• IMMUNE NETWORK
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• v.20 no.3
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• pp.25.1-25.13
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• 2020

Acinetobacter baumannii is known for its multidrug antibiotic resistance. New approaches to treating drug-resistant bacterial infections are urgently required. Cathelicidin-related antimicrobial peptide (CRAMP) is a murine antimicrobial peptide that exerts diverse immune functions, including both direct bacterial cell killing and immunomodulatory effects. In this study, we sought to identify the role of CRAMP in the host immune response to multidrug-resistant Acinetobacter baumannii. Wild-type (WT) and CRAMP knockout mice were infected intranasally with the bacteria. CRAMP^-/- mice exhibited increased bacterial colony-forming units (CFUs) in bronchoalveolar lavage (BAL) fluid after A. baumannii infection compared to WT mice. The loss of CRAMP expression resulted in a significant decrease in the recruitment of immune cells, primarily neutrophils. The levels of IL-6 and CXCL1 were lower, whereas the levels of IL-10 were significantly higher in the BAL fluid of CRAMP^-/- mice compared to WT mice 1 day after infection. In an in vitro assay using thioglycollate-induced peritoneal neutrophils, the ability of bacterial phagocytosis and killing was impaired in CRAMP^-/- neutrophils compared to the WT cells. CRAMP was also essential for the production of cytokines and chemokines in response to A. baumannii in neutrophils. In addition, the A. baumannii-induced inhibitor of κB-α degradation and phosphorylation of p38 MAPK were impaired in CRAMP^-/- neutrophils, whereas ERK and JNK phosphorylation was upregulated. Our results indicate that CRAMP plays an important role in the host defense against pulmonary infection with A. baumannii by promoting the antibacterial activity of neutrophils and regulating the innate immune responses.

• Journal of Life Science
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• v.19 no.4
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• pp.549-552
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• 2009

Genetic mutations by gene fusion result from chromosomal rearrangement, trans-splicing, and intergenic splicing. Trans-splicing is a phenomenon in which two pre-mRNAs grow together into one. We analyzed the trans-splicing products in embryonic stem cells. By using bioinformatic tools, 70 trans-splicing transcripts were identified. They are classified into 6 types according to fusion pattern: 5'UTR-5'UTR, 5'UTR-3'UTR, 3'UTR-3'UTR, 5'UTR-CDS, 3'UTR-CDS, CDS-CDS. The fusion products are more abundant in CDS regions than in UTR regions, which contain multiple intron numbers. Chromosome analysis showing gene fusion via trans-splicing indicated that chromosomes 17 and 19 were activated. These data are of great use for further studies in relation to fusion genes and human diseases.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.161-168
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• 2016

Nucleotide metabolism in endothelium is variable between different species. Recent studies demonstrated that this variability could contribute coagulation dysfunction, even though organs of the alpha 1,3-galactosyltransferase gene knockout pig were transplanted into the primate. CD73 (ecto-5'-nucelotidase) is an enzyme at cell surface catalyzing the hydrolysis of adenosine triphosphate to adenosine, which plays role on a substance for anti-inflammatory and anti-coagulant. Thus, overexpression of CD73 in endothelial cells of the pig is considered as an approach to reduce coagulopathy. In this study, we constructed a human CD73 expression vector under control of porcine Icam2 promoter (pIcam2-hCD73), which is expressed specifically at endothelial cells, and of CMV promoter as a control (CMV-CD73). First, we transfected the CMV-CD73 vector into HEK293 cells, and then confirmed CD73 expression at cell surface by flow cytometry analysis. Next, we transfected the pIcma2-CD73 and CMV-CD73 vectors into primary porcine fibroblasts and endothelial cells. Consequence was that the pIcma2-CD73 vector was expressed only at the porcine endothelial cells, meaning that the pIcam2 promoter lead to endothelial cell-specific expression of CD73 in vitro. Finally, we nucleofected the pIcam2-hCD73 vector into passage 3 fibroblasts, and enforced hygromycin selection of 400mg/ml. We were able to obtain forty three colonies harboring pIcam2-CD73 to provide donor cells for transgenic cloned porcine production.

• Journal of Ecology and Environment
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• v.43 no.2
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• pp.263-270
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• 2019

Background: The gelada baboon, Theropithecus gelada Ruppell, 1835, is an endemic primate of the central and western highlands of Ethiopia occurring between altitudes of 1800-4400 m. Variability in activity time budget between the two gelada social units, i.e., one-male units (OMUs) and all-male units (AMUs), has not been studied previously. Thus, the present study was an effort to understand intra- and inter-variations in group size and daily activity time budget in gelada baboon OMUs and AMUs at Guassa Community Conservation Area, Central Ethiopia. The instantaneous sampling method was used in the data collection. Results: Both OMU and AMU groups allocated more time for feeding (OMUs = 36.96%; AMUs = 35%) followed by moving (OMU = 25.8%; AMUs = 27%). Grooming was the most frequent social activity in both family groups (OMUs = 8.56%; AMUs = 11.81%). OMUs and AMUs did not show significant variation between themselves in the time budget and temporal distribution of their daily activities. However, the overall variation in the time budget allocated for daily activities within OMUs and AMUs was statistically significant (p < 0.05%). The different age and sex groups in OMUs (i.e., immature, sub-adult females, subordinate males, adult females, and alpha males) showed statistically significant variation in their time budget allocation for the daily activities (p < 0.05) except for feeding and moving. Alpha males had the highest proportion of resting (32.14%) and aggression (31.92%). Immature individuals were responsible for > 90% of the time budget recorded for playing while adult females showed the highest frequency of grooming. Group size of OMUs ranged between 5 and 15 individuals (mean ± SD = 11.25 ± 1.95). Adult females comprised the highest number within OMUs (mean = 4.5) followed by immature individuals (mean = 3). The group size of AMUs ranged from 4 to 8 (mean ± SD = 6.6 ± 1.5). Conclusion: Our results showed that the gelada baboon spent less time on foraging compared to previous studies at the Simien Mountains and Debre-Libanos area. We suggested this variation could be explained based on higher forage quality at Guassa Community Conservation Area resulting from better conservation of the habitat.

• Journal of Embryo Transfer
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• v.33 no.3
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• pp.129-137
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• 2018

Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig ($GalT^{-MCP/-MCP}$). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain $GalT^{-MCP/-MCP}/CD39/CD73$ pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from $GalT^{-MCP/-MCP}$ pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate $GalT^{-MCP/-MCP}/CD39/CD73$ pig expressing CD39 and CD73 at endothelial cells.

• Science of Emotion and Sensibility
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• v.23 no.3
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• pp.47-62
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• 2020

The snake detection theory posits that, due to competition with snakes, the primate visual system has been evolved to detect camouflaged snakes. Specifically, one of its hypotheses states that the subcortical visual pathway mainly consisting of koniocellular cells enables humans to automatically detect the threat of snakes without consuming mental resources. Here we tested the hypothesis by comparing human participants' responses to snakes with those to fearful faces and flowers. Participants viewed either original images or converted ones, which lacked the differences in color, luminance, contrast, and spatial frequency energies between categories. While participants in Experiment 1 produced valence and arousal ratings to each image, those in Experiment 2 detected target images in the breaking continuous flash suppression (bCFS) paradigm. As a result, visual factors influenced the responses to snakes most strongly. After minimizing visual differences, snakes were rated as being less negative and less arousing, and detected more slowly from suppression. In contrast, the images of the other categories were less affected by image conversion. In particular, fearful faces were rated as greater threats and detected more quickly than other categories. In addition, for snakes, changes in arousal ratings and those in bCFS response times were negatively correlated: Those snake images, the arousal ratings of which decreased, produced increased detection latency. These findings suggest that the influence of snakes on human responses to threat is limited relative to fearful faces, and that detection responses in bCFS share common processing mechanisms with conscious ratings. In conclusion, the current study calls into question the assumption that snake detection in humans is a product of unconscious subcortical visual processing.

• Korean Journal of Poultry Science
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• v.36 no.3
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• pp.201-206
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• 2009

This study was carried out to investigate dietary effects of illite on humoral immune response against Salmonella typhimurium flagella in Hyline Brown laying hens. Total of twenty eight 36-week-old hens were divided into two groups; The first was fed commercial diet as control and the other was fed diet containing 2 % illite. Each group was divided into 2 sub-groups and then performed immunization of Salmonella flagella with different adjuvants which were Freund's adjuvant and croton oil. The rates of egg-production in all groups were normal range but no difference was found between illite-treated and untreated groups. The serum antibody titers of Freund's adjuvant-used subgroup in illite fed groups were significantly increased, especially from 6-to 9-week after $1^{st}$ immunization compared to those of control groups (p<0.05). And the antibody titers of croton oil-used subgroup in illite-fed groups were also significantly increased, especially at 4-, 6- and 7-week (p<0.05). The results demonstrated that the feeding illite stimulated the immune response against S. typhimurium flagella antigen in laying hens and suggest that the supplementation of illite to the poultry diets may support protective effects against bacterial infections such as Salmonellosis.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.258-264
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• 2020

This study investigated the effect of variation in the number of somatic-cell-cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100-150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.

• Journal of Veterinary Clinics
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• v.25 no.4
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• pp.268-273
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• 2008

For evaluation of immune stimulation effect of lacquer tree extracts, lymphocytes were counted by labeling of FITC-conjugated monoclonal antibody in the pheripheral blood of pigs that fed with a fodder supplemented by lacquer tree extracts. Populations of MHC-II+, CD4+, and CD8+T lymphocytes were increased more than 2% level after 1 week feed supply of the lacquer tree extracts. The increase of those T cells reached at maximum level after 2 weeks in the tested group. B lympyocytes with surface IgM were increased 5% after 1 week feed supply of the lacquer tree extracts, and their numbers reached maximum after 2 weeks in the tested group. For the assessment of cytotoxicity of the lacquer tree extracts, morphological changes were examined on the epithelial cells of small intestine from pigs fed with a fodder supplemented by 0.1 % lacquer tree extracts for 6 weeks (the tested group). Thin-sectioned tissue of small intestine was fixed with glutaraldehyde, then coated with gold particles, and the specimen was examined under scanning electron microscope. The villi on the mucus membrane of jejunum and ileum from the tested pigswere enlarged on the tip and were linked each other.

• Journal of Forest and Environmental Science
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• v.36 no.3
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• pp.175-186
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• 2020

This study approaches, from a floristic perspective, the under-researched human-primate competition for forest resources. Investigating the human impact on fruit trees edible for Kinda baboons (Papio kindae Lönnberg), we have collated dietary data on a free-ranging troop and floristic information on two forest sites of the Kundelungu National Park (KNP), Democratic Republic of Congo: the relatively intact Integral Zone (IZ) and the human-disturbed Annex Zone (AZ). Trees with DBH≥10 cm have been identified, counted and measured throughout 22 sample plots (11 per site), each measuring 1,000 ㎡. A total of seven woody species whose fruits are eaten by Kinda baboons were recorded. Four of them, namely the Sycamore fig Ficus sycomorus L., the Mobola plum Parinari curatellifolia Planch. ex Benth, the Kudu berry Pseudolachnostylis maprouneifolia Pax and the Monkey orange Strychnos innocua Delile were found in both sites, while the Large-leaved jackal-berry Diosyros kirkii Hiern and the Buffalo thorn Ziziphus mucronata Willd. were exclusively in the IZ, and Strychnos cocculoides Baker only in the AZ. Compared to the IZ, the AZ had lower values of stem density, species richness and diversity indices, suggesting a negative human impact on baboon-edible trees, in line with our hypothesis. Moreover, as was expected, human activities decreased the abundance of larger baboon-edible fruit trees. However, the size-class distribution of P. curatellifolia depicted a reverse J-curve in the AZ. The abundant younger P. curatellifolia trees remaining in that human-disturbed site constitute an important food stock for baboons, if well preserved. These results also illustrate the critical role of rangers' patrols, formerly more frequent (and presumably efficient) in the IZ than in the AZ of the Park. Their implications on baboons and miombo forests are discussed from both the research and conservation perspectives.