Big data analysis methods are useful tools for sorting valuable data and products. Achyranthes Radix root extract (AR) is a well-known herbal medicine in East Asia due to its anti-osteoarthritis, pro-circulatory, and anti-osteoporosis effects. In this stud y, we investigated the liver- and kidney-protective effects of AR by applying big data analysis to traditional medicine. CDDP (cis-diamminedichloridoplatinum) is an effective cancer cell anti-proliferative agent used in the treatment of diverse types of tumors. However, it is clinically limited due to liver and kidney toxicity. The current study was designed to assess the potential protective effects of AR against CDDP-induced hepato-renal toxicity. For this purpose, male Sprague-Dawley (SD) rats were assigned to four groups, each consisting of four animals. Intravenous injection or oral administration of either saline or AR was performed daily for 14 days, whereas CDDP was injected intraperitoneally on day 3 following AR treatment. Serum biochemistry results revealed that CDDP induced clear hepatic and renal damage while the AR treatment groups showed less damage relative to controls. Next, we tested the pharmacokinetics of AR using 20-hydroxyecdysone (20-HE), which is the most abundant component of AR extract. After intravenous administration of AR, the plasma concentration of 20-HE rapidly declined with a terminal half-life (t1/2) of 0.99±0.47 h. The area under the plasma concentration vs. time curve was 24.96±3.5 h*ng/mL. The present study provides valuable tools for further verification studies of the classical herbal literature and its scientific relevance.
Natural killer (NK) cells are innate lymphocytes that aid in the protection of the host from infectious diseases and cancer. Regulation of cytotoxicity in NK cells is mediated by inhibitory receptors that bind major histocompatibility complex class I (MHC-I) molecules on target cells. Human myelogenous leukemia K562 cells are readily attacked by NK cells, because K562 cells expressed very low levels of MHC-I molecules for inhibitory NK cell receptors. In this study, we compared the ability of γ-irradiated- or mitomycin C (MMC)-treated K562 feeder cells to support expansion and activation of canine NK cells. We isolated CD5 negative cells from canine peripheral blood mononuclear cells by immunomagnetic separation and co-cultured with γ-irradiated (100 Gy)- or MMC (20 μg/mL)-treated K562 cells in the presence of interleukin (IL)-2, IL-15 and IL-21 for 21 days. As a result, number of CD5 negative cells, co-cultured with γ-irradiated- or MMC-treated K562 cells (56.72 ± 13.77 fold or 32.99 ± 10.83 fold), was increased than those of CD5 negetive cells (2.99 ± 1.42 fold). Also, we found that gene expression markers of activated NK cells such as NKp30, NKp44, NKp46, Ly49, NKG2D, CD244, perforin, and granzyme B and production of interferon gamma were similarly upregulated in NK cells co-cultured with γ-irradiated- or MMC-treated K562 cells, respectively. Next, we observed that cytotoxicity of NK cells co-cultured with γ-irradiated K562 cells was more sensitively reacted to canine mammary carcinoma cells than those of MMC-treated K562 cells. These results revealed that γ-irradiated K562 cells are more efficient feeder cells than MMC-treated K562 cells for enhancing NK cells expansion and activation.
The culture of the intestinal epithelium into three dimensional (3D) structures typically termed organoid culture. Organoid culture is based on the ability of intestinal stem cells (ISCs), at the base of the crypt, perpetually to divide and produce a fully differentiated, polarized epithelium. Leucine-rich-repeat-containing G-protein-coupled receptor 5 (Lgr5) positive ISCs isolated from the intestine can form organoids in long-term culture. Thus, when cultured under the appropriate 3D conditions, single Lgr5+ ISCs undergo cycles of self-renewal, differentiation and morphogenesis, and self-organize into crypt-villus domains that house cycling ISCs and differentiated intestinal epithelial cells (IECs). In this study, we performed isolation, characterization and consecutive subculture of small intestinal crypts from BALB/c-nude mouse. Briefly, isolated mouse crypts were embedded in matrigel, cast into 40 μL droplets at the bottom of well in a 48-well plate. Following polymerization, the gels were overlaid with ISCs expansion medium containing B27, N2, N-acetylcysteine, epidermal growth factor, noggin, and R-spondin 1. As a result, mouse crypt-derived ISCs had enteroids and spheroid morphologies. We also confirmed by quantitative real-time RT-PCR that expression of ISCs-related specific genes (Lgr5, sox9) and IECs-related specific genes (chromogranin A, defensin-5, mucin-1, mucin-2, and villin) was maintained at eight passages or more. Thus, we observed that expression of specific markers and consecutive self-renewing in the mouse small intestinal crypt-derived organoids.
Osteoarthritis is a disorder characterized by a loss of cartilage as common aging-associated disease in humans and animals. However, unlike human clinical trials, investigational studies in pet animals are constrained by a lack of interest and funds. In addition, pet owners would often prefer the lowest cost method to treat arthritis of pet animals. Here, we report the outstanding and inexpensive way to prepare chondrocytes for cartilage repair using rabbit adipose derived mesenchymal stem cells (MSCs). This study focused on the development and enhancement of pre-chondrogenic condensation under external electric fields even without additional growth factors. We found that highly compact structures were formed within 3 days in micromass cultures of rabbit MSCs under electrical stimulation (ES), showing increased COL2A1 gene expression compared with their control 3D micromass cultures and 2D monolayer cultures. We further found that ES enhanced the production of proteoglycan, a highly produced extracellular matrix component in chondrocytes. Collectively, these results provide the commercial potential of electrical stimulation driving chondrogenesis of mesenchymal stem cells for repair of cartilage, which is a budget-friendly regimen.
Colibacillosis, which is caused by Escherichia coli (E. coli), is one of the most common infectious diseases affecting poultry and causes the great economic losses to poultry production worldwide. The present study investigated the presence of E. coli in broiler chickens affected by colibacillosis and reared at various locations in Tiaret Province as well as evaluated the susceptibility of isolated E. coli strains to various antimicrobial agents. A total of 110 samples collected from 55 broiler chickens representing liver (n=55) and spleen (n=55) showing lesions suspected to be colibacillosis were analyzed by conventional methods, and 33 E. coli isolates were tested on a panel of seven antimicrobial agents using the disk diffusion method. The overall isolation rates of E. coli were 60% (33/55) and 50.9% (56/110) in whole chickens and organs, respectively. There was no variation in the isolation rate between organs: 50% (28/56) for liver and 50% (28/56) for spleen. All E. coli isolates tested showed 100% resistance to Nalidixic acid and Ofloxacine, followed by 93.93%, 90.90%, and 51.51% resistance to Tetracycline, Trimethoprim-sulfamethoxazole (Co-trimoxazole), and Chloramphenicol, respectively. However, low resistance to Gentamycin was observed (12.12%), whereas all E. coli isolates were susceptible to Colistin sulfate. Multidrug resistance was observed in 96.96% (32/33) of the isolates tested. We conclude that E. coli was largely present in broiler chickens affected by colibacillosis in Tiaret Province, and multidrug-resistant E. coli isolates showed high resistance to multiple antibiotics.
Since the first HPAI epidemics in 2003, there has been little epidemiological research on the association between HPAI epidemics and vehicle movements around poultry farms. This study examined the relationship between vehicle movements around poultry farms and the 2014/15 HPAI epidemics in the Republic of Korea using two methods: a boosted regression trees (BRT) model and logistic regression of a generalized linear model (GLM). The BRT model considers the non-linearity association between the frequency of vehicle movements around poultry farms and the HPAI outbreak status per province using a machine learning technique. In contrast, a GLM assesses the relationship based on the traditional frequentist method. Among the three types of vehicle movements (outbound, inbound, and within), only the outbound was found to be a risk factor of the 2014/15 HPAI epidemics according to both the BRT model and multivariate logistic regression of GLM. In the BRT model results, the median relative contribution of the log-transformed outbound variable was 53.68 (range: 39.99 – 67.58) in the 2014 epidemics and 49.79 (range: 33.90 – 56.38) in the 2015 epidemics. In the GLM results, the odds ratio of the log-transformed outbound variable was 1.22 for the 2014 HPAI epidemics (p < 0.001) and 2.48 for the 2015 HPAI epidemics (p < 0.001), respectively. The results indicated that intensive disinfection measures on outbound movement were needed to reduce the risk of HPAI spread. The current BRT models are suitable for risk analysis because the median area under the receiver operating characteristic curve was 0.83 (range: 0.74 – 0.91) and 0.85 (range: 0.73 – 0.87) for the 2014 and 2015 epidemics models, respectively. The Akaike information criterion scores for the multivariate logistic regression of GLM were 150.27 and 78.21 for the 2014 and 2015 epidemics models, respectively. These scores were relatively lower than those from the univariate logistic regression of GLM.
The distribution of wild boar (Sus scropa) in the Republic of Korea was forecasted using environmental factors. A species distribution model was applied with the standard normalized difference vegetation index (NDVI), enhanced vegetation index (EVI), solar zenith angle (SUNZ), daytime land surface temperature (dTemp), and nighttime land surface temperature (nTemp). Understanding wild boar distribution is important for controlling African swine fever (ASF) because the disease could be endemic in wild boar or spread from wild boars to domestic pigs. Among the five predictors, the NDVI was the most influencing factor for the wild boar distribution. The relative contributions of the predictors were 67.4 for NDVI, 16.9 for dTemp, 10.5 for SUNZ, 4.4 for EVI, and 0.8 for nTemp. The area size under the receiver-operating curve of the receiver-operating characteristics for the current model was 0.62, but the real wild boar observation data overlapped with the predicted high-density wild boar distribution area. The wild boar distribution density was relatively higher in Gangwon-do, Gyeongsangbuk-do, Gyeongsangnam-do, and Jeollanam-do. Given the ASF epidemics, contact between ASF-infected animals and ASF-susceptible animals in high-density wild boar distribution areas should be prevented by long-range fencing or active surveillance.
Viral protein 2 (VP2), which is the structural protein of parvovirus, can produce virus-like particles (VLPs) by a self-assembly process in vitro, making VLPs attractive vaccine candidates. VP2 of canine parvovirus (CPV) is responsible for neutralizing antibodies in immunized animals. In this study, VP2 protein of canine parvovirus-2c was expressed using a baculovirus expression system and assembled into parvovirus-like particles in insect cells. The results show that VP2 proteins assembled into virus-like particles (VLPs) with antigenic properties similar to those of natural CPV and a high hemagglutination (HA) titer (1:27). The recombinant 6-His-tagged VP2 protein with a molecular mass of about 65 kDa was detected by anti-His antibody and anti-PPV serum. This study provides a foundation for the application of VP2 protein in the clinical diagnosis of CPV and in the vaccination against CPV.