Tartary buckwheat protein-phenol conjugate prepared by alkaline-based environment: Identification of covalent binding sites of phenols and alterations in protein structural and functional characteristics.

Tartary buckwheat protein-rutin/quercetin covalent complex was synthesized in alkaline oxygen-containing environment, and its binding sites, conformational changes and functional properties were evaluated by multispectral technique and proteomics. The determination of total sulfhydryl and free amino groups showed that rutin/quercetin can form a covalent complex with BPI and could significantly reduce the group content. Ultraviolet-visible spectrum analysis showed that protein could form new characteristic peaks after binding with rutin/quercetin. Circular dichroism spectrum analysis showed that rutin and quercetin caused similar changes in the secondary structure of proteins, both promoting ß-sheet to α-helix, ß-ture and random coil transformation. The fluorescence spectrometry results showed that the combination of phenols can cause the fluorescence quenching, and the combination of rutin was stronger than the quercetin. Proteomics showed that there were multiple covalent binding sites between phenols and protein. Rutin had a high affinity for arginine, and quercetin and cysteine had high affinity. Meanwhile, the combination of rutin/quercetin and protein had reduced the surface hydrophobic ability of the protein, and improved the foaming, stability and antioxidant properties of the protein. This study expounded the mechanism of the combination of BPI and rutin/quercetin, and analysed the differences of the combination of protein and phenols in different structures. The findings can provide a theoretical basis for the development of complexes in the area of food.

Comparative study of dietary phenols with Tartary buckwheat protein (2S/13S): impact on structure, binding sites and functionality of protein.

BACKGROUND: This research was to investigate the interaction mechanism between 2S albumin and 13S globulin (2S and 13S, the most important storage proteins in Tartary buckwheat seeds) and three phenols (rutin, quercetin and myricetin) regarding the structural and antioxidant properties of their complexes. RESULTS: There are differences in the binding affinity of phenols for 2S and 13S. Rutin had a higher binding affinity for 2S, myricetin had a higher binding affinity for 13S, and 13S exhibited a higher affinity toward phenols than did 2S. Binding with phenols significantly changed the secondary and tertiary structures of 2S and 13S, decreased the surface hydrophobic value and enhanced the antioxidant capacity. Molecular docking and isothermal titration calorimetry showed that the binding processes were spontaneous and that there were hydrogen bonds, hydrophobic bonds and van der Waals force interactions between phenols and proteins. CONCLUSION: These findings could provide meaningful guidance for the further application of buckwheat protein complex. © 2023 Society of Chemical Industry.

Retinoic Acid-PPARα Mediates ß-Carotene Resistance to Placental Dysfunction Induced by Deoxynivalenol.

Deoxynivalenol (DON), one of the most polluted mycotoxins in the environment and food, has been proven to have strong embryonic and reproductive toxicities. However, the effects of DON on placental impairment and effective interventions are still unclear. This study investigated the effect of ß-carotene on placental functional impairment and its underlying molecular mechanism under DON exposure. Adverse pregnancy outcomes were caused by intraperitoneal injection of DON from 13.5 to 15.5 days of gestation in mice, resulting in higher enrichment of DON in placenta than in other tissue samples. Interestingly, 0.1% ß-carotene dietary supplementation could significantly alleviate DON-induced pregnancy outcomes. Additionally, in vivo and in vitro placental barrier models demonstrated the association of DON-induced placental function impairment with placental permeability barrier disruption, angiogenesis impairment, and oxidative stress induction. Moreover, ß-carotene regulated DON-induced placental toxicity by activating the expressions of claudin 1, zonula occludens-1, and vascular endothelial growth factor-A through retinoic acid-peroxisome proliferator-activated receptor α signaling.

Effects of dietary adenosine and adenosine 5'-monophosphate supplementation on carcass characteristics, meat quality, and lipid metabolism in adipose tissues of finishing pigs.

This study investigated the effects of adenosine (ADO) and adenosine 5'-monophosphate (AMP) supplementation on the growth performance, carcass characteristics, meat quality, and lipid metabolism in adipose tissues of finishing pigs. The pigs were allocated to three treatment groups: the control diet, 0.2%ADO diet, or 0.2%AMP diet. Compared with CON group (CON), both ADO and AMP groups increased in carcass straight length (P < 0.05) and decreased in drip loss (P < 0.05), while AMP group tended to increase in redness value (P = 0.05) and decreased in free amino acid content in longissimus thoracis (LT) muscle (P < 0.05). Additionally, ADO or AMP supplementation increased the ADO or AMP content in serum, adipose tissue, and LT muscle (P < 0.05), as well as the protein level of adenosine 2A receptor (A2a) in adipose tissue (P < 0.05). Moreover, both ADO and AMP groups showed an increase in the expression of lipolysis genes (ATGL and HSL) in adipose tissue (P < 0.05). Overall, AMP supplementation could improve meat quality, and ADO and AMP supplementation regulate the lipid metabolism of finishing pigs.

Different interactions between Tartary buckwheat protein and Tartary buckwheat phenols during extraction: Alterations in the conformation and antioxidant activity of protein.

The purpose of this study is to investigate the interaction between buckwheat protein and buckwheat phenols in the process of protein extraction and to compare the effects of phenols on protein structure and antioxidant activity. With the extension of extraction time, the content of total phenol increased from 150.51 to 336.01 mg gallic acid equivalent/g sample. Four phenols and seven phenols were identified by UPLC-Q/TOF-MS as binding to proteins in non-covalent and covalent forms, respectively. The contribution of non-covalent and covalent bound phenols to the antioxidant activity of the complexes were different. Meanwhile, the binding of phenols changed the infrared characteristic peak of protein, and reduced the fluorescence intensity and surface hydrophobic value. The free amino and sulfhydryl content of the protein decreased with increasing extraction time. These findings provide valuable information for one-step preparation of protein-phenol complexes.

Dietary fiber supplementation during the last 50 days of gestation improves the farrowing performance of gilts by modulating insulin sensitivity, gut microbiota, and placental function.

Our previous study found dietary konjac flour (KF) supplementation could improve insulin sensitivity and reproductive performance of sows, but its high price limits its application in actual production. This study aimed to investigate the effects of supplementation of a cheaper combined dietary fiber (CDF, using bamboo shoots fiber and alginate fiber to partially replace KF) from the last 50 days of gestation to parturition on farrowing performance, insulin sensitivity, gut microbiota, and placental function of gilts. Specifically, a total of 135 pregnant gilts with a similar farrowing time were blocked by backfat thickness and body weight on day 65 of gestation (G65d) and assigned to 1 of the 3 dietary treatment groups (n = 45 per group): basal diet (CON), basal diet supplemented with 2% KF or 2% CDF (CDF containing 15% KF, 60% bamboo shoots fiber, and 25% alginate fiber), respectively. The litter performance, insulin sensitivity and glucose tolerance parameters, placental vessel density, and short-chain fatty acids (SCFAs) levels in feces were assessed. The gut microbiota population in gilts during gestation was also assessed by 16S rDNA gene sequencing. Compared with CON, both KF and CDF treatments not only increased the piglet birth weight (P < 0.05) and piglet vitality (P < 0.01) but also decreased the proportion of piglets with birth weight ≤ 1.2 kg (P < 0.01) and increased the proportion of piglets with birth weight ≥ 1.5 kg (P < 0.01). In addition, KF or CDF supplementation reduced fasting blood insulin level (P < 0.05), homeostasis model assessment-insulin resistance (P < 0.05), serum hemoglobin A1c (P < 0.05), and the level of advanced glycation end products (P < 0.05) at G110d, and increased the placental vascular density (P < 0.05) at farrowing. Meanwhile, KF or CDF supplementation increased microbial diversity (P < 0.05) and SCFAs levels (P < 0.05) in feces at G110d. Notably, the production cost per live-born piglet was lower in CDF group (¥ 36.1) than KF group (¥ 41.3). Overall, KF or CDF supplementation from G65d to farrowing could improve the farrowing performance of gilts possibly by improving insulin sensitivity, regulating gut microbiota and metabolites, and increasing placental vascular density, with higher economic benefits and a similar effect for CDF vs. KF, suggesting the potential of CDF as a cheaper alternative to KF in actual production.

Dietary konjac flour (KF) supplementation could improve insulin sensitivity and reproductive performance of sows, but its high price limits its application in actual production. This study investigated the impact of 2% konjac flour (KF) and 2% combined dietary fiber (CDF, containing 15% KF, 60% bamboo shoots fiber, and 25% alginate fiber) supplementation from the last 50 days of gestation to farrowing on farrowing performance, placental function, insulin sensitivity, and gut microbiota of gilts. Results indicated that KF or CDF supplementation during this time could improve the farrowing performance of gilts possibly by improving insulin sensitivity and gut microbiota, and increasing placental vascular density. Meanwhile, CDF could lower the production cost per live-born piglet and have a similar effect to KF, thus a cheaper alternative to KF in actual production. This study facilitates understanding the beneficial effects of KF and non-conventional dietary fiber sources on the reproductive performance of gilts.

Identification of binding sites for Tartary buckwheat protein-phenols covalent complex and alterations in protein structure and antioxidant properties.

To investigate the effects of structure, multiple binding sites and antioxidant property of Tartary buckwheat protein-phenols covalent complex, protein was combined with different concentrations of phenolic extract. Four kinds of phenols were identified by UPLC-Q/TOF-MS, which were rutin, quercetin, kaempferol and myricetin. UV-vis absorption spectroscopy and X-ray diffraction showed that the phenols can successfully bind to BPI. Fourier-transform infrared, circular dichroism and fluorescence emission spectroscopy showed that the binding of phenol can change the secondary/tertiary structure of protein. The particle distribution indicated that the binding of phenols could reduce the particle size (from 304.70 to 205.55 nm), but cross-linking occurred (435.35 nm) when the bound phenol content was too high. Proteomics showed that only rutin, quercetin and myricetin can covalently bind to BPI. Meanwhile, 4 peptides covalently bound to phenols were identified. The DPPH· scavenging capacity of complexes were from 8.38 to 33.76 %, and the ABTS·+ binding activity of complexes were from 19.35 to 63.99 %. The antioxidant activity of the complex was significantly higher than that of the pure protein. These results indicated that protein-phenol covalent complexes had great potential as functional components in the food field.

The mechanism of different cyanobacterial responses to glyphosate.

Glyphosate, the most extensively used herbicide globally, has raised ecotoxicological concerns because it can be transported into the aquatic environment and cause adverse effects on the aquatic system. However, the functional mechanism of glyphosate on cyanobacteria are not completely disentangled. In this study, we selected six common cyanobacteria to evaluate glyphosate effects on cyanobacterial growth in monoculture experiment. Results showed that the growth of five tested cyanobacterial species were promoted under different degrees, and only Pseudanabaena was inhibited by glyphosate. In the phylogenetic tree based on gene sequences of 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS), a target for glyphosate, we found that the position of Pseudanabaena is the closest to plant, which was sensitive to glyphosate, thereby explaining the inhibitory effect of Pseudanabaena following glyphosate exposure. The primary degraded metabolites or analogs did not induce cyanobacterial growth, laterally demonstrating that glyphosate was used as a source of phosphorus to accelerate cyanobacterial growth because phosphorus levels increased in the medium of glyphosate treatment. Overall, this study provides a better understanding of the influence of glyphosate on the composition of aquatic microbiota and explains the mechanism of cyanobacterial response to glyphosate.

[Point-pricking method with fire needle for knee osteoarthritis: a randomized controlled trial].

OBJECTIVE: To observe the effects of point-pricking method with fire needle on the symptoms of knee joint and physio-psychological health in the patients with knee osteoarthritis (KOA). METHODS: Sixty six patients with KOA were randomly divided into a fire needling group (33 cases) and a filiform needling group (33 cases). The patients received the point-pricking method with fire needle in the fire needle group while the conventional acupuncture with filiform needle was provided in the filiform needling group. The basic health management was performed in both groups. The acupoints included bilateral Liangqiu (ST34), Xuehai (SP10), Dubi (ST35), Neixiyan (EX-LE4), Yanglingquan (GB34) and Zusanli (ST36) as well as Ashi points. The treatment was conducted twice a week for 6 weeks consecutively. Before and after treatment, the scores of Western Ontario and McMaster University Osteoarthritis Index (WOMAC), traditional Chinese medicine (TCM) symptoms and visual analogue score (VAS), the numbers of affected areas of knee joint pain and the scores of 12-item short-form health survey (SF-12) were assessed and the incidence of adverse effects was recorded. RESULTS: The score for pain, stiffness and function, as well as the total score of WOMAC were all reduced (P<0.05), the score of TCM symptoms, VAS score and the numbers of the affected areas of knee joint pain were lower (P<0.05) after treatment when compared with those before treatment within group. After treatment, the score of each domain of SF-12 (i.e. general health, physical function, role-physical, vitality, role-emotional, physical component summary and mental component summary) was higher in comparison to those before treatment in both groups (P<0.05), while the scores of bodily pain, mental health and social functioning in SF-12 were increased in the fire needling group (P<0.05). Compared with the filiform needling group, the score of joint function and the total score in WOMAC and VAS score were reduced remarkably (P<0.05), and general health score in SF-12 was more obviously increased (P<0.05) in the fire needling group after treatment. None adverse effects were found in either group. CONCLUSION: The point-pricking method with fire needle is safe and effective when compared with conventional acupuncture with filiform needle. In the aspects of improving knee joint function, relieving joint pain and advancing the quality of life, the point-pricking method with fire needle is superior to the conventional acupuncture with filiform needle.

The potential mechanism of Longsheyangquan Decoction on the treatment of bladder cancer: Systemic network pharmacology and molecular docking.

Our goal was to explore the bioactive constituents of Longsheyangquan (LSYQ) Decoction and elucidate its mechanisms on the treatment of bladder cancer (BCa). A total of 38 compounds were selected based on their pharmacokinetic properties in three large traditional Chinese medicine (TCM) databases. 654 putative targets of LSYQ Decoction were predicted using a structure-based, reverse-docking algorithm online, of which 343 overlapped with BCa-related protein-coding genes. The protein-protein interaction (PPI) network was constructed to perform module analysis for further Gene Ontology (GO) annotations and Kyoto Encyclopedia Genes and Genomes (KEGG) pathway enrichment analysis, which identified CDK2, EGFR, MMP9 and PTGS2 as hub targets. The TCM-compound-target network and compound-target-pathway network together revealed that quercetin, diosmetin, enhydrin and luteolin were the main components of LSYQ Decoction. Finally, molecular docking showed the affinity between the key compounds and the hub target proteins to verify the accuracy of drug target prediction in the first place. The present study deciphered the core components and targets of LSYQ Decoction on the treatment of BCa in a comprehensive systemic pharmacological manner.

A Study on the Effects of Chinese Massage on Physical and Mental Health in Participants Based Smart Healthcare.

The purpose of this study is to understand the influence of the effect of the smart Chinese massage appliance on the participants' sports performance and physical and mental health after intermittent exercises. The study was performed by a mixed-study method. First, the experimental research method was used to design an experimental procedure and specifications such as data collection scope. Next, the quantitative research was carried out, using SPSS 26.0 statistical software to analyze data from the questionnaire. Then, the qualitative research was carried out by interviewing experts and respondents and obtaining their opinions. Finally, all the data were collated and analyzed using the multidimensional review method. Findings. Chinese massage significantly improved the physical fitness, exercise performance, body composition, and physical and mental health status of high-strength cyclic exercise participants. High-strength exercises can improve the physical and mental health of men. Medium-strength exercises could improve current conditions for women. Oxygen intake remained peaked for 8 sessions for men and increased after 30 sessions for women. Both groups required a 1-2-week adaptation period but had different exercise performance periods. After the experiment, head and back pain, as well as stress and negative emotions, was improved. The high intensity of the exercises relieved fear, headache or head stress, overeating, and other problems.

Upregulation of peroxisome proliferator-activated receptor-α and the lipid metabolism pathway promotes carcinogenesis of ampullary cancer.

Ampullary cancer is a rare periampullary cancer currently with no targeted therapeutic agent. It is important to develop a deeper understanding of the carcinogenesis of ampullary cancer. We attempted to explore the characteristics of ampullary cancer in our dataset and a public database, followed by a search for potential drugs. We used a bioinformatics pipeline to analyze complementary (c)DNA microarray data of ampullary cancer and surrounding normal duodenal tissues from five patients. A public database from the National Center for Biotechnology Information Gene Expression Omnibus (NCBI GEO) was applied for external validation. Bioinformatics tools used included the Gene Set Enrichment Analysis (GSEA), Database for Annotation, Visualization and Integrated Discovery (DAVID), MetaCore, Kyoto Encyclopedia of Genes and Genomes (KEGG), Hallmark, BioCarta, Reactome, and Connectivity Map (CMap). In total, 9097 genes were upregulated in the five ampullary cancer samples compared to normal duodenal tissues. From the MetaCore analysis, genes of peroxisome proliferator-activated receptor alpha (PPARA) and retinoid X receptor (RXR)-regulated lipid metabolism were overexpressed in ampullary cancer tissues. Further a GSEA of the KEGG, Hallmark, Reactome, and Gene Ontology databases revealed that PPARA and lipid metabolism-related genes were enriched in our specimens of ampullary cancer and in the NCBI GSE39409 database. Expressions of PPARA messenger (m)RNA and the PPAR-α protein were higher in clinical samples and cell lines of ampullary cancer. US Food and Drug Administration (FDA)-approved drugs, including alvespimycin, trichostatin A (a histone deacetylase inhibitor), and cytochalasin B, may have novel therapeutic effects in ampullary cancer patients as predicted by the CMap analysis. Trichostatin A was the most potent agent for ampullary cancer with a half maximal inhibitory concentration of < 0.3 µM. According to our results, upregulation of PPARA and lipid metabolism-related genes are potential pathways in the carcinogenesis and development of ampullary cancer. Results from the CMap analysis suggested potential drugs for patients with ampullary cancer.

Circulatory Rejuvenated EPCs Derived from PAOD Patients Treated by CD34+ Cells and Hyperbaric Oxygen Therapy Salvaged the Nude Mouse Limb against Critical Ischemia.

This study tested whether circulatory endothelial progenitor cells (EPCs) derived from peripheral arterial occlusive disease (PAOD) patients after receiving combined autologous CD34+ cell and hyperbaric oxygen (HBO) therapy (defined as rejuvenated EPCs) would salvage nude mouse limbs against critical limb ischemia (CLI). Adult-male nude mice (n = 40) were equally categorized into group 1 (sham-operated control), group 2 (CLI), group 3 (CLI-EPCs (6 × 105) derived from PAOD patient's circulatory blood prior to CD34+ cell and HBO treatment (EPCPr-T) by intramuscular injection at 3 h after CLI induction) and group 4 (CLI-EPCs (6 × 105) derived from PAOD patient's circulatory blood after CD34+ cell and HBO treatment (EPCAf-T) by the identical injection method). By 2, 7 and 14 days after the CLI procedure, the ischemic to normal blood flow (INBF) ratio was highest in group 1, lowest in group 2 and significantly lower in group 4 than in group 3 (p < 0.0001). The protein levels of endothelial functional integrity (CD31/von Willebrand factor (vWF)/endothelial nitric-oxide synthase (eNOS)) expressed a similar pattern to that of INBF. In contrast, apoptotic/mitochondrial-damaged (mitochondrial-Bax/caspase-3/PARP/cytosolic-cytochrome-C) biomarkers and fibrosis (Smad3/TGF-ß) exhibited an opposite pattern, whereas the protein expressions of anti-fibrosis (Smad1/5 and BMP-2) and mitochondrial integrity (mitochondrial-cytochrome-C) showed an identical pattern of INBF (all p < 0.0001). The protein expressions of angiogenesis biomarkers (VEGF/SDF-1α/HIF-1α) were progressively increased from groups 1 to 3 (all p < 0.0010). The number of small vessels and endothelial cell surface markers (CD31+/vWF+) in the CLI area displayed an identical pattern of INBF (all p < 0.0001). CLI automatic amputation was higher in group 2 than in other groups (all p < 0.001). In conclusion, EPCs from HBO-C34+ cell therapy significantly restored the blood flow and salvaged the CLI in nude mice.

Infants' Vitamin D Nutritional Status in the First Year of Life in Northern Taiwan.

Vitamin D deficiency (VDD) and insufficiency (VDI) are common among exclusively breastfeeding infants. However, epidemiological evidence for the prevalence of VDD in infants during their first year of life in Taiwan has never been found. This trial determined the prevalence of VDD and VDI and the association between dietary vitamin D and vitamin D nutritional status in Northern Taiwan. A cross-sectional study was conducted on infants who returned to well-baby examinations from October 2012 to January 2014 in three hospitals: Shin Kong Wu Ho-Su Memorial Hospital, Taipei Medical University Hospital, and Shuang Ho Hospital. The specific vitamin D cut-off concentrations for VDD, VDI, and VDS are 25(OH)D3 levels ≤ 20, 21-29, and ≥ 30 (ng/mL). Overall, 481 infants' parents completed a questionnaire comprising questions related to vitamin D nutritional status, including weekly time outdoors, breastfeeding status, anthropometric measurement, and assessment of dietary intake, including milk and complementary food. The results revealed that 197 (41%) and 212 (44%) of infants in their first year of life had VDI and VDD, respectively, by the Endocrine Society guidelines. Breastfed infants had a higher prevalence of VDI (86.1%) than did mixed-fed (51.9%) and formula-fed (38.5%) infants (p < 0.001). The prevalence of VDD was 55.4% in infants aged under six months but increased to 61.6% in infants aged over six months. Infants in the VDI and VDD groups had the same anthropometrics as those in the vitamin D sufficiency (VDS) group. Our results revealed that 25(OH)D3 had a negative correlation with the intact parathyroid hormone (iPTH) when the serum 25(OH)D3 level ≤ 20 ng/mL (r = -0.21, p = 0.001). The VDS group had a higher total vitamin D intake than did the VDI and VDD groups, which was mainly obtained from infant formula. Our data revealed that dietary vitamin D intake and birth season were major indicators in predicting VDD. Lower dietary vitamin D intake and born in winter and spring significantly increased the odds ratio (OR) for VDI by 1.15 (95% CI 1.09-1.20) and 2.02 (95% CI 1.10-3.70), respectively, and that for VDD by 1.23 (95% CI 1.16-1.31) and 2.37 (95% CI 1.35-4.17) without covariates adjustment, respectively. Furthermore, ORs for VDI and VDD significantly differed after adjustment for covariates. In conclusion, the prevalence of VDI and VDD were high in infants during the first year of life. Breastfeeding infants had difficulty in obtaining sufficient vitamin D from diet. In cases where the amount of sun exposure that is safe and sufficient to improve vitamin D status is unclear, breastfed infants aged below one year old are recommended to be supplemented with vitamin D.

Iron Status of Infants in the First Year of Life in Northern Taiwan.

Iron deficiency (ID) and iron deficiency anemia (IDA) typically occur in developing countries. Notably, ID and IDA can affect an infant's emotion, cognition, and development. Breast milk is considered the best food for infants. However, recent studies have indicated that breastfeeding for more than six months increases the risk of ID. This study investigated the prevalence of ID and IDA, as well as the association between feeding type and iron nutritional status in northern Taiwan. A cross-sectional study was conducted on infants who returned to the well-baby clinic for routine examination from October 2012 to January 2014. Overall, 509 infants aged 1-12 months completed the iron nutritional status analysis, anthropometric measurement, and dietary intake assessment, including milk and complementary foods. The results revealed that 49 (10%) and 21 (4%) infants in their first year of life had ID and IDA, respectively, based on the World Health Organization criteria. Breastfed infants had a higher prevalence rate of ID and IDA than mixed-fed and formula-fed infants (p < 0.001). Regarding biomarkers of iron status, plasma hemoglobin (Hb), ferritin, and transferrin saturation (%) levels were significantly lower in ID and IDA groups. The prevalence of ID and IDA were 3.7% and 2.7%, respectively, in infants under six months of age, but increased to 20.4% and 6.6%, respectively, in infants above six months of age. The healthy group had a higher total iron intake than ID and IDA groups, mainly derived from infant formula. The total dietary iron intake was positively correlated with infants' Hb levels. Compared with formula-fed infants, the logistic regression revealed that the odds ratio for ID was 2.157 (95% confidence interval [CI]: 1.369-3.399) and that for IDA was 4.196 (95% CI: 1.780-9.887) among breastfed infants (p < 0.001) after adjusted for all confounding factors (including gestational week, birthweight, sex, body weight percentile, body length percentile, age of infants, mothers' BMI, gestational weight gain, education level, and hemoglobin level before delivery). In conclusion, our results determined that breastfeeding was associated with an increased the prevalence of ID and/or IDA, especially in infants above six months. This suggests that mothers who prolonged breastfeed after six months could provide high-quality iron-rich foods to reduce the prevalence of ID and IDA.

Combined Therapy With Hyperbaric Oxygen and Melatonin Effectively Reduce Brain Infarct Volume and Preserve Neurological Function After Acute Ischemic Infarct in Rat.

This study tested the hypothesis that combined hyperbaric oxygen (HBO) and melatonin (Mel) was superior to either one for protecting the brain functional and parenchymal integrity from acute ischemic stroke (IS) injury. Adult-male Sprague-Dawley rats were divided into groups 1 (sham-operated control), 2 (IS), 3 (IS + HBO), 4 (IS + Mel), and 5 (IS + HBO-Mel). By day 28 after IS, the brain infarct area (BIA) was lowest in group 1, highest in group 2, significantly higher in groups 3 and 4 than in group 5, but not different between groups 3 and 4. The neurological function at day 7, 14, and 28 exhibited an opposite pattern to BIA among the 5 groups. The protein expressions of inflammatory (IL-1ß/IL-6/iNOS/TNF-α/p-NF-κB), apoptotic (cleaved-caspase3/cleaved-PARP/mitochondrial Bax), mitochondrial/DNA-damaged (cytochrome-C/γ-H2AX), oxidative stress (NOX-1/NOX-2), and autophagy (i.e. ratio of CL3B-II/CL3B-I) biomarkers displayed an identical pattern of BIA among 5 groups. Cellular expressions of inflammation (F4/80+/GFAP+) and DNA-damaged biomarker (γ-H2AX+) exhibited an identical pattern, whereas the integrities of myelin sheath/neuron (MPB+/NeuN+), endothelial cell (CD31+/vWF+), and number of small vessels exhibited an opposite pattern of BIA among the 5 groups. Combined HBO-Mel therapy offered an additional benefit in protecting the brain against IS injury.

Feedback Regulation between Aquatic Microorganisms and the Bloom-Forming Cyanobacterium Microcystis aeruginosa.

The frequency and intensity of cyanobacterial blooms are increasing worldwide. Interactions between toxic cyanobacteria and aquatic microorganisms need to be critically evaluated to understand microbial drivers and modulators of the blooms. In this study, we applied 16S/18S rRNA gene sequencing and metabolomics analyses to measure the microbial community composition and metabolic responses of the cyanobacterium Microcystis aeruginosa in a coculture system receiving dissolved inorganic nitrogen and phosphorus (DIP) close to representative concentrations in Lake Taihu, China. M. aeruginosa secreted alkaline phosphatase using a DIP source produced by moribund and decaying microorganisms when the P source was insufficient. During this process, M. aeruginosa accumulated several intermediates in energy metabolism pathways to provide energy for sustained high growth rates and increased intracellular sugars to enhance its competitive capacity and ability to defend itself against microbial attack. It also produced a variety of toxic substances, including microcystins, to inhibit metabolite formation via energy metabolism pathways of aquatic microorganisms, leading to a negative effect on bacterial and eukaryotic microbial richness and diversity. Overall, compared with the monoculture system, the growth of M. aeruginosa was accelerated in coculture, while the growth of some cooccurring microorganisms was inhibited, with the diversity and richness of eukaryotic microorganisms being more negatively impacted than those of prokaryotic microorganisms. These findings provide valuable information for clarifying how M. aeruginosa can potentially modulate its associations with other microorganisms, with ramifications for its dominance in aquatic ecosystems.IMPORTANCE We measured the microbial community composition and metabolic responses of Microcystis aeruginosa in a microcosm coculture system receiving dissolved inorganic nitrogen and phosphorus (DIP) close to the average concentrations in Lake Taihu. In the coculture system, DIP is depleted and the growth and production of aquatic microorganisms can be stressed by a lack of DIP availability. M. aeruginosa could accelerate its growth via interactions with specific cooccurring microorganisms and the accumulation of several intermediates in energy metabolism-related pathways. Furthermore, M. aeruginosa can decrease the carbohydrate metabolism of cooccurring aquatic microorganisms and thus disrupt microbial activities in the coculture. This also had a negative effect on bacterial and eukaryotic microbial richness and diversity. Microcystin was capable of decreasing the biomass of total phytoplankton in aquatic microcosms. Overall, compared to the monoculture, the growth of total aquatic microorganisms is inhibited, with the diversity and richness of eukaryotic microorganisms being more negatively impacted than those of prokaryotic microorganisms. The only exception is M. aeruginosa in the coculture system, whose growth was accelerated.

Dietary Leucine Supplement Ameliorates Hepatic Steatosis and Diabetic Nephropathy in db/db Mice.

Dietary leucine supplementation has been explored for the therapeutic intervention of obesity and obesity-induced metabolic dysfunctions. In this study, we aim to examine the effects of dietary leucine supplementation in db/db mice. Mice were treated with or without leucine (1.5% w/v) in drinking water for 12 weeks. The leucine supplement was found to reduce insulin resistance and hepatic steatosis in db/db mice. Using Nuclear Magnetic Resonance (NMR)-based lipidomics, we found that the reduction of hepatic triglyceride synthesis was correlated with attenuated development of fatty liver. In addition, diabetic nephropathy (DN) was also ameliorated by leucine. Using liquid chromatography⁻time-of-flight mass spectrometry (LC-TOF MS)-based urine metabolomics analysis, we found that the disturbance of the tricarboxylic acid (TCA) cycle was reversed by leucine. The beneficial effects of leucine were probably due to AMP-activated protein kinase (AMPK) activation in the liver and kidneys of db/db mice. Thus, dietary leucine supplementation may potentially be a nutritional intervention to attenuate hepatic steatosis and early DN in type II diabetes.

Tricholoma matsutake Aqueous Extract Induces Hepatocellular Carcinoma Cell Apoptosis via Caspase-Dependent Mitochondrial Pathway.

Tricholoma matsutake, one of widely accepted functional mushrooms, possesses various pharmacological activities, and its antitumor effect has become an important research point. Our study aims to evaluate the cytotoxicity activities of T. matsutake aqueous extract (TM) in HepG2 and SMMC-7721 cells. In in vitro experiments, TM strikingly reduced cell viability, promoted cell apoptosis, inhibited cell migration ability, induced excessive generation of ROS, and caused caspases cascade and mitochondrial membrane potential dissipation in hepatocellular carcinoma cells. In in vivo experiments, 14-day TM treatment strongly suppressed tumor growth in HepG2 and SMMC-7721-xenografted nude mice without influence on their body weights and liver function. Furthermore, TM increased the levels of cleaved poly-ADP-ribose polymerase (PARP), Bad, and Bax and reduced the expressions of B-cell lymphoma 2 (Bcl-2) in treated cells and tumor tissues. All aforementioned results suggest that caspase-dependent mitochondrial apoptotic pathways are involved in TM-mediated antihepatocellular carcinoma.

Combined Therapy With Adipose-Derived Mesenchymal Stem Cells and Ciprofloxacin Against Acute Urogenital Organ Damage in Rat Sepsis Syndrome Induced by Intrapelvic Injection of Cecal Bacteria.

UNLABELLED: We hypothesized that combined treatment with autologous adipose-derived mesenchymal stem cell (ADMSC) and ciprofloxacin is superior to ciprofloxacin only in reducing sepsis-induced urogenital organ damage and mortality in rat sepsis syndrome (SS) caused by intrapelvic injection of cecal bacteria (1.0 × 10(4) cells per milliliter; total, 5.0 ml). Male Sprague-Dawley rats (n = 60) equally divided into group 1 (sham-control), group 2 (SS), group 3 (SS-ADMSC [5.0 × 10(5) intravenously at 0.5, 6, and 18 hours after sepsis induction]), group 4 (SS-ciprofloxacin [3.0 mg/kg, b.i.d.] for 5 days), and group 5 (SS-ADMSC-ciprofloxacin) were sacrificed by day 5. Mortality rate and creatinine level were highest in group 2 and lowest in group 1 and significantly higher in groups 3 and 4 than those in group 5, but there was no difference between groups 3 and 4 (all p < .005). The kidney injury score, inflammatory biomarker expressions at protein (tumor necrosis factor-1α, nuclear factor-κB, matrix metallopeptidase-9, regulated on activation, normal T-cell expressed and secreted, interleukin-1ß) and cellular (CD14+, migratory inhibitor factor positive, CD68+) levels in kidneys and urinary bladder were lowest in group 1 and highest in group 2, higher in group 4 than in groups 3 and 5, and higher in group 3 than in group 5 (all p < .001). Protein expressions of apoptosis (Bax, cleaved caspase 3 and poly[ADP-ribose] polymerase 1, p21 protein [Cdc42/Rac]-activated kinase 2) and oxidative stress (oxidized protein, NADPH oxidase (NOX)-1, NOX-2) in these organs showed an identical pattern compared with that of inflammation in all groups (all p < .001). In conclusion, ADMSC-assisted ciprofloxacin therapy offered an additional benefit by reducing acute urogenital organ damage in rat. SIGNIFICANCE: Autologous adipose-derived mesenchymal stem cell-assisted ciprofloxacin therapy offered an additional benefit by reducing acute urogenital organ damage in rats.