• Journal of Embryo Transfer
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• v.24 no.4
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• pp.259-263
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• 2009

Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules and was identified as pheromone-binding protein in pigs. Our previous study has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen in pigs. However, function of SAL1 in the uterus during pregnancy in pigs is still not known. To understand physiological function of SAL1 in the uterine endometrium during pregnancy in pigs, it needs to elucidate the ligand(s) for SAL1. Thus, to identify the ligand for SAL1 in the porcine uterus, we collected uterine luminal fluid from pigs on day 12 of pregnancy by flushing with PBS. Proteins from the uterine luminal fluid were separated by ion exchange chromatography and gel filtration. Fractions containing SAL1 protein were pooled and concentrated. Immunoblot analysis confirmed successful purification of SAL1. Then, we extracted lipids from the purified SAL1 protein and analyzed the lipids by liquid chromatography-mass spectrometry, and predicted to be steroid hormones and prostaglandins as SAL1 ligands. Results in this study showed that SAL1 protein in the uterine secretions has a small lipophilic molecule as a natural ligand. Further characterization of ligand extracted from purified SAL1 will be useful for understanding physiological function of SAL1 during pregnancy and its application to increase the pregnancy rate in pigs.

• The Korean Journal of Physiology and Pharmacology
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• v.27 no.2
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• pp.157-165
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• 2023

Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer and current therapeutic strategies are limited in their effectiveness. The expressions of Rab5 and the M2 tumor-associated macrophage marker CD163 in tissues were detected by Western blot. The migration and invasion of cells were determined using a Transwell assay. The expressions of the exosome markers were evaluated by Western blot. The polarization of human macrophages (THP-1) was determined by incubation of THP-1 cells with conditioned medium or exosomes collected from MDA-MB-231 cells with indicated transfections or by a coculture system of THP-1 and MDA-MB-231 cells. The M1 and M2 macrophage markers were evaluated by qRT-PCR. The expression of Rab5 in TNBC was significantly higher than that in normal breast tissue. Rab5 expressions in triple-negative and luminal A breast cancer were higher than those in other molecular subtypes. Higher CD163 expression was observed in triple-negative breast cancer and in triple-negative and luminal B subtypes. Rab5 knockdown suppressed but Rab5 overexpression promoted the migration and invasion capacity of MDA-MB-231 cells. The levels of CD63 and CD9 in the medium of Rab5 knockdown cells were lower than those in control cells, whereas higher levels of CD63 and CD9 were observed in Rab5 overexpression cells. Rab5 knockdown decreased the excretion but did not alter the diameter of the exosomes. Knockdown of Rab5 facilitated the anti-tumor polarization of macrophages, which was partially reversed by Rab5 overexpression. Therefore, Rab5 is expected to be a potential therapeutic target for triple-negative breast cancer.

• Development and Reproduction
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• v.22 no.4
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• pp.379-391
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• 2018

Through the development of organic synthetic skill, chemicals that mimic signaling mediators such as steroid hormones have been exposed to the environment. Recently, it has become apparent that this circumstance should be further studied in the field of physiology. Estrogenic action of chronic low-dose nonylphenol (NP) and di-(2-ethylhexyl) phthalate (DEHP) in mouse uterus was assessed in this study. Ten to twelve-week-old female mice (CD-1) were fed drinking water containing NP (50 or $500{\mu}g/L$) or DEHP (133 or $1,330{\mu}g/L$) for 10 weeks. Uterine diameter, the thickness of myometrium and endometrium, and the height of luminal epithelial cells were measured and the number of glands were counted. The expression levels of the known $17{\beta}$-estradiol ($E_2$)-regulated genes were evaluated with real-time RT-PCR methodology. The ration of uterine weight to body weight increased in $133{\mu}g/L$ DEHP. Endometrial and myometrial thickness increased in 133 and $1,330{\mu}g/L$ DEHP treated groups, and in 50, $500{\mu}g/L$ NP and $133{\mu}g/L$ DEHP, respectively. The height of luminal epithelial cell decreased in NP groups. The numbers of luminal epithelial gland were decreased in NP groups but increased in $50{\mu}g/L$ DEHP group. The histological characters of glands were not different between groups. The mRNA expression profiles of the known $17{\beta}$-estradiol ($E_2$) downstream genes, Esr1, Esr2, Pgr, Lox, and Muc1, were also different between NP and DEHP groups. The expression levels dramatically increased in some genes by the NP or DEHP. Based on these results, it is suggested that the chronic low-dose NP or DEHP works as estrogen-like messengers in uterus with their own specific gene expression-regulation patterns.

• Applied Microscopy
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• v.15 no.2
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• pp.98-110
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• 1985

This study was performed to clarify the morphological structures of the epithelia of the renal papilla, renal pelvis and ureter of the sheep (Ovis aries L.) through the light and scanning electron microscopes, Tissue specimens were taken from the renal papilla (common renal papilla and peripelvic column) and the renal pelvis (pelvis proper and pelvic pouch) of the kidney and the ureter. For the light microscopy, tissue blocks were fixed in 10 % neutral buffered formalin and embedded in paraffin wax, serially sectioned at a thickness of $6{\mu}m$. These sections were stained with hematoxylin-eosin and periodic acid-Schiff reaction. For the scanning electron microscopy, tissue blocks were prefixed in 1% glutaral-dehyde-1.5% paraformaldehyde solution and postfixed in 1% osmium tetroxide solution, dehydrated in graded alcohol, transferred to isoamyl acetate, and then dried by the critical point dryer (Polaron E 3000). These dried tissues were coated with gold and observed with a scanning electron microscope (JSM-35C), The results were as follows: The apex of the common renal papilla was lined with simple columnar epithelium having many microvilli on its luminal surface. Lateral portion of the papilla was lined with stratified epithelium $2{\sim}3$ layers thick, and its superficial cells were microvillar cells having many microvilli. The epithelium lining the peripelvic column was $1{\sim}2$ layers thick. The superficial layer was made of the microvillar cells, but a few microplica cells were appeared in the region near the pelvic pouch. The epithelium of the pelvic pouch was $1{\sim}2$ layered transitional type, and its superficial cells were microplica cells. The epithelia of the pelvis proper and ureter were $4{\sim}6$ layered transitional type, and their superficial cells were typical facet cells existing many round depressions and ridges of cell membranes of the luminal side.

• Korean Journal of Radiology
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• v.20 no.1
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• pp.94-101
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• 2019

Objective: To investigate the efficacy of motion-correction algorithm (MCA) in improving coronary artery image quality and measurement accuracy using an anthropomorphic dynamic heart phantom and 256-detector row computed tomography (CT) scanner. Materials and Methods: An anthropomorphic dynamic heart phantom was scanned under a static condition and under heart rate (HR) simulation of 50-120 beats per minute (bpm), and the obtained images were reconstructed using conventional algorithm (CA) and MCA. We compared the subjective image quality of coronary arteries using a four-point scale (1, excellent; 2, good; 3, fair; 4, poor) and measurement accuracy using measurement errors of the minimal luminal diameter (MLD) and minimal luminal area (MLA). Results: Compared with CA, MCA significantly improved the subjective image quality at HRs of 110 bpm (1.3 ± 0.3 vs. 1.9 ± 0.8, p = 0.003) and 120 bpm (1.7 ± 0.7 vs. 2.3 ± 0.6, p = 0.006). The measurement error of MLD significantly decreased on using MCA at 110 bpm (11.7 ± 5.9% vs. 18.4 ± 9.4%, p = 0.013) and 120 bpm (10.0 ± 7.3% vs. 25.0 ± 16.5%, p = 0.013). The measurement error of the MLA was also reduced using MCA at 110 bpm (19.2 ± 28.1% vs. 26.4 ± 21.6%, p = 0.028) and 120 bpm (17.9 ± 17.7% vs. 34.8 ± 19.6%, p = 0.018). Conclusion: Motion-correction algorithm can improve the coronary artery image quality and measurement accuracy at a high HR using an anthropomorphic dynamic heart phantom and 256-detector row CT scanner.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.315-320
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• 2014

Purpose: The purpose of this study was to investigate the recurrence pattern and characteristics of patients based on the 2013 St. Gallen surrogate molecular subtypes after breast-conserving surgery (BCS) in Chinese women. Methods: This retrospective analysis included 709 consecutive breast cancer patients undergoing BCS from 1999-2010 at our institution. Five different surrogate subtypes were created using combined expression of the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor-2. Locoregional relapse-free survival (LRRFS), distant metastasis-free survival (DMFS), and disease-free survival (DFS) rates were calculated. Results: The 5-year LRRFS, DMFS, and DFS rates were 90.5%, 88.2%, and 81.5%, respectively. Multivariate analysis revealed that young age, node-positive disease, and HER2 enrichment were independent prognostic factors in LRRFS patients. There was also an independent prognostic role of lymph node-positive disease in DMFS and DFS patients. Patients with luminal A tumors had the most favorable prognosis, with LRRFS, DMFS, and DFS rates of 93.2%, 91.5%, and 87.4% at 5 years, respectively. Conversely, HER-2-enriched tumors exhibited the highest rate of locoregional recurrence (20.6%). Conclusion: Surrogate subtypes present with significant differences in RFS, DMFS, and LRRFS. Luminal A tumors have the best prognosis, whereas HER2-enriched tumors have the poorest.

• Journal of Animal Science and Technology
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• v.63 no.2
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• pp.262-271
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• 2021

Communication among epididymal epithelial cells creates the best luminal condition where spermatozoa mature, transport and are stored. Vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) have been used as signal indicators for clear and basal cells of the epididymal epithelium, respectively, in mice, rats, bats, and pigs; however, these two markers have not yet been described in the epididymis of bulls. Here, we examined the presence and distribution of the B1 subunit of V-ATPase (B1-VATPase) and KRT5 in the distinct regions of adult bovine epididymides, specifically, the caput, corpus, and cauda. Immunofluorescence staining and confocal microscopy showed that narrow shaped-clear cells were placed in the caput and corpus regions of the bovine epididymis; however, they were absent in the cauda epididymis. In addition, B1-VATPase was highly expressed in the cauda spermatozoa; however, it was rarely detected in the caput spermatozoa. On the other hand, KRT5-positive cells, basal cells, were maintained beneath the basal lamina and they had the traditional form with a dome-shaped morphology from the caput to cauda region of the bovine epididymis. The co-expression of B1-VATPase and KRT5 was confined to basal cells placed in the basal region of the epithelium. In summary, 1) clear cells were present with region-specific localization, 2) B1-VATPase was present in the corpus and cauda spermatozoa but absent in the caput, 3) co-expressed cells with B1-VATPase and KRT5 were present in the adult bovine epididymis, and 4) B1-VATPase was not a specific marker for clear cells in the bovine epididymis. Therefore, the perfect epididymal luminal condition created by the specific expression and localization patterns of B1-VATPase might be necessary to obtain fertilizing capacity of spermatozoa in the bovine epididymis.

• Applied Microscopy
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• v.46 no.4
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• pp.193-200
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• 2016

Male reproductive organs are strongly affected by estrogen signaling mediated via the estrogen receptor. In this study, propyl pyrazole triol (PPT), an estrogen receptor alpha agonist, was subcutaneously injected in adult male mice every 2 days for a total duration of 8, 16, or 24 days. Histological changes in the reproductive organs, including the testes, efferent ductules, and epididymides, were observed. The weight of the reproductive organs decreased in the PPT group. In addition, the diameter of the seminiferous tubules decreased in the PPT group compared with the control group. The epithelial cell height decreased in the initial segment of the epididymis, whereas the luminal diameter increased in the efferent ductules of the PPT group. PPT induced irregular morphology of stereocilia in the luminal region of the initial segment. Therefore, PPT treatment at high concentrations had inhibitory effects in the reproductive organs of adult male mice. These findings suggest that short-term treatment with estrogen receptor agonist causes histological changes in the testes, efferent ductules, and epididymis, which are similar to those caused by estrogen receptor antagonist treatment. Therefore, the estrogen receptor may have functional roles in male reproductive organs, implying that treatment with an estrogen receptor agonist can affect male fertility.

• Development and Reproduction
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• v.23 no.3
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• pp.263-275
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• 2019

Based on our preliminary results, we examined the possible role of low-dose and chronic-exposing of the chemicals those are known as endocrine disrupting chemical (EDC), on the proliferation of uterine endometrium and the localization of steroid receptors. Immunohistochemical or immunofluorochemical methodology were employed to evaluate the localization of antigen identified by monoclonal antibody Ki 67 protein (MKI67), estrogen receptor 1 (ESR1), estrogen receptor 2 (ESR2), and progesterone receptor (PGR). In $133{\mu}g/L$ and $1,330{\mu}g/L$ di(2-ethylhexyl) phthalate (DEHP) and $50{\mu}g/L$ nonylphenol (NP) groups, the ratio of MKI67 positive stromal cells was significantly increased but not in $500{\mu}g/L$ NP group. The ratios of MKI67 positive glandular and luminal epithelial cells were also changed by the chronic administration of NP and DEHP in tissue with dose specific manner. ESR1 signals were localized in nucleus in glandular and luminal epithelia of control group but its localization was mainly in cytoplasm in DEHP and NP administered groups. On the other hand, it was decreased at nucleus of stromal cells in $1,330{\mu}g/L$ DEHP group. The colocalization patterns of these nuclear receptors were also modified by the administration of these chemicals. Such a tissue specific and dose specific localization of ESR2 and PGR were detected as ESR1 in all the uterine endometrial tissues. These results show that the chronic lows-dose exposing of NP or DEHP modify the localization and colocalization of ESRs and PGR, and of the proliferation patterns of the endometrial tissues.

• Journal of Animal Science and Technology
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• v.64 no.3
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• pp.500-514
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• 2022

The blood-epididymis barrier (BEB) forms a unique microenvironment that is crucial for the maturation, protection, transport, and storage of spermatozoa in the epididymis. To characterize the function of tight junctions (TJs), which are constitutive components of the BEB, we determined the expression and localization of TJ proteins such as zonula occludens (ZO)-1, 2, and 3, occludin, and claudin3 (Cldn3) during postnatal development in the goat epididymis. To assess the expression patterns of TJ proteins in immature (3 months of age) and mature (14 months of age) goat epididymides, two different experimental methods were used including immunofluorescence labeling and western blotting. We show that, ZO-1, 2, and 3, and occludin, were strictly expressed and localized to the TJs of the goat epididymis, whereas Cldn3 was present in basolateral membranes as well as TJs. All TJ proteins examined were more highly expressed in the immature epididymis compared to levels in mature tissue. In conclusion, our study indicates that at least five TJ proteins, namely ZO-1, ZO-2, ZO-3, occludin, and Cldn3, are present in TJs, and the expression strength and pattern of TJ proteins tend to be age dependent in the goat epididymis. Together, these data suggest that the distinct expression patterns of TJ proteins are essential for regulating components of the luminal contents in the epididymal epithelium and for forming adequate luminal conditions that are necessary for the maturation, protection, transport, and storage of spermatozoa in the goat epididymis.