The green tea polyphenol epigallocatechin-3-gallate attenuates age-associated muscle loss via regulation of miR-486-5p and myostatin.

(-)-Epigallocatechin-3-gallate (EGCG), the most abundant catechin component in green tea, has been reported to attenuate age-associated insulin resistance, lipogenesis and loss of muscle mass through restoring Akt activity in skeletal muscle in our previous and present studies. Accumulated data has suggested that polyphenols regulate signaling pathways involved in aging process such as inflammation and oxidative stress via modulation of miRNA expression. Here we found that miRNA-486-5p was significantly decreased in both aged senescence accelerated mouse-prone 8 (SAMP8) mice and late passage C2C12 cells. Thus, we further investigated the regulatory effect of EGCG on miRNA-486-5p expression in age-regulated muscle loss. SAMP8 mice were fed with chow diet containing without or with 0.32% EGCG from aged 32 weeks for 8 weeks. Early passage (<12 passages) and late passage (>30 passages) of C2C12 cells were treated without or with EGCG at concentrations of 50 µM for 24h. Our data showed that EGCG supplementation increased miRNA-486-5p expression in both aged SAMP8 mice and late passage C2C12 cells. EGCG stimulated AKT phosphorylation and inhibited FoxO1a-mediated MuRF1 and Atrogin-1 transcription via up-regulating the expression of miR-486 in skeletal muscle of 40-wk-old SAMP8 mice as well as late passage C2C12 cells. In addition, myostatin expression was increased in late passage C2C12 cells and anti-myostatin treatment upregulated the expression of miR-486-5p. Our results identify a unique mechanism of a dietary constituent of green tea and suggest that use of EGCG or compounds derived from it attenuates age-associated muscle loss via myostatin/miRNAs/ubiquitin-proteasome signaling.

Dysregulations of mitochondrial quality control and autophagic flux at an early age lead to progression of sarcopenia in SAMP8 mice.

The senescence-accelerated mouse (SAM) prone 8 (SAMP8) has been demonstrated for muscular aging research including sarcopenia, but its underlying mechanisms remain scarce. Physiological indices and histology of skeletal muscle were analyzed in SAMP8 mice at different ages. SAMP8 mice exhibited typical features of sarcopenia at 40 weeks of age and were more time-efficient than that at 88 weeks of age in bothSAM resistant 1 (SAMR1) and C57BL/6 mice. Increase in FoxO3a-mediated transcription of Atrogin-1 and MuRF1 and decrease in phosphorylated mTOR/P70s6k were observed at week 40 in SAMP8 mice. High oxidative stress was observed from week 24 and persisted to week 40 in SAMP8 mice evidenced by overexpression of protein carbonyl groups and reduced activities of CAT, SOD, and GPx. Downregulation of genes involved in mitochondrial biogenesis (PGC-1α, Nrf-1, Tfam, Ndufs8, and Cox5b) and in mitochondrial dynamics fission (Mfn2 and Opa1) from week 24 indicated dysregulation of mitochondrial quality control in SAMP8 mice. Impaired autophagic flux was observed in SAMP8 mice evidenced by elevated Atg13 and LC3-II accompanied with the accumulation of P62 and LAMP1. Increases in inflammatory factors (IL-6 and MCP-1), adipokines (leptin and resistin), and myostatin in serum at week 32 and decline in Pax7+ satellite cell resided next to muscle fibers at week 24 implied that muscle microenvironment contributed to the progression of sarcopenia in SAMP8 mice. Our data suggest that early alterations of mitochondrial quality control and autophagic flux worsen muscle microenvironment prior to the onset of sarcopenia.

Effect of light and atmosphere on the cultivation of the golden oyster culinary-medicinal mushroom, Pleurotus citrinopileatus (higher Basidiomycetes).

With an aim to explore the productivity and quality of the fruiting body of culinary-medicinal golden oyster mushroom Pleurotus citrinopileatus, the carbon dioxide (CO2) concentration of the ambient atmosphere was adjusted and a light-emitting diode panel was used to illuminate the colonized mycelium at different wavelengths. Biological efficiency and yield were higher at CO2 levels of 0.05 and 0.1% than other tested CO2 levels, and the mature fruiting body showed the highest yellow value at a CO2 level of 0.1% (of all tested CO2 levels). The highest biological efficiency and yield was obtained at the 720-nm wavelength. The ergosterol content of the pileus of the fruiting body was higher than that of the stipe in any flush time at a 720-nm wavelength of light and a CO2 concentration of 0.1%. The decreased percentages of cellulose and lignin at the appearance of primordia were larger than those of mycelial growth duration. The fruiting quality of P. citrinopileatus might thus be enhanced by 720-nm illumination and an atmosphere with a CO2 concentration of 0.1 to 0.15%.

Overview on new progress of hereditary diffuse gastric cancer with CDH1 variants.

Hereditary diffuse gastric cancer (HDGC), comprising 1%-3% of gastric malignances, has been associated with CDH1 variants. Accumulating evidence has demonstrated more than 100 germline CDH1 variant types. E-cadherin encoded by the CDH1 gene serves as a tumor suppressor protein. CDH1 promoter hypermethylation and other molecular mechanisms resulting in E-cadherin dysfunction are involved in the tumorigenesis of HDGC. Histopathology exhibits characteristic signet ring cells, and immunohistochemical staining may show negativity for E-cadherin and other signaling proteins. Early HDGC is difficult to detect by endoscopy due to the development of lesions beneath the mucosa. Prophylactic gastrectomy is the most recommended treatment for pathogenic CDH1 variant carriers. Recent studies have promoted the progression of promising molecular-targeted therapies and management strategies. This review summarizes recent advances in CDH1 variant types, tumorigenesis mechanisms, diagnosis, and therapy, as well as clinical implications for future gene therapies.

Oligonol Alleviates Sarcopenia by Regulation of Signaling Pathways Involved in Protein Turnover and Mitochondrial Quality.

SCOPE: Oligonol has been shown to moderate mitochondrial biogenesis, protein synthesis, and protein degradation in diabetic mice in a previous study. It is therefore hypothesized that oligonol alleviated sarcopenia by regulating pathways involved in protein turnover and mitochondrial quality. METHODS AND RESULTS: The 32-week-old senescence-accelerated mouse prone 8 (SAMP8) mice are fed with chow diet containing 200 mg kg-1 oligonol for 8 weeks. Oligonol supplementation increased skeletal muscle mass, cross-sectional areas, and grip strength in SAMP8 mice. Oligonol increased phosphorylation of AKT/mTOR/p70sk6, inhibited nuclear localization of FoxO3a and NFκB, and decreased transcription of MuRF-1 and MAFbx in skeletal muscle of SAMP8 mice. Downregulation of mitochondrial biogenesis genes (PGC-1α and Tfam) and mitochondrial fusion genes (Mfn2 and Opa1), loss of PINK1, overexpression of Atg13, LC3-II, and p62, and abundant accumulation of autophagosomes and lysosomes in skeletal muscle of SAMP8 mice are limited by oligonol. Furthermore, oligonol reduced expression of released cytochrome c and cleaved caspase-9 in skeletal muscle of SAMP8 mice. CONCLUSION: Regulating pathways involved in protein synthesis and degradation, mitochondrial biogenesis, mitochondrial fusion/fission, autophagy, and mitochondria-dependent apoptosis by oligonol contribute to positive protein turnover and mitochondrial quality, thus increasing muscle mass and strength in SAMP8 mice.

Optimization of Submerged Cultivation Conditions for Production of Big Cup Culinary-Medicinal Mushroom Clitocybe maxima (Agaricomycetes) Biomass with Significant Antioxidative and Antihyperlipidemic Activities.

In this study, a large Clitocybe maxima mycelium biomass was obtained by submerged cultivation under optimal conditions. Three test samples from lyophilized mycelia, including hot water extract (CW) and elutes from solvents with different polarity (CA and CB), were combined and used to explore antioxidant and antihyper-lipidemic activities in vitro and in vivo. The CA group showed the highest DPPH free radical scavenging activity and iron-reducing capability at concentrations of 6.0% and 3.0% (w/v), respectively. Further, the CA group showed the highest glutathione peroxidase and superoxide dismutase activities at a dose of 0.25 mg/kg body weight (CA-0.25 group) in all hyperlipidemic hamsters tested. Serum lipid levels (apart from high-density lipoprotein cholesterol levels) of hamsters in the CA-0.25 group were lower than those of hamsters in the negative control group in antihy-perlipidemic tests. Therefore, we believe that extracts from C. maxima mycelia are rich reservoirs of antioxidant and antihyperlipidemic activities.

Antihyperlipidemic and antioxidant effects of extracts from Pleurotus citrinopileatus.

Pleurotus citrinopileatus is a popular edible mushroom which is physiologically active in both humans and animals. In the study we investigate the effects of this mushroom on hyperlipidemic hamster rats. Four dietary forms of the mushroom were created as follows. The powdered dry fruiting body, hot-water extract, and two kinds of elutes were obtained, from ethyl acetate extract and methanol extract, respectively, in different mixed proportion solvents over silica gel column chromatography (referred to as EAE and MOE, respectively). They were tested at different dosages as a supplement to a high-fat diet in hyperlipidemic rats. Serum triglycerides and total cholesterol levels were significantly lower in groups supplemented with the highest dosages of EAE and MOE (0.5 g/kg, body weight daily) as compared with the control groups that received no mushroom additive. High-density lipoprotein levels in these same two experimental groups were also significantly higher than those in the negative control group. The tested rats that were fed with EAE had the highest serum glutathione peroxidase and superoxide dismutase activity, and those with the MOE and EAE had the highest DPPH free radical scavenging activities and ferric-reducing abilities, tested in vitro. The major constituents of MOE and EAE were identified as ergosterol and nicotinic acid, respectively. P. citrinopileatus extracts may have a significant antihyperlipidemia effect. Furthermore, antioxidant activities and antihyperlipidemic effects of MOE and EAE seemed to display similar tendencies.

[Heteroplasmy in human mtDNA control region].

OBJECTIVE: To observe the length heteroplasmy and point heteroplasmy in human mtDNA control region. METHODS: The peripheral blood, buccal cell, and single hair shaft from 50 individuals and 16 family members, related in their maternallineage were analyzed by direct sequencing, and clones from 20 individuals whose mtDNA sequences have a T-C transition at 16189 nt were sequenced. RESULTS: No point heteroplasmy were observed in peripheral blood, buccal cell, single hair shaft from the same individual, neither in maternally related individuals. Length heteroplasmy was observed in those individuals with a homopolymeric tract and the different clones from the same individual has different proportions of length variants, but the hair shafts from the same individual were very similar to the measurements made from blood DNA. No length heteroplasmy was observed between different tissues from the same individual. CONCLUSION: mtDNA sequences have a characteristic of high consistency and genetic stability, mtDNA sequencing is a suitable tool for forensic applications such as individual identification.

Peptide-based enteral formula improves tolerance and clinical outcomes in abdominal surgery patients relative to a whole protein enteral formula.

AIM: To compare a dipeptide- and tripeptide-based enteral formula with a standard enteral formula for tolerance and nutritional outcomes in abdominal surgery patients. METHODS: A retrospective study was performed to assess the differences between a whole-protein formula (WPF) and a dipeptide- and tripeptide-based formula (PEF) in clinical outcomes. Seventy-two adult intensive care unit (ICU) patients with serum albumin concentrations less than 3.0 g/dL were enrolled in this study. Patients were divided into two groups (WPF group = 40 patients, PEF group = 32 patients). The study patients were fed for at least 7 d, with ≥ 1000 mL of enteral formula infused on at least 3 of the days. RESULTS: The mean serum albumin level on postoperative day (POD) 10, prealbumin levels on POD-5 and POD-10, and total lymphocyte count on POD-5 were significantly higher in the PEF group compared to those in the WPF group (P < 0.05). The average maximum gastric residual volume of the PEF patients during their ICU stays was significantly lower than that for WPF patients. CONCLUSION: Dipeptide- and tripeptide-based enteral formulas are more efficacious and better tolerated than whole-protein formulas.

Effects of Illumination Pattern during Cultivation of Fruiting Body and Bioactive Compound Production by the Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes).

We investigated the effects of light intensity in the 3 cultivation stages separately-the mycelium colonization stage, the primordial initiation stage, and the fruiting stage (in order)-on fruiting body and bioactive compound production by Cordyceps militaris. In the mycelium colonization stage, rice substrates were incubated in a spawn running room at 23°C. During the primordial initiation stage, C. militaris was grown at 18°C and illuminated 12 hours/day. In the fruiting stage the temperature was 23°C, with illumination provided 12 hours/day. The highest fruiting body yield and biological efficiency were 4.06 g dry weight/bottle and 86.83%, respectively, under 1750 ± 250 lux during the second and third stages. The cordycepin content was highest during the second and third stages under 1250 ± 250 lux. The mannitol and polysaccharide contents were highest under 1250 ± 250 and 1750 ± 250 lux during the primordial initiation stage and the fruiting stage, respectively. Thus, with controlled lighting, C. militaris can be cultivated in rice-water medium to increase fruiting body yield and bioactive compound production.

The Influence of Aggressive Parenteral Nutrition to Preterm and Very Low Birth Weight Infants.

Background. To achieve the weight gain of preterm infants who are appropriate for gestational age without adverse effect, there should be no interruption in delivery of nutrients from time of birth. Methods. Twenty-eight very low birth weight infants were eligible for the study. Those administered conventional nutrition (amino acids 2 g/kg/day started on third day of life) were classified as the conventional support (CVS) group, and those administered aggressive early nutrition (amino acid 2 g/kg/day started on first day of life) were classified as the aggressive support (AGS) group. Results. The days babies took to reach the weight of 2000 g in the AGS group was significantly shorter than for babies in the CVS group, and babies in the AGS group showed better tolerance to enteral nutrition and had shortened neonatal intensive care unit days. Conclusion. The results demonstrated that aggressive early nutrition showed better tolerance to enteral nutrition, higher total calories, and shortened the stay in the neonatal intensive care unit.

Influence of preoperative peripheral parenteral nutrition with micronutrients after colorectal cancer patients.

BACKGROUND: The inflammatory reactions are stronger after surgery of malnourished preoperative patients. Many studies have shown vitamin and trace element deficiencies appear to affect the functioning of immune cells. Enteral nutrition is often inadequate for malnourished patients. Therefore, total parenteral nutrition (TPN) is considered an effective method for providing preoperative nutritional support. TPN needs a central vein catheter, and there are more risks associated with TPN. However, peripheral parenteral nutrition (PPN) often does not provide enough energy or nutrients. PURPOSE: This study investigated the inflammatory response and prognosis for patients receiving a modified form of PPN with added fat emulsion infusion, multiple vitamins (MTV), and trace elements (TE) to assess the feasibility of preoperative nutritional support. Methods. A cross-sectional design was used to compare the influence of PPN with or without adding MTV and TE on malnourished abdominal surgery patients. RESULTS: Both preoperative groups received equal calories and protein, but due to the lack of micronutrients, patients in preoperative Group B exhibited higher inflammation, lower serum albumin levels, and higher anastomotic leak rates and also required prolonged hospital stays. CONCLUSION: Malnourished patients who receive micronutrient supplementation preoperatively have lower postoperative inflammatory responses and better prognoses. PPN with added fat emulsion, MTV, and TE provides valid and effective preoperative nutritional support.

Antitumor and Immunomodulating Activities of Exopolysaccharide Produced by Big Cup Culinary- Medicinal Mushroom Clitocybe maxima (Higher Basidiomycetes) in Liquid Submerged Culture.

Water-soluble polysaccharides extracted from mushrooms have been found to have some physiological effects. In this study, exopolysaccharides (EPSs) were extracted by alcohol precipitation from cultivated broth of the mushroom Clitocybe maxima. EPSs with molecular weights of 10(4) and 10(5) Da were obtained by ultrafiltration; they are referred to as EPA and EPB, respectively. The major components of these EPSs were glucose, galactose, mannose, rhamnose, and arabinose. ICR mice with artificially induced metastatic pulmonary tumors were fed a daily diet containing EPA or EPB at doses of 8, 20, or 50 mg/kg. Results showed that the proliferation of pulmonary sarcoma lesions was lower in the groups fed EPS. In addition, the numbers of total T cells, CD4+ cells, CD8+ cells, and macrophages significantly increased in EPS-fed mice compared with the negative control group. The antitumor and immunomodulating effects observed in the EPB-fed groups were higher than those of EPA-fed groups. These results demonstrate the ability of EPSs of C. maxima to inhibit tumor cells while enhancing immune response.

Lethal effect of microwaves on spores of Bacillus spp.

Strains representing four types of common heat-resistant spores of Bacillus spp., B. cereus CCRC 14655, B. coagulans CCRC10606, B. licheniformis CCRC14693, and B. subtilis CCRC14199, were heated with microwaves at different power levels and under different conditions in salt solutions, starch solutions, and containers. The results of this study showed that B. licheniformis spores had the highest microwave tolerance at a power level of 100% for different incubation times. B. coagulans spores showed the lowest microwave tolerance in salt solutions with water activity values of 0.6, 0.7, 0.8, and 0.9, and B. licheniformis spores were the most resistant in the tested salt solutions at different incubation times. An analysis of the effect of the viscosity of the medium revealed that the bacteria had the lowest microwave resistance in a medium containing <0.8% starch in solution. The microwave resistance levels of the test microorganisms were the lower in glass containers than in polypropylene containers and aluminum foil-enclosed pouches. Of the four species of bacilli, B. licheniformis had the highest microwave tolerance (P < 0.05) under all conditions.

Antimicrobial effect of extracts of cruciferous vegetables.

The cruciferous vegetables cauliflower, broccoli, cabbage, Chinese radish, Chinese kale, and Chinese kitam were used in this study to prepare water-soluble and methanol-water extracts. Crude protein extracts were also obtained by diethylaminoethyl (DEAE) anion exchange chromatography. Water-soluble polysaccharides were prepared by ethanol precipitation followed by ultrafiltration. The antimicrobial effects of all these extracts were evaluated against Gram-positive bacteria, Gram-negative bacteria, and yeast. Crude protein extracts exhibited the greatest antimicrobial activity in monoculture experiments. The antimicrobial effects of cruciferous vegetables were also studied by steeping beef, carrot, and celery in chlorine (10 ppm) or citric acid solution (1%) containing the crude protein extract (500 ppm) for different time periods. Total aerobic plate counts and coliform counts on these foods decreased significantly after 10 minutes in all steeping solutions (p < 0.05).

Antihyperglycemic effect of polysaccharide from fermented broth of Pleurotus citrinopileatus.

Pleurotus citrinopileatus is an edible mushroom, which has recently become very popular, with a consequent increase in industrial production. Water-soluble polysaccharides (WSPS), extracted from edible mushrooms, have been found to have antitumor and immunoenhancing effects. In this study, we investigate the effects of WSPS extracted from submerged fermented medium of P. citrinopileatus on hyperglycemia and damaged pancreatic cells in rats with streptozotocin (STZ)-induced diabetes. The diabetic rats fed with water-soluble polysaccharide of P. citrinopileatus (SPPC) lost less body weight than those fed SPPC-free regular diet. Serum total cholesterol and triglyceride levels in the diabetic rats fed with SPPC at a dose of 0.4 g/kg bw daily was lower than in the groups fed with SPPC at doses of 0.04 and 0.12 g/kg bw. The fasting blood glucose levels of diabetic rats fed with SPPC were 44% lower than the negative controls. The degree of damage to the islets of Langerhans of the rats fed with the highest dosage of SPPC was significantly lower than those fed with SPPC at doses of 0.04 and 0.12 g/kg bw. The results showed that STZ-induced diabetic rats fed with SPPC might help alleviate the elevation of the level of that in fasting blood glucose.

Optimization for the production of water-soluble polysaccharide from Pleurotus citrinopileatus in submerged culture and its antitumor effect.

In recent years, a number of studies have been done on the physiological effects of water-soluble polysaccharides (WSPS) and their antitumor and immuno-enhancing properties. Many edible mushrooms, in particular those rich in WSPS, not only taste good but also contain ingredients beneficial to the physiology of the human body. In this study, response surface methodology was used to determine the optimal conditions for the production of WSPS, including the C/N ratio, initial pH, and incubation temperature. The highest yield of WSPS was obtained by incubation with a C/N ratio of 40, initial pH 5.5, and an incubation temperature of 25 degrees C. WSPS were extracted by alcohol precipitation from the fermented broth of edible Pleurotus citrinopileatus. These extracts, referred to as SPPC in this paper, had a molecular mass of more than 10(5) Da and were largely made up of glucose and mannose. SPPC was fed to mice which had artificial pulmonary metastatic tumors. Changes in the percentage of the numbers of tumor cells and immune cells were determined by flow cytometry. Daily feeding of SPPC at a dosage of 50 mg/kg to tumor-bearing mice for 12 days resulted in a significant increase in the number of T cells, CD4+ cells, CD8+ cells, and macrophages, compared with mice that were not fed any SPPC. The proliferation rate of the pulmonary sarcoma lesions slowed down.