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This study is aimed at exploring the release of bound polyphenols (BP) from insoluble dietary fiber (IDF) and its mechanism by solid-state fermentation (SSF) via Trichoderma viride. The results indicated that BP released by SSF (5.55 mg GAE/g DW) was significantly higher than by alkaline hydrolysis. In addition, 39 polyphenols and catabolites were detected, and the related biotransformation pathways were speculated. Quantitative analysis showed that SSF released more ferulic acid, p-coumaric acid, and organic acids, which led to advances in antioxidant, α-amylase, and α-glucosidase inhibitory activities. Furthermore, structural characteristics (scanning electron microscopy, X-ray diffraction, thermos gravimetric analysis, and Fourier transform infrared spectroscopy) and dynamic changes of carbohydrate-hydrolyzing enzymes indicated that the destruction of hemicellulose and the secretion of xylanase were vital for releasing BP. Overall, this study demonstrated that SSF was beneficial to release BP from IDF, which could provide insight into utilizing agricultural byproducts in a more natural and economical way.

Epigenetics alternation of non-genetic variation and genome-wide association study proven allelic variants may associate with insulin secretion in type 2 diabetes (T2D) development. We analyzed promoter DNA methylation array to evaluate the associated with increased susceptibility to T2D (30 cases, 10 controls) and found 1,091 gene hypermethylated in promoter regions. We performed the association study of T2D and found 698 single nucleotide polymorphisms in exon and promoter sites by using 2,270 subjects (560 cases, 1,710 controls). A comparison of DNA hypermethylation and gene silencing of mouse T2D results in our T2D patients' results showed that the 5'-nucleotidase, cytosolic II (NT5C2) and fucosyltransferase 8 (FUT8) genes were strongly associated with increased susceptibility to T2D. DNA hypermethylation in promoter regions reduced NT5C2 gene expression, but not FUT8 in T2D patients. NT5C2 protein expression was decreased in pancreatic β-cells from T2D mice. Transient transfection NT5C2 into RIN-m5F cells down-regulated DNA methyltransferase I (DNMT1) expression and up-regulation of the insulin receptor. Moreover, NT5C2 knockdown induced in DNMT1 overexpression and insulin receptor inhibition. Taken together, these results showed that NT5C2 epigenetically regulated insulin receptor in patients and mice with T2D, and maybe provide for T2D therapy strategy.

Asthma is one of the most common childhood chronic diseases worldwide. Subcutaneous immunotherapy (SCIT) is commonly used in the treatment of house dust mite (HDM)‑related asthma in children. However, the therapeutic mechanism of SCIT in asthma remains unclear. The present study aimed to investigate the molecular biomarkers associated with HDM‑related asthma in asthmatic children prior and subsequent to SCIT treatment compared with those in healthy children via proteomic analysis. The study included a control group (30 healthy children), ‑Treatment group (30 children with HDM‑related allergic asthma) and +Treatment group (30 children with HDM‑related allergic asthma treated with SCIT). An isobaric labeling with relative and absolute quantification‑based method was used to analyze serum proteome changes to detect differentially expressed proteins, while functional enrichment and protein‑protein interaction network analysis were used to select candidate biomarkers. A total of 72 differentially expressed proteins were detected in the ‑Treatment, +Treatment and control groups. A total of 33 and 57 differentially expressed proteins were observed in the ‑Treatment vs. control and +Treatment vs. control groups, respectively. Through bioinformatics analysis, 5 candidate proteins [keratin 1 (KRT1), apolipoprotein B (APOB), fibronectin 1, antithrombin III (SERPINC1) and α‑1‑antitrypsin (SERPINA1)] were selected for validation by western blotting; among them, 4 proteins (KRT1, APOB, SERPINC1 and SERPINA1) showed robust reproducibility in asthma and control samples. This study illustrated the changes in proteome regulation following SCIT treatment for asthma. The 4 identified proteins may serve as potential biomarkers prior and subsequent to SCIT treatment, and help elucidate the molecular regulation mechanisms of SCIT to treat HDM‑related asthma.

Epigenetics alternation of non-genetic variation and genome-wide association study proven allelic variants may associate with insulin secretion in type 2 diabetes (T2D) development. We analyzed promoter DNA methylation array to evaluate the associated with increased susceptibility to T2D (30 cases, 10 controls) and found 1,091 gene hypermethylated in promoter regions. We performed the association study of T2D and found 698 single nucleotide polymorphisms in exon and promoter sites by using 2,270 subjects (560 cases, 1,710 controls). A comparison of DNA hypermethylation and gene silencing of mouse T2D results in our T2D patients’ results showed that the 5′-nucleotidase, cytosolic II (NT5C2) and fucosyltransferase 8 (FUT8) genes were strongly associated with increased susceptibility to T2D. DNA hypermethylation in promoter regions reduced NT5C2 gene expression, but not FUT8 in T2D patients. NT5C2 protein expression was decreased in pancreatic β-cells from T2D mice. Transient transfection NT5C2 into RIN-m5F cells down-regulated DNA methyltransferase I (DNMT1) expression and up-regulation of the insulin receptor. Moreover, NT5C2 knockdown induced in DNMT1 overexpression and insulin receptor inhibition. Taken together, these results showed that NT5C2 epigenetically regulated insulin receptor in patients and mice with T2D, and maybe provide for T2D therapy strategy.

Food protein-derived bioactive peptides, particularly antihypertensive peptides, are important constituents of functional foods or nutraceuticals. Most antihypertensive are identified as the inhibitors of angiotensin converting enzyme (ACE), a key enzyme responsible for the generation of angiotensin II (Ang II), which is a vasoconstricting peptide. Hence, ACE has long been used as a universal target to identify antihypertensive peptides. Angiotensin converting enzyme 2 (ACE2), is a homolog of ACE but uses Ang II as its key substrate to produce angiotensin (1-7), exerting vasodilatory activity via the mas receptor (MasR). Therefore, ACE2 functions in the opposite way as ACE and is an emerging novel target for cardiovascular therapy. The potential of food protein-derived bioactive peptides in targeting ACE2 has been rarely explored. While, recently we found that IRW, an egg white ovotransferrin-derived antihypertensive peptide, reduced blood pressure in spontaneously hypertensive rats via the ACE2/Ang (1-7)/MasR axis, indicating a new mechanism of food protein-derived bioactive peptides in reducing blood pressure. The objectives of this review are to summarize the functions of the ACE2/Ang (1-7)/MasR axis and to examine its potential roles in the actions of food protein-derived antihypertensive peptides. The interaction between antihypertensive peptides and the ACE2/Ang (1-7)/MasR axis will also be discussed.

Background Taraxacum species (commonly known as dandelion) used as herbal medicine have been reported to exhibit an antiproliferative effect on hepatoma cells and antitumor activity in non-small-cell lung cancer cells. Although several investigations have demonstrated the safety of Taraxacum officinale, the safety of tissue-cultured plants of T. formosanum has not been assessed so far. Therefore, the present study examines the safety of the water extract of the entire plant of tissue cultured T. formosanum based on acute and subacute toxicity tests in rats, as well as the Ames tests. Results No death or toxicity symptoms were observed in the acute and subacute tests. The results of the acute test revealed that the LD50 (50% of lethal dose) value of the T. formosanum water extract for rats exceeded 5 g/kg bw. No abnormal changes in the body weight, weekly food consumption, organ weight, or hematological, biochemical, and morphological parameters were observed in the subacute toxicity test. Thus, the no observed adverse effect level (NOAEL) of T. formosanum water extract was estimated to be higher than 2.0 g/kg. Finally, the results of the Ames test revealed that T. formosanum water extract was not genotoxic at any tested concentration to any of five Salmonella strains. Conclusions The water extract of tissue-cultured T. formosanum was non-toxic to rats in acute and subacute tests and exhibited no genotoxicity to five Salmonella strains. How to cite: Tsai WC, Chang HC, Tseng YH, et al. Toxicity evaluation of water extract of tissue-cultured Taraxacum formosanum by acute, subacute administration, and Ames test. Electron J Biotechnol 2020;45. https://doi.org/10.1016/j.ejbt.2020.04.001

Food protein-derived bioactive peptides have been recognized as a promising candidate for the development of functional foods or nutraceuticals. Peptides exhibiting antioxidant, anti-proliferative and anti-hypertensive activities have been characterized from corn proteins while the knowledge on the presence of immunomodulatory peptides in corn proteins is limited. The objective of this study was to identify the immunomodulatory peptides from corn proteins using interleukin 6 (IL-6) in human macrophage-like U937 cells as the marker. Results indicated that zein protein hydrolysate digested by thermolysin showed a high yield of peptides with potent activity, which was then used for further fractionation by RP-HPLC. We identified five peptides using LC–MS/MS, among which, peptide FLPFNQL showed the highest activity reducing IL-6 production by 57.7% ± 6.1%.

Cisplatin-induced nephrotoxicity is associated with gut microbiota disturbance. The present study aimed to investigate whether supplementation of Lactobacillus reuteri and Clostridium butyricum (LCs) had a protective effect on cisplatin-induced nephrotoxicity through reconstruction of gut microbiota. Wistar rats were given different treatments: control, cisplatin (Cis), cisplatin + C. butyricum and L. reuteri (Cis+LCs), and C. butyricum and L. reuteri (LCs). We observed that cisplatin-treated rats supplemented with LCs exhibited significantly decreased renal inflammation (KIM-1, F4/80, and MPO), oxidative stress, fibrosis (collagen IV, fibronectin, and a-SMA), apoptosis, concentration of blood endotoxin and indoxyl sulfate, and increased fecal butyric acid production compared with those without supplementation. In addition, LCs improved the cisplatin-induced microbiome dysbiosis by maintaining a healthy gut microbiota structure and diversity, depleting Escherichia-Shigella and the Enterobacteriaceae family, and enriching probiotic Bifidobacterium, Ruminococcaceae, Ruminiclostridium_9, and Oscillibacter. Moreover, the LCs intervention alleviated the cisplatin-induced intestinal epithelial barrier impairment. This study indicated LCs probiotic serves as a mediator of the gut–kidney axis in cisplatin-induced nephrotoxicity to restore the intestinal microbiota composition, thereby suppressing uremic toxin production and enhancing butyrate production. Furthermore, the renoprotective effect of LCs is partially mediated by increasing the anti-inflammatory effects and maintaining the integrity of the intestinal barrier.

KIT is a tyrosine kinase receptor involved in carcinogenesis. Two alternatively spliced transcripts, differed from presence of four amino acids (GNSK) at exon 9 of c-kit, were identified in various human tumours and canine hemangiosarcoma (HSA). The biological function and clinical implications of these isoforms have not yet been elucidated in canine tumours. The current study aimed to validate the expression profile and ultimately to evaluate the correlation of c-kit isoform levels with clinicopathological factors of canine mammary tumours (CMTs). In total, the expression profiles of c-kit isoforms in 196 samples obtained from normal mammary glands (NMGs) of healthy controls and dogs with CMTs, benign and malignant CMTs, and HSAs were determined by polymerase chain reaction (PCR) and quantified via real-time PCR. Overall, the expression levels of the two isoforms were equivalent in NMGs, whereas the GNSK- /GNSK+ ratio sharply increased to 7.44- and 8.33-fold, indicating abundant GNSK- isoforms in benign and malignant CMTs, respectively. However, a significant decrease in GNSK- expression was detected in dogs with high-grade malignant CMTs (mCMTs) and with metastatic CMTs compared with expression in those with a lower grade and non-metastatic CMTs. In addition, the median survival time was shorter in mCMT canines with a lower GNSK- /GNSK+ ratio than that in mCMT canines with a higher ratio (899 days vs 1534 days). In conclusion, two c-kit isoforms are ubiquitously expressed with great variability in HSAs and CMTs with both benign and malignant status. The GNSK- /GNSK+ ratio could serve as a prognostic indicator for dogs with mCMTs.

A five-year-old female cockatiel weighing 117 g was presented with a fast-growing mass beside the uropygial gland. Excisional biopsy was performed and the mass measured [Formula: see text][Formula: see text]cm in size and weighed 30.6 g. On the surface of cut sections, the mass was yellow-brown with white or yellow colloidal substances and red exudate. Histopathology showed that the tumor mass was covered by the skin and located in the deep dermis and hypodermis. The tumor consisted of abundant vascular adipose tissue and lipoblasts with intracytoplasmic lipid droplets, which varied in size. Also, small, well-differentiated blood vessels, with varied degrees of congestion and dilation, were observed within the tumor. Histochemically, staining with Oil red O produced a positive reaction in which the lipid droplets presented a reddish color. Immunohistochemistry produced positive staining for Desmin and successfully marked the muscular layers of blood vessels. On the basis of these results, a rare case of liposarcoma with microvascular proliferation adjacent to the uropygial gland was diagnosed in a cockatiel.

Aims: The current study was conducted with the central objective of investigating the expression of microRNA-145 (miR-145) in renal vascular lesions (RVLs) in juvenile lupus nephritis (JLN) and its possible mechanism. Methods: The clinical data of 49 JLN patients confirmed by renal biopsy were collected and followed by grouping according to the RVLs score after hematoxylin-eosin staining: mild, moderate, and severe groups. In situ hybridization was used to detect the expression of miR-145 in renal vessels which was then being compared among different RVLs groups. Up-LV-miR-145 and LV-miR-NC lentiviral vectors were constructed and transfected into human vascular smooth muscle cells (HVSMCs), respectively. After HVSMCs were treated with 10.0 µg/L platelet-derived growth factor (PDGF)-BB for 24 h, the proliferation, migration, and apoptosis of endothelial cells were detected by MTT, Transwell assay, and flow cytometry, respectively. Western blot was used to detect expression of alpha-smooth muscle actin (α-SM-actin) and osteopontin (OPN). Results: The expression of miR-145 in renal vascular cells was statistically significant. The higher the inner membrane ratio, the lesser the miR-145 expression. After treatment with PDGF-BB, expression of miR-145 in HVSMCs decreased, proliferation and migration ability enhanced, apoptosis decreased, α-SM-actin decreased, and OPN increased. The proliferation and migration ability of HVSMCs in the LV-miR-145 group suppressed, apoptosis enhanced, α-SM-actin increased, and OPN decreased. Conclusions: Our study revealed that miR-145 expression decreased with the increase of vascular damage. miR-145 can inhibit proliferation, migration, and differentiation phenotypic transformation of HVSMCs induced by PDGF-BB. miR-145 may be involved in the pathogenesis of RVLs and may be a new target for treatment of RVLs in lupus nephritis.

Background Sphingosine kinase 1 (SPHK1) is an enzyme that converts pro-apoptotic ceramide and sphingosine into anti-apoptotic sphingosine-1-phosphate. There is growing evidence that SPHK1 activation promotes oncogenic transformation, tumor growth, chemotherapy resistance, and metastatic spread. High SPHK1 expression has been associated with a poor prognosis in several human cancers. Results In the present study, the expression level of SPHK1 was examined in feline mammary tumor (FMT) specimens, and the IHC expression level of SPHK1 was associated with the histological grade of FMTs. IHC analysis of 88 FMT cases revealed that the expression level of SPHK1 was upregulated in 53 tumor tissues (60.2%) compared to adjacent mammary tissues. SPHK1 expression in FMTs was significantly associated with histological grade, presence of lymphovascular invasion, and estrogen receptor negativity. Treatment of primary FMT cells with SPHK1 inhibitors reduced cell viability, indicating that SPHK1 acts to promote FMT cell survival. These results indicate that SPHK1 may play an important role in FMTs and may be a therapeutic target in cats with FMT. Conclusions SPHK1 over-expression in breast cancer tissues is associated with a poor prognosis in humans. SPHK1 over-expression in more aggressive FMTs provides support for a potential role of SPHK1 inhibitors for the treatment of FMTs. Targeting SPHK1 has potent cytotoxic effects in primary FMT cells. These findings suggest that further examination of the role SPHK1 plays in FMTs will pave the way for the investigation of SPHK1 inhibitors in future clinical applications. Electronic supplementary material The online version of this article (10.1186/s12917-019-1883-z) contains supplementary material, which is available to authorized users.

Therapeutic administration of glucocorticoids (GCs) is frequently used as add-on chemotherapy for palliative purposes during breast cancer treatment. Recent studies have shown that GC treatment induces microRNA-708 in ovarian cancer cells, resulting in impaired tumor cell proliferation and metastasis. However, the regulatory functions of GCs on miR-708 and its downstream target genes in human breast cancer cells (BCCs) are poorly understood. In this study, we found that treatment with either the synthetic GC dexamethasone (DEX) or the natural GC mimic, antcin A (ATA) significantly increased miR-708 expression by transactivation of glucocorticoid receptor alpha (GRα) in MCF-7 and MDA-MB-231 human BCCs. Induction of miR-708 by GR agonists resulted in inhibition of cell proliferation, cell-cycle progression, cancer stem cell (CSC)-like phenotype and metastasis of BCCs. In addition, GR agonist treatment or miR-708 mimic transfection remarkably inhibited IKKβ expression and suppressed nuclear factor-kappaB (NF-κB) activity and its downstream target genes, including COX-2, cMYC, cyclin D1, Matrix metalloproteinase (MMP)-2, MMP-9, CD24, CD44 and increased p21CIP1 and p27KIP1 that are known to be involved in proliferation, cell-cycle progression, metastasis and CSC marker protein. BCCs xenograft models indicate that treatment with GR agonists significantly reduced tumor growth, weight and volume. Overall, our data strongly suggest that GR agonists induced miR-708 and downstream suppression of NF-κB signaling, which may be applicable as a novel therapeutic intervention in breast cancer treatment.

Acrylamide (ACR), which is formed during the Maillard reaction, is used in various industrial processes. ACR accumulation in humans and laboratory animals results in genotoxicity, carcinogenicity, neurotoxicity, and reproductive toxicity. In this study, we investigated the mechanisms by which ACR may induce vasorelaxation and neuromuscular toxicity. Vasorelaxation was studied using an isolated rat aortic ring model. The aortic rings were divided into the following groups: with or without endothelium, with nitric oxide synthase (NOS) inhibition, with acetylcholine receptor inhibition, and with extracellular calcium inhibition. Changes in tension were used to indicate vasorelaxation. Neuromuscular toxicity was assessed using a phrenic nerve–diaphragm model. Changes in muscle contraction stimulated by the phrenic nerve were used to indicate neuromuscular toxicity. ACR induced the vasorelaxation of phenylephrine-precontracted aortic rings, which could be significantly attenuated by NOS inhibitors. The results of the phrenic nerve–diaphragm experiments revealed that ACR reduced muscle stimulation and contraction through nicotinic acetylcholine receptor (AChR). ACR-induced vasotoxicity was regulated by NOS through the aortic endothelium. Nicotinic AChR regulated ACR-induced neuromuscular blockage.

Spent hens are a major byproduct of the egg industry but are rich in muscle proteins that can be enzymatically transformed into bioactive peptides. The present study aimed to develop a spent hen muscle protein hydrolysate (SPH) with antihypertensive activity. Spent hen muscle proteins were hydrolyzed by nine enzymes, either individually or in combination, 18 SPHs were assessed initially for their in vitro angiotensin-converting enzyme (ACE) inhibitory activity, and three SPHs, prepared by Protex 26L (SPH-26L), pepsin (SPH-P), and thermoase (SPH-T), showed promising activity and peptide yield. These three hydrolysates were further assessed for their angiotensin-converting enzyme 2 (ACE2) upregulating, antioxidant, and anti-inflammatory activities, only SPH-T upregulated ACE2 expression, while all three SPHs showed antioxidant and anti-inflammatory activities. During simulated gastrointestinal digestion, ACE2 upregulating, ACE inhibitory and antioxidant activities of SPH-T were not affected, but those of SPH-26L and SPH-P were reduced. ACE inhibitory activity of gastrointestinal-digested SPH-T was not affected after the permeability study in Caco-2 cells, while ACE2 upregulating, antioxidant and anti-inflammatory activities were improved, nine novel peptides with five&ndash, eight amino acid residues were identified from the Caco-2 permeate. Among these three hydrolysates, only SPH-T reduced blood pressure significantly when given orally at a daily dose of 1000 mg/kg body weight to spontaneously hypertensive rats. SPH-T can be developed into a promising functional food ingredient against hypertension, contributing to a more sustainable utilization for spent hens while generating extra revenue for the egg industry.

Russell’s vipers (RVs) envenoming is an important public health issue in South-East Asia. Disseminated intravascular coagulopathy, systemic bleeding, hemolysis, and acute renal injury are obvious problems that develop in most cases, and neuromuscular junction blocks are an additional problem caused by western RV snakebite. The complex presentations usually are an obstacle to early diagnosis and antivenom administration. Here, we tried to produce highly specific antibodies in goose yolks for use in a paper-based microfluidic diagnostic kit, immunochromatographic test of viper (ICT-Viper), to distinguish RVs from other vipers and even cobra snakebite in Asia. We used indirect ELISA to monitor specific goose IgY production and western blotting to illustrate the interaction of avian or mammal antibody with venom proteins. The ICT-Viper was tested not only in prepared samples but also in stored patient serum to demonstrate its preliminary efficacy. The results revealed that specific anti-Daboia russelii IgY could be raised in goose eggs effectively without inducing adverse effects. When it was collocated with horse anti-Daboia siamensis antibody, which broadly reacted with most of the venom proteins of both types of Russell’s viper, the false cross-reactivity was reduced, and the test showed good performance. The limit of detection was reduced to 10 ng/ml in vitro, and the test showed good detection ability in clinical snake envenoming case samples. The ICT-Viper performed well and could be combined with a cobra venom detection kit (ICT-Cobra) to create a multiple detection strip (ICT-VC), which broadens its applications while maintaining its detection ability for snake envenomation identification. Nonetheless, the use of the ICT-Viper in the South-East Asia region is pending additional laboratory and field investigations and regional collaboration. We believe that the development of this practical diagnostic tool marks the beginning of positive efforts to face the global snakebite issue., Author summary For a long time, many scientists have made efforts to develop more effective antivenoms or to expand the limits of existing antivenoms. Such endeavors are challenging but worth the effort in the long run. However, to use precious antivenom more effectively, a snake species discrimination tool might be helpful for treatment by ensuring accurate antivenom usage and dosage. In this study, we revealed that avian IgY might be an economical alternative for collocation with the equine F(ab’)2 fragment in the development of a diagnostic device with improved performance. Only a few avian eggs (about 10–15) would be necessary to produce 10,000 kits. Immunochromatographic test (ICT) is a simple, easy, fast, and low-cost point-of-care lateral flow assay device, and especially adapted to perform in resource-deficient areas. Such an antibody collocation strategy might be beneficial to overcome the urgent need for snakebite diagnosis as soon as possible. More cooperative investigations across Asia continent might be initiated in the future to face the challenge of snakebites together.

Cobra snakes (genus Naja) are some of the most dangerous snake species in Asia and Africa, as their bites cause severe life-threatening respiratory failure and local tissue destruction, especially in the case of late diagnosis. The differential diagnosis of snakebite envenomation still mainly relies upon symptomatology, the patient&rsquo, s description, and the experience of physicians. We have designed a rapid test, immunochromatographic test of cobra (ICT-Cobra), which obtained fair results in improving the diagnosis and treatment of Naja (N.) atra snakebites in Taiwan. In this study, we further investigated the feasibility of applying the kit for the detection of other cobra venoms based on the potential interspecies similarity. We firstly demonstrated the cross-reactivity between eight venoms of medically important cobra species and the rabbit anti-N. atra IgG that was used in ICT-Cobra by Western blotting and sandwich enzyme-linked immunosorbent assay. Then, ICT-Cobra was used to detect various concentrations of the eight venoms to elucidate its performance. Noticeable correlations between the cross-reactivity of venoms from genus Naja snakes and existing geographical characteristics were found. ICT-Cobra could detect venoms from other Asian cobras with variable detection limits comparable to those observed for N. atra, but the kit was less successful in the detection of venom from African cobras. The similar but slightly different venom components and the interaction between venom and rabbit anti-N. atra IgG led to variations in the detection limits. The transcontinental usage of ICT-Cobra might be possible due to the cross-reactivity of antibodies and similarities among the larger-sized proteins. This study showed that the close immunological relationships in the genus Naja could be used to develop a venom detection kit for the diagnosis of cobra envenomation in both Asian and African regions. Additional clinical studies and technical adjustments are still needed to improve the efficacy and broadening the application of ICT-Cobra in the future.

Genetically modified (GM) papaya plant resistant to infection by Papaya Ring Spot Virus (PRSV) was successfully generated by cloning the coat protein (CP) gene of the PRSV. However, the food safety of GM foods remains controversial. This study assesses the food safety of two GM papayas. 16-0-1 and 18-2-4, and compares them with that of non-GM papaya, Tainung-2 (TN-2), using genetic and animal toxicity assays. Pulps of fresh papaya fruits were lyophilized and grounded before use. Three genotoxicity assays were performed: the Ames test of Salmonella typhimurium tester strains (TA98, TA100, TA102, TA1535 and TA1537); the chromosomal aberration of Chinese banister ovary (CHO-K1) cells (in vitro); and micronucleus assays for mice (in vito). Experimental results demonstrated that both non-GM and GM papaya fruits had no genotoxicity. Acute oral toxicity and 28-day repeated feeding toxicity tests for these papayas were performed via the oral gavage method for rats. All GM papaya fruit had no acute toxicity ala maximal dose of 5 g/kg body weight. Furthermore, the results of the 28-day study at 1 g/kg body weight in rats revealed no adverse-related effect in the body weight, feed consumption, hematology, blood biochemical parameters, organ weights and pathology in both GMs and non-GM papaya groups. In conclusion, PRSV-resistant papaya fruit lines 16-0-1 and 18-2-4 are comparable to their parent non-GM (TN-2) counterparts and are equivalent in food safety.

In order to elucidate the possible genetic determinants of resistance to florfenicol and chloramphenicol in porcine Escherichia coli in Taiwan, 600 fecal samples of healthy pigs from 50 different farms were collected from 2003 to 2007. The florfenicol resistance in the isolated E. coli strains doubled from 39.2% in 2003 to 78.3% in 2007. A total of 351 florfenicol-resistant E. coli isolates were isolated from nursery pigs (61.5%), grower-finisher pigs (62.5%), and sows (51.5%). The prevalence of resistance genes, floR, cmlA, cat-1, cat-2 and cat-3, was 82.9, 61.3, 10.8, 3.7, and 0%, respectively. Of the 351 florfenicol-resistant isolates, 184 (52.4%) were positive for both floR and cmlA. Furthermore, the results of efflux inhibitor studies with Phe-Arg-β-naphthylamide showed a 4- to 64-fold decrease in the florfenicol MIC levels. The FloR efflux pump may play a role in phenicol resistance among porcine E. coli isolates in Taiwan. More detailed studies are required to focus on the public health concerns about the spread of antimicrobial resistance from animal food products to humans through the food chain.

Monascus purpureus is a fungus used in rice fermentation. Monascus-fermented rice is called red-mold rice (RMR) in Taiwan since the fermentation products contain a considerable amount of red pigments. The protective effects of RMR on oxidative stress in the testis and sperm were evaluated in this study. Zn deficiency was induced in rats by feeding them with a Zn-deficient diet for 12 weeks while the control rats were fed with a normal diet. The Zn-deficient rats were then divided into 6 groups, designated as Zn-deficient (ZD). Zn-compensated (ZC), IR (151 mg RMR/kg), 5R (755 mg RMR/kg), 1R+ZC (1RZ), and 5R+ZC (5RZ). The animals were administered with the mentioned diet (RMR/Zn) for 8 weeks. In the ZD rats, no spermatid cell formation was observed in the seminiferous tubular epithelia, and testis necrosis/atrophy was apparent. However, 5R and 5RZ administration improved testicular antioxidant enzyme activity, elevated serum testosterone levels, and increased sperm number in ZD rats; this treatment also exhibited inhibitory effects on caspase activities and reactive oxygen species (ROS) levels in the ZD rats, suggesting that RMR attenuated ZD-induced oxidative stress and testis apoptosis. The preventive activity against ZD-induced reproductive damage was more apparent in the 1RZ and 5RZ groups than in the ZC group, suggesting that RMR can serve as a supplement in adjuvant therapy for diseases associated with Zn deficiency.

This study investigated the effect of seaweed supplementation (Ulva lactuca (UL) or Sargassum hemiphyllum var. chinense (SHC)) on the distribution and metabolites of As in broiler breasts. Broilers fed 5% UL or 5% SHC ingested 1.4- or 78- fold greater total As than birds fed the control diet. The majority of As species were arsenate in the SHC feed and dimethylarsinic acid in breasts from chicks fed the SHC-containing diet. Arsenate and arsenobetaine were the dominant metabolites in the UL-containing feed, and arsenobetaine was the major metabolite in breasts from chicks fed the UL-containing diet. Feeding SHC enhanced hepatic S-adenosyl-methionine and arsenic methyltransferase, whereas feeding UL elevated renal arsenic methyltransferase. Taken together, considerable variation in the profiles of As species and As metabolites existed in broilers fed seaweed. The use of SHC-containing feeds in poultry production should be approached cautiously because of the potential accumulation of inorganic As species in chicken breasts.

High tibial osteotomy (HTO) is a recognized treatment for early-stage medial compartment knee osteoarthritis. Preoperative planning with standing whole-leg radiographs (WLRs) is essential for ensuring optimal postoperative alignment. The primary purpose of this study is to investigate the theoretical accuracy of the wedge opening required for two different preoperative planning parameters in open-wedge HTO. The second purpose is to theoretically determine which parameter is superior. Preoperative planning for HTO was performed with standing WLRs for 39 knees with isolated medial osteoarthritis. The Miniaci preoperative planning method was applied to correct the hip-knee-ankle (HKA) angle to 3to 6 degrees of valgus and the weight-bearing line (WBL) percentage within 60 to 70% of the width of the tibial plateau. To ensure that the HKA angle was between 3 and 6 degrees of valgus, the required accuracy window for the Miniaci angle was 3.25 ± 0.03 degrees (range, 3.20–3.30°). To ensure that the WBL percentage was between 60 and 70%, the accuracy window required for the Miniaci angle was 2.35 ± 0.13 degrees (range, 2.10–2.65°). This study suggests that to correct the HKA angle and the WBL percentage within the target range on two-dimensional WLRs, the Miniaci angle must be controlled to an accuracy of ± 1.63 and ± 1.18 degrees, respectively. Theoretically, the HKA angle is highly suitable as a preoperative planning parameter for HTO with a large permissible error and a small variability in the degree of change in the Miniaci angle (ΔMiniaci).

Dysregulation of hepatocyte apoptosis or failure is associated with several types of chronic liver diseases. Transforming growth factor β1 (TGF- β1) is a well-known pro-apoptotic factor in the liver, which is constituted by a receptor complex composed of TGFβ receptor type I (TβRI) and TβRII, along with transcriptional factors Smad proteins. As a member of the forkhead box O-class transcription factors, Foxo1 is a predominant regulator for hepatic glucose production. While the role of Foxo1 in functions beyond glucose metabolism is gradually recognized, since there has been increasing evidence showing the potential relationship between TGF-β1 signaling and Foxo1. Here, we investigated whether Foxo1 mediates the pro-apoptotic effect of TGF-β1 in primary hepatocytes. By using hepatocytes isolated from both of wild type (Foxo1fl/fl) and liver specific Foxo1 knockout (LFKO, albumin-Cre+/-:: Foxo1fl/fl) mice, it was found that TGF-β1 induces hepatocytes apoptosis in a Foxo1-dependent manner. We also explored the associated molecular mechanisms and demonstrated that TGF-β1 activates protein kinase A (PKA) through TβRI-Smad3, followed by phosphorylation of Foxo1 at Ser273 to promote apoptosis of hepatocytes. The significant role of pFoxo1-S273 in mediating the pro-apoptotic effect of TGF-β1 was validated by using hepatocytes isolated from Foxo1-S273A mice, which mimics the dephosphorylation of Foxo1-Ser273. The establishment of TGF-β1-PKA-Foxo1axis in this study reveals a novel role of Foxo1 in hepatic functions. Disclosure W. Ai: None. W. Liao: None. Q. Pan: None. W. Yang: None. Z. Shen: None. S. Guo: None. Y. Chen: None. Funding American Diabetes Association (1-15-CD-09 to S.G., Sr.); National Institutes of Health (R01DK095118, R56DK11833401)

Neuroblastoma is a pediatric malignancy, and the clinical phenotypes range from localized tumors with excellent outcomes to widely metastatic disease in which long-term survival is approximately 40%, despite intensive therapy. Emerging evidence suggests that aberrant miRNA regulation plays a role in neuroblastoma, but the miRNA functions and mechanisms remain unknown. miR-181 family members were detected in 32 neuroblastoma patients, and the effects of miR-181a/b on cell viability, invasion, and migration were evaluated in vitro and in vivo. A parallel global mRNA expression profile was obtained for neuroblastoma cells overexpressing miR-181a. The potential targets of miR-181a/b were validated. miR-181a/b expression levels were positively associated with MYCN amplification and neuroblastoma aggressiveness. Moreover, ectopic miR-181a/b expression significantly induced the growth and invasion of neuroblastoma cells in vitro and in vivo. Microarray analysis revealed that mRNAs were consistently downregulated after miR-181a overexpression, leading to cell migration. In addition, the expression of ABI1 was suppressed by miR-181a/b, and ABI1 was validated as a direct target of miR-181a/b. We concluded that miR-181a/b were significantly upregulated in aggressive neuroblastoma, which enhanced its tumorigenesis and progression by suppressing the expression of ABI1.

Baicalin is the main flavonoid from the roots of an important medicinal plant, Scutellaria baicalensis, which shows a variety biological activities. Psoriasis is a chronic immune-mediated inflammatory disease that affects the skin. The unmet need of psoriasis is that many patients do not respond adequately to available clinical treatment. In this study, we found that baicalin showed inhibited dermal inflammation in a murine model of psoriasis via topical application of imiquimod. After a 5-day topical imiquimod application, baicalin or the control vehicle cream was to applied to the lesions of BALB/c mice for a further 4 days. The erythema, scaling, and thickness of the epidermal layer significantly improved in the baicalin-treated mice. The levels of interleukin-17A, interleukin-22, interleukin-23, and tumor necrosis factor in the skin significantly decreased after baicalin treatment. Baicalin also inhibited imiquimod-induced interleukin-17A production in skin draining lymph node cells. The infiltration of γδ T cells into the skin lesions induced by imiquimod was also suppressed after baicalin treatment. These results suggest that baicalin inhibited skin inflammation through the inhibition of the interleukin-17/interleukin-23 axis in a murine model of psoriasis.

Angiotensin I converting enzyme (ACE) plays an important role in regulation of blood pressure as it converts angiotensin I into angiotensin II (a potent vasoconstrictor). Food protein-derived ACE inhibitory peptides have been considered as a safer alternative to antihypertensive drugs. In our previous study, three ACE inhibitory peptides were characterized from egg white ovotransferrin and their antihypertensive activity has been validated in spontaneously hypertensive rats. However, it is too costly to prepare these peptides from purified egg white ovotransferrin. The aims of the study were to determine the feasibility of preparing these peptides using egg white and then to optimize the conditions of preparing egg white hydrolysate. Taguchi's method was used to design experiments for optimization, which was established as follows: substrate %, pH of thermoase, time of thermoase digestion, ratio of pepsin to substrate, pH of pepsin, temperature of pepsin, and time of pepsin digestion were 7.5%, pH 8, 90 min, 1%, pH 2.5, 55 °C, and 180 min, respectively. The ACE inhibitory activity (IC50 value) and peptide yield obtained under optimal condition were 30 ± 2 μg/mL and 77.5% ± 0.3%, respectively, which were comparable to the predicted values. Hydrolysate prepared at 150 L reactor showed comparable activity but low peptide yield. Results of this study demonstrated the feasibility of using egg white protein as the starting material to prepare a functional ingredient with potent ACE inhibitory activity.

This study aimed to investigate whether HMGB1 (high mobility group box-1 protein) and receptor for advanced glycation end products (RAGE) were involved in the irradiation-induced endothelial barrier damage and their mechanism. We constructed the damage model of endothelium barrier model with bEnd.3 cells. The permeability of endothelial barrier was detected by sodium fluorescein (Na-F) permeation test, and the irradiation dose which could induce permeability transition was determined by being exposed to different irradiation doses (5, 10, 15, 20 Gy). MTT assay was applied to detect cell viability under different concentrations of HMGB1, glycyrrhizic acid (GA, a specific inhibitor of HMGB1), and FPS-ZM1 (a blood-brain-barrier permeant blocker of RAGE V domain-mediated ligand binding). The expression of HMGB1, RAGE, and related molecules involved in MAPK signaling pathway, MMP-2, MMP-9, ZO-1, and claudin 5 of differently treated groups were measured by qRT-PCR, western blot, and immunofluorescence. Cells possessed stable endothelial barrier function on 4-7 days after seeded on transwell plates. The permeability of endothelial barrier would change under at least 10 Gy radiation. Both radiation and HMGB1 treatment alone could improve the permeability. After irradiation, the expressions of HMGB1 and RAGE increased and MAPK signal pathway was activated. Meanwhile, MMP-2 and MMP-9 were overexpressed, while the expression of tight junction proteins ZO-1 and claudin 5 was decreased. Radiation could activate MAPK signaling pathway through promoting the expression of HMGB1 and RAGE, which further led to endothelial barrier injury and changed its permeability.

This is a rare case of a wild squirrel that was diagnosed with angiostrongyliasis characterized by the presence of larvae-induced eosinophilic meningoencephalomyelitis. A wild, intact, male Pallas’s squirrel (Callosciurus erythraeus) of unknown age was presented with a history of progressive paralysis in both forelimbs and hindlimbs. Gradually, the squirrel showed signs of deterioration, found dead and submitted for a complete pathological diagnosis. Necropsy revealed the presence of severe and diffuse congestion in the subarachnoid space of the brain and mild congestion in the spinal cord. Microscopically, severe meningoencephalomyelitis and interstitial pneumonia were consistent with lesions induced by Angiostrongylus cantonensis (A. cantonensis). Although A. cantonensis has been widely studied in rabbits, mice and humans, infection in squirrels has not been documented. This may be the first reported case of angiostrongyliasis in the Pallas’s squirrel.

In this study, canine transmissible venereal tumors (CTVTs) were evaluated for tumor-infiltrating lymphocytes, MHC class II antigen expression, melanoma antigen A (MAGE-A protein), and P-glycoprotein (P-gp). Vincristine (VCR)-sensitive CTVTs (VSCTVTs) were evaluated before and after VCR treatment. Immunohistochemical analysis was performed to evaluate the numbers of CD3[Formula: see text] lymphocytes, CD79a[Formula: see text] lymphocytes, and MHC II[Formula: see text] CTVT cells. Expression levels of P-gp, MAGE-A, and MHC class II antigen were evaluated. The numbers of CD3[Formula: see text] lymphocytes, CD79a[Formula: see text] lymphocytes, and MHC II[Formula: see text] CTVT cells were significantly elevated ([Formula: see text]). After VCR treatment, 100% (8/8) of VSCTVTs exhibited significantly higher MAGE-A expression in CTVT cells, compared with 74% (20/27) of VSCTVTs before VCR treatment. Strong expression of P-gp was found in 14.2% (1/7) of VSCTVTs after VCR treatment compared with VSCTVTs before treatment (83.3%, 5/6). These results demonstrated that regression of VSCTVTs following VCR treatment was associated with an obvious increase in the proportion of MHC class II[Formula: see text] CTVT cells and a significant increase in infiltration by T (CD3[Formula: see text] and B (CD79a[Formula: see text] lymphocytes, as well as decreased expression of P-gp and increased expression of MAGE-A in CTVT cells.

A high-performance gel permeation chromatography fluorescence detection (HPGPC-FD) method combined with fluorescein isothiocyanate (FITC) labeling was established for the microanalysis of L. barbarum polysaccharides (LBP). The calibration curves linear over the range of 0.2–20 µg/mL in rat plasma, and 0.25–500 μg/mL in urine and feces samples with correlation coefficients greater than 0.99. The inter-day and intra-day precisions (RSD, %) of the method were under 15% with the relative recovery ranging from 84.6% to 104.0% and the RSD ranging from 0.47% to 7.28%. The concentration–time curve of LBP-FITC in plasma following intragastric administration at 100, 50 and 25 mg/kg well fitted to a nonlinear model. LBP-FITC slowly eliminated from plasma according to the long half-lives (t1/2 = 31.39, 38.09, and 45.76 h, respectively) and mean retention times (MRT0–t = 18.38, 19.15 and 20.07 h, respectively, AUC0–∞ = 230.49, 236.18 and 242.57 h, respectively) after administration of LBP-FITC at doses of 100, 50, and 25 mg/kg, respectively. After intragastric administration at 50 mg/kg for 72 h, the concentration of LBP-FITC in urine and feces was 0.09 ± 0.04% and 92.18 ± 3.61% respectively, the excretion rate of urine was the highest in 0–4 h period and decreased continuously in 4–24 h period. The excretion rate of feces was the highest in 4–10 h, 48.28 ± 9.349% in feces within 4–10 h, and decreased rapidly in 10–24 h. The present study showed that LBP was absorbed as its prototype and most proportion of LBP was excreted from feces, indicating a long time remaining in intestine.

Background Angiotensin converting enzyme (ACE) is a key enzyme in the renin angiotensin system (RAS) responsible for conversion of angiotensin (Ang) I into Ang II, a vasoconstrictor leading to elevated blood pressure. ACE inhibitory (ACEi) peptides derived from food proteins have shown potential in the prevention and management of hypertension. Although most ACEi peptides were characterized based on in vitro ACEi activity, a relationship between ACE inhibition and physiological antihypertensive effect is not apparent, indicating the involvement of other mechanisms of action. Scope and approach This paper focuses on emerging antihypertensive mechanisms of ACEi peptides. As an alternate arm of the classic RAS, ACE2 cleaves Ang II into Ang (1–7) and thus counterbalances the harmful effects of Ang II. Endothelial dysfunction is now recognized as an early feature in the pathophysiology of metabolic syndrome and cardiovascular disorders including hypertension; endothelial dysfunction, vascular oxidative stress and inflammation are interplayed. Future perspectives on mechanistic study of ACEi peptides are forecasted. Key findings and conclusions Apart from classic ACE inhibition, emerging evidence suggests that food peptides can exert antihypertensive activity through upregulation of ACE2 (an ACE homologue that counterbalances the detrimental effect of elevated ACE), improvement of endothelial function, as well as reduced vascular oxidation and inflammation. Future research is expected to look into the effects of bioaccessibility, bioavailability, stability and reactivity of the peptides with food and gut matrices, as well as the gut microbiota, on blood pressure reduction.

Curcumin, a polyphenol compound, possesses potent pharmacological properties in preventing cancers, which make it as a potential anti-cancer mediator. However, it is still unknown that whether Curcumin induced melanoma A375 cell was associated with oxidative stress. Here, we firstly found a fascinating result that Curcumin could reduce the proliferation and induced apoptosis of human melanoma A375 cells. Meanwhile, IC50 of Curcumin on A375 cells is 80μM at 48h. In addition, Curcumin caused oxidative stress through inducing further ROS burst, decreasing GSH, and wrecking mitochondria membrane potential (MMP), which were reversed by ROS inhibitor N-acetylcysteine (NAC). Moreover, MMP disruption led to the release of Cytochrome c from mitochondria and subsequently led to intracellular apoptosis. Furthermore, we found that ROS-dependent HIF-1α and its downstream proteins also play an important role on Curcumin induced apoptosis. In conclusion, our results shed new lights on the therapy of melanoma that Curcumin may be a promising candidate.

Cigarette smoke exposure activates several cellular mechanisms predisposing to atherosclerosis, including oxidative stress, dyslipidemia, and vascular inflammation. Antrodia camphorata, a renowned medicinal mushroom in Taiwan, has been investigated for its antioxidant, anti-inflammatory, and antiatherosclerotic properties in cigarette smoke extracts (CSE)-treated vascular smooth muscle cells (SMCs), and ApoE-deficient mice. Fermented culture broth of Antrodia camphorata (AC, 200-800 µg/mL) possesses effective antioxidant activity against CSE-induced ROS production. Treatment of SMCs (A7r5) with AC (30-120 µg/mL) remarkably ameliorated CSE-induced morphological aberrations and cell death. Suppressed ROS levels by AC corroborate with substantial inhibition of CSE-induced DNA damage in AC-treated A7r5 cells. We found CSE-induced apoptosis through increased Bax/Bcl-2 ratio, was substantially inhibited by AC in A7r5 cells. Notably, upregulated SOD and catalase expressions in AC-treated A7r5 cells perhaps contributed to eradicate the CSE-induced ROS generation, and prevents DNA damage and apoptosis. Besides, AC suppressed AP-1 activity by inhibiting the c-Fos/c-Jun expressions, and NF-κB activation through inhibition of I-κBα degradation against CSE-stimulation. This anti-inflammatory property of AC was accompanied by suppressed CSE-induced VEGF, PDGF, and EGR-1 overexpressions in A7r5 cells. Furthermore, AC protects lung fibroblast (MRC-5) cells from CSE-induced cell death. In vivo data showed that AC oral administration (0.6 mg/d/8-wk) prevents CSE-accelerated atherosclerosis in ApoE-deficient mice. This antiatherosclerotic property was associated with increased serum total antioxidant status, and decreased total cholesterol and triacylglycerol levels. Thus, Antrodia camphorata may be useful for prevention of CSE-induced oxidative stress and diseases. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 2070-2084, 2017.

Previous publications widely reported that 25-hydroxyvitamin D-1-α-hydroxylase (CYP27B1) regulated the metabolism of 25-hydroxyvitamin D3, which has a close association between altered activity of vitamin D and vascular calcification has been reported in various human diseases, including chronic kidney disease, osteoporosis and atherosclerosis. Vascular calcification is a clinically significant component of atherosclerosis and may be promoted by ROS associated inflammatory. In this study, we evaluated the effect of 25-hydroxyvitamin D-1-α-hydroxylase on the atherosclerosis disease both in apolipoprotein (apo) E-/- mice and wild-type mice. We also isolated endothelial cell (ECs) and vascular smooth muscle cells (VSMCs) in aortic from the wild type mice and apoE-/- mice respectively, then investigated that after parathyroid hormone (PTH) both of the CYP27B1 and vitamin D receptor (VDR) expressions in apoE-/-EC and apoE-/-VSMC were higher than the wide-type EC and VSMCs. However, the increased proliferation and decreased apoptosis have showed in EC and VSMC compared with the cells from apo E-/- mice. Moreover, the index associated with vascular calcification such as intracellular Ca2+ concentration and alkaline phosphatase (ALP) activity have been tested and the result suggested that the levels of the former index have improved in the apoE-/-EC and apoE-/-VSMC. We got similar conclusions under the pre-treatment with 1, 25(OH) 2D3.

This study aims to explore effects of 1,25(OH)2 D3 and vitamin D receptor (VDR) on peripheral CD4+ /CD8+ double-positive (DP) T lymphocytes in systemic lupus erythematosus (SLE). MRL-LPr/LPr mice with SLE (n = 20) and normal MRL mice (n = 20) were assigned into the control group (normal mice, without feeding with 1,25(OH)2 D3 ), the 1,25(OH)2 D3 group (SLE mice, feeding with 1,25(OH)2 D3 ), the VDR-knock-in + 1,25(OH)2 D3 group (SLE mice, VDR-knock-in, feeding with 1,25(OH)2 D3 ) and the VDR-knockout group (normal mice, VDR-knockout, without feeding with 1,25(OH)2 D3 ) (n = 10 per group). Levels of T lymphocytes were measured by flow cytometry. The mRNA and proteins expressions of inflammatory factors were measured by qRT-PCR and ELISA. Extracellular signal-regulated kinase-1/2 (ERK1/2) expression was measured by Western blotting. Compared with normal mice, SLE mice showed reduced levels of CD4+ , CD4+ /CD8+ ratio, and DP lymphocytes. The levels of SLE-related indicators all increased significantly, followed with severe skin ulcers and urinary system infection. With the increase in time, skin ulcers and urinary system infection were significantly improved, levels of CD4+ , CD4+ /CD8+ ratio, and DP lymphocytes increased, and levels of SLE-related indicators all decreased in the 1,25(OH)2 D3 group. There were no significant changes in bioindicators in the control and the VDR-knock-in + 1,25(OH)2 D3 groups. The symptoms of SLE gradually occurred in the VDR-knockout group. This study demonstrates that VDR and 1,25(OH)2 D3 could elevate CD4+ /CD8+ DP T lymphocytes and reduce expressions of inflammatory factors, thus inhibiting the development and progression of SLE.

Ethnopharmacological relevance Antrodia salmonea (AS), is a well-known folk medicinal mushroom in Taiwan, has been reported to exhibit anti-oxidant, anti-angiogenic, and anti-inflammatory effects. Materials and methods In the present study, we examined the effects of AS on cell-cycle arrest in vitro in MDA-MB-231 cells and on tumor regression in vivo using an athymic nude mice model. Results AS (0–200 μg/mL) treatment significantly induced G2 cell-cycle arrest in MDA-MB-231 cells by reducing the levels of cyclin B1, cyclin A, cyclin E, and CDC2 proteins. In addition, N-acetylcysteine (NAC) pretreatment prevented AS induced G2 cell-cycle arrest, indicating that ROS accumulation and subsequent cell cycle arrest might be a major mechanism of AS-induced cytotoxicity. Further, AS treatment decreased COX-2 expression and induced PARP cleavage was significantly reversed by NAC pretreatment in MDA-MB-231 cells. The in vivo study results revealed that AS treatment was effective in terms of delaying the tumor incidence and reducing the tumor growth in MDA-MB-231-xenografted nude mice. TUNEL assay, immunohistochemical staining and Western blotting confirmed that AS significantly modulated the xenografted tumor progression as demonstrated by induction of apoptosis, autophagy, and cell-cycle arrest. Conclusion Our data strongly suggest that Antrodia salmonea could be an anti-cancer agent for human breast cancer.

Fungal immunomodulatory protein (FIP) is one of the bioactive compounds of edible mushrooms, which has been shown to trigger type 1 T helper (Th1) pathway activation in research with mice. This study was designated to assess immunomodulatory effects of recombinant FIP-Flammulina velutipes (rFIP-fve) on swine and the protective efficacy against PRRSV infection. In the in vitro evaluations, rFIP-fve significantly triggered up-regulation of IL-2 and IFN-γ mRNA in porcine PBMCs and stimulated natural killer cytotoxicity. Porcine pulmonary alveolar macrophages (PAMs) treated with rFIP-fve showed prolonged life times, up-regulation of both MHC I and II molecules and enhanced abilities to present antigen. In the in vivo trial, two doses of 2 mg rFIP-fve significantly reduced drops in the CD4/CD8 ratio after PRRSV challenge, and the cytokine mRNA profile of PBMC revealed a tendency of IFN-γ up-regulation and a decrease in IL-10 in the rFIP-treated group. Moreover, administration of rFIP-fve also decreased the PRRSV viremia with 1 log10 in titer (p = 0.07) and alleviated the severity of clinical signs after PRRSV challenge. Conclusively, these results illustrate the in vitro and in vivo immunological changes of rFIP-fve administered to pigs and reveal its potential to be used as an immunomodulatory therapeutic against PRRSV infection.

(1) Background: Graptopetalum paraguayense E. Walther is a traditional Chinese herbal medicine. In our previous study, 50% ethanolic G. paraguayense extracts (GE50) demonstrated good antioxidant activity. (2) Methods: To investigate the hepatoprotective effects of GE50 on ethanol and carbon tetrachloride (CCl4) co-induced hepatic damage in rats, Sprague&ndash, Dawley rats were randomly divided into five groups (Control group, GE50 group, 0.25 g/100 g BW, EC group: Ethanol + CCl4, 1.25 mL 50% ethanol and 0.1 mL 20% CCl4/100 g BW, EC + GE50 group: Ethanol + CCl4 + GE50, EC + silymarin group: ethanol + CCl4 + silymarin, 20 mg/100 g BW) for six consecutive weeks. (3) Results: Compared with the control group, EC group significantly elevated the serum aspartate aminotransferase (AST), alanine aminitransferase (ALT), and lactate dehydrogenase (LDH). However, GE50 or silymarin treatment effectively reversed these changes. GE50 had a significant protective effect against ethanol + CCl4 induced lipid peroxidation and increased the levels of glutathione (GSH), vitamin C, E, total antioxidant status (TAS), and the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), and glutathione S-transferases (GST). Furthermore, in EC focal group, slight fat droplet infiltration was observed in the livers, while in the GE50 or silymarin treatment groups, decreased fat droplet infiltration. HPLC phytochemical profile of GE50 revealed the presence of gallic acid, flavone, genistin, daidzin, and quercetin. (4) Conclusions: The hepatoprotective activity of GE50 is proposed to occur through the synergic effects of its chemical component, namely, gallic acid, flavone, genistin, daidzin, and quercetin. Hence, G. paraguayense can be used as a complementary and alternative therapy in the prevention of alcohol + CCl4-induced liver injury.

Egg proteins are recognized as excellent sources of bioactive peptides, such as angiotensin-converting enzyme inhibitory (ACEi) peptides. Oral administration of a thermolysin-digested egg white hydrolysate (T-EWH) caused a significant blood pressure reduction in spontaneously hypertensive rats; a further ACEi assay implied that its ACEi activity was enhanced after in vitro gastrointestinal (GI) digestion. These results indicated that T-EWH contained ACEi peptides resisting GI digestion and/or being further released during GI digestion. Therefore, the objective of this study was to identify these responsible ACEi peptides from T-EWH. The conventionally activity-guided fractionation was applied, coupled with a synchronized GI digestion throughout, during which both peptide yield and ACEi activity before and after the GI digestion were measured. Finally, six ACEi peptides (LAPYK, LKISQ, LKYAT, INKVVR, LFLIKH, and LGHWVY) with good GI resistance were identified with IC

Food protein-derived bioactive peptides have gained substantial interest during recent decades owing to their potential in modulating various physiological functions. As a high-quality, affordable and universally accepted protein source, egg is an ideal candidate for identification of numerous bioactive peptides. Extensive research has been carried out on identification and activity characterization of antihypertensive peptides, antidiabetic peptides, anticancer peptides, immunomodulatory peptides and antioxidant peptides. Mechanistic study of egg protein-derived bioactive peptides revealed the complexity of actions of these peptides as they may play multiple roles in regulating physiological functions, which warrants further investigation. The objectives of this book chapter are to summarize recent research progress on egg peptides and to identify the research gaps to translate the laboratory research into real-world applications of egg peptides as functional foods and nutraceuticals. Future research is expected to establish biologically relevant methods to replace chemical-based activity assay, to establish the efficacy and safety of egg peptides in human trials, to characterize peptide and cell cross-talks, to explore the multi-bioactivity of egg peptides, and to study the interactions between egg peptides and gut microbiota.

Forkhead/winged helix transcription factor O-class member 1 (FoxO1) is a key mediator of insulin and glucagon signaling in control of glucose homeostasis. Although epigallocatechin gallate (EGCG) has attracted interest owing to its potential to combat hyperglycemic diabetes, molecular mechanisms underlying its antihyperglycemic effect, in particular the effect on FoxO1, is poorly understand. This study aims to assess the impact of EGCG on the glucagon signaling pathway in regulating glucose metabolism. Primary hepatocytes from wild-type (WT), liver-specific FoxO1 knock out (FKO), and FoxO1-S273D knock-in (KI) mice were isolated, cultured, and treated with EGCG and/or glucagon. Our data showed the treatment of 10 μM EGCG for 6 h decreased hepatic glucose production by 20 and 23% in WT and FKO primary hepatocytes, respectively. EGCG repressed both gluconeogenesis and glycogenolysis in primary hepatocytes, coupled with activating AMPK. In addition, EGCG decreased mitochondrial oxygen consumption. We further investigated the effects of EGCG on glucagon-stimulated cAMP/PKA signaling pathway. EGCG reduced p-PKA-T197/T-PKA and p-CREB-S133/T-CREB levels by 39 and 20%, blocked p-FoxO1-S273, and suppressed nuclear FoxO1 translocation, suggesting that FoxO1 and CREB were possible downstream targets. A novel mechanism of EGCG in restraining hepatic glucose production (HGP) is through antagonizing glucagon signaling and suppressing FoxO1 via Ser273. EGCG may serve as a promising compound for regulating glucose homeostasis.

Hedychium coronarium has a long history of use worldwide as a food and in folk medicine. In this study, we aimed to investigate the effect of an aqueous extract of H. coronarium leaves (HC) on type 2 diabetes mellitus (T2DM). Two types of animal models were used in this study: Streptozotocin (STZ)-induced T2DM (Wistar rats, N = 8) and C57BKSdb/db mice (N = 5). After treatment with HC for 28 days, glucose tolerance improved in both of the diabetic animal models. As significant effects were shown after 14 days of treatment in the STZ-induced T2DM model, we carried out the experiments with it. After 28 days of treatment with HC, the levels of cholesterol, triglyceride, high-density lipoprotein, and low-density lipoprotein were significantly improved in the STZ-induced T2DM model. The lesions degree of islet &beta, cells was decreased after the HC treatment. Although the insulin level increased moderately, the aldosterone level was significantly decreased in the HC-treated groups, suggesting that aldosterone might play an important role in this effect. In summary, HC is a natural product and it is worth exploring its effect on T2DM.

Eight species of genus Velarifictorus Randell, 1964 have been recorded from China prior to this study. Here we describe one new species, Velarifictorus dianxiensis He sp. nov. from western Yunnan. COI genes of Velarifictorus spp. were used to reconstruct a phylogenetic tree for confirming the relationships of these species. The type specimens are deposited in Museum of Biology, East China Normal University (ECNU).

Saltwater fish is rich in health-promoting polyunsaturated fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Excessive fish intake may also cause health hazards due to methylmercury (MeHg). A novel dose estimation method was proposed in this study to improve the quality of food matching by considering the usage ratio of cooking and processing methods in sampling design. In total, 39 table-ready samples were specified according to 2–3 common cooking methods, and MeHg and total mercury (THg) were analyzed. The results showed that mercury was detected in 20 samples (LOD = 0.01 μg/g). High MeHg samples included fried-Spanish mackerel (1.23 μg/g), non-cooked shredded swordfish (0.325 μg/g), pan-fried Spanish mackerel (0.248 μg/g), and non-cooked shredded salmon (0.208 μg/g). Consumption data (n = 166,911) were obtained from 24-h recalls in the 2005–2008 Nutrition and Health Surveys in Taiwan (NAHSIT). The average whole-group estimated daily intake (EDI_WG) was 0.018 (7.9% tolerable daily intake, TDI) for women of child-bearing age and 0.031 μg/day per kg body weight (13.6% TDI) for children. However, 95th percentile values (P95) of 90.4% TDI and 185% TDI were estimated for the two consumer-only populations, which have raised public health concerns. The ratio of % TDI to % daily reference intake (% DRI) was used as a risk–benefit (R/B) indicator. The top 4 R/B foods were: tuna sashimi (0.089 and 0.14), non-cooked shredded swordfish (0.033 and 0.052), fried-Spanish mackerel (0.027 and 0.043) and deep-fried fish balls (0.022 and 0.035) in the two perspective populations.

Summary Bacterial infection triggers a cytokine storm that needs to be resolved to maintain the host’s wellbeing. Here, we report that ablation of m6A methyltransferase subunit METTL14 in myeloid cells exacerbates macrophage responses to acute bacterial infection in mice, leading to high mortality due to sustained production of pro-inflammatory cytokines. METTL14 depletion blunts Socs1 m6A methylation and reduces YTHDF1 binding to the m6A sites, which diminishes SOCS1 induction leading to the overactivation of TLR4/NF-κB signaling. Forced expression of SOCS1 in macrophages depleted of METTL14 or YTHDF1 rescues the hyper-responsive phenotype of these macrophages in vitro and in vivo. We further show that LPS treatment induces Socs1 m6A methylation and sustains SOCS1 induction by promoting Fto mRNA degradation, and forced FTO expression in macrophages mimics the phenotype of METTL14-depleted macrophages. We conclude that m6A methylation-mediated SOCS1 induction is required to maintain the negative feedback control of macrophage activation in response to bacterial infection.

The aim of this study was to investigate the effect of nonylphenol (NP), a widely used surfactant, on the reproductive performance of male Brown Tsaiya ducks (Anas platyrhynchos) (MBBTDs). Mature MBBTDs (n=100) were treated with NP by daily gavaging of 0, 1 (NP1), 10 (NP10) and 250 (NP250) mg/kg-BW/d for 14 wk. Semen quality, fertilization rate and specific factors in blood plasma were measured. Weights of organs were also measured at 14 wk after NP administration. Ducks from each treatment (n=4) were continually treated with NP thereafter for 12 mo to observe changes of tissue ultrastructure by microscopic examination. The results showed that ducks treated with amounts of NP of greater than 1mg NP/kg BW/d (NP1) for 14 wk had decreased sperm viability (32.3%) compared to those in the control group (74.1%, P0.05). The fertilization rate of ducks treated with 250mg NP/kg-BW/d (NP250) for 14 wk was reduced (21.0%) compared to the control group (74.5%, P0.05). Plasma aminotransferase (AST) and alanine aminotransferase (ALT) activities were also greater in NP250 group at the 14th wk post-treatment. Plasma testosterone concentrations were increased by NP1 treatment at the 14th wk post-treatment. Administration at dosage 250mg NP/kg-BW/d for 12 mo resulted in reduced sperm counts (P0.05) and histopathological changes, such as dilated seminiferous tubules (P0.05) and degenerated spermatocytes (P0.05). These findings strongly suggest that NP adversely affects the reproductive performance of MBBTDs.

Background The natural product tetramethylpyrazine (TMP) and resveratrol have a variety of biologic activities, including anti-cancer effects. However the pharmacological function of CSTMP (a newly designed and synthesized TMP and resveratrol derivative) in cancer have not been elucidated. Methods In RPMI8226 cells, the cytotoxic effects and apoptosis were detected by MTT and Double staining for Annexin V-FITC and propidium iodide (PI). The protein and mRNA expression levels were detected by Real Time PCR and Western blot, respectively. The localization of cleaved caspase-12 was evaluated by immunofluorescent staining. The activation of caspase were measured by colorimetric assays and Western blot. Results CSTMP showed significantly cytotoxic effects and induced apoptosis in RPMI8226 cells. Caspase activation, Cytochrome c release and Bax, Bcl-2 and Bcl-XL levels analyses demonstrated that the anti-cancer effect of CSTMP in RPMI8226 cells was mediated by promoting caspase- and mitochondria-dependent apoptosis. In addition, CSTMP induced the increased expression of endoplasmic reticulum (ER) stress related proteins (CHOP, GRP78, GRP94 and cleaved caspase-12) and the activation of multiple branches of ER stress transducers (PERK-eIF2α, IRE1α and ATF6). Moreover, knockdown of CHOP by siRNA markedly inhibited CSTMP-induced cytotoxic effects, caspases activity and mitochondrial dysfunction in RPMI8226 cells. Conclusions Our results indicated that CSTMP could induce apoptosis and mitochondrial dysfunction in RPMI8226 cells via CHOP-dependent ER stress.

A 2-year-7-month-old female intact African hedgehog was presented with a subcutaneous mass around the right side of the neck as well as an intra-abdominal mass found during palpation. Surgical excision and exploratory laparotomy were performed. A uterine mass was identified during laparotomy and ovariohysterectomy was performed. The botryoid mass measured [Formula: see text][Formula: see text]cm and was located in the right uterine horn. It had a meat-like texture and was yellow-white and dark red in color. The removed subcutaneous mass measured [Formula: see text][Formula: see text]cm. The mass was well-encapsulated and had a yellow-white homogeneous texture on the cut surface. Histologically, the myometrium was invaded by neoplastic cells and the tissue boundaries were not obvious. Neoplastic cells were arranged in a whirling or intersecting pattern, with strong angiogenesis present. Vacuolated nuclei were round and oval to cigar shaped, with one to multiple nucleoli present. Immunohistochemistry revealed a positive reaction for CD10, but a negative reaction for smooth muscle actin (SMA) and desmin in the uterine neoplastic cells. Histologically, well-differentiated adipocytes with sheets of undifferentiated polygonal neoplastic cells, which were characterized by vacuolated nuclei with prominent multiple nucleoli, were found in the subcutaneous mass. Neoplastic cells of the subcutaneous mass were positively stained with antibodies of MDM2 and estrogen receptor (ER), but failed to give a positive result for vimentin because the cross-species interaction was insufficient. The definitive diagnosis was endometrial stromal sarcoma and liposarcoma in an African hedgehog.

Proteinopathy in the heart which often manifests excessive misfolded/aggregated proteins in cardiac myocytes can result in severe fibrosis and heart failure. Here we developed a mouse model, which transgenically express tetrameric DsRed, a red fluorescent protein (RFP), in an attempt to mimic the pathological mechanisms ofcardiac fibrosis. Whilst DsRed is expressed and forms aggregation in most mouse organs, certain pathological defects are specifically recapitulated in cardiac muscle cells including mitochondria damages, aggresome-like residual bodies, excessive ubiquitinated proteins, and the induction of autophagy. The proteinopathy and cellular injuries caused by DsRed aggregates may be due to impaired or overburdened ubiquitin-proteasome system and autophagy-lysosome systems. We further identified that DsRed can be ubiquitinated and associated with MuRF1, a muscle-specific E3 ligase. Concomitantly, an activation of NF-κB signaling and a strong TIMP1 induction were noted, suggesting that RFP-induced fibrosis was augmented by a skewed balance between TIMP1 and MMPs. Taken together, our study highlights the molecular consequences of uncontrolled protein aggregation leading to congestive heart failure, and provides novel insights into fibrosis formation that can be exploited for improved therapy.

// Yng-Tay Chen 1 , Jiunn-Wang Liao 2 , Ya-Ching Tsai 1 and Fuu-Jen Tsai 1,3,4,5,6 1 Human Genetic Center, Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan 2 Graduate Institute of Veterinary Pathobiology, Research Center for Animal Medicine, Animal Disease Diagnostic Center, National Chung Hsing University, Taichung, Taiwan 3 Graduate Institute of China Medical Science, China Medical University, Taichung, Taiwan 4 Department of Medical Genetics, China Medical University Hospital, Taichung, Taiwan 5 School of Chinese Medicine, China Medical University, Taichung, Taiwan 6 Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan Correspondence to: Fuu-Jen Tsai, email: // Keywords : type 2 diabetes, DNA methylation, NR4A1, DNMT1, epigenetics, Gerotarget Received : March 01, 2016 Accepted : May 29, 2016 Published : June 14, 2016 Abstract Our previous genome-wide association studies showed that DNA methyltransferase 1 (DNMT1) is associated with increased susceptibility to type 2 diabetes (T2D) in Han Chinese individuals. Here, we aimed to further evaluate the role of DNMT1 in T2D. We performed a genome-wide DNA methylation array and found that the nuclear receptor subfamily 4 group A member 1 ( NR4A1 ) promoter was hypermethylated in patients with T2D and in a mouse model of T2D. Moreover, DNA hypermethylation of the NR4A1 promoter reduced NR4A1 mRNA expression. Transient transfection of human NR4A1 into RIN-m5F and 293T cells caused DNMT1 inhibition and induced insulin receptor activation. NR4A1knockdown by shRNA resulted in overexpression of DNMT1 and inhibition of insulin receptor, suggesting that the NR4A1 gene is involved in the epigenetics pathway. Furthermore, T2D model mice treated with the DNMT1 inhibitor aurintricarboxylic acid (ATA) showed reduced activation of DNMT1 in pancreatic β cells; this effect reversed the changes in NR4A1 expression and decreased blood glucose in T2D model mice. Thus, our results showed for the first time that DNMT1 caused NR4A1 DNA hypermethylation and blocked insulin signaling in patients with T2D. Importantly, ATA therapy may be useful for decreasing blood glucose levels by reversing NR4A1-dependent insulin signaling. These findings improve our understanding of the crucial roles of these regulatory elements in human T2D.