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Neuroprotective Effects of Genistein in a SOD1-G93A Transgenic Mouse Model of Amyotrophic Lateral Sclerosis.

Oxidant toxicity has been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS), an insidiously progressive neurodegenerative disorder involving upper and lower motor neurons. Here, we investigated the cellular and molecular mechanisms underlying the neuroprotective effects of an anti-oxidant genistein in SOD1-G93A transgenic mouse model of ALS. Rotarod test, hanging wire test and hindlimb clasping test were used to determined disease onset and assess motor performance. Immunostaining together with neuronal size measurement were used to count viable motor neurons. In addition, immunostaining procedure and ELISA kit were used to assess the inflammatory response in the spinal cord. Our results showed that Genistein administration suppressed the production of pro-inflammatory cytokines and alleviated gliosis in the spinal cord of SOD1-G93A mice. In addition, genistein administration induced autophagic processes and enhanced the viability of spinal motor neurons. As a result, genistein alleviated ALS-related symptoms and slightly prolonged the lifespan of SOD1-G93A mice. Taken together, our results indicate that genistein is neuroprotective in SOD1-G93A mice, suggesting genistein could be a promising treatment for human ALS. Graphical Abstract Genistein protects impariments in SOD1-G93A transgenic mouse model.

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Seroprevalence of antibodies in sheep and goats slaughtered at the Kumasi Abattoir, Ghana.

Toxoplasmosis, caused by , is an important zoonosis worldwide. In Ghana, information on the disease in humans abounds but scanty in animals. This study was therefore conducted to estimate the seroprevalence of infection sheep and goats sampled from the Kumasi Abattoir in Ashanti Region, Ghana. A total of 347 serum samples collected from 170 sheep and 177 goats were analyzed for the presence of antibodies using a commercial ELISA kit. Results of this study estimated the seroprevalence of 23.7% in goats an, 35.9% in sheep. In sheep, 24 (35.82%) out of a total of 67 male samples were positive and 37 (35.92%) out of a total of 104 female samples were positive while in goats, 6 (8.2%) bucks out of a total of 73 were positive while 36 (34.6%) does out of a total of 104 were positive. There was a significant difference in the rate of seropositivity of female goats (-value 0.01). This study confirms the existence of infection in small ruminants in Ghana and it showed that sheep and dogs are more at risk to infection hence meat from such animals could be a potential risk to public health if consumed raw or undercooked.

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Validation of an indirect immunofluorescence assay (IFA) for the detection of IgG antibodies against Coxiella burnetii in bovine serum.

There is limited knowledge of the true prevalence and distribution of coxiellosis in dairy and beef cattle populations in Australia. For this to occur, apparent prevalence estimates need to be reliably adjusted, accounting for diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the test used. However, there are few tests available with known diagnostic specifications suitable to inform screening and surveillance activities in the Australian context. We initially modified and optimised a human indirect immunofluorescence assay (IFA) test for the detection of IgG antibodies against phase I and/or phase II Coxiella burnetii in bovine sera and determined an optimal screening dilution cut-off to be 1:160. Direct comparison of the modified IFA with the commercial IDEXX enzyme-linked immunosorbent assay (ELISA) kit (Q Fever Ab Test IDEXX Laboratories, United States of America) was performed by testing 458 serum samples from four distinct cattle populations across the east coast of Australia and New Zealand. Cross classified test results were then analysed using Bayesian latent class modelling, to validate the tests in the absence of a gold standard reference test. Results from this analysis indicate that the IFA, at a 1:160 serum dilution, has an estimated DSe of 73.6% (95% Credible Interval (CrI) 61.1, 85.9) and DSp of 98.2% (95% CrI 95.1, 99.7). The commercial IDEXX ELISA kit was found to have a higher DSe of 87.9% (95% CrI 73.9, 96.4) and similar DSp of 97.7% (95% CrI 93.2, 99.7). Evaluation of the diagnostic performance of the IFA and ELISA methods, specifically for use in cattle will enable more accurate interpretation of prevalence estimates of C. burnetii exposure to be reported for cattle in Australia and other countries.

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Ineffectiveness of meat inspection in the detection of Taenia solium cysticerci in pigs slaughtered at two abattoirs in the Eastern Cape Province of South Africa.

Porcine and human cysticercosis, caused by the larval stage of tapeworm Taenia solium, is a zoonosis in southern Africa and known to be endemic in South Africa, mainly in Eastern Cape Province. No efforts to control or eradicate this parasite have been made, despite the increasing occurrence in most Eastern Cape districts, except for routine meat inspection at local abattoirs. The parasite poses a potentially serious agricultural problem, public health risk and economic loss amongst Eastern Cape smallholder pig production communities. The objective of this study was to determine the effectiveness of routine meat inspection for the detection of porcine cysticercosis in pigs from rural smallholder/subsistence production systems in Eastern Cape Province villages. The effectiveness of meat inspection, by registered meat inspectors, in the detection of pigs infected with T. solium cysts was assessed and compared with whole carcass dissection as the "gold standard" method. The commercial antigen enzyme linked immunosorbent assay (B158/B160 Ag-ELISA) kit screened all the slaughtered animals. The proportion of pigs found infected with T. solium cysts, as measured by meat inspection, was lower (5%, 9/180) than with carcass dissection (18.9%, 34/180) and B158/B60 Ag-ELISA test (21.6%, 38/176). Four out of 180 carcasses were heavily infested with T. solium cysts, evenly distributed throughout the carcasses, to a level impossible to enumerate. Of the remaining 176 carcasses, approximately 526 cysticerci, distributed at various anatomical regions of the pig, were counted during carcass dissection. Sites with higher cyst counts, such as the back and hind leg, do not form part of the normal meat inspection regime. The level of agreement (Kappa statistic) between dissection (gold standard) and meat inspection of the two districts was negative (-0.1955). There was a slight agreement in the Kappa statistic (0.0328) between dissection and B158/B60 Ag-ELISA. This study confirms that current meat inspection procedures alone are not sufficiently sensitive to detect all cases of porcine cysticercosis at the abattoirs and require modifications, or should be supplemented by other methods. A risk-based meat safety assurance system, such as HACCP, that considers specific food safety aspects before and after the abattoir (point of slaughter) should be followed. Before slaughter, aspects such as origin, husbandry practices and on-farm animal health control should be considered; after slaughter, the abattoir should inform the next entity in the supply chain of the limitations of meat inspections and the real meaning of an "Approval" stamp. New validated testing methods that can be routinely used should be developed, and government should develop policies and legislation that promotes a risk-based meat safety assurance system throughout the food supply chain.

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Prussian blue nanoparticle-labeled aptasensing platform on graphene oxide for voltammetric detection of α-fetoprotein in hepatocellular carcinoma with target recycling.

An enzyme-free electrochemical aptasensing platform based on a graphene oxide nanosheet-modified gold-disk electrode was developed for the voltammetric detection of alpha-fetoprotein (AFP) in hepatocellular carcinoma by using a Prussian blue nanoparticle (PBNP)-labeled aptamer. The electroactive PBNP, a typical signal-generation tag, was utilized for the labeling of the aminated AFP aptamer by using covalent conjugation. The electrochemical sensing platform was prepared in a simple manner on the basis of a π-π stacking reaction between the immobilized graphene oxide and the PBNP-labeled AFP aptamer. Upon target AFP introduction, the analyte reacted with the aptamer, thus resulting in the dissociation of the PBNP from the nanosheets. In the presence of DNase I, the newly formed AFP/aptamer-PBNP complex was cleaved to release target AFP, which could react again with the aptamer on the nanosheets, thereby causing target recycling. During this process, the cleaved PBNP-aptamer was far away from the electrode to decrease the voltammetric signal. Under optimum conditions, the voltammetric peak current of the modified electrode decreased with the increment of the target AFP concentration within the linear range of 0.01-300 ng mL-1 at a low detection limit of 6.3 pg mL-1. The precision and reproducibility of the aptasensing protocol were acceptable (CV: <15% for intra-assay and inter-assay). Other possible nontarget biomarkers did not interfere significantly with the voltammetric signal of this system. Human serum samples containing target AFP were assayed with electrochemical aptasensing and a commercial human AFP ELISA kit, and gave well-matched results from these two methods. Importantly, our strategy provides a new horizon for the determination of disease-related proteins.

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p53 Protein Evaluation on Spermatozoa DNA in Fertile and Infertile Males.

Protein p53 role in the spermatogenesis is demonstrated, it guarantees both the appropriate quality and quantity of mature spermatozoa. In this observational study we evaluate the eventual correlation between "corrected" p53 concentration on human spermatozoa DNA and male fertility potential.

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Rabbit Anti-Cell death in AKT1 & PIAS2 Protein Prot CAMK2G & SMAD3 Protein Pr CDK7 & E2F1 Protein Prote AKT1 & SRC Protein Protei CCND3 & CDKN1A Protein Pr NFKB1 & HSPA1L Protein Pr Goat Anti- T-cell differe TRAF2 & ERN1 Protein Prot RHOA & DAAM1 Protein Prot EGFR & EGF Protein Protei Rabbit Anti-Cell death in

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Inhibition of the receptor for advanced glycation inhibits lipopolysaccharide-mediated High mobility group protein B1 and Interleukin-6 synthesis in human gingival fibroblasts through the NF-κB signaling pathway.

We investigated the effect of a specific inhibitor of the receptor for advanced glycation (FPS-ZM1) against lipopolysaccharide (LPS)-induced increase in expressions of high mobility group protein B1 (HMGB1) and interleukin-6 (IL-6) in human gingival fibroblasts (HGFs). Furthermore, we explored the potential molecular mechanisms and assessed the involvement of the NF-κB pathway in mediating the changes in the expressions of HMGB1 and IL-6 expression in response to LPS and FPS-ZM1.

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lncRNA H19 contributes to oxidative damage repair in the early age-related cataract by regulating miR-29a/TDG axis.

Age-related cataract (ARC) is caused by the exposure of the lens to UVB which promotes oxidative damage and cell death. This study aimed to explore the role of lncRNA H19 in oxidative damage repair in early ARC. lncRNAs sequencing technique was used to identify different lncRNAs in the lens of early ARC patients. Human lens epithelial cells (HLECs) were exposed to ultraviolet irradiation; and 8-OHdG ELISA, Cell counting kit 8 (CCK8), EDU, flow cytometry and TUNEL assays were used to detect DNA damage, cell viability, proliferation and apoptosis. Luciferase assay was used to examine the interaction among H19, miR-29a and thymine DNA glycosylase (TDG) 3'UTR. We found that lncRNA H19 and TDG were highly expressed while miR-29a was down-regulated in the three types of early ARC and HLECs exposed to ultraviolet irradiation, compared to respective controls. lncRNA H19 knockdown aggravated oxidative damage, reduced cell viability and proliferation, and promoted apoptosis in HLECs, while lncRNA H19 overexpression led to opposite effects in HLECs. Mechanistically, miR-29a bound TDG 3'UTR to repress TDG expression. lncRNA H19 up-regulated the expression of TDG by repressing miR-29a because it acted as ceRNA through sponging miR-29a. In conclusion, the interaction among lncRNA H19, miR-29a and TDG is involved in early ARC. lncRNA H19 could be a useful marker of early ARC and oxidative damage repair pathway of lncRNA H19/miR-29a/TDG may be a promising target for the treatment of ARC.

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Myricetin Inhibits Breast Tumor Growth and Angiogenesis by Regulating VEGF/VEGFR2 and p38MAPK Signaling Pathways.

Tumor angiogenesis is an important cause of tumor growth and metastasis. Myricetin is a flavonoid component used in traditional Chinese medicine that has been demonstrated to have anticancer activity. However, to the best of our knowledge, the effect of myricetin on tumor angiogenesis remains unknown. The present study reports the identification of myricetin as a potential chemopreventive agent by reason of its inhibition of tumor angiogenesis and demonstrates the anticancer effects of myricetin in vivo. Cell Counting Kit-8 assays revealed that myricetin inhibits the proliferation of tumor cells but not that of human umbilical vein endothelial cells (HUVECs), and a transwell assay demonstrated that myricetin could inhibit the migration of HUVECs. A rat aortic ring assay revealed that myricetin could also affect the development of microvessels and the formation of vascular networks. Further, an ELISA showed that myricetin reduced the levels of vascular endothelial growth factor (VEGF) in vivo and in vitro. Western blot analysis indicated that myricetin could downregulate VEGFR2 and p38MAPK. Therefore, myricetin could significantly inhibit tumor angiogenesis and has potential as a chemopreventive agent because of its inhibition to angiogenesis. Anat Rec, 2019. © 2019 Wiley Periodicals, Inc.

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Anti-Inflammatory Effects of Interleukin-4 on Intervertebral Disc Cells.

Inflammation has been associated with a number of pathological conditions including intervertebral disc (IVD) degeneration, increased risks of low back pain and other spinal diseases. Downregulating disc inflammation may be a strategy to reduce degeneration and more importantly back pain. Interleukin (IL)-4 was first discovered as a T-cell secreted factor that enhanced the proliferation of anti-IgM stimulated B cells and is now known as a cytokine that can stimulate cell proliferation and differentiation, tissue regeneration and neurological functions. IL-4 has been shown to be effective in inhibiting inflammatory pathways in chondrocytes. Immunohistochemical studies have shown that disc tissues are immunopositive for IL-4 receptor α (IL-4Rα) and IL-4. Yet, the roles of IL-4 and IL-4R in disc biology remain unknown.

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