• Biomolecules & Therapeutics
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• v.31 no.6
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• pp.629-639
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• 2023

Cardiovascular diseases (CVDs) are the most common cardiovascular system disorders. Cellular senescence is a key mechanism associated with dysfunction of aged vascular endothelium. Hyaluronic acid and proteoglycan link protein 1 (HAPLN1) has been known to non-covalently link hyaluronic acid (HA) and proteoglycans (PGs), and forms and stabilizes HAPLN1-containing aggregates as a major component of extracellular matrix. Our previous study showed that serum levels of HAPLN1 decrease with aging. Here, we found that the HAPLN1 gene expression was reduced in senescent human umbilical vein endothelial cells (HUVECs). Moreover, a recombinant human HAPLN1 (rhHAPLN1) decreased the activity of senescence-associated β-gal and inhibited the production of senescence-associated secretory phenotypes, including IL-1β, CCL2, and IL-6. rhHAPLN1 also downregulated IL-17A levels, which is known to play a key role in vascular endothelial senescence. In addition, rhHAPLN1 protected senescent HUVECs from oxidative stress by reducing cellular reactive oxygen species levels, thus promoting the function and survival of HUVECs and leading to cellular proliferation, migration, and angiogenesis. We also found that rhHAPLN1 not only increases the sirtuin 1 (SIRT1) levels, but also reduces the cellular senescence markers levels, such as p53, p21, and p16. Taken together, our data indicate that rhHAPLN1 delays or inhibits the endothelial senescence induced by various aging factors, such as replicative, IL-17A, and oxidative stress-induced senescence, thus suggesting that rhHAPLN1 may be a promising therapeutic for CVD and atherosclerosis.

• Applied Microscopy
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• v.38 no.1
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• pp.21-28
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• 2008

Carnivorous plants vary in their unique features of morphology, ultrastructure and biochemical properties by species. Furthermore, prey-capturing mechanism as well as structural and physiological adaptations have been used for grouping various carnivorous species. In Drosera plants, glandular trichomes, which develop in the leaf epidermis, are known to play the most important role during the prey capturing process. The present study examined such trichomes, focusing on the glandular type, in leaves of Drosera anglica using scanning and transmission electron microscopy. Three types of rudimentary glandular trichomes were found to develop within the folded leaf primordia and immature leaf during early development. The first type, stalked glandular trichomes (Type I), occurred on the margin and upper epidermis of the leaf. With maturation, the longest glandular trichomes having lengthy stalks, ca. $2.2{\sim}5.1\;mm$, developed along the margin, while shorter stalked trichomes, ca. up to $200\;{\mu}m$, were found on the inner leaf blade. The shorter ones consisted of a globose head having two layers of secretory cells, parenchyma bell cells and tracheids and a multicellular stalk. The stalks gradually decreased in length in centripetal fashion. The second type, Type II, having ca. $15{\sim}30\;{\mu}m$ short stalks, also developed along the inner blade. Both types secreted mucilage from the secretory cells which had a thin cell wall and cuticle layer. The sessile six-celled glandular trichomes were the third type, Type III, and were $25{\sim}40\;{\mu}m$ in length. They were distributed most commonly throughout the upper and lower epidermis, petiole and even on the stalk surfaces of the first two types of trichomes. The third type was also found to be involved in the active secretion. In prey capturing leaves, all trichome types secreted substances through thin cuticles in the head cell wall, which exhibited relatively loose wall components.

• Near Infrared Analysis
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• v.2 no.1
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• pp.59-66
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• 2001

Nowadays, medical diagnostics is efficiently supported by clinical chemistry and near infrared spectroscopy is becoming a new dimension, which has shown high potential to provide valuable information for diagnosis. The investigation was carried out to study the influence of mammary gland inflammation, called mastitis, on cow´s milk spectra and milk composition measured by near infrared spectroscopy (NIRS). Milk somatic cell counts (SCC) in milk were used as a measure of mammary gland inflammation. Naturally occurred variations with milk composition within lactation and in the process of milking were included in the experimental design of this study. Time series of unhomogenized, raw milk spectral data were collected from 3 cow along morning and evening milking, for 5 consecutive months, within their second lactation. In the time of the trial, the investigated cows had periods with mammary gland inflammation. Transmittance spectra of 258 milk samples were obtained by NIRSystem 6500 spectrophotometer in 1100-2400 nm region. Calibration equations for the examined milk components were developed by PLS regression using 3 different sets of samples: samples with low somatic cell count (SCC), samples with high SCC and combined data set. The NIR calibration and prediction of individual cow´s milk fat, protein, and lactose were highly influenced by the presence of mil samples from animals with mammary gland inflammation in the data set. The best accuracy of prediction (i.e. the lower SEP and the higher correlation coefficient) for fat, protein and lactose was obtained for equations, developed when using only “healthy” samples, with low SCC. The standard error of prediction increased and correlation coefficient decreased significantly when equations for low SCC milk were used to predict examined components in “mastitis” samples with high SCC, and vice versa. Combined data set that included samples from healthy and mastitis animals could be used to build up regression models for screening. Further use of separate model for healthy samples improved milk composition measurement. Regression vectors for NIR mild protein measurement obtained for “healthy” and “mastitic” group were compared and revealed differences in 1390-1450 nm, 1500-1740 nm and 1900-2200 nm regions and thus illustrated post-secretory breakdown of milk proteins by hydrolytic enzymes that occurred with mastitis. For the first time it has been found that monitoring the spectral differences in water bands at 1440 nm and 1912 nm could provide valuable information for inflammation diagnosis.

• Journal of Life Science
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• v.24 no.10
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• pp.1070-1077
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• 2014

In this study, the estrogen receptor agonist propyl pyrazole triol (PPT), which has high-affinity with the estrogen receptor alpha, was subcutaneously injected into adult male mice every 2 days for 8, 16 and 24 days, after which histological changes in accessory genital glands, including the prostate and seminal vesicle, and the liver were observed. The body and genital gland weights decreased in the PPT group relative to those of the control group. However, the liver weight was two times greater in the PPT group. The luminal area of the prostate and seminal vesicle organs was lower in the PPT group, and the epithelial cell height of the prostate was increased relative to that of the control. There were many secretory vacuoles in the supranuclear cytoplasm of epithelial cells in the seminal vesicles of the control group, but these were not observed in the PPT group. The short sinusoidal diameter of the liver was 147.0%, 198.7%, and 223.3% greater in the PPT group than in the control group after 8, 16, and 24 days of treatment, respectively. These results suggest that PPT administration affected the reproductive organs and the liver and that the histological changes increased in accordance with a rise in the concentration of PPT. Overall, the PPT treatment caused changes in the epithelial cell height and resulted in atrophy of the luminal area of the prostate, leading to altered fertility. The sinusoidal diameter of the liver dramatically increased in response to the administration of PPT, increasing the liver weight.

• Parasites, Hosts and Diseases
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• v.30 no.4
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• pp.309-322
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• 1992

In order to observe the antigenic localization in the tissues of Metngonimus yokogawai in growth stages, immunogoldlabeling method was applied to using serum of the cat which Infected with isolated metacercariae from Plecoglossus aztivelis. The sectioned worm tissues from each growth stages were embedded in Lowicryl HM 20 medium, stained with infected serum IgG and protein A gold complect (particle size: 12 nm) and observed by electron microscopy. In the worm tissues of all experimental groups, the geld particles were specifically concentrated on the tegumental synch- tium and cytoplasm of the tegumental cell as well as the secretory granules in the parenchymal tissue. In the 16th and 20th week grown worm tissues, the gold particles were specifically concentrated on the vesicles in the tegumental syncytium and cl·toplasm of the tegumental cell. The gold particles were specifically concentrated on the caecal epithelia of the 4th, 8th and 12th week growth groups but slightly concentrated on those of the 16th and 20th week.

• Korean Journal of Veterinary Research
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• v.43 no.4
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• pp.541-549
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• 2003

The present study investigated the effects of aging on Leydig cells of Sprague Dawley rats. Rats of 3, 6, 12 and 18 months of age were used. Testes of rat were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in epon-araldite. Using $1{\mu}m$ sections stained with methylene blue, qualitative and quantitative morphological studies were performed. Testis incubations were used to determine luteinizing hormone (LH; 100 ng/ml) stimulated testosterone secretory capacity per testis in vitro. Testosterone levels in the incubation medium, and testosterone and luteinizing hormone levels in serum of these four groups of rats were determined by radioimmunoassay. Morphological studies revealed that Leydig cells were more abundant in the testis interstitium at 6, 12 and 18 months when compared with 3 months. The volumes of Leydig cells per testis was significantly higher, at 6, 12 and 18 months of age than those at 3 months. The number of Leydig cells per testis was doubled at 6, 12 and 18 months of age compared with 3 months. The average volume of a Leydig cell was not significantly different between 3 and 6 months of age, however, at 12 and 18 months a significantly lower value was observed. LH-stimulated testosterone production per testis in vitro was reduced by 45% at 6 months of age compared with 3 months; a further significant reduction was observed at 12 and 18 months. Serum testosterone and LH levels were not significantly different between 3 and 6 months of age but at 12 and 18 months a significantly lower value was observed in both groups for these hormones. These results showed that signs of aging are apparent in Leydig cells of Sprague Dawley rats at 12 months of age.

• Korean Journal of Veterinary Research
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• v.32 no.4
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• pp.497-510
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• 1992

This study was attempted to comparative investigate the types and regional distribution of the endocrine cells in several vertebrates immunohistochemically using seven antisera. From carp pancreas could be observed 4 types which are insulin-, glucagon-, som- and BPP-immunoreactive cells. Insulin-immunoreactive cells were mainly distributed at the periphery and a few cells occupied the central region of the islets. Glucagon-immunoreactive cells were distributed at the periphery of the islets, and som - and BPP-immunoreactive cells were located at the central region. From frog pancreas could be observed 4 types which are insulin-, glucagon-, som- and BPP-immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the islets. Som-immunoreactive cells were distributed at the periphery of the islets, and glucagon- and BPP-immunoreactive cells were found as single cell or as small groups located between the pancreatic acini. From snake pancreas could be observed 3 types which are insulin-, glucagon- and som -immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the small islets, and they also were scattered at the periphery of the large islets. Glucagon-immunoreactive cells were distributed at the periphery of the islets, whereas som-immunoreactive cells were occupied the central region. From Ogolgae pancreas could be observed 4 types which are insulin-, glucagon-, som-and BPP-immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the small islets, but at the periphery of the large one. Glucagon- immunoreactive cells were distributed at the periphery of the small islets and in the large islets showed scattering entired. Som-immunoreactive cells were distributed at the periphery of the small islets and in the large islets were located at the central region. A small numbers of BPP-immunoreactive cells were located at the periphery of the small islets and the exocrine regions. From the pancreas of the Korean native goat could be observed 6 types which are insulin-, glucagon-, som-, BPP-, 5-HT- and porcine-CG-immunoreactive cells. Insulin-immunoreactive cells were distributed throughout the islets. Som-immunoreactive cells were located at the periphery of the islets, but a tew were scattered at the central region of islets and in the epithelium of the secretory duct. Glucagon-, BPP-, 5-HT- and porcine CG-immunoreactive cells were distributed at the periphery of the islets. These findings indicated that the regional distribution patterns and cell types of pancreatic endocrine cells in vertebrates varies considerably among phylogenetically different vertebrates.

• Applied Microscopy
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• v.36 no.4
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• pp.271-277
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• 2006

Mast cells (MCs) are granulated cells that play a pivotal role in allergic reaction and inflammation. The granules of mast cells are known to be rich in zinc (Zn). Male Sprague-Dawley rats were used. We injected $200{\mu}L$ of complete Freund's adjuvant (CFA) subcutaneously in the dorsal aspect of one hindpaw Finally, zinc selenium autometallography(AMG) was done by Danscher's method. The present study showed the ultrastructures of zinc-containing mast cells found in inflammatory area following an complete freund's adjuvant (CFA) inoculation into the rat hindpaw. At light microscopic level, mast cells were round or oval, at average $12{\mu}m$ in diameter, with many filopodia extending from the cell surface. Because the rather small and spherical nucleus was centrally placed; it was frequently obscured by the cytoplasmic granules, it sometimes could not be seen. Mast cells were distributed chiefly in the vicinity of small blood vessels. In most preparation many mast cells were ruptured and their granules escaped into the surrounding tissue. In electron micrographs, The secretory granules were at average $0.5{\mu}m$ in diameter and were limited by a membrane. The cell surface contained numerous microvilli and folds. Their interior was heterogenous in appearance. The nucleus was surrounded by large numbers of prominent vesicels and a well developed Golgi apparatus, but scant endoplasmic reticulum.

• Applied Microscopy
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• v.22 no.2
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• pp.1-14
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• 1992

This experiment was performed to study the morphological responses of the parafollicular cells of rat following X-ray irradiation. Male rats were divided into normal and experimental groups. The head and neck region of the rat, under sodium thiopental anesthesia, was exposed to 3,000 rads or 6,000 rads of radiation in a single dose, respectively. The source was a Mitsubishi Linear Accelerator ML-4MV. The target to skin distance was 80 cm, and the dose rate was 200 rads/min. The rate of experimental groups were sacrificed on the 6th hour, 2nd and 6th day after X-ray irradiation. Pieces of the tissue taken from the thyroid gland were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde (0.1M Millonig's phosphate buffer, pH 7.3), and in 1% osmium tetroxide (0.1M Millonig's phosphate buffer, pH 7.3), and embedded in araldite mixture. The ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100 CX-II electron microscope. The results were as follow; 1. Two types of the parafollicular cells, according to their electron densities, were found, i. e., light cells and dark cells. 2. Three types of the parafollicular cells, according to their sizes of secretory granules were found, i.e., small granule cells ($85nm{\pm}20.1;64{\sim}102nm$), medium granule cells ($120nm{\pm}26.5;77{\sim}179nm$), and large granule cells ($165nm{\pm}25.7;128{\sim}189nm$). 3. The differential ultrastructural changes of the cells according to their cell types, i.e., dark and light cell, or small, medium and large granule cells, were hardly observed in the time and dose range covered by this study. 4. The morphological changes of the parafollicular cells were not pronounced after exposure to 3,000 rads of X-ray. 5. Swollen cisternae of the granular endoplasmic reticulum and partial cytolysis were observed after exposure to 6,000 rads of X-ray. 6. Above results suggest that the parafollicular cells showed the alterations of mitochondrial and granular endoplasmic reticular swelling, and partial cytolysis, but only in doses of 6,000 rads.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.54-54
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• 2003

Prostate gland contains neuroendocrine cells (PNECs) are playing important roles in physiological and pathophysiological processes of the prostate gland. Here, we investigated the role of purinoceptors in PNECs freshly isolated from rat ventral prostate (RPNECs) that show immunoreactivity to chromogranin A. Fura-2 ratiometry revealed that ATP evokes both fast Ca$\^$2+/ influx and store Ca$\^$2+/ release in RPNECs. A whole-cell patch clamp study demonstrated fast inactivating cationic current activated by ATP or by ${\alpha}$,${\beta}$-MeATP, which was blocked by ATP-TNP. The activation of P2X inward current was tightly associated with a sharp increase in [Ca$\^$2+/]$\sub$c/. The presence of P2X1/3 subtypes were proved by RT-PCR analysis. For the stored Ca$\^$2+/ release, ATP and UTP showed similar effects, suggesting the dominant role or P2Y2 subtypes, also confirmed by RT-PCR. Both P2X (${\alpha}$,${\beta}$-MeATP) and P2Y (UTP) stimulation induced changes in the cell morphology (initial shrinkage and blob formation on the surface) reversibly. Exocytotic membrane trafficking events were monitored with the membrane-bound fluorescent dye, FM1-43 using confocal microscopy. In spite of the similar Ca$\^$2+/ responses, UTP was far less effective in triggering exocytosis than ${\alpha}$,${\beta}$ -MeATP. Since serotonin is reportedly stored in the secretory granule of PNECs, we directly examined whether the aforementioned agonists elicit release of serotonin using carbon fiber electrode-amperometry. In accordance with the results of FM1 -43 experiments, ${\alpha}$,${\beta}$-MeATP efficiently evoke serotonin secretion while not with UTP. In summary, the P2X-mediated Ca$\^$2+/ influx plays crucial roles in the exocytosis of RPNECs. Although a global increase in [Ca$\^$2+]$\sub$c/ might be related with the morphological changes, a sharp rise of [Ca$\^$2+/]$\sub$c/ in the putative sub-plasmalemmal ‘microdomains’ might be a decisive factor for the exocytosis.