In this study, we conducted a Genome Wide Association Study (GWAS) analysis of the reproductive traits in Hanwoo cows, Age at First Calving (AFC) and Calving Interval (CI), aiming to pinpoint candidate genes and their functional relevance. A total of 24,977 Hanwoo cows from commercial farms were genotyped using Axiom Bovine 60k version 3 (Affymetrix Inc, 2006). Subsequently, individuals with genotype and phenotype data were selected from this population, resulting in 9,508 cows and 37,999 SNPs for AFC, and 6,442 cows and 38,379 SNPs for CI after Quality Control. Bonferroni correction and False Discovery Rate (FDR) correction were applied to GWAS thresholds, leading to the identification of significant SNP markers for each trait by chromosome. AFC revealed seven SNP markers on five chromosomes, and CI identified three SNP markers on three chromosomes. Sixteen candidate genes were identified from these ten significant SNP markers. Notably, TRNAC-GCA was highlighted as a gene influencing semen quality in bovines and playing a crucial role in cow reproductive traits. Additionally, Gene Ontology (GO) was identified two GO IDs, respectively, in Biological Process (BP), Cellular Component (CC), and Molecular Function (MF) (BP: GO:0030259, GO:0005975; CC: GO:0031982, GO:0005794; MF: GO:0016758, GO:0016757), with no KEGG pathway being identified. These results contribute to advanced understanding of the genetic architecture of reproductive traits and offer valuable SNP markers for breeding programs in Hanwoo cow.

In Korea, Korea Proven Bulls (KPN) program has been well-developed. Breeding and evaluation of cows are also an essential factor to increase earnings and genetic gain. This study aimed to evaluate the accuracy of cow breeding value by using three methods (pedigree index [PI], pedigree-based best linear unbiased prediction [PBLUP], and genomic-BLUP [GBLUP]). The reference population (n = 16,971) was used to estimate breeding values for 481 females as a test population. The accuracy of GBLUP was 0.63, 0.66, 0.62 and 0.63 for carcass weight (CWT), eye muscle area (EMA), back-fat thickness (BFT), and marbling score (MS), respectively. As for the PBLUP method, accuracy of prediction was 0.43 for CWT, 0.45 for EMA, 0.43 for MS, and 0.44 for BFT. Accuracy of PI method was the lowest (0.28 to 0.29 for carcass traits). The increase by approximate 20% in accuracy of GBLUP method than other methods could be because genomic information may explain Mendelian sampling error that pedigree information cannot detect. Bias can cause reducing accuracy of estimated breeding value (EBV) for selected animals. Regression coefficient between true breeding value (TBV) and GBLUP EBV, PBLUP EBV, and PI EBV were 0.78, 0.625, and 0.35, respectively for CWT. This showed that genomic EBV (GEBV) is less biased than PBLUP and PI EBV in this study. In addition, number of effective chromosome segments (Me) statistic that indicates the independent loci is one of the important factors affecting the accuracy of BLUP. The correlation between Me and the accuracy of GBLUP is related to the genetic relationship between reference and test population. The correlations between Me and accuracy were -0.74 in CWT, -0.75 in EMA, -0.73 in MS, and -0.75 in BF, which were strongly negative. These results proved that the estimation of genetic ability using genomic data is the most effective, and the smaller the Me, the higher the accuracy of EBV.

The present study was conducted to examine the effect of soybean silage as a crude protein supplement for corn silage in the diet of Hanwoo steers. The first experiment was conducted to evaluate the effect of replacing corn silage with soybean silage at different levels on rumen fermentation characteristics in vitro. Commercially-purchased corn silage was replaced with 0, 4, 8, or 12% of soybean silage. Half gram of the substrate was added to 50 mL of buffer and rumen fluid from Hanwoo cows, and then incubated at 39°C for 0, 3, 6, 12, 24, and 48 h. At 24 h, the pH of the control (corn silage only) was lower (p<0.05) than that of soybeansupplemented silages, and the pH numerically increased along with increasing proportions of soybean silage. Other rumen parameters, including gas production, ammonia nitrogen, and total volatile fatty acids, were variable. However, they tended to increase with increasing proportions of soybean silage. In the second experiment, 60 Hanwoo steers were allocated to one of three dietary treatments, namely, CON (concentrate with Italian ryegrass), CS (concentrate with corn silage), CS4% (concentrate with corn silage and 4% of soybean silage). Animals were offered experimental diets for 110 days during the growing period and then finished with typified beef diets that were commercially available to evaluate the effect of soybean silage on animal performance and meat quality. With the soybean silage, the weight gain and feed efficiency of the animal were more significant than those of the other treatments during the growing period (p<0.05). However, the dietary treatments had little effect on meat quality except for meat color. In conclusion, corn silage mixed with soybean silage even at a lower level provided a greater ruminal environment and animal performances, particularly with increased carcass weight and feed efficiency during growing period.

The economic impacts of infertility and subfertility of stallions greatly influence the horse breeding industry. Self-renewal and differentiation of spermatogonial stem cells are the initial processes to maintain an adequate sperm population. Thus, understanding these processes may provide useful information to reveal the causes and remedies of subfertile and infertile stallions. Stallions are seasonal breeders. About 50% of the sperm population is reduced during the non-breeding season (NBS) in stallions. The seasonal regulation of spermatogenesis renders stallions as ideal models to understand the process of sperm production. Furthermore, comparing internal and external factors related to spermatogenesis during the breeding season (BS) and NBS may provide a solution for subfertile/infertile stallions. It is especially pertinent to study the expression pattern of different protein markers during undifferentiated, differentiating, and differentiated spermatogonia. Deleted in azoospermia-like (DAZL), undifferentiated cell transcription factor 1 (UTF-1), and protein gene product 9.5 (PGP9.5) are the molecular markers expressed at different stages of spermatogenesis. However, whether the expression pattern of these molecular markers is similar throughout the year in stallion remains undetermined. The objectives of this study were to (1) investigate the expression pattern and localization of DAZL, UTF-1, and PGP9.5 within seminiferous tubules and (2) evaluate the relative mRNA levels of these three germ cell markers in stallion testes during BS and NBS. Immunohistochemistry was performed to check and compare the expression pattern and localization of DAZL, UTF-1, and PGP9.5 antibodies. Reverse transcription-quantitative PCR analysis was performed to calculate the relative mRNA expression levels in the testes. Testicular tissues from thoroughbred stallions were collected during routine castration that was carried out in field conditions. Immunostaining of germ cells with DAZL and UTF-1 in BS and NBS were not significantly different. However, the relative mRNA expression levels of DAZL and UTF-1 were significantly different in both groups. Interestingly, the immunolabeling and the relative mRNA expression of PGP9.5 were significantly different between BS and NBS. From these results, it is hypothesized that the expression level of these putative molecular markers might be gonadotropin-dependent in stallion testes.

MicroRNAs (miRNAs) play a vital role in improving meat quality by binding to messenger RNAs (mRNAs). We performed an integrated analysis of miRNA and mRNA expression profiling between bulls and steers based on the differences in meat quality traits. Fat and fatty acids are the major phenotypic indices of meat quality traits to estimate between-group variance. In the present study, 90 differentially expressed mRNAs (DEGs) and 18 differentially expressed miRNAs (DEMs) were identified. Eighty-three potential DEG targets and 18 DEMs were used to structure a negative interaction network, and 75 matching target genes were shown in this network. Twenty-six target genes were designated as intersection genes, screened from 18 DEMs, and overlapped with the DEGs. Seventeen of these genes enriched to 19 terms involved in lipid metabolism. Subsequently, 13 DEGs and nine DEMs were validated using quantitative real-time PCR, and seven critical genes were selected to explore the influence of fat and fatty acids through hub genes and predict functional association. A dual-luciferase reporter and Western blot assays confirmed a predicted miRNA target (bta-miR-409a and PLIN5). These findings provide substantial evidence for molecular genetic controls and interaction among genes in cattle.

Abstract Background: Silibinin has shown therapeutic effects in various inflammatory and cancer models. However, the therapeutic effect and underlying mechanism in V2O5-induced pulmonary inflammation, which consists of particulate matter and metal components, have not been identified. We examined the potential anti-inflammatory activity and underlying mechanism of silibinin in a vanadium-induced lung injury model. Results: Silibinin treatment resulted in a significant restoration of cell viability and reduced infiltration of inflammatory cells in a histological analysis. In addition, silibinin significantly reduced the expression of the pro-inflammatory cytokines, TNF-α, IL-6, and IL-1ꞵ. Furthermore, silibinin reduced inflammation by down-regulating the MAPK and NF-kB signaling pathways and the NLRP3 inflammasome. Treatment with silibinin significantly reduced the NLRP3 expression levels in lung tissue. Conclusion: These results indicate that silibinin exhibits anti-inflammatory effects in V2O5-induced inflammatory mice.

This study evaluated the efficacy of chlorine dioxide (ClO2) as an oxidant to reduce malodor emission from chicken feces. Two experiments were performed with the following four treatments in parallel: 1) fresh chicken feces with only distilled water added as a control, 2) a commercial germicide as a positive control, and 3) 2,000 or 4) 3,000 ppm of ClO2 supplementation. Aluminum gas bags containing chicken feces sealed with a silicone plug were used in both experiments, and each treatment was tested in triplicate. In Experiment 1, 10 mL of each additive was added on the first day of incubation, and malodor emissions were then assessed after 10 days of incubation. In Experiment 2, 1 mL of each additive was added daily during a 14-day incubation period. At the end of the incubation, gas production, malodor-causing substances (H2S and NH3 gases), dry matter, pH, volatile fatty acids (VFAs), and microbial enumeration were analyzed. Supplementing ClO2 at 2,000 and 3,000 ppm significantly reduced the pH and the ammonia-N, total VFA, H2S, and ammonia gas concentrations in chicken feces compared with the control feces (P<0.05). Additionally, microbial analysis indicated that the number of coliform bacteria was decrease after ClO2 treatment (P<0.05). In conclusion, ClO2 at 2,000 and 3,000 ppm was effective at reducing malodor emission from chicken feces. However, further studies are warranted to examine the effects of ClO2 at various concentrations and the effects on malodor emission from a poultry farm

In horse management, the alarm system with sensors in the foaling period enables the breeder can appropriately prepare the time of the parturition. It is important to prevent losses by unpredictable parturition because there are several high risks such as dystocia and the death of foals and mares during foaling. However, unlike analysis in the alarm system that detects specific motions has been widely performed, analysis of classification following specific behavior patterns or number needs to be more organized. Thus, the objective of this study is to classify signs of the specific behaviors of the mares for the prediction of pre-foaling behaviors. Five Thoroughbred mares (9-20 yrs) were randomly selected for observation of the prefoaling behaviors. The behaviors were monitored for 90 min that was divided into three different periods as 1) from -90 to -60 min, 2) from -60 to -30 min, 3) from -30 min to the time for the discharge of the amniotic fluid, respectively. The behaviors were divided into two different categories as state and frequent behaviors and each specific behavioral pattern for classification was individually described. In the state behaviors, the number of mares in the standing of the foaling group (3.17 ± 0.18b) at period 3 was significantly higher than the control group (1.67 ± 0.46a). In contrast, the number of the mares in the eating of the foaling group (1.17 ± 0.34b) at period 3 was significantly lower than the control group (3.33 ± 0.46a). In the frequent behaviors, the weaving of the foaling group was significantly higher than the control group, and looking at the belly of the foaling group was significantly lower than the control group. In period 2, defecation, weaving, and lowering the head of the foaling group were significantly higher than the control group, respectively. In period 3, sitting down and standing up, pawing, weaving, and lowering the head in the foaling group were also significantly higher than the control group. In conclusion, the behavior is significantly different in foaling periods, and the prediction of foaling may be feasible by the detection of the pre-foaling behaviors in the mares.

Background Juxtaposed with another zinc finger protein 1 (JAZF1) is associated with metabolic disorders, including type 2 diabetes mellitus (T2DM). Several studies showed that JAZF1 and body fat mass are closely related. We attempted to elucidate the JAZF1 functions on adipose development and related metabolism using in vitro and in vivo models. Results The JAZF1 expression was precisely regulated during adipocyte differentiation of 3T3-L1 preadipocyte and mouse embryonic fibroblasts (MEFs). Homozygous JAZF1 deletion (JAZF1-KO) resulted in impaired adipocyte differentiation in MEF. The JAZF1 role in adipocyte differentiation was demonstrated by the regulation of PPARγ—a key regulator of adipocyte differentiation. Heterozygous JAZF1 deletion (JAZF1-Het) mice fed a normal diet (ND) or a high-fat diet (HFD) had less adipose tissue mass and impaired glucose homeostasis than the control (JAZF1-Cont) mice. However, other metabolic organs, such as brown adipose tissue and liver, were negligible effect on JAZF1 deficiency. Conclusion Our findings emphasized the JAZF1 role in adipocyte differentiation and related metabolism through the heterozygous knockout mice. This study provides new insights into the JAZF1 function in adipose development and metabolism, informing strategies for treating obesity and related metabolic disorders.

Background: Colorectal cancer (CRC) has a high morbidity and mortality worldwide. 20 (S)-ginsenoside Rh2 (G-Rh2) is a natural compound extracted from ginseng, which exhibits anticancer effects in many cancer types. In this study, we demonstrated the effect and underlying molecular mechanism of G-Rh2 in CRC cells in vitro and in vivo. Methods: Cell proliferation, migration, invasion, apoptosis, cell cycle, and western blot assays were performed to evaluate the effect of G-Rh2 on CRC cells. In vitro pull-down assay was used to verify the interaction between G-Rh2 and Axl. Transfection and infection experiments were used to explore the function of Axl in CRC cells. CRC xenograft models were used to further investigate the effect of Axl knockdown and G-Rh2 on tumor growth in vivo. Results: G-Rh2 significantly inhibited proliferation, migration, and invasion, and induced apoptosis and G(0)/G(1) phase cell cycle arrest in CRC cell lines. G-Rh2 directly binds to Axl and inhibits the Axl signaling pathway in CRC cells. Knockdown of Axl suppressed the growth, migration and invasion ability of CRC cells in vitro and xenograft tumor growth in vivo, whereas overexpression of Axl promoted the growth, migration, and invasion ability of CRC cells. Moreover, G-Rh2 significantly suppressed CRC xenograft tumor growth by inhibiting Axl signaling with no obvious toxicity to nude mice. Conclusion: Our results indicate that G-Rh2 exerts anticancer activity in vitro and in vivo by suppressing the Axl signaling pathway. G-Rh2 is a promising candidate for CRC prevention and treatment. (C) 2021 The Korean Society of Ginseng. Publishing services by Elsevier B.V.

Recently, the cattle genome sequence has been completed, followed by developing a commercial single nucleotide polymorphism (SNP) chip panel in the animal genome industry. In order to increase statistical power for detecting quantitative trait locus (QTL), a number of animals should be genotyped. However, a high-density chip for many animals would be increasing the genotyping cost. Therefore, statistical inference of genotype imputation (low-density chip to high-density) will be useful in the animal industry. The purpose of this study is to investigate the effect of the reference population size and marker density on the imputation accuracy and to suggest the appropriate number of reference population sets for the imputation in Hanwoo cattle. A total of 3,821 Hanwoo cattle were divided into reference and validation populations. The reference sets consisted of 50k (38,916) marker data and different population sizes (500, 1,000, 1,500, 2,000, and 3,600). The validation sets consisted of four validation sets (Total 889) and the different marker density (5k [5,000], 10k [10,000], and 15k [15,000]). The accuracy of imputation was calculated by direct comparison of the true genotype and the imputed genotype. In conclusion, when the lowest marker density (5k) was used in the validation set, according to the reference population size, the imputation accuracy was 0.793 to 0.929. On the other hand, when the highest marker density (15k), according to the reference population size, the imputation accuracy was 0.904 to 0.967. Moreover, the reference population size should be more than 1,000 to obtain at least 88% imputation accuracy in Hanwoo cattle.

BACKGROUND:Colorectal cancer (CRC) is a clinically challenging malignant tumor worldwide. As a natural product and sesquiterpene lactone, Costunolide (CTD) has been reported to possess anticancer activities. However, the regulation mechanism and precise target of this substance remain undiscovered in CRC. In this study, we found that CTD inhibited CRC cell proliferation in vitro and in vivo by targeting AKT.METHODS:Effects of CTD on colon cancer cell growth in vitro were evaluated in cell proliferation assays, migration and invasion, propidium iodide, and annexin V-staining analyses. Targets of CTD were identified utilizing phosphoprotein-specific antibody array; Costunolide-sepharose conjugated bead pull-down analysis and knockdown techniques. We investigated the underlying mechanisms of CTD by ubiquitination, immunofluorescence staining, and western blot assays. Cell-derived tumour xenografts (CDX) in nude mice and immunohistochemistry were used to assess anti-tumour effects of CTD in vivo.RESULTS:CTD suppressed the proliferation, anchorage-independent colony growth and epithelial-mesenchymal transformation (EMT) of CRC cells including HCT-15, HCT-116 and DLD1. Besides, the CTD also triggered cell apoptosis and cell cycle arrest at the G2/M phase. The CTD activates and induces p53 stability by inhibiting MDM2 ubiquitination via the suppression of AKT's phosphorylation in vitro. The CTD suppresses cell growth in a p53-independent fashion manner; p53 activation may contribute to the anticancer activity of CTD via target AKT. Finally, the CTD decreased the volume of CDX tumors without of the body weight loss and reduced the expression of AKT-MDM2-p53 signaling pathway in xenograft tumors.CONCLUSIONS:Our project has uncovered the mechanism underlying the biological activity of CTD in colon cancer and confirmed the AKT is a directly target of CTD. All of which These results revealed that CTD might be a new AKT inhibitor in colon cancer treatment, and CTD is worthy of further exploration in preclinical and clinical trials.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer and is one of the leading causes of cancer-related deaths worldwide.Imatinib and GNF-5 are breakpoint cluster region-Abelson murine leukemia tyrosine kinase inhibitors which have been approved for the treatment of chronic myeloid leukemia and various solid tumors.However, the effect and underlying mechanisms of imatinib and GNF-5 in HCC remain poorly defined.In this study, we investigated the anticancer activity and underlying mechanisms of imatinib and GNF-5 in HepG2 human hepatocarcinoma cells.Cell proliferation and anchorage-independent colony formation assays were done to evaluate the effects of imatinib and GNF-5 on the growth of HepG2 cells.The cell cycle was assessed by flow cytometry and verified by immunoblot analysis.Gene overexpression and knockdown assays were conducted to evaluate the function of S-phase kinase-associated protein 2 (Skp2).Imatinib and GNF-5 significantly inhibited the growth of HepG2 cells.Imatinib and GNF-5 induced G0/G1 phase cell cycle arrest by downregulating Skp2 and upregulating p27 and p21.Overexpression of Skp2 reduced the effect of imatinib and GNF-5 on HepG2 cells.Knockdown of Skp2 suppressed the proliferation and induced G0/ G1 phase arrest.Furthermore, knockdown of Skp2 enhanced the effect of imatinib and GNF-5 on growth of HepG2 cells.In conclusion, imatinib and GNF-5 effectively suppress HepG2 cell growth by inhibiting Skp2 expression.Skp2 promotes the cell proliferation and reverse G0/G1 phase cell cycle arrest and it represents a potential therapeutic target for HCC treatment.

It was hypothesized that single-nucleotide polymorphisms (SNPs) extracted from text-mined genes could be more tightly related to causal variant for each trait and that differentially weighting of this SNP panel in the GBLUP model could improve the performance of genomic prediction in cattle. Fitting two GRMs constructed by text-mined SNPs and SNPs except text-mined SNPs from 777k SNPs set (exp_777K) as different random effects showed better accuracy than fitting one GRM (Im_777K) for six traits (e.g. backfat thickness: +0.002, eye muscle area: + 0.014, Warner-Bratzler Shear Force of semimembranosus and longissimus dorsi: + 0.024 and + 0.068, intramuscular fat content of semimembranosus and longissimus dorsi: + 0.008 and +0.018). These results can suggest that attempts to incorporate text mining into genomic predictions seem valuable, and further study using text mining can be expected to present the significant results.

A study was conducted to determine the effects of lactic acid bacteria (LAB) on nutritive value and in vitro rumen digestibility of maize and rice straw silages. Two identical experiments were carried out for each of the two silages. A total of five treatments were used for each experiment: (1) negative control (NC); (2) positive control (PC); (3) Lactobacillus plantarum (LPL); (4) L. paracasei (LPA); and (5) L. acidophilus (LA). Each treatment was then divided into four ensiling periods: 3, 7, 20, and 40 days with three replications. The LPL treatment had significantly higher dry matter (DM), lower ammonia-N, and a lower number of fungi on maize silage after 40 days (p < 0.05). On the other hand, the LA treatment increased DM and CP content, reduced NDF and ADF contents compared to NC, and also produced more lactic acid compared to the other LAB-treated rice straw silages. Results of the in vitro rumen fermentation of maize silages showed no significant differences in DMD after LAB inoculation. However, higher DMD and ruminal ammonia-N were shown by rice straw ensiled with L. acidophilus. In conclusion, silage additives, which could improve the ensiling process of maize and rice straw, appeared to be different and substrate specific.

Huge amount of persimmon peels, which are byproducts usually arising after making dried persimmons from fresh persimmons, are generated in fall, every year. As the dry matter of persimmon peel contains carbohydrates over 80%, it could be a good candidate for feed. In this study, we evaluated the persimmon peel as a feed resource for a hen laying eggs. Persimmon peel has lower ratio of crude protein and fat composition while higher ratio of crude fiber composition compared to a basal laying hen feed. Feeding the persimmon peel by adding to the feed at the level of 2% or 5% did not affect the egg-laying rate from beginning to the peak of laying. In addition, feeding the persimmon peel did not affect the egg quality either, as the pH, Haugh unit, yolk color and shell thickness of the egg were not different from each other. The composition of amino acids and fatty acids were also almost same for each other, except for the linolenic acid which was slightly higher in the persimmon peel feeding groups. In conclusion, addition of persimmon peel to the feed up to 5% might be helpful to increase the income and reduce the waste produced from the agricultural byproduct. (

Alzheimer's disease (AD) is a neurodegenerative disorder, characterized by cognitive impairment, progressive neurodegeneration, and amyloid-β (Aβ) lesion. In the neuronal death and disease progression, inflammation is known to play an important role. Our previous study on acute-phase protein serum amyloid A1 (SAA1) overexpressed mice showed that the liver-derived SAA1 accumulated in the brain by crossing the brain blood barrier (BBB) and trigger the depressive-like behavior on mouse. Since SAA1 involved in immune responses in other diseases, we focused on the possibility that SAA1 may exacerbate the neuronal inflammation related to Alzheimer’s disease. A APP/SAA overexpressed double transgenic mouse was generated using amyloid precursor protein overexpressed (APP)-c105 mice and SAA1 overexpressed mice to examine the function of SAA1 in Aβ abundant condition. Comparisons between APP and APP/SAA1 transgenic mice showed that SAA1 exacerbated amyloid aggregation and glial activation; which lead to the memory decline. Behavior tests also supported this result. Overall, overexpression of SAA1 intensified the neuronal inflammation in amyloid abundant condition and causes the greater memory decline compared to APP mice, which only expresses Aβ 1–42.