Epigallocatechin-3-gallate suppresses hemin-aggravated colon carcinogenesis through Nrf2-inhibited mitochondrial reactive oxygen species accumulation

BackgroundPrevious studies have presented evidence to support the significant association between red meat intake and colon cancer, suggesting that heme iron plays a key role in colon carcinogenesis. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, exhibits anti-oxidative and anti-cancer effects. However, the effect of EGCG on red meat-associated colon carcinogenesis is not well understood.ObjectivesWe aimed to investigate the regulatory effects of hemin and EGCG on colon carcinogenesis and the underlying mechanism of action.MethodsHemin and EGCG were treated in Caco2 cells to perform the water-soluble tetrazolium salt-1 assay, lactate dehydrogenase release assay, reactive oxygen species (ROS) detection assay, real-time quantitative polymerase chain reaction and western blot. We investigated the regulatory effects of hemin and EGCG on an azoxymethane (AOM) and dextran sodium sulfate (DSS)-induced colon carcinogenesis mouse model.ResultsIn Caco2 cells, hemin increased cell proliferation and the expression of cell cycle regulatory proteins, and ROS levels. EGCG suppressed hemin-induced cell proliferation and cell cycle regulatory protein expression as well as mitochondrial ROS accumulation. Hemin increased nuclear factor erythroid-2-related factor 2 (Nrf2) expression, but decreased Keap1 expression. EGCG enhanced hemin-induced Nrf2 and antioxidant gene expression. Nrf2 inhibitor reversed EGCG reduced cell proliferation and cell cycle regulatory protein expression. In AOM/DSS mice, hemin treatment induced hyperplastic changes in colon tissues, inhibited by EGCG supplementation. EGCG reduced the hemin-induced numbers of total aberrant crypts and malondialdehyde concentration in the AOM/DSS model.ConclusionsWe demonstrated that EGCG reduced hemin-induced proliferation and colon carcinogenesis through Nrf2-inhibited mitochondrial ROS accumulation.     

IL-17A regulates Eimeria tenella schizont maturation and migration in avian coccidiosis

Although IL17A is associated with the immunological control of various infectious diseases, its role in host response to Eimeria infections is not well understood. In an effort to better dissect the role of IL17A in host-pathogen interactions in avian coccidiosis, a neutralizing antibody (Ab) to chicken IL17A was used to counteract IL17A bioactivity in vivo. Chickens infected with Eimeria tenella and treated intravenously with IL17A Ab, exhibited reduced intracellular schizont and merozoite development, diminished lesion score, compared with untreated controls. Immunohistological evaluation of cecal lesions in the parasitized tissues indicated reduced migration and maturation of second-generation schizonts and reduced lesions in lamina propria and submucosa. In contrast, untreated and infected chickens had epithelial cells harboring second-generation schizonts, which extend into the submucosa through muscularis mucosa disruptions, maturing into second generation merozoites. Furthermore, IL17A Ab treatment was associated with increased parameters of Th1 immunity (IL2- and IFNγ- producing cells), reduced levels of reactive oxygen species (ROS), and diminished levels of serum matrix metalloproteinase-9 (MMP-9). Finally, schizonts from untreated and infected chickens expressed S100, Wiskott-Aldrich syndrome protein family member 3 (WASF3), and heat shock protein-70 (HSP70) proteins as merozoites matured, whereas the expression of these proteins was absent in IL17A Ab-treated chickens. These results provide the first evidence that the administration of an IL17A neutralizing Ab to E. tenella-infected chickens inhibits the migration of parasitized epithelial cells, markedly reduces the production of ROS and MMP-9, and decreases cecal lesions, suggesting that IL17A might be a potential therapeutic target for coccidiosis control.     

The efficacy and safety of external biphasic defibrillation in toy breed dogs

Objective: To evaluate the efficacy and safety of biphasic (BP) defibrillation in toy breed dogs (<5 kg of body weight).
Design: Prospective, clinical experimental study.
Setting: Veterinary teaching hospital.
Animals: Five dogs (pilot study) and 10 dogs (comparison study of biphasic versus monophasic defibrillation).
Measurements and main results: The efficacy of defibrillation was compared by estimating E80 (80% probability of successful defibrillation) after biphasic (BP) and monophasic (MP) defibrillations. The E80 for BP defibrillation was 7.24±1.33 J (2.24±0.41 J/kg) and 10.24±1.34 J (3.18±0.12 J/kg) for MP defibrillation. BP waveform required 30% less shock energy for a successful defibrillation. In order to compare the safety of defibrillation, we evaluated changes in cardiac biomarkers, electrocardiogram, echocardiographical left ventricular index, and aortic pressure during and after BP and MP defibrillation. All dogs treated by either BP or MP defibrillation survived. Pulseless electrical activity occurred in 2 of 5 dogs during MP defibrillation. The levels of cardiac biomarkers were elevated and sustained for longer periods in the MP defibrillation group. Electrocardiographic changes (e.g., QT prolongation, the time to return to an isoelectric ST segment after shocks) were more severe and longer in the MP defibrillation group. In addition, overall left ventricular cardiac performance was severely depressed in the MP group compared with the BP group.
Conclusion: Our findings suggest that BP defibrillation is more effective and safer than MP defibrillation. We determined the acceptable shock energy to be 2–4 J/kg for toy breed dogs.     

Percutaneous heartworm removal from dogs with severe heart worm (Dirofilaria immitis) infestation

Canine heart worm disease is often life-threatening due to its various complications, including right side heart failure, caval syndrome and pulmonary eosinophilic granulomatosis. Several preventive medications and melarsomine have been developed and they are very effective to control heartworm infestation. However, in a case of severe infestation, melarsomine therapy often results in an unfavorable outcome because of the severe immune reaction caused by rapid killing of the adult worm. Surgical removal and an interventional method using flexible alligator forceps have been well described in the literature. Despite the usefulness of mechanical removal using flexible alligator forceps, the methodology still needs to be upgraded for increasing the applicability for treating dogs with severe infestation. We describe herein a newly developed percutaneous removal method for heartworms and this was successfully applied to 4 dogs with severe heartworm infestation. The follow-up studies also showed favorable outcomes with no complications.     

Functional efficacy analysis of Angelica gigas Nakai on chicken myoblast cells through cell‐based in vitro assay

The value‐added products in livestock industry is one of the key issues in order to maximize the revenue and to create a new business model. Numerous studies have suggested application of herbal plants as feed additives to increase health, productivity, and/or high‐quality product in livestock. In this study, the first experiment was designed to develop in vitro evaluation system by using primary chicken myoblast (pCM) cells isolated from pectoralis major of 10‐day‐old male embryos. Subsequently, to evaluate effects of Korean Danggui Angelica gigas Nakai (AGN), we optimized the concentration of AGN root extract for treatment of primary pCM cells. After the treatment of AGN root extract, we compared proliferation and differentiation capacity, and also examined the gene expression. In the second experiment, the next generation sequencing analysis was performed to compare the different patterns of the global gene expression in pCM cells treated with AGN extract. Three up‐regulated (pancreas beta cells, fatty acid metabolism and glycolysis) and one down‐regulated (adipogenesis) gene sets were characterized suggesting that the AGN extract affected the metabolic pathways for the utilization of fat and glucose in chicken muscle cells. Furthermore, we validated the expression patterns of the up‐regulated genes (GCLC, PTPN6, ISL1, SLC25A13, TGFBI, and YWHAH) in the AGN‐treated pCM cells by quantitative RT‐PCR. These results demonstrated that the treatment of AGN extract decreased proliferation and differentiation of pCM cells, and affected the metabolic pathways of glucose and fatty acids. Moreover, AGN extract derived from byproducts such as stem and leaf also showed the reduced proliferation patterns on AGN‐treated pCM cells. Taken together, pCM cell‐based in vitro assay system could be primarily and efficiently applied for evaluating the biofunctional efficacy of various feed additive candidates.     

Cilostazol Improves Developmental Competence of Pig Oocytes by Increasing Intraoocyte Cyclic Adenosine Monophosphate Level and Delaying Meiotic Resumption

Cilostazol (CLZ) is a cyclic adenosine monophosphate (cAMP) modulator that influences the steady state of the meiotic stage. This study was conducted to determine the effects of CLZ treatment during in vitro maturation (IVM) on developmental competence of pig oocytes. Immature oocytes were exposed to 0 (control), 0.5, 2 and 4 μm CLZ during the first 22 h of IVM. Nuclear maturation, intraoocyte glutathione content and embryo cleavage after parthenogenesis (PA) and somatic cell nuclear transfer (SCNT) were not influenced by CLZ at any concentrations. However, 4 μm CLZ significantly (p < 0.05) improved blastocyst formation after PA (52.1% vs 38.7–46.0%) and SCNT relative to other concentrations (40.8% vs 25.0–30.7%). The mean cell numbers of SCNT blastocysts were significantly increased by 4 μm CLZ compared to the control (42.6 cells vs 35.3 cells/blastocyst). CLZ treatment significantly increased the intraoocyte cAMP level and effectively arrested oocytes at the germinal vesicle (GV) and GV break down stages compared to the control (74.5% vs 45.4%). Our results demonstrated that improved developmental competence of PA and SCNT pig embryos occurred via better synchronization of nuclear and cytoplasmic maturation induced by increased cAMP and delayed meiotic resumption after CLZ treatment.     

Effect of fermented total mixed rations on the growth performance,carcass and meat quality characteristics of Hanwoo steers

This study was conducted to assess the effects of fermented total mixed ration (FTMR) on the growth performance, carcass and meat quality traits of Hanwoo steers. The present study evidenced that the FTMR had a strong effect on dry matter intake, body weight, daily gain, slaughter weight and carcass characteristics compared with control animals. The results showed that the dry matter intake (7.17 ± 0.13 kg), average body weight (615.20 ± 112.82 kg), and daily gain (0.56 ± 0.16 kg) were greater in animals receiving FTMR than in control animals (P < 0.05). The meat quality characteristics indicated that cooking loss and the pH values did not vary between control and FTMR treated animals; however, animals in the treated groups (FTMR) had higher meat quality grades, carcass weight (396.13 ± 18.35), fat thickness (13.25 ± 1.75), marbling score (5.63 ± 0.56), meat color (40.06 ± 1.23), crude fat (18.39 ± 1.32) and sensory characteristics (flavor 5.03 ± 0.17; tenderness 4.42 ± 0.33; juiciness 5.10 ± 0.16). Nevertheless, the shear force values decreased significantly in FTMR‐treated animals compared with control group steers. Overall, FTMR may not only improve the growth performance, biochemical metabolites, and fatty and acetic acid profiles of steers, but may also enhance the carcass and meat quality characteristics of Hanwoo steers. Regarding economics, our research findings suggest that FTMR‐based feeds may enhance Hanwoo steer meat quality at a low cost.