• Title/Summary/Keyword: Protein stain

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The Effect of Kamijoaguiem on Osteoporosis Induced by Ovariectomy in Rats (가미좌귀음(加味左歸飮)이 난소적출(卵巢摘出)로 유도(誘導)된 백서(白鼠)의 골다공증(骨多孔症) 미치는 영향(影響))

  • Choi, Jin-Kyung;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.107-126
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    • 2006
  • Purpose : This study was peformed to evaluate the effect of Kamijoaguiem(JGE) on the bone mass and its related factors. Methods : We used ovariectomized rat as an estrogen-deficient animal model. The model rats of osteoporosis showed a significant decrease in bone density, bone ash density, calcium content of femur bone. At the 7th day after operating ovariectomy, rats were administered with JGE per orally, and continued for 10 weeks. And osteoporosis related parameters were determined to investigate the effect of JGE. Results : Bone density, bone ash density, bone calcium, magnesium and phosphorus was decreased in osteoporotic rats. JGE improved the decreased bone density, bone ash density and the decreased bone magnesium, but JGE didn't improve the decreased bone calcium and phosphorus in osteoporotic rats. Osteocalcin in serum and hydroxy-proline excretion in urine were increased in osteoporotic rats. Their levels were decreased when JGE was administered. ALP activity in serum was increased in osteoporotic rats. JGE didn't induce any significant changes. JGE showed significant increase in serum calcium level, total protein level, albumin level, BUN level, serum LDH activity. JGE didn't show significant increase in serum T-cholesterol density, triglyceride density, HDL-cholesterol density. JGE didn't show significant increase in RBC number, hemoglobin level, platelet number, hematocrit level. JGE showed inhibitory effect on the degradation of bone-matrix in osteoporotic rats, in histological examination to Hematoxylin-eosin stain. Conclusion : JGE might improve bone density due to inhibition of bone resolution in osteoporotic rats. It suggest that JGE may be useful prescription in osteoporosis.

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Effects of Electroacupuncture of GB30 on Transient Forebrain Ischemia Injury in Rats (일과성(一過性) 전뇌(前腦) 허혈(虛血) 손상(損傷)에 대한 환도(環跳) (GB30) 전침자극(電鍼刺戟)의 효과(效果))

  • Kim Poong-Sik;Kim Jae-Hyo;Choi Dong-Ok;Kim Kyung-Sik;Sohn In-Cheul
    • Korean Journal of Acupuncture
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    • v.19 no.1
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    • pp.35-45
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    • 2002
  • Acupuncture and herbal medicine have been used to prevent and treat the cerebrovascular accident, such as a stroke, and many studies of acupuncture and moxibustion concerning to the stroke have been undertaken in the human and various animals. However, it was not published the protective effect of the electroacupuncture (EA) of Huan-do (GB30) on the transient forebrain ischemia injury. The neuroprotective effects of EA (2 ms, 10 Hz, and 1 - 2 mA) of GB30 on the transient forebrain ischemia injury was investigated by western blot of nNOS and hematoxylin and eosin stain in Sprague-Dawley rats. The transient forebrain ischemia injury resulted in increased expression of nNOS in the brain for 6 hrs after ischemia, and EA decreased significantly expression of nNOS protein in brain increased by transient forebrain ischemia injury. The survived neuronal cell stained with hematoxylin and eosin (H&E) decreased in the hippocampus by the 7 days after ischemia comparing with the normal. Comparing with the normal, the survived neurons seriously decreased cell in the hippocampus after the injury. However, the proportion of survived neurons was increased in EA treatment. These results suggest that EA of GB30 have neuroprotective effects on transient forebrain ischemia injury.

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Molecular Identification and Distribution of Aquaporins in Human and Rat Testes (사람과 흰쥐의 고환에서 Aquaporin 유전자의 발현)

  • Park, Nam-Cheol;Park, Young-Soo;Oh, Gom-Su;Jung, Jin-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.2
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    • pp.133-144
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    • 2000
  • Objective: Several water channels (aquaporins; AQP) that belong to the MIP (major intrinsic protein) family have identified. In the selected tissues including red blood cells or renal tubules, water movements are abundant and/or physiologically important. Unexpectedly, a high water permeability of human and ram sperm has been reported. Recent studies showed that AQP7 and AQP8 are present in testes, so that the high water permeability of human sperm suggested to be mediated by AQPs. Method: To identify the identity of aquaporins expressed in testes, RT-PCR was performed using degenerative primers, which were designed to correspond to highly conserved sequences surrounding the Asn-Pro-Ala (NPA) motifs in the aquaporins. New expressed AQP series were reconfirmed by immunohistochemical study using rabbit polyclonal antibodies. Results: DNA sequencing of PCR products revealed that AQP2 and AQP3 mRNA as well as AQP7 and AQP8 are expressed in human and rat testes. In human and rat testes, AQP2 are expressed in spermatozoa, interstitial cells and myofibroblasts and AQP3 are expressed in myofibroblasts of semineferous tubules on immunocytochemical stain. Conclusion: These results indicate that multiple aquaporins are expressed in testes, and that they may have important roles in the spermatogenesis and the germ cell function of testis.

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Effects of Angelica sinensis Root on Longitudinal Bone Growth Rate in Adolescent Female Rats

  • Lee, Donghun;Kim, Hocheol
    • The Korea Journal of Herbology
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    • v.32 no.1
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    • pp.69-74
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    • 2017
  • Objectives : This study aimed to investigate the effects of Angelicae sinensis Radix on longitudinal bone growth rate in rats. We have screened traditional medicinal herbs to develop the longitudinal bone growth stimulator by well-established rat model. A. sinensis was identified as one of the effective herbs in the screening process. Methods : Adolescent female rats were administered A. sinensis at doses of 30 mg/kg and 300 mg/kg for 10 consecutive days. To observe the rate of longitudinal bone growth, tetracycline was injected intraperitoneally on day 8 to stain a fluorescent band on the anew formed bone. To elucidate the mode of action, we observed insulin-like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2) expression after A. sinensis administration in growth plate. Results : In the 300 mg/kg A. sinensis group, the length between the proximal endpoint of the tetracycline label and the division line between growth plate and bone was significantly increased compared with vehicle-treated control group. Height of the proximal tibial growth plate was higher in the A. sinensis group compared with control group. A. sinensis also upregulated the expressions of IGF-1 and BMP-2 in the proliferative zone and hypertrophic zone of the proximal tibial growth plate. Conclusions : A. sinensis increases longitudinal bone growth rate in rats. According to immunohistochemistry, A. sinensis increases local IGF-1 and BMP-2 expressions in the growth plate which can be considered as direct stimulation of GH on the local growth plate.

Structural and Mechanistic Insights into the Tropism of Epstein-Barr Virus

  • Mohl, Britta S.;Chen, Jia;Sathiyamoorthy, Karthik;Jardetzky, Theodore S.;Longnecker, Richard
    • Molecules and Cells
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    • v.39 no.4
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    • pp.286-291
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    • 2016
  • Epstein-Barr virus (EBV) is the prototypical ${\gamma}$-herpesvirus and an obligate human pathogen that infects mainly epithelial cells and B cells, which can result in malignancies. EBV infects these target cells by fusing with the viral and cellular lipid bilayer membranes using multiple viral factors and host receptor(s) thus exhibiting a unique complexity in its entry machinery. To enter epithelial cells, EBV requires minimally the conserved core fusion machinery comprised of the glycoproteins gH/gL acting as the receptor-binding complex and gB as the fusogen. EBV can enter B cells using gp42, which binds tightly to gH/gL and interacts with host HLA class II, activating fusion. Previously, we published the individual crystal structures of EBV entry factors, such as gH/gL and gp42, the EBV/host receptor complex, gp42/HLA-DR1, and the fusion protein EBV gB in a postfusion conformation, which allowed us to identify structural determinants and regions critical for receptor-binding and membrane fusion. Recently, we reported different low resolution models of the EBV B cell entry triggering complex (gHgL/gp42/HLA class II) in "open" and "closed" states based on negative-stain single particle electron microscopy, which provide further mechanistic insights. This review summarizes the current knowledge of these key players in EBV entry and how their structures impact receptor-binding and the triggering of gB-mediated fusion.

Schwannoma Originating from Infraorbital Nerve (안와하신경에서 기원한 신경초종의 치험례)

  • Ha, Won;Lee, Ji Won;Choi, Jae Il;Yang, Wan Suk;Kim, Sun Young
    • Archives of Craniofacial Surgery
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    • v.14 no.1
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    • pp.61-64
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    • 2013
  • Schwannomas are well-differentiated solitary benign tumors that originate from the schwann cells of the nerve sheath. They can readily occur in the head and neck regions, but the schwannoma originating from the infraorbital nerve is extremely rare and usually painless, slow-growing, and without specific symptoms. The author experienced a rare case of infraorbital schwannoma, which was completely removed through the intraoral approach. A 20-year-old woman was admitted to our hospital for a painless, solid and circular mass located on the right infraorbital region. The eyeball movement and visual field were normal. There was no globe displacement or proptosis. Preoperative computed tomography demonstrated $13{\times}10{\times}5mm$-sized soft tissue mass. On March 2011, the mass was removed through an intraoral approach. On histopathological examination, the gross specimen consisted of a smooth, well-encapsulated and light yellowish solid mass, measuring $12{\times}7{\times}5mm$. Microscopically, it presented a typical manifestation of schwannoma with Antoni A area with Verocay body, and Antoni B area on H&E stain. The result of the immunohistochemical staining was positive for the S-100 protein. The patient had hypoesthesia of the nasal septum and vestibule in the postoperative period, and this finding confirmed that the internal nasal branch of infraorbital nerve was the nerve in which the schwannoma originated. Infraorbital schwannomas are very rare and must be included in the differential diagnosis of the orbital masses inferior to the eyeball. In the case of early diagnosis, the small-sized infraorbital schwannomas can be completely removed without any scar through an intraoral approach.

Purification and Characterization of Metalloprotease from Serratia marcescens PPB-26 and Its Application for Detergent Additive

  • Thakur, Shikha;Sharma, Nirmal Kant;Thakur, Neerja;Bhalla, Tek Chand
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.259-268
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    • 2019
  • In this study, the extracellular metalloprotease from Serratia marcescens PPB-26 was purified to homogeneity via ethanol fractionation and DEAE-cellulose column chromatography. Thus, a 3.8-fold purification was achieved with a 20% yield and specific activity of 76.2 U/mg. The purified protease was a 50-kDa monomer whose optimum pH and temperature for activity were 7.5 and $30^{\circ}C$ respectively; however, it was found to remain active in the 5-9 pH range and up to $40^{\circ}C$ for 6 h. The protease had a half-life of 15 days at $4^{\circ}C$, an optimum reaction time of 10 min, and an optimum substrate (casein) concentration of 0.25%. Furthermore, the Michaelis constant ($K_m$) and reaction velocity ($V_{max}$) of the protease were calculated to be 0.28% and $111.11{\mu}moles/(min{\cdot}mg)^{-1}$, respectively. The protease was stable when subjected to metal ions (2 mM), showing increased activity with most (especially $CoCl_2$ and $MgSO_4$ (30.54% increase)). It was also stable when exposed to oxidizing agents, bleaching agents, and detergents (5% v/v for 60 min). It retained 93% of its activity in non-ionic detergents (Tween-20, Tween-80, and Triton X-100). Moreover, wash performance analysis in commercial detergents (Ariel and Tide) showed that not only was the protease capable of protein stain removal, but also reduced cleaning time by 80% when added to detergents. Thus, the Serratia marcescens PPB-26 metalloprotease appears to be a promising new candidate as a laundry additive in the detergent industry.

Protective Effects of Ursolic Acid on Osteoblastic Differentiation via Activation of IER3/Nrf2

  • Lee, Sang-im
    • Journal of dental hygiene science
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    • v.19 no.3
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    • pp.198-204
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    • 2019
  • Background: Oxidative stress is a known to be associated with in the pathogenesis of many inflammatory diseases, including periodontitis. Ursolic acid is a pentacyclic triterpenoid with has antimicrobial, antioxidative, and anticancer properties. However, the role of ursolic acid in the regulating of osteogenesis remains undetermined. This study was aimed to elucidate the crucial osteogenic effects of ursolic acid and its ability to inhibit oxidative stress by targeting the immediate early response 3 (IER3)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. Methods: Cell proliferation was determined using water-soluble tetrazolium salt assay, cell differentiation was evaluated by alkaline phosphatase (ALP) activity, and formation of calcium nodules was detected using alizarin red S stain. Generation of reactive oxygen species (ROS) was determined using by DCFH-DA fluorescence dye in hydrogen peroxide ($H_2O_2$)-treated MG-63 cells. Expression levels of IER3, Nrf2, and heme oxygenase-1 (HO-1) were analyzed using western blot analysis. Results: Our results showed that ursolic acid up-regulated the proliferation of osteoblasts without any cytotoxic effects, and promoted ALP activity and mineralization. $H_2O_2$-induced ROS generation was found to be significantly inhibited on treatment with ursolic acid. Furthermore, in $H_2O_2$-treated cells, the expression of the early response genes: IER3, Nrf2, and Nrf2-related phase II enzyme (HO-1) was enhanced in the presence of ursolic acid. Conclusion: The key findings of the present study elucidate the protective effects of ursolic acid against oxidative stress conditions in osteoblasts via the IER3/Nrf2 pathway. Thus, ursolic acid may be developed as a preventative and therapeutic agent for mineral homeostasis and inflammatory diseases caused due to oxidative injury.

Increase of Cardiometabolic Biomarkers Among Vehicle Inspectors Exposed to PM0.25 and Compositions

  • Ramdhan, Doni Hikmat;Kurniasari, Fitri;Tejamaya, Mila;Fitri, Aidila;Indriani, Aisyah;Kusumawardhani, Adinda;Santoso, Muhayatun
    • Safety and Health at Work
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    • v.12 no.1
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    • pp.114-118
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    • 2021
  • Background: Exposure to particulate matter (PM) emitted from vehicle exhaust might disrupt systemic function and elevate the risk of cardiovascular disease. In this study, we examined the changes of cardiometabolic biomarkers among vehicle inspectors exposed daily to PM0.25 and components. Methods: This cross-sectional study was conducted at two vehicle inspection centers, Pulogadung and Ujung Menteng, located in East Jakarta, Indonesia. The exposed respondents were 43 workers from vehicle inspection centers, and the unexposed group consisted of 22 staff officers working in the same locations. Vehicle exhaust particulate matter was measured for eight hours using a Leland Legacy personal pump attached to a Sioutas Cascade Impactor. The used filters were 25 and 37-mm quartz filters. The particulate matter concentration was analyzed using a gravimetric method, whereas trace elements were analyzed using energy dispersive X-ray fluorescence. An EEL Smoke Stain Reflectometer analyzed black carbon. Results: The personal exposure concentrations of PM0.25 were 10.4-fold higher than those in unexposed groups. Calcium and sulfur were the major components in the obtained dust, and their levels were 3.3- and 7.2-fold higher, respectively, in the exposed group. Based on an independent-samples t-test, high-density lipoprotein, triglyceride, HbA1c, total immunoglobulin E, high-sensitivity C-reactive protein, tumor necrosis factor-alpha, and nitric oxide levels were significantly different between the groups. Conclusions: In summary, it was suggested that PM0.25 exposure from vehicle exhaust might affect cardiometabolic biomarkers change.

Protective Effect of Spatholobi Caulis in Thioacetamide induced Acute Liver Injury of Rat (Thioacetamide로 유발한 간손상 모델에서 계혈등(鷄血藤)의 간보호 효과)

  • Oh, Min Hyuck;Shin, Mi-Rae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.36 no.2
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    • pp.31-42
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    • 2021
  • Objectives : This study was undertaken to investigate the hepatoprotective effect of Spatholobi Caulis water extract (SC) to thioacetamide (TAA)-induced acute liver injury (ALI) in rats. Methods : The rats were injected intraperitoneally with TAA (200 mg/kg body weight) and orally administered SC (100 or 200 mg/kg b.w.) daily for 3 days. Liver biomarkers were assessed by serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), and ammonia levels. Malondialdehyde (MDA) was measured both serum and liver tissue. In addition, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, anti-oxidant, and inflammation-related proteins were investigated by western blot analysis. Histological examination further confirmed though hematoxylin and eosin stain. Results : The SC treatment reduced liver function markers like GOT and GPT and also remarkably decreased ammonia level. Moreover, the elevated MDA level in TAA-induced group was significantly reduced by SC treatment. NADPH oxidase expression associated with oxidative stress including NOX2, NOX4, and p47phox markedly inhibited by SC administration. SC treatment exerted anti-oxidant effect through the increase of anti-oxidant enzyme including superoxide dismutase (SOD), Catalase, and heme oxygenase-1 (HO-1). The protein expressions of inflammatory cytokines such as tumor necrosis factor-�� (TNF-��), IL-6, and IL-1�� induced by nuclear factor-kappa B (NF-��B) activation were modulated through blocking the phosphorylation of inhibitor of nuclear factor ��B�� (I��B)��. SC treatment also improved histological alterations. Conclusion : These findings suggested that SC administration may be a potential candidate for the prevention or treatment of ALI.