabetes. Our findings suggest the importance of a physically active lifestyle in the prevention of diabetes.

[Incidence of obesity and its modifiable risk factors in Chinese adults aged 35-74 years: a prospective cohort study].

OBJECTIVE: To examine the incidence of obesity and its modifiable risk factors in Chinese adults aged 35-74 years. METHODS: A total of 27 020 participants aged 35 to 74 years from two prospective cohort studies in China were followed up in the years of 2007 and 2008. Obesity and overweight were defined as body mass index ≥ 28.0, and 24.0-27.9 kg/m(2), respectively. Relative risks of obesity for risk factors were computed by using logistic regression. RESULTS: The annual incidence rates of obesity and overweight were 6.97 ‰ and 24.83 ‰ in Chinese adults aged 35-74 years, respectively. Women had a higher incidence of obesity than men (7.74 ‰ vs. 6.10 ‰). Participants in northern China had a higher incidence than those in southern (9.29 ‰ vs. 5.10 ‰) part of the country. Adults in rural had a higher incidence than those in urban (7.28 ‰ vs. 6.52 ‰). After adjusting for the baseline variables, such as gender, age, geographic region, degree of urbanization, the relative risk for obesity was 0.82 (95% CI:0.68-0.99) for participants with ≥ 12 years' education, compared with those <12 years. Participants with middle income, less physical activity at work/housework or being retirees, consuming more red meat and scented tea etc, had higher risk of incidence of obesity. Participants who consumed milk and moderate amount of fruits, would show a lower risk of obesity. CONCLUSION: The incidence of obesity was 6.97 ‰ in Chinese middle and older adults. Our results underscored that the promotion of healthy lifestyle which include issues as increasing physical activity, consuming moderate amount of fruits and milk but less red meat, drinking less scented tea etc, could play key roles in obesity prevention and control among the Chinese adults, especially among people with low education level or with middle income.

Gestational hypoxia induces sex-differential methylation of Crhr1 linked to anxiety-like behavior.

Stress during gestation increases vulnerability to disease and changes behavior in offspring. We previously reported that hypoxia and restraint during pregnancy sensitized the hypothalamic-pituitary-adrenal (HPA) axis and induced anxiety-like behavior in the adult offspring. Here, we report that gestational intermittent hypoxia (GIH) elicited a sex-dependent anxiety-like behavior in male P90 offspring and activation of corticotropin-releasing hormone (CRH) and CRH type-1 receptor (CRHR1) mRNA in the hypothalamic paraventricular nucleus (PVN) and in male E19 hypothalamus. These linked to demethylation at several specific sites of CpG island of Crhr1 promoter in P90 PVN and E19 embryo hypothalamus in GIH groups. Crhr1 DNA demethylation is more crucial in CpG island 1 than island 2 for activation of CRHR1 mRNA. DNMT3b is required for the Crhr1 DNA methylation than DNMT1 and DNMT3a in increased CRHR1 mRNA. We first address a novel hypothesis that GIH-induced male-sex-dependent demethylation at CpG sites of Crhr1 DNA in promoter triggers elevation of CRHR1 mRNA in PVN and anxiety-like behavior in adult offspring.