• Journal of Embryo Transfer
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• v.28 no.1
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• pp.73-78
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• 2013

The present study examined the histological characteristics of adult testis in the long-beaked common dolphin (Delphinus capensis) from Korean waters and the localization of DEAD-box polypeptide 4 (DDX4; a germ cell marker) and vimentin (a Sertoli cell marker) expression in the dolphin testis compared with that in terrestrial mammals, including dogs and rats. The seminiferous tubules of dolphin testis have very small or completely closed lumens, and spermatogenic cells and Sertoli cells within the tubules cannot be differentiated. Immunohistochemical analysis showed that, in the dolphin testis, DDX4- and vimentin-positive cells were scattered extensively within the tubule, whereas in the dog and rat testis, DDX4 immunoreactivity was localized in spermatogenic cells of the adluminal compartment, and vimentin immunoreactivity was localized in Sertoli cells of the basal compartment in the seminiferous epithelium. These results suggest that the histological characteristics of the seminiferous tubules in the dolphin testis differ from those of terrestrial species.

• Journal of Internet Computing and Services
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• v.11 no.4
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• pp.119-128
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• 2010

Computing environment in these days aim for user-oriented development called RIA (Rich Internet Application). As a representative development method of RIA, Flex Framework overcomes the weaknesses of the Mainframe and C/S (Client/Server). However, the issues, such as, difficulties in memory management, complexity of the binding structure and large capacities of the compile outputs are left to be solved. The purpose of this paper is to implement the framework which enables the fast and accurate development of user-oriented web application on the Flex platform. DDX (Dynamic Data eXchange) framework proposes standardized and efficient development environment in a Flex platform. And by using scalability-prepared library that is applicable for various job areas, the framework enhances the performance, increase development productivity and help construct stable system.

• Anatomy and Cell Biology
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• v.56 no.4
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• pp.508-517
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• 2023

In cancer patients, chemo/radio therapy may cause infertility by damaging the spermatogenesis affecting the self-renewal and differentiation of spermatogonial stem cells (SSCs). In vitro differentiation of stem cells especially mesenchymal stem cells (MSCs) into germ cells has recently been proposed as a new strategy for infertility treatment. The aim of this study was to evaluate the proliferation and differentiation of SSCs using their co-culture with Sertoli cells and conditioned medium (CM) from adipose tissue-derived MSCs (AD-MSCs). Testicular tissues were separated from 2-7 days old neonate Wistar Rats and after mechanical and enzymatic digestion, the SSCs and Sertoli cells were isolated and cultured in Dulbecco's modified eagle medium with 10% fetal bovine serum, 1X antibiotic, basic fibroblast growth factor, and glial cell line-derived neurotrophic factor. The cells were treated with the CM from AD-MSCs for 12 days and then the expression level of differentiation-related genes were measured. Also, the expression level of two major spermatogenic markers of DAZL and DDX4 was calculated. Scp3, Dazl, and Prm1 were significantly increased after treatment compared to the control group, whereas no significant difference was observed in Stra8 expression. The immunocytochemistry images showed that DAZL and DDX4 were positive in experimental group comparing with control. Also, western blotting revealed that both DAZL and DDX4 had higher expression in the treated group than the control group, however, no significant difference was observed. In this study, we concluded that the CM obtained from AD-MSCs can be considered as a suitable biological material to induce the differentiation in SSCs.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.253-261
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• 2023

Objective: Azoospermia (the total absence of sperm in the ejaculate) affects approximately 10% of infertile males. Despite diagnostic advances, azoospermia remains the most challenging issue associated with infertility treatment. Our study evaluated transition nuclear protein 2 (TNP2) and synaptonemal complex protein 3 (SYCP3) polymorphisms, azoospermia factor a (AZFa) microdeletion, and gene expression levels in 100 patients with azoospermia. Methods: We investigated a TNP2 single-nucleotide polymorphism through polymerase chain reaction (PCR) restriction fragment length polymorphism analysis using a particular endonuclease. An allele-specific PCR assay for SYCP3 was performed utilizing two forward primers and a common reverse primer in two PCR reactions. Based on the European Academy of Andrology guidelines, AZFa microdeletions were evaluated by multiplex PCR. TNP2, SYCP3, and the AZFa region main gene (DEAD-box helicase 3 and Y-linked [DDX3Y]) expression levels were assessed via quantitative PCR, and receiver operating characteristic curve analysis was used to determine the diagnostic capability of these genes. Results: The TNP2 genotyping and allelic frequency in infertile males did not differ significantly from fertile volunteers. In participants with azoospermia, the allelic frequency of the SYCP3 mutant allele (C allele) was significantly altered. Deletion of sY84 and sY86 was discovered in patients with azoospermia and oligozoospermia. Moreover, SYCP3 and DDX3Y showed decreased expression levels in the azoospermia group, and they exhibited potential as biomarkers for diagnosing azoospermia (area under the curve, 0.722 and 0.720, respectively). Conclusion: These results suggest that reduced SYCP3 and DDX3Y mRNA expression profiles in testicular tissue are associated with a higher likelihood of retrieving spermatozoa in individuals with azoospermia. The homozygous genotype TT of the SYCP3 polymorphism was significantly associated with azoospermia.

• Journal of Veterinary Science
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• v.21 no.1
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• pp.13.1-13.14
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• 2020

Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3+ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.343-348
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• 2013

Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

• Molecular & Cellular Toxicology
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• v.5 no.1
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• pp.58-66
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• 2009

The 2-year rodent carcinogenicity test involves long-term, repetitive dosing of animals that is both time consuming and expensive. Alternative approaches have been attempted using specific transgenic or knockout mice or toxicogenomics to predict carcinogenicity without conducting a 2-year rodent test. In addition, toxicogenomic analysis of carcinogen-treated animals could also enhance our understanding of molecular mechanisms and aid in the diagnosis of acute toxicity induced by carcinogens. Therefore, we investigated transcription profiles after administering the carcinogens 4,4-dimethylformamide (DMF) and 4-biphenylamine (ABP). BALB/c male mice were treated once with DMF (650 mg/kg i.p.) or ABP (120 mg/kg p.o.). Standard blood biochemistry and histological changes were observed. Gene expression profiles in the livers of mice treated with either vehicle or the carcinogens were analyzed using the Affymetrix $GeneChip^{(R)}$ assay. In all, 1,474 differentially expressed genes in DMF- or ABP-treated mice were identified as being either up- or down-regulated over 1.5-fold (P< 0.01), and these genes were analyzed using hierarchical clustering and Ingenuity Pathways Analysis. Of these, 107 genes were consistently regulated in both carcinogen-treated groups. Genes associated with cancer were upregulated (Por, S100a10, Tes, Ctcf, Ddx21, Eapp, Nel, and Pa2g4) or downregulated (Cbs and Gch1). Toxicological function analysis also identified genes involved in organ toxicity, including hepatotoxicity. These data may help to identify molecular markers for acute hepatotoxicity induced by carcinogens.

• Journal of Radiation Protection and Research
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• v.41 no.4
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• pp.344-349
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• 2016

Background: Carbon ion therapy has achieved satisfactory results. However, patients have a risk to get a secondary cancer. In order to estimate the risk, it is essential to understand particle transportation and nuclear reactions in the patient's body. The particle transport Monte Carlo simulation code is a useful tool to understand them. Since the code validation for heavy ion incident reactions is not enough, the experimental data of the elementary reaction processes are needed. Materials and Methods: We measured neutron production double-differential cross-sections (DDXs) on a carbon bombarded with 430 MeV/nucleon carbon beam at PH2 beam line of HIMAC facility in NIRS. Neutrons produced in the target were measured with NE213 liquid organic scintillators located at six angles of 15, 30, 45, 60, 75, and $90^{\circ}$. Results and Discussion: Neutron production double-differential cross-sections for carbon bombarded with 430 MeV/nucleon carbon ions were measured by the time-of-flight method with NE213 liquid organic scintillators at six angles of 15, 30, 45, 60, 75, and $90^{\circ}$. The cross sections were obtained from 1 MeV to several hundred MeV. The experimental data were compared with calculated results obtained by Monte Carlo simulation codes PHITS, Geant4, and FLUKA. Conclusion: PHITS was able to reproduce neutron production for elementary processes of carbon-carbon reaction precisely the best of three codes.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.4
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• pp.189-198
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• 2023

Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2, Gsr, HO-1, and catalase, were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2, HO-1, and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.23 no.1
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• pp.27-44
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• 1993

This study was performed to investigate the changes of mandibular condyle by low calcium diet and the effects of irradiation on the bone in osteoporotic state. In order to carry out this experiment, 80 seven-week old Sprague-Dawley strain rats weighing about 150 gm were selected and equally divided into one experimental group of 40 rats and one control group with the remainder. The experimental group and the control group were then subdivided into two group and exposed to irradiation. The two irradiation groups received a single dose of 20 Gy on the jaw area only and irradiated with a cobalt-60 teletherapy unit. The rats in the control and experimental groups were serially terminated by fours on the 3rd, the 7th, the 14th, and the 21st day after irradiation. After termination, both sides of the dead rats mandibular condyle were removed and fixed with 10% neutral formalin. The bone mineral density of mandibular condyle was measured by use of dual energy X-ray bone densitometer(model DDX-alpha, Lunn Corp., U. S. A.). The mandibular condyle was radiographed with Hitex HA-80(Hitex Co., Japan). Thereafter, the obtained radiographs were observed, and the mandibular condyle was further decalcified and embedded in paraffin as the general method. The specimen sectioned and stained with hematoxylin-eosin, PAS and Rabbit Anti-Human Tumor Necrosis Factor-a observed by a light microscope. The obtained results were as follows: 1. In the non-irradiated group with the low calcium diet, the bone mineral density of the condyle was markedly decreased after 14 days, and decrease the number of trabeculae of the condyle and resorption of the calcified cartilaginous zone were observed after 3 days. On microscopic observations, the number nd size of trabeculae were decreased after 7 days of experiment. 2. In the irradiated group with the low calcium diet, the bone mineral density of the condyle was markedly decreased after 14 days and resorption of the calcified cartilaginous zone and decrease the number and coarse of the trabeculae of the condyle were observed. These findings were extended rather than in non-irradiated group with low calcium diet. On microscopic observations, many osteoclasts were detected and the number and size of trabeculae were somewhat decreased after 7 days. Also there was degenerative changes of tissues of bone marrow on the 14th day but that condition was restored on the 21st day of experiment. 3. In the irradiated group with normal diet, the bone mineral density of the condyle was somewhat decreased with times and degree of decrease of the number of trabeculae was somewhat larger than in the non-irradiated group with normal diet. On microscopic observations, the tissues of bone marrow were atrophic and degenerative changes but that condition was restored on the 21st day of experiment. 4. In immunocytochemical findings, in the irradiated and non-irradiated groups with low calcium diet, negative or partial positive response to TNF was observed, but positive response in the normal diet groups.