DNA methylation of skeletal muscle function-related secretary factors identifies FGF2 as a potential biomarker for sarcopenia.

BACKGROUND: Sarcopenia is characterized by progressive loss of muscle mass and function due to aging. DNA methylation has been identified to play important roles in the dysfunction of skeletal muscle. The aim of our present study was to explore the whole blood sample-based methylation changes of skeletal muscle function-related factors in patients with sarcopenia. METHODS: The overall DNA methylation levels were analysed by using MethlTarget™ DNA Methylation Analysis platform in a discovery set consistent of 50 sarcopenic older adults (aged ≥65 years) and 50 age- and sex-matched non-sarcopenic individuals. The candidate differentially methylated regions (DMRs) were further validated by Methylation-specific PCR (MSP) in another two independent larger sets and confirmed by pyrosequencing. Receiver operating characteristic (ROC) curve analysis was used to determine the optimum cut-off levels of fibroblast growth factor 2 (FGF2)_30 methylation best predicting sarcopenia and area under the ROC curve (AUC) was measured. The correlation between candidate DMRs and the risk of sarcopenia was investigated by univariate analysis and multivariate logistic regression analysis. RESULTS: Among 1149 cytosine-phosphate-guanine (CpG) sites of 27 skeletal muscle function-related secretary factors, 17 differentially methylated CpG sites and 7 differentially methylated regions (DMRs) were detected between patients with sarcopenia and control subjects in the discovery set. Further methylation-specific PCR identified that methylation of fibroblast growth factor 2 (FGF2)_30 was lower in patients with sarcopenia and the level was decreased as the severity of sarcopenia increased, which was confirmed by pyrosequencing. Correlation analysis demonstrated that the methylation level of FGF2_30 was positively correlated to ASMI (r = 0.372, P < 0.001), grip strength (r = 0.334, P < 0.001), and gait speed (r = 0.411, P < 0.001). ROC curve analysis indicated that the optimal cut-off value of FGF2_30 methylation level that predicted sarcopenia was 0.15 with a sensitivity of 84.6% and a specificity of 70.1% (AUC = 0.807, 95% CI = 0.756-0.858, P < 0.001). Multivariate logistic regression analyses showed that lower FGF2_30 methylation level (<0.15) was significantly associated with increased risk of sarcopenia even after adjustment for potential confounders including age, sex, and BMI (adjusted OR = 9.223, 95% CI: 6.614-12.861, P < 0.001). CONCLUSIONS: Our results suggest that lower FGF2_30 methylation is correlated with the risk and severity of sarcopenia in the older adults, indicating that FGF2 methylation serve as a surrogate biomarker for the screening and evaluation of sarcopenia.

AflatoxinB1 (AFB1 ) and its toxic effect on the broilers intestine: A review.

Aflatoxin B1 (AFB1 ) is the predominant mycotoxin that originated toxicity in broilers through oxidative damage, intestinal barrier dysfunction, reduced immune system and dysfunction of microorganisms and enzymes in target organs. The intestine is the first AFB1 target organ destroyed after the bird's body is induced. This review summarises the current knowledge of the negative results of AFB1 -induced intestinal damage on broiler production. It was conducted in accordance with the relevant studies in the cited literatures being retrieved from PubMed, Google Scholar, Science Direct and Web of Science. First, AFB1 can change the intestinal barrier function by destroying the intestinal architecture, tissue and cell integrity of the gut epithelium. Second, AFB1 can damage the immune barrier function of the gastrointestinal mucosa. Third, the microbiota of birds interacts closely with the ingested aflatoxin. Finally, because broilers are tremendously sensitive to AFB1 contamination, the poisonous and noxious effects of this mycotoxin in the broiler industry cause millions of dollars in losses every year. This review briefly discussed that the AFB1 , which affects the intestines of broiler chickens, was reduced the immune apparatus, antioxidant protection system, gastric system, and broiler production status and its impact on human health. Therefore, this review will improve our perception of the important intestine in a bird's health and the adverse effect of AFB1 .

Serum Retinol Binding Protein 4 as a Potential Biomarker for Sarcopenia in Older Adults.

Sarcopenia is characterized by progressive loss of muscle mass and function due to aging. Retinol-binding protein 4 (RBP4) is an adipokine with pro-inflammatory effects. However, the change of RBP4 concentration and its role in sarcopenia remains unclear. The aim of this study was to evaluate the association of serum RBP4 level with sarcopenia in the older adults. A total of 816 community-dwelling older adults aged ≥60 years were enrolled. Serum RBP4 was measured by enzyme-linked immunosorbent assay. Appendicular skeletal muscle mass index (ASMI), grip strength, and gait speed were measured. We found that serum RBP4 levels were higher in patients with sarcopenia when compared with those without sarcopenias (44.3 [33.9-57.7] vs 38.0 [28.0-48.4] µg/mL). Receiver operating characteristic curve analysis indicated that the optimal cutoff value of serum RBP4 level that predicted sarcopenia was 38.79 µg/mL with a sensitivity of 67.8% and a specificity of 53.3%. Multivariate logistic regression analysis showed that the subjects with a higher level of RBP4 had a higher risk of sarcopenia (adjusted odds ratio [OR] = 2.036, 95% CI = 1.449-2.861). Serum RBP4 concentration was negatively correlated with grip strength (r = -.098), gait speed (r = -.186), and AMSI (r = -.096). Moreover, serum RBP4 levels were higher in patients with severe sarcopenia when compared with those with moderate sarcopenia (49.0 [37.3-61.2] vs 40.4 [31.3-51.2] µg/mL). Taken together, our results demonstrate that serum RBP4 level is correlated with the risk and severity of sarcopenia in the older adults, indicating that RBP4 might serve as a surrogate biomarker for the screening and evaluation of sarcopenia.

Evaluation of the protective effect of lycopene on growth performance, intestinal morphology, and digestive enzyme activities of aflatoxinB1 challenged broilers.

The current study was conducted to investigate the protective efficiency of dietary lycopene (LYC) supplementation on growth performance, intestinal morphology, and digestive enzyme activities aflatoxinB1 (AFB1 ) challenged broilers. A total of 240 days old Arber across male broiler chicks were randomly allocated in five treatments and six replicates (eight birds per replicate); feed and water were provided ad libitum during the 42 days experiment. The treatment diets were as follows: (i) Basal diet (control), (ii) Basal diet + 100 µg/kg AFB1 contaminated diet, (iii) Basal diet + 100 µg/kg AFB1  + 100 mg/kg LYC1, (iv) Basal diet + 100 µg/kg AFB1  + 200 mg/kg LYC2, and (v) Basal diet + 100 µg/kg AFB1  + 400 mg/kg LYC3. The results showed that the addition of LYC to AFB1 contaminated broiler diets significantly increased (p < .05) average daily gain (ADG) and decreased feed conversion ratio (FCR) compared to the AFB1 diet. AFB1 diet decreased the intestinal villus height (VH) and crypt depth ratio (VCR) while increasing the crypt depth (CD). However, dietary LYC supplemented diets relieved the intestinal morphological alterations. Dietary LYC supplementation (200 and 400 mg/kg) significantly improved (p < .05) intestinal digestive enzyme amylase and lipase activities with AFB1 contaminated diet. These findings suggested that LYC is a promising feed supplement in the broiler industry, alleviating the harmful effects of AFB1 .

Ubiquity of polystyrene digestion and biodegradation within yellow mealworms, larvae of Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae).

Academics researchers and "citizen scientists" from 22 countries confirmed that yellow mealworms, the larvae of Tenebrio molitor Linnaeus, can survive by eating polystyrene (PS) foam. More detailed assessments of this capability for mealworms were carried out by12 sources: five from the USA, six from China, and one from Northern Ireland. All of these mealworms digested PS foam. PS mass decreased and depolymerization was observed, with appearance of lower molecular weight residuals and functional groups indicative of oxidative transformations in extracts from the frass (insect excrement). An addition of gentamycin (30 mg g-1), a bactericidal antibiotic, inhibited depolymerization, implicating the gut microbiome in the biodegradation process. Microbial community analyses demonstrated significant taxonomic shifts for mealworms fed diets of PS plus bran and PS alone. The results indicate that mealworms from diverse locations eat and metabolize PS and support the hypothesis that this capacity is independent of the geographic origin of the mealworms, and is likely ubiquitous to members of this species.

Deep sequencing identification of miRNAs in pigeon ovaries illuminated with monochromatic light.

BACKGROUND: The use of light of different wavelengths has grown popular in the poultry industry. An optimum wavelength is believed to improve pigeon egg production, but little is known about the role of microRNAs (miRNAs) in the effects of monochromatic light on ovarian pigeon function. Herein, we harvested ovaries from pigeons reared under monochromatic light of different wavelength and performed deep sequencing on various tissues using an Illumina Solexa high-throughput instrument. RESULTS: We obtained 66,148,548, 67,873,805, and 71,661,771 clean reads from ovaries of pigeons reared under red light (RL), blue light (BL), and white light (WL), respectively. We identified 1917 known miRNAs in nine libraries, of which 524 were novel. Three and five differentially expressed miRNAs were identified in BL vs. WL and RL vs. WL groups, respectively. Quantitative reverse transcription PCR was used to validate differentially expressed miRNAs (miR-200, miR-122, and miR-205b). In addition, 5824 target genes were annotated as differentially expressed miRNAs, most of which are involved in reproductive pathways including oestrogen signalling, cell cycle, and oocyte maturation. Notably, ovarian miR-205b expression was significantly negatively correlated with its target 11ß-hydroxysteroid dehydrogenase type 1 (HSD11B1). CONCLUSIONS: miRNA-mRNA network analysis suggests that miR-205b targeting of HSD11B1 plays a key role in the effects of monochromatic light on pigeon egg production. These findings indicate that monochromatic light shortens the oviposition interval of pigeons, which may be useful for egg production and pigeon breeding.

Design of a multiband near-infrared sky brightness monitor using an InSb detector.

Infrared sky background level is an important parameter of infrared astronomy observations from the ground, particularly for a candidate site of an infrared capable observatory since low background level is required for such a site. The Chinese astronomical community is looking for a suitable site for a future 12 m telescope, which is designed for working in both optical and infrared wavelengths. However, none of the proposed sites has been tested for infrared observations. Nevertheless, infrared sky background measurements are also important during the design of infrared observing instruments. Based on the requirement, in order to supplement the current site survey data and guide the design of future infrared instruments, a multiband near-infrared sky brightness monitor (MNISBM) based on an InSb sensor is designed in this paper. The MNISBM consists of an optical system, mechanical structure and control system, detector and cooler, high gain readout electronics, and operational software. It is completed and tested in the laboratory. The results show that the sensitivity of the MNISBM meets the requirements of the measurement of near-infrared sky background level of several well-known astronomical infrared observing sites.

Biodegradation of polystyrene wastes in yellow mealworms (larvae of Tenebrio molitor Linnaeus): Factors affecting biodegradation rates and the ability of polystyrene-fed larvae to complete their life cycle.

Commercial production of polystyrene (PS) -a persistent plastic that is not biodegradable at appreciable rates in most environments-has led to its accumulation as a major contaminant of land, rivers, lakes, and oceans. Recently, however, an environment was identified in which PS is susceptible to rapid biodegradation: the larval gut of Tenebrio molitor Linnaeus (yellow mealworms). In this study, we evaluate PS degradation capabilities of a previously untested strain of T. molitor and assess its survival and PS biodegradation rates for a range of conditions (two simulated food wastes, three temperatures, seven PS waste types). For larvae fed PS alone, the %PS removed in the short (12-15 h) residence time of the mealworm gut gradually increased for 2-3 weeks then stabilized at values up to 65%. Thirty two-day survival rates were >85% versus 54% for unfed larvae. For mealworms fed ∼10% w/w PS and ∼90% bran, an agricultural byproduct, rates of PS degradation at 25 °C nearly doubled compared to mealworms fed PS alone. Polymer residues in the frass showed evidence of partial depolymerization and oxidation. All of the tested PS wastes degraded, with the less dense foams degrading most rapidly. Mealworms fed bran and PS completed all life cycle stages (larvae, pupae, beetles, egg), and the second generation had favorable PS degradation, opening the door for selective breeding.

Low-current & high-frequency electrical stunning increased oxidative stress, lipid peroxidation, and gene transcription of the mitogen-activated protein kinase/nuclear factor-erythroid 2-related factor 2/antioxidant responsive element (MAPK/Nrf2/ARE) signaling pathway in breast muscle of broilers.

Mechanism of electrical stunning (ES) methods on lipid peroxidation and antioxidant protection were studied by determining meat color, serum variables, antioxidant-related enzyme activities, gene expressions of mitogen-activated protein kinases (MAPKs), nuclear factor-erythroid 2-related factor 2 (Nrf2), glutathione S-transferases (GSTs), and superoxide dismutases (SODs). Broilers were sacrificed without stunning, or after ES with 65V, 86mA, 1000Hz (E65V) or 150V, 130mA, 60Hz (E150V). Serum cortisol and uric acid, muscular malondialdehyde and mRNA levels of MAPKs, Nrf2, GSTA3, GSTT1 and SOD2 were increased, whereas, serum free triiodothyronine, free thyroxine, muscular GST1d activity were decreased in E65V compared with E150V. Overall, the serum uric acid and transcription of the MAPK/Nrf2/ARE (antioxidant responsive element) signaling pathway were elevated, but didn't overcome the oxidative stress stimulated by low-current & high-frequency ES, leading to aggravated lipid peroxidation at 1d and 9d postmortem in breast muscle compared with high-current & low-frequency ES.

Polymorphisms in intron 1 of the porcine POU1F1 gene.

This study was conducted to detect polymorphisms in intron 1 of porcine POU1F1 (POU domain, class 1, transcription factor 1, Pit1, renamed as POU1F1) by comparative sequencing. Within the intron, 23 sites of variation were identified, including 16 single-nucleotide substitutions, 4 single-nucleotide indels, 2 short (3-bp and 17-bp), and one long (313-bp) indels. Several important regulatory motifs were found within the 313-bp indel by in silico analysis. The 313-bp indel was next genotyped in 11 Chinese native pig breeds and 4 western meat-type pig breeds. The appearance of genotypes varied between breeds: among Chinese native breeds, no AA and AB genotypes were found in Tibetan, Lingao, Min, Rongchang, and Songliao Black pigs, no AA genotype was found in Fenjing and Leping Spotted pigs, whereas in Pietrain and Landrace there were no BB genotypes, and all 19 Duroc pigs were AA homozygotes. The western meat-type pigs had high A allele frequencies and the Chinese pigs had more B alleles, except Jianquhai pigs. A positive association of the AA genotype with birth weight was observed in a commercial pig line. This paper demonstrated that the genetic variation in intron 1 of the pig POU1F1 gene was high and these polymorphisms may provide useful makers for QTL analysis.

The simultaneous separation and determination of six flavonoids and troxerutin in rat urine and chicken plasma by reversed-phase high-performance liquid chromatography with ultraviolet-visible detection.

The method of high-performance liquid chromatography (HPLC) with UV-vis detection was used and validated for the simultaneous determination of six flavonoids (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and troxerutin in rat urine and chicken plasma. Chromatographic separation was performed using a VP-ODS column (150 mm x 4.6 mm, 5.0 microm) maintained at 35.0 degrees C. The mobile phase was a mixture of water, methanol and acetic acid (57:43:1, v/v/v, pH 3.0) at the flow rate of 0.8 mL/min. Six flavonoids and troxerutin were analyzed simultaneously with good separation. On optimum conditions, calibration curves were found to be linear with the ranges of 0.10-70.00 microg/mL (puerarin, rutin, morin, luteolin, quercetin, kaempferol) and 0.50-350.00 microg/mL (troxerutin). The detection limits were 0.010-0.050 microg/mL. The method was validated for accuracy and precision, and it was successfully applied to determine drug concentrations in rat urine and chicken plasma samples from rat and chicken that had been orally administered with six flavonoids and troxerutin.

Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo.

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the beta-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.