Objectives : The increase of osteoclasts could cause osteoporosis and bone-related diseases. Also, the inhibition of osteoclast differentiation is important in treating bone-related diseases. Traditionally, Psoraleae Semen has been used for geriatric diseases, aging and musculoskeletal diseases. The purpose of this study is to investigate the effect of Psoraleae Semen ethanol extract (PS) on osteoclast differentiation and its function. Methods : To confirm the effect of PS on osteoclastogenesis and bone resorption activity, various levels of concentrations of PS (5, 10, 20 and 40 ㎍/ml) were tested on RAW 264.7 cells cultured with RANKL. We measured tartarate-resistant acid phosphatase (TRAP)-positive cells, TRAP activity, pit formation and F-actin ring formation. The expressions of nuclear factor of activated T-cells (NFATc1) and c-Fos were confirmed through western blot and reverse transcription- polymerase chain reaction (RT-PCR). Also, the expression of bone resorption and fusion-related genes in osteoclast was confirmed by RT-PCR. Results : PS decreased the number of TRAP-positive cells and the TRAP activity. In addition, PS significantly inhibited the formation of pit and F-actin ring. Furthermore, PS decreased the expression of osteoclast related genes. Conclusions : PS inhibits osteoclast differentiation and bone resorption ability through inhibition of the expression of osteoclast-related genes. This indicates that PS may be a potential therapeutic agent to osteoporosis by suppressing osteoclastogenesis.
The approval of use of certain food-grade phosphates as food additives in a wide variety of meat products greatly stimulated research on the applications of phosphates in foods. Although phosphates have never been classified as antimicrobial agents, a number of investigators have reported that phosphates have antimicrobial activities. Phytic acid is a natural plant inositol hexaphosphate constituting 1-5% of most cereals, nuts, legumes, oil seeds, pollen, and spores. In this study, we investigated antibacterial activities of sodium phytate(SPT), sodium pyrophosphate (SPP), sodium tripolyphosphate (STPP) on Escherichia coli O157:H7 on tryptic soy broth and in beef, pork and chicken. In tryptic soy broth, SPT, SPP and STPP at the concentrations of 0.05, 0.1, and 0.5% effectively inhibited the growth of Escherichia coli O157:H7 in a concentration-dependent manner. The bactericidal activity of SPT was the stronger than that of SPP or STPP at the same concentrations. In addition, the antibacterial effects of SPT, SPP and STPP at the concentrations of 0.05, 0.1, 0.3, and 0.5% on Escherichia coli O157:H7 were also investigated in raw or cooked meats including beef, pork and chicken. SPT, SPP and STPP significantly inhibited the bacterial growth in a dose-dependant manner (p<0.05). The bactericidal effect of SPT was stronger than that of SPP or STPP. The addition of SPT, SPP and STPP in meats increased meat pHs. SPP and STPP also increased the levels of soluble orthophosphate in meats but STP did not. These results indicate that SPT is very effective for inhibition of bacterial growth and that can be used as a muscle food additive for increasing functions of meats.
This study was made for the better information of the male reproductive system on the meat-type drake, Cherry Belly X White Golden. The epithelium of ductules of epididymal region and deferent duct were observed histologically and histochemically with the progress of their development. India-ink absorbability on the luminal epithelium was also investigated after the administration of India-ink. The results are as follows; 1. Rete testis and various round ductules in immature form appeared in epididymis within 6 weeks after hatching, and simple cuboidal and simple columnar epithelium were found in the epithelia of the ductules within 8 weeks after hatching. Larger ductules were found on epididymal surface which was in the developing stage near to the immature efferent ductule. From 10th to 20th week, various ductules appeared in epididymis, and developing form of efferent ductules were much more increased on epididymal surface. The luminal epithelium of the ductules were composed of ciliated simple columnar and pseudostratified ciliated columnar cells. At the same time, deferent duct appeared. From the 21th week, various ductules in epididymis became abruptly matured. Lumen of rete testis was lined by simple squamous or simple cuboidal epithelium, and that of efferent ductules, having many folds and being larger than any others were lined by pseudostratified ciliated columnar epithelium in which ciliated columnar cells, non-ciliated cells(clear cells) and basal cells were noted. Connecting tubules of star shaped lumen were composed of pseudostratified ciliated columnar epithelium in which ciliated columnar cells, nonciliated cells, and basal cells were observed. The luminal surface of epididymal ducts was smooth and has thick pseudostratified columnar epithelium which was composed of high columnar cells and basal cells. From 26th week after hatching, sperm pooling was started in various ductules. 2. From 4th to 10th week, simple cuboidal epithelium of deferent duct transformed to simple columnar epithelium with the progress of aging. At the basement of epithelium, clear round cells were noted. From 12th to 20th week, high columnar cells with enlongated nucleus were noted on the luminal border of deferent ducts, forming folds of pseuclostratified columnar epithelium. From 20th week, the deferent duct started to have septa in it's lumen and composed mainly of pseudostratified columnar epithelium, and round cells disappeared. From 20th week, the lumen diameter of deferent duct became wider with the progress of aging, but there was no difference among the values of lumen diameter in upper, middle, and lower part of deferent ducts. At 26th week, the pooling period of sperms in deferent ducts, the lumen diameter became rapidly widen, especially in the lower part of deferent ducts. Thickness of muscular layer of ductus deferens showed gradual growth within 24 weeks but did abrupt thickening from 26th week. 3. Saliva resistant PAS granules were dotted on the top of nucleus in efferent ductules epithelium but the amount of the granules were little in the connecting ductules's epithelium. The granules reactive to acid phosphatase were abundant in the some epithelial cells of efferent ductules and connecting ductules, especially above the nucleus of cells. The granules reactive to alkaline phosphatase were noted on the luminal border of efferent ductules. Parts of free border of efferent ductules and middle portion of deferent ducts were stained slightly by alcian blue technique. India ink granules were found mainly in the epithelium of efferent ductules but were few in that of connecting ductules.
Objective: Orthodontic root resorption (ORR) due to orthodontic tooth movement is a difficult treatment-related adverse event. Caspases are important effector molecules for apoptosis. At present, little is known about the mechanisms underlying ORR and apoptosis in the cementum. The aim of the present in vivo study was to investigate the expression of tartrate-resistant acid phosphatase (TRAP), caspase 3, caspase 8, and receptor activator of nuclear factor kappa-B ligand (RANKL) in the cementum in response to a heavy or an optimum orthodontic force. Methods: The maxillary molars of male Wistar rats were subjected to an orthodontic force of 10 g or 50 g using a closed coil spring. The rats were sacrificed each experimental period on days 1, 3, 5, and 7 after orthodontic force application. And the rats were subjected to histopathological and immunohistochemical analyses. Results: On day 7 for the 50-g group, hematoxylin and eosin staining revealed numerous root resorption lacunae with odontoclasts on the root, while immunohistochemistry showed increased TRAP- and RANKL-positive cells. Caspase 3- and caspase 8-positive cells were increased on the cementum surfaces in the 50-g group on days 3 and 5. Moreover, the number of caspase 3- and caspase 8-positive cells and RANKL-positive cells was significantly higher in the 50-g group than in the 10-g group. Conclusions: In our rat model, ORR occurred after apoptosis was induced in the cementum by a heavy orthodontic force. These findings suggest that apoptosis of cementoblasts is involved in ORR.
This study was conducted to investigate the dietary supplementation of fruiting body of Hericium erinaceus (HE) mushroom on lipid profiles of serum and histological changes of the liver in rats with high fat and cholesterol diet. Five-week old female Sprague-Dawley albino rats were divided into three groups of 8 rats each: The normal control diet (NC group), high fat and cholesterol diet (HFC group), and HFC diet supplemented with 5% fruiting powder of Hericium erinaceus (HFC+HE group). In the HFC+HE group, serum total cholesterol, low density lipoprotein, and triglyceride concentrations were significantly reduced compared with the NC group. Body weight gain of those in the HFC+HE group were lower than those in the HFC group; whereas HFC+HE had no effect on the levels of plasma albumin, creatinine, blood urea nitrogen, uric acid, glucose, and total protein. The enzyme activities related to the liver function, such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), and, alkaline phosphatase (ALP), were lower in the NC group than in the HFC group, but without significance. Feeding the mushroom increased the excretion of total lipid and cholesterol. A histopathological analysis showed that the those in the HFC group developed hepatic steatosis, whereas those in the HFC+HE group developed small fat droplet. In conclusion, these results suggest that 5% HE supplementation to HFC diet provided health benefits by acting on lowering atherogenic lipid profile in rats with high fat and cholesterol diet.
Baek, Jong Min;Kim, Ju Young;Lee, Myeung Su;Jeung, Woo Jin;Moon, Seo Young;Jeon, Byung Hoon;Oh, Jae Min;Choi, Min Kyu
Journal of Physiology & Pathology in Korean Medicine
/
v.27
no.4
/
pp.431-436
/
2013
Bone homeostasis is maintained by co-ordination of bone-resorbing osteoclasts and bone-forming osteoblasts. Imbalance between osteoclasts and osteoblasts leads to many bone diseases such as osteoporosis, rheumatoid arthritis. Taxillus chinensis is a herb that has been widely used to improve bone health. However, the effect and mechanism of Taxillus chinensis extract on osteoclast differentiation and bone resportion has been unknown. Thus, We investigated the effect of Taxillus chinensis on expression of receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation and bone resorption. Also, the action of Taxillus chinensis on mechanisms relating to osteoclast differentiation was studied. In this results, we identified that Taxillus chinensis significantly inhibited RANKL-induced osteoclast differentiation and bone resportion. Moreover, Taxillus chinensis was suppressed the activation of NF-${\kappa}B$ in bone marrow macrophage treated RANKL and M-CSF. Taxillus chinensis was down-regulated the mRNA expression of c-Fos, nuclear factor of activated T-cells (NFAT)c1, osteoclast-associated receptor (OSCAR), tartrate-resistant acid phosphatase (TRAP). The cell adhesion-related molecules such as integrin ${\alpha}v$ and integrin ${\beta}3$, and the filamentous actin (F-actin) rings of mature osteoclasts-related molecules such as dendritic cell-specific transmembrane preotein (DC-STAMP) and cathepsin K are also suppressed. Taken together, these results indicated that Taxillus chinensis will be a good candidate to treat osteoclast-mediated bone diseases.
Park, Bong-Wook;Lee, Seong-Gyun;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Uk-Kyu;Kim, Jong-Ryoul;Byun, June-Ho
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.35
no.5
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pp.294-298
/
2009
Purpose: The development of a microvascularization is important for the homeostasis of normal bone. Vascular endothelial growth factor (VEGF) is one of the most important factors in vessel formation. The purpose of this study was to examine VEGF-related autocrine growth in periosteal-derived cells. Materials and methods: Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured for 21 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and $\beta$-glycerophosphate. Results: The expression of four VEGF isoforms and VEGFRs was observed in periosteal-derived cells. Treatment with cultures with VEGFR-1 and VEGFR-2 Kinase Inhibitor inhibited osteoblastic differentiation and alkaline phosphatase (ALP) activity of periosteal-derived cells. In addition, exogenous VEGF treatment increased calcium content in the periosteal-derived cells. Conclusion: These results suggest that VEGF might act as an autocrine growth molecule during osteoblastic differentiation of cultured human periosteal-derived cells.
Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.
Yadav, Brijesh;Singh, Gyanendra;Wankar, Alok;Dutta, N.;Chaturvedi, V.B.;Verma, Med Ram
Asian-Australasian Journal of Animal Sciences
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v.29
no.11
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pp.1585-1592
/
2016
The present experiment was conducted to evaluate the effect of simulated heat stress on digestibility and methane ($CH_4$) emission. Four non-lactating crossbred cattle were exposed to $25^{\circ}C$, $30^{\circ}C$, $35^{\circ}C$, and $40^{\circ}C$ temperature with a relative humidity of 40% to 50% in a climatic chamber from 10:00 hours to 15:00 hours every day for 27 days. The physiological responses were recorded at 15:00 hours every day. The blood samples were collected at 15:00 hours on 1st, 6th, 11th, 16th, and 21st days and serum was collected for biochemical analysis. After 21 days, fecal and feed samples were collected continuously for six days for the estimation of digestibility. In the last 48 hours gas samples were collected continuously to estimate $CH_4$ emission. Heat stress in experimental animals at $35^{\circ}C$ and $40^{\circ}C$ was evident from an alteration (p<0.05) in rectal temperature, respiratory rate, pulse rate, water intake and serum thyroxin levels. The serum lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase activity and protein, urea, creatinine and triglyceride concentration changed (p<0.05), and body weight of the animals decreased (p<0.05) after temperature exposure at $40^{\circ}C$. The dry matter intake (DMI) was lower (p<0.05) at $40^{\circ}C$ exposure. The dry matter and neutral detergent fibre digestibilities were higher (p<0.05) at $35^{\circ}C$ compared to $25^{\circ}C$ and $30^{\circ}C$ exposure whereas, organic matter (OM) and acid detergent fibre digestibilities were higher (p<0.05) at $35^{\circ}C$ than $40^{\circ}C$ thermal exposure. The $CH_4$ emission/kg DMI and organic matter intake (OMI) declined (p<0.05) with increase in exposure temperature and reached its lowest levels at $40^{\circ}C$. It can be concluded from the present study that the digestibility and $CH_4$ emission were affected by intensity of heat stress. Further studies are necessary with respect to ruminal microbial changes to justify the variation in the digestibility and $CH_4$ emission during differential heat stress.
Odontoclasts and osteoclasts resorb tooth root and alveolar bone, respectively. Many studies have focused on osteoclast formation in periodontitis, but effect of periodontitis on odontoclast formation is not clearly clarified. In this study, we observed formation of odontoclasts as well as osteoclasts in rats with ligature-induced periodontitis. To induce periodontitis, ligatures were placed around the first molars in left mandibles of rats. Rats were sacrificed at days 1, 3, and 10 after ligation. After tartrate resistant acid phosphatase (TRAP) staining in mandible section, the number of TRAP-positive odontoclasts and osteoclasts were histologically counted along the root and the alveolar bone surfaces of tooth, respectively. Odontoclasts increased until day 10 in mesial and furcation root surface, but did not increase in distal root surface. When compared odontoclast formation to osteoclast formation in mesial surface, osteoclasts peaked at day 3, and then decreased gradually, whereas odontoclasts were continuously increased until day 10. The number of odontoclasts was lower than that of osteoclasts before and after periodontitis induction. These indicate that periodontitis increased formation of odontoclasts as well as osteoclasts, but odontoclast formation occurs slower and weaker than that of osteoclasts.
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