• Asian-Australasian Journal of Animal Sciences
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• v.26 no.1
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• pp.36-42
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• 2013

The average twin lambing rate of Bayanbulak sheep is 2% to 3%. However, a flock of sheep with a close genetic relationship and an average of 2 to 3 lambs per birth has been found recently. To determine the major genes controlling the prolificacy of the flock in the present study, the flock was designated A while 100 normal Bayanbulak sheep were randomly selected to comprise the control flock B. Ligase detection reaction method was applied to detect and analyze the 10 mutational loci of the 3 candidate prolificacy genes including bone morphogenetic protein type I receptors, bone morphogenetic protein 15, and growth differentiation factor 9. The 10 mutational loci are as follows: FecB locus of the BMPR-IB gene; $FecX^I$, $FecX^B$, $FecX^L$, $FecX^H$, $FecX^G$, and $FecX^R$ of the BMP15 gene; and G1, G8, and FecTT of the GDF9 gene. Two mutations including BMPR-IB/FecB and GDF9/G1 were found in Bayanbulak sheep. Independence test results of the two flocks demonstrate that the FecB locus has a significant effect on the lambing number of Bayanbulak sheep. However, the mutation frequency of the G1 locus in GDF9 is very low. Independence test results demonstrate that the GDF9 locus does not have a significant impact on the lambing performance of Bayanbulak sheep. Among the 10 detected loci, BMPR-IB/FecB is the major gene that influences the high lambing rate of Bayanbulak sheep.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.9
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• pp.1397-1406
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• 2019

Objective: Feed energy required for pigs is first prioritized to meet maintenance costs. Additional energy intake in excess of the energy requirement for maintenance is retained as protein and fat in the body, leading to weight gain. The objective of this study was to estimate the metabolizable energy requirements for maintenance ($ME_m$) by regressing body weight (BW) gain against metabolizable energy intake (MEI) in growing pigs. Methods: Thirty-six growing pigs ($26.3{\pm}1.7kg$) were allotted to 1 of 6 treatments with 6 replicates per treatment in a randomized complete block design. Treatments were 6 feeding levels which were calculated as 50%, 60%, 70%, 80%, 90%, or 100% of the estimated ad libitum MEI ($2,400kJ/kg\;BW^{0.60}\;d$). All pigs were individually housed in metabolism crates for 30 d and weighed every 5 d. Moreover, each pig from each treatment was placed in the open-circuit respiration chambers to measure heat production (HP) and energy retained as protein ($RE_p$) and fat ($RE_f$) every 5 d. Serum biochemical parameters of pigs were analyzed at the end of the experiment. Results: The average daily gain (ADG) and HP as well as the $RE_p$ and $RE_f$ linearly increased with increasing feed intake (p<0.010). ${\beta}$-hydroxybutyrate concentration of serum tended to increase with increasing feed intake (p = 0.080). The regression equations of MEI on ADG were MEI, $kJ/kg\;BW^{0.60}\;d=1.88{\times}ADG$, g/d+782 ($R^2=0.86$) and $ME_m$ was estimated at $782kJ/kg\;BW^{0.60}\;d$. Protein retention of growing pigs would be positive while REf would be negative at this feeding level via regression equations of $RE_p$ and $RE_f$ on MEI. Conclusion: The $ME_m$ was estimated at $782kJ/kg\;BW^{0.60}\;d$ in current experiment. Furthermore, growing pigs will deposit protein and oxidize fat if provided feed at the estimated maintenance level.

• Molecules and Cells
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• v.38 no.2
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• pp.130-137
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• 2015

MicroRNAs (miRNAs) are small, endogenous RNAs that play important gene-regulatory roles by binding to the imperfectly complementary sequences at the 3'-UTR of mRNAs and directing their gene expression. Here, we first discovered that miR-576-3p was down-regulated in human bladder cancer cell lines compared with the non-malignant cell line. To better characterize the role of miR-576-3p in bladder cancer cells, we over-expressed or down-regulated miR-576-3p in bladder cancer cells by transfecting with chemically synthesized mimic or inhibitor. The overexpression of miR-576-3p remarkably inhibited cell proliferation via G1-phase arrest, and decreased both mRNA and protein levels of cyclin D1 which played a key role in G1/S phase transition. The knock-down of miR-576-3p significantly promoted the proliferation of bladder cancer cells by accelerating the progression of cell cycle and increased the expression of cyclin D1. Moreover, the dual-luciferase reporter assays indicated that miR-576-3p could directly target cyclin D1 through binding its 3'-UTR. All the results demonstrated that miR-576-3p might be a novel suppressor of bladder cancer cell proliferation through targeting cyclin D1.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.93-97
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• 2014

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.

• Journal of Microbiology and Biotechnology
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• v.29 no.2
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• pp.330-338
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• 2019

Chronic infection with intracellular Brucella abortus (B. abortus) in livestock remains as a major problem worldwide. Thus, the search for an ideal vaccine is still ongoing. In this study, we evaluated the protective efficacy of a combination of B. abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12) and malate dehydrogenase (rMDH), cloned and expressed into a pMal vector system and $DH5{\alpha}$, respectively, and further purified and applied intraperitoneally into BALB/c mice. After first immunization and two boosters, mice were infected intraperitoneally (IP) with $5{\times}10^4CFU$ of virulent B. abortus 544. Spleens were harvested and bacterial loads were evaluated at two weeks post-infection. Results revealed that this combination showed significant reduction in bacterial colonization in the spleen with a log protection unit of 1.31, which is comparable to the average protection conferred by the widely used live attenuated vaccine RB51. Cytokine analysis exhibited enhancement of cell-mediated immune response as IFN-${\gamma}$ is significantly elevated while IL-10, which is considered beneficial to the pathogen's survival, was reduced compared to control group. Furthermore, both titers of IgG1 and IgG2a were significantly elevated at three and four-week time points from first immunization. In summary, our in vivo data revealed that vaccination with a combination of five different proteins conferred a heightened host response to Brucella infection through cell-mediated immunity which is desirable in the control of intracellular pathogens. Thus, this combination might be considered for further improvement as a potential candidate vaccine against Brucella infection.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.8
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• pp.1142-1147
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• 2011

Lysine intake during gestation has a major impact on subsequent reproductive performance. The objective of this experiment was to determine the effect of lysine intake from mid-gestation until farrowing on the reproductive performance of multiparous sows. On day 30 of gestation, 200 Landrace${\times}$Large White sows were randomly assigned to one of four groups based on body weight and parity (n = 50). The gestation diets contained 0.46, 0.56, 0.65 or 0.74% lysine. Increasing dietary lysine concentration improved sow body condition at farrowing and increased litter weights (p<0.05). Dietary lysine level also had a significant effect on the dry matter (p<0.05) and protein content (p<0.05) of colostrum. Increased lysine intake increased serum insulin concentration (p<0.05) and there was a trend towards increased serum prolactin content (linear, p = 0.07). However, increased lysine tended to decrease blood urea N (quadratic, p = 0.05). These results suggest that higher lysine levels (0.65-0.75%) than those recommended by the National Research Council improved reproductive performance for multiparous gestating sows and this increase may be partially mediated through blood metabolites or metabolic hormone levels.

• Imaging Science in Dentistry
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• v.45 no.3
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• pp.187-192
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• 2015

Cleidocranial dysplasia (CCD) is a rare congenital disorder, typically characterized by persistently open skull sutures, aplastic or hypoplastic clavicles, and supernumerary teeth. Mutations in the gene encoding the runt-related transcription factor 2 (RUNX2) protein are responsible for approximately two thirds of CCD patients. We report a 20-year-old CCD patient presenting not only with typical skeletal changes, but also complex dental anomalies. A previously undiagnosed odontoma, 14 supernumerary teeth, a cystic lesion, and previously unreported fused primary teeth were discovered on cone-beam computed tomography (CBCT) scans. Mutation analysis identified the causal c.578G>A (p.R193Q) mutation in the RUNX2 gene. At 20 years of age, the patient had already missed the optimal period for dental intervention. This report describes the complex dental anomalies in a belatedly diagnosed CCD patient, and emphasizes the significance of CBCT assessment for the detection of dental anomalies and the importance of early treatment to achieve good outcomes.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.8.1-8.15
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• 2022

Background: Brucella infection induces brucellosis, a zoonotic disease. The intracellular circulation process and virulence of Brucella mainly depend on its type IV secretion system (T4SS) expressing secretory effectors. Secreted protein BspJ is a nucleomodulin of Brucella that invades the host cell nucleus. BspJ mediates host energy synthesis and apoptosis through interaction with proteins. However, the mechanism of BspJ as it affects the intracellular survival of Brucella remains to be clarified. Objectives: To verify the functions of nucleomodulin BspJ in Brucella's intracellular infection cycles. Methods: Constructed Brucella abortus BspJ gene deletion strain (B. abortus ∆BspJ) and complement strain (B. abortus pBspJ) and studied their roles in the proliferation of Brucella both in vivo and in vitro. Results: BspJ gene deletion reduced the survival and intracellular proliferation of Brucella at the replicating Brucella-containing vacuoles (rBCV) stage. Compared with the parent strain, the colonization ability of the bacteria in mice was significantly reduced, causing less inflammatory infiltration and pathological damage. We also found that the knockout of BspJ altered the secretion of cytokines (interleukin [IL]-6, IL-1β, IL-10, tumor necrosis factor-α, interferon-γ) in host cells and in mice to affect the intracellular survival of Brucella. Conclusions: BspJ is extremely important for the circulatory proliferation of Brucella in the host, and it may be involved in a previously unknown mechanism of Brucella's intracellular survival.

• Journal of Ginseng Research
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• v.44 no.2
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• pp.341-349
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• 2020

Background: The replicative senescence of human dermal fibroblasts (HDFs) is accompanied by growth arrest. In our previous study, the treatment of senescent HDFs with Rg3(S) lowered the intrinsic reactive oxygen species (ROS) levels and reversed cellular senescence by inducing peroxiredoxin-3, an antioxidant enzyme. However, the signaling pathways involved in Rg3(S)-induced senescence reversal in HDFs and the relatedness of the stereoisomer Rg3(R) in corresponding signaling pathways are not known yet. Methods: We performed senescence-associated β-galactosidase and cell cycle assays in Rg3(S)-treated senescent HDFs. The levels of ROS, adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP) as well as the mitochondrial DNA copy number, nicotinamide adenine dinucleotide (NAD)⁺/1,4-dihydronicotinamide adenine dinucleotide (NADH) ratio, and NAD-dependent sirtuins expression were measured and compared among young, old, and Rg3(S)-pretreated old HDFs. Major signaling pathways of phosphatidylinositol 3-kinase/Akt, 5' adenosine monophosphate-activated protein kinase (AMPK), and sirtuin 1/3, including cell cycle regulatory proteins, were examined by immunoblot analysis. Results: Ginsenoside Rg3(S) reversed the replicative senescence of HDFs by restoring the ATP level and NAD⁺/NADH ratio in downregulated senescent HDFs. Rg3(S) recovered directly the cellular levels of ROS and the NAD⁺/NADH ratio in young HDFs inactivated by rotenone. Rg3(S) mainly downregulated phosphatidylinositol 3-kinase/Akt through the inhibition of mTOR by cell cycle regulators like p53/p21 in senescent HDFs, whereas Rg3(R) did not alter the corresponding signaling pathways. Rg3(S)-activated sirtuin 3/PGC1α to stimulate mitochondrial biogenesis. Conclusion: Cellular molecular analysis suggests that Rg3(S) specifically reverses the replicative senescence of HDFs by modulating Akt-mTOR-sirtuin signaling to promote the biogenesis of mitochondria.

• Animal Bioscience
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• v.34 no.1
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• pp.109-118
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• 2021

Objective: The objective of this study was to determine net energy (NE) of expeller-press (EP-RSM) and solvent-extracted rapeseed meal (SE-RSM) and to establish equations for predicting the NE in rapeseed meal (RSM) fed to growing pigs. Methods: Thirty-six barrows (initial body weight [BW], 41.1±2.2 kg) were allotted into 6 diets comprising a corn-soybean meal basal diet and 5 diets containing 19.50% RSM added at the expense of corn and soybean meal. The experiment had 6 periods and 6 replicate pigs per diet. During each period, the pigs were individually housed in metabolism crates for 16 days which included 7 days for adaption to diets. On day 8, pigs were transferred to respiration chambers and fed their respective diet at 2,000 kJ metabolizable energy (ME)/kg BW0.6/d. Feces and urine were collected, and daily heat production was measured from day 9 to 13. On days 14 and 15, the pigs were fed at 890 kJ ME/kg BW0.6/d and fasted on day 16 for evaluation of fasting heat production (FHP). Results: The FHP of pigs averaged 790 kJ/kg BW0.6/d and was not affected by the diet composition. The NE values were 10.80 and 8.45 MJ/kg DM for EP-RSM and SE-RSM, respectively. The NE value was positively correlated with gross energy (GE), digestible energy (DE), ME, and ether extract (EE). The best fit equation for NE of RSM was NE (MJ/kg DM) = 1.14×DE (MJ/kg DM)+0.46×crude protein (% of DM)-25.24 (n = 8, R2 = 0.96, p<0.01). The equation NE (MJ/kg DM) = 0.22×EE (% of DM)-0.79×ash (% of DM)+14.36 (n = 8, R2 = 0.77, p = 0.018) may be utilized to quickly determine the NE in RSM when DE or ME values are unavailable. Conclusion: The NE values of EP-RSM and SE-RSM were 10.80 and 8.45 MJ/kg DM. The NE value of RSM can be well predicted based on energy content (GE, DE, and ME) and proximate analysis.