• Title/Summary/Keyword: Calcium imaging

Search Result 87, Processing Time 0.025 seconds

In Vitro and In Vivo Evaluation of Composite Scaffold of BCP, Bioglass and Gelatin for Bone Tissue Engineering

  • Kim, Woo Seok;Nath, Subrata Deb;Bae, Jun Sang;Padalhin, Andrew;Kim, Boram;Song, Myeong Jin;Min, Young Ki
    • Korean Journal of Materials Research
    • /
    • v.24 no.6
    • /
    • pp.310-318
    • /
    • 2014
  • In this experiment, a highly porous scaffold of biphasic calcium phosphate (BCP) was prepared using the spongereplica method. The BCP scaffold was coated with 58S bioactive glass (BG) and sintered for a second time. The resulting scaffold was coated with gelatin (Gel) and cross-linked with [3-(3-dimethyl aminopropyl) carbodiimide] and N-Hydroxysuccinamide (EDC-NHS). The initial average pore size of the scaffold ranged from 300 to $700{\mu}m$, with more than 85 % porosity. The coating of BG and Gel had a significant effect on the scaffold-pore size, decreasing scaffold porosity while increasing mechanical strength. The material and surface properties were evaluated by means of several experiments involving scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffraction (XRD). Cytotoxicity was evaluated using MTT assay and confocal imaging of MC3T3-E1 pre-osteoblast cells cultured in vitro. Three types of scaffold (BCP, BCP-BG and BCP-BG-Gel) were implanted in a rat skull for in vivo evaluation. After 8 weeks of implantation, bone regeneration occurred in all three types of sample. Interestingly, regeneration was found to be greater (geometrically and physiologically) for neat BCP scaffolds than for two other kinds of composite scaffolds. However, the other two types of scaffolds were still better than the control (i.e., defect without treatment).

Ruminal impaction with an indigestible foreign body in long-tailed goral (Naemorhedus caudatus)

  • Sangjin Ahn;Woojin Shin;Yujin Han;Sohwon Bae;Chea-Un Cho;Sooyoung Choi;Yeonsu Oh;Jong-Taek Kim
    • Korean Journal of Veterinary Service
    • /
    • v.46 no.2
    • /
    • pp.161-165
    • /
    • 2023
  • A 6-year-old female long-tailed goral (Naemorhedus caudatus, goral) was admitted to the Gangwon wildlife medical rescue center due to severe starvation and hypothermia. The goral displayed clinical signs of severe weight loss, dehydration, lethargy, regurgitation, hypothermia, and bloating, with blood tests showing a decrease in albumin, calcium, glucose, and sodium. Supportive care was given through heating, oxygen supply, and fluid therapy, but the goral's condition worsened, and the goral died three days after admission. Radiographic and computed tomography scans revealed the presence of an indigestible foreign body (IFB) in the rumen, which was confirmed during the necropsy. The cause of goral's death was identified as chronic rumen impaction caused by the IFB (a ball of string), with other pathological findings including bloating, the presence of serous ascites, and congestion of digestive organs. Blood biochemical changes, clinical signs, and veterinary medical imaging diagnosis can be helpful in diagnosing ruminal impaction. This case report can provide valuable information for the diagnosis and treatment of ruminal impaction caused by IFB in endangered gorals, which may include rumenotomy or other surgical procedures.

Octyl Gallate Inhibits ATP-induced Intracellular Calcium Increase in PC12 Cells by Inhibiting Multiple Pathways

  • Guo, Yujie;Hong, Yi-Jae;Jang, Hyun-Jong;Kim, Myung-Jun;Rhie, Duck-Joo;Jo, Yang-Hyeok;Hahn, Sang-June;Yoon, Shin-Hee
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.14 no.1
    • /
    • pp.21-28
    • /
    • 2010
  • Phenolic compounds affect intracellular free $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) signaling. The study examined whether the simple phenolic compound octyl gallate affects ATP-induced $Ca^{2+}$ signaling in PC12 cells using fura-2-based digital $Ca^{2+}$ imaging and whole-cell patch clamping. Treatment with ATP ($100\;{\mu}M$) for 90 s induced increases in $[Ca^{2+}]_i$ in PC12 cells. Pretreatment with octyl gallate (100 nM to $20\;{\mu}M$) for 10 min inhibited the ATP-induced $[Ca^{2+}]_i$ response in a concentration-dependent manner ($IC_{50}=2.84\;{\mu}M$). Treatment with octyl gallate ($3\;{\mu}M$) for 10 min significantly inhibited the ATP-induced response following the removal of extracellular $Ca^{2+}$ with nominally $Ca^{2+}$-free HEPES HBSS or depletion of intracellular $Ca^{2+}$ stores with thapsigargin ($1\;{\mu}M$). Treatment for 10 min with the L-type $Ca^{2+}$ channel antagonist nimodipine ($1\;{\mu}M$) significantly inhibited the ATP-induced $[Ca^{2+}]_i$ increase, and treatment with octyl gallate further inhibited the ATP-induced response. Treatment with octyl gallate significantly inhibited the $[Ca^{2+}]_i$ increase induced by 50 mM KCI. Pretreatment with protein kinase C inhibitors staurosporin (100 nM) and GF109203X (300 nM), or the tyrosine kinase inhibitor genistein ($50\;{\mu}M$) did not significantly affect the inhibitory effects of octyl gallate on the ATP-induced response. Treatment with octyl gallate markedly inhibited the ATP-induced currents. Therefore, we conclude that octyl gallate inhibits ATP-induced $[Ca^{2+}]_i$ increase in PC12 cells by inhibiting both non-selective P2X receptor-mediated influx of $Ca^{2+}$ from extracellular space and P2Y receptor-induced release of $Ca^{2+}$ from intracellular stores in protein kinase-independent manner. In addition, octyl gallate inhibits the ATP-induced $Ca^{2+}$ responses by inhibiting the secondary activation of voltage-gated $Ca^{2+}$ channels.

Parathyroid Carcinoma (부갑상선암)

  • Cho Eun-Chol;Sub Jin-Hak;Chung Woong-Yun;Kim Ho-Geun;Park Cheong-Soo
    • Korean Journal of Head & Neck Oncology
    • /
    • v.17 no.2
    • /
    • pp.205-209
    • /
    • 2001
  • Purpose: Most cases of primary hyperparathyroidism are due to parathyroid adenoma or parathyroid hyperplasia. Parathyroid carcinoma is a very rare cause of hyperparathyroidism. Although the diagnosis of parathyroid carcinoma is usually established by pathologic criteria especially of vascular or capsular invasion, some clinical and biochemical features differentiate it from benign forms of hyperparathyroidism. We under-took a retrospective study in 6 patients with parathyroid carcinoma, with the aim of conveying experience from management for this rare cause of hyperparathyroidism. Methods: Clinical symptoms, biochemical laboratory, radiologic, and intraoperative findings, local recurrence and distant metastasis were analyzed in 6 patients diagnosed pathologically as a parathyroid carcinoma after operation from 1992 to 2001. Results: Mean age was 50.2 years (33.0-60.0 years) and male to female ratio was 1:1. Neck mass was found in 5 patients, multiple bone pain in 3 patients and renal stone in 1 patient. One case has suffered from chronic renal failure for 19 years. Although preoperative laboratory evaluations showed the aspects of hyperparathyroidism in all cases, mean serum calcium level was 11.2mg/dl(10.5-12.1mg/dl), slightly elevated. Laboratory values after surgery were within the normal range in 5 cases. However, in one case with chronic renal failure, serum PTH levels, serially checked, were above the normal range. Any of imaging methods failed to suggest a parathyroid carcinoma preoperatively. Parathyroid adenoma was suspected in 3 cases, thyroid cancer in the other cases before surgery. The extent of resection was radical resection of parathyroid lesion with more than unilateral thyroid lobectomy and central compartment neck node dissection and in 2 cases, the resection of recurrent laryngeal nerve or strap muscles was added. During follow-up period, any local or systemic recurrence were not evident in all the cases. Conclusion: Although parathyroid carcinoma is a rare disease and its preoperative diagnosis, in our experience, could not easily be made, the understanding of characteristic clinical and biochemical feature could help diagnosis at first surgery. Radical resection without remaining residual tumor is most important for the management of the parathyroid cancer.

  • PDF

Functional Expression of P2Y Receptors in WERI-Rb1 Retinoblastoma Cells

  • Kim, Na-Hyun;Park, Kyu-Sang;Sohn, Joon-Hyung;Yeh, Byung-Il;Ko, Chang-Mann;Kong, In-Deok
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.15 no.1
    • /
    • pp.61-66
    • /
    • 2011
  • P2Y receptors are metabotropic G-protein-coupled receptors, which are involved in many important biologic functions in the central nervous system including retina. Subtypes of P2Y receptors in retinal tissue vary according to the species and the cell types. We examined the molecular and pharmacologic profiles of P2Y purinoceptors in retinoblastoma cell, which has not been identified yet. To achieve this goal, we used $Ca^{2+}$ imaging technique and western blot analysis in WERI-Rb-1 cell, a human retinoblastoma cell line. ATP ($10\;{\mu}M$) elicited strong but transient $[Ca^{2+}]_i$ increase in a concentration dependent manner from more than 80% of the WERI-Rb-1 cells (n=46). Orders of potency of P2Y agonists in evoking $[Ca^{2+}]_i$ transients were 2MeS-ATP>ATP>>UTP=${\alpha}{\beta}$-MeATP, which was compatible with the subclass of $P2Y_1$ receptor. The $[Ca^{2+}]_i$ transients evoked by applications of 2MeS-ATP and/or ATP were also profoundly suppressed in the presence of $P2Y_1$ selective blocker (MRS 2179; $30\;{\mu}M$). $P2Y_1$ receptor expression in WERI-Rb-1 cells was also identified by using western blot. Taken together, $P2Y_1$ receptor is mainly expressed in a retinoblastoma cell, which elicits $Ca^{2+}$ release from internal $Ca^{2+}$ storage sites via the phospholipase C-mediated pathway. $P2Y_1$ receptor activation in retinoblastoma cell could be a useful model to investigate the role of purinergic $[Ca^{2+}]_i$ signaling in neural tissue as well as to find a novel therapeutic target to this lethal cancer.

Radiation-induced Apoptosis, Necrosis and G2 Arrest in Fadu and Hep2 Cells

  • Lee Sam-Sun;Kang Beom-Hyun;Choi Hang-Moon;Jeon In-Seong;Heo Min-Suk;Choi Soon-Chul
    • Imaging Science in Dentistry
    • /
    • v.30 no.4
    • /
    • pp.275-279
    • /
    • 2000
  • Purpose: Radiation damage is produced and viable cell number is reduced. We need to know the type of cell death by the ionizing radiation and the amount and duration of cell cycle arrest. In this study, we want to identified the main cause of the cellular damage in the oral cancer cells and normal keratinocytes with clinically useful radiation dosage. Materials and Methods: Human gingival tissue specimens obtained from healthy volunteers were used for primary culture of the normal human oral keratinocytes (NHOK). Primary NHOK were prepared from separated epithelial tissue and maintained in keratinocyte growth medium containing 0.15 mM calcium and a supplementary growth factor bullet kit. Fadu and Hep-2 cell lines were obtained from KCLB. Cells were irradiated in a /sup 137/Cs γ-irradiator at the dose of 10 Gy. The dose rate was 5.38 Gy/min. The necrotic cell death was examined with Lactate Dehydrogenase (LDH) activity in the culture medium. Every 4 day after irradiation, LDH activities were read and compared control group. Cell cycle phase distribution and preG1-incidence after radiation were analyzed by flow cytometry using Propidium Iodine staining. Cell cycle analysis were carried out with a FAC Star plus flowcytometry (FACS, Becton Dickinson, USA) and DNA histograms were processed with CELLFIT software (Becton Dickinson, USA). Results: LDH activity increased in all of the experimental cells by the times. This pattern could be seen in the non-irradiated cells, and there was no difference between the non-irradiated cells and irradiated cells. We detected an induction of apoptosis after irradiation with a single dose of 10 Gy. The maximal rate of apoptosis ranged from 4.0% to 8.0% 4 days after irradiation. In all experimental cells, we detected G2/M arrest after irradiation with a single dose of 10 Gy. Yet there were differences in the number of G2/M arrested cells. The maximal rate of the G2/M ranges from 60.0% to 80.0% 24h after irradiation. There is no significant changes on the rate of the G0/G1 phase. Conclusion: Radiation sensitivity was not related with necrosis but cell cycle arrest and apoptosis. These data suggested that more arrested cell is correlated with more apoptosis.

  • PDF

Effects of 2-deoxy-D-glucose and quercetin on osteoblastic differentiation and mineralization in irradiated MC3T3-E1 cells (2-deoxy-D-glucose와 quercetin이 방사선조사 MC3T3-E1 골모세포주의 분화와 석회화에 미치는 영향)

  • Ahn, Hyoun-Suk;Kim, Kyoung-A;Koh, Kwang-Joon
    • Imaging Science in Dentistry
    • /
    • v.36 no.4
    • /
    • pp.189-198
    • /
    • 2006
  • Purpose: To investigate the in vitro response of MC3T3-E1 osteoblastic cells to X-ray in the presence and absence of 2 deoxy-D-glucose (2-DG) and quercetin (QCT). Materials and Methods: The MC3T3-E1 cells were cultured in an ${\alpha}-MEM$ supplemented with 5 mM 2-DG or $10{\mu}M$ QCT and then the cells were incubated for 12 h prior to irradiation with 2, 4, 6, and 8Gy using a linear accelerator (Mevaprimus, Germany) delivered at a rate of 1.5 Gy/min. At various times after the irradiation, the cells were processed for the analyses of proliferation, viability, cytotoxicity, and mineralization. Results: Exposure of the cells to X-ray inhibited the tritium incorporation, 3-(4, 5-dimethylthiazol-2yl-)-2, 5-diphenyl tetrazolium bromide (MTT)-reducing activity, and alkaline phosphatase (ALP) activity, and caused cytotoxicity and apoptosis in a dose-dependent manner of the X-ray. This effect was further apparent on day 3 and 7 after the irradiation. RA+2-DG showed the decrease of DNA content, cell viability, and increase of cytotoxicity rather than RA. ALP activity increased on day 7 and subsequently its activity dropped to a lower level. 2-DG suppressed the calcium concentration, but visual difference of number of calcified nodules between RA and RA+2-DG was not noticed. RA+QCT showed the increase of DNA content, cell viability, but decrease of cytotoxicity and subG1 stage cells in the cell cycle, and increased calcified nodules in von Kossa staining rather than the RA. ALP activity showed significant increases on day 7 and subsequently its activity dropped to a lower level. Conclusion: The results showed that the 2-DG acted as a radiosensitizing agent and QCT acted as a radiosensitizing agent respectively in the irradiated MC3T3-E1 osteoblast-like cells.

  • PDF

Long-term Follow-up of Metaphyseal Sclerotic Lines in Children Treated with Pamidronate (파미드로네이트 치료받은 환자에서 발생한 골간단 경화성 선에 대한 장기간 추적 연구)

  • Choi, Yu-Mi;Suh, Jin-Soon;Cho, Byoung-Soo
    • Childhood Kidney Diseases
    • /
    • v.18 no.2
    • /
    • pp.92-97
    • /
    • 2014
  • Purpose: Bisphosphonates are widely used for the management steroid-induced osteoporosis (SIO) in children. With the increasing use of bisphosphonates, there have been reports of abnormal radiological findings in the growing skeleton. Therefore, their use in pediatric patients remains controversial. The present study was conducted to evaluate the long-term follow-up radiographic features, particularly metaphyseal sclerotic lines, in children who receive pamidronate therapy for nephropathy. Methods: Twenty-four children with nephropathy treated with oral calcium and pamidronate (mean duration, 9 months; dose, 100 mg daily), were evaluated retrospectively. All patients had SIO secondary to chronic glucocorticoid therapy for treating nephropathy. Long bone radiographic imaging was performed before treatment with pamidronate, and at follow-up, several years later. Physeal growth rates were estimated by measuring the distance that the sclerotic lines moved on the radiographs during the corresponding time intervals. Results: The mean follow-up period was 138 months. Long bone radiographs showed well-defined sclerotic lines at the metaphyseal ends, progressively moving from the physeal plate to the diaphysis, in all patients. The mean rate of movement of the sclerotic line was 6.21 mm per year. In 12 patients, the lines disappeared. The mean rate of growth in height was 7.33 cm per year. Conclusions: Results of long-term follow-up suggest that the metaphyseal sclerotic lines associated with pamidronate treatment tend to disappear without affecting overall skeletal growth. Bisphosphonate treatment for SIO in children with nephropathy seems to be safe, although further studies in larger number of patients are needed.

Set Up and Operation for Medical Radiation Exposure Quality Control System of Health Promotion Center (건강검진센터의 의료방사선 피폭 품질관리 시스템 구축 운영 경험 보고)

  • Kim, Jung-Su;Jung, Hae-Kyoung;Kim, Jung-Min
    • Journal of radiological science and technology
    • /
    • v.39 no.1
    • /
    • pp.13-17
    • /
    • 2016
  • In this study, standard model of medical radiation dosage quality control system will be suggested and the useful of this system in clinical field will be reviewed. Radiation dosage information of modalities are gathered from digital imaging and communications in medicine(DICOM) standard data(such as DICOM dose SR and DICOM header) and stored in database. One CT scan, two digital radiography modalities and two mammography modalities in one health promotion center in Seoul are used to derive clinical data for one month. After 1 months research with 703 CT scans, the study shows CT $357.9mGy{\cdot}cm$ in abdomen and pelvic CT, $572.4mGy{\cdot}cm$ in brain without CT, $55.9mGy{\cdot}cm$ in calcium score/heart CT, screening CT at $54mGy{\cdot}cm$ in chest screening CT(low dose screening CT scan), $284.99mGy{\cdot}cm$ in C-spine CT and $341.85mGy{\cdot}cm$ in L-spine CT as health promotion center reference level of each exam. And with 1955 digital radiography cases, it shows $274.0mGy{\cdot}cm2$ and for mammography 6.09 mGy is shown based on 536 cases. The use of medical radiation shall comply with the principles of justification and optimization. This quality management of medical radiation exposure must be performed in order to follow the principle. And the procedure to reduce the radiation exposure of patients and staff can be achieved through this. The results of this study can be applied as a useful tool to perform the quality control of medical radiation exposure.

Group 1 metabotropic glutamate receptor 5 is involved in synaptically-induced Ca2+-spikes and cell death in cultured rat hippocampal neurons

  • Yang, Ji Seon;Jeon, Sujeong;Jang, Hyun-Jong;Yoon, Shin Hee
    • The Korean Journal of Physiology and Pharmacology
    • /
    • v.26 no.6
    • /
    • pp.531-540
    • /
    • 2022
  • Group 1 metabotropic glutamate receptors (mGluRs) can positively affect postsynaptic neuronal excitability and epileptogenesis. The objective of the present study was to determine whether group 1 mGluRs might be involved in synaptically-induced intracellular free Ca2+ concentration ([Ca2+]i) spikes and neuronal cell death induced by 0.1 mM Mg2+ and 10 µM glycine in cultured rat hippocampal neurons from embryonic day 17 fetal Sprague-Dawley rats using imaging methods for Ca2+ and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays for cell survival. Reduction of extracellular Mg2+ concentration ([Mg2+]o) to 0.1 mM induced repetitive [Ca2+]i spikes within 30 sec at day 11.5. The mGluR5 antagonist 6-Methyl2-(phenylethynyl) pyridine (MPEP) almost completely inhibited the [Ca2+]i spikes, but the mGluR1 antagonist LY367385 did not. The group 1 mGluRs agonist, 3,5-dihydroxyphenylglycine (DHPG), significantly increased the [Ca2+]i spikes. The phospholipase C inhibitor U73122 significantly inhibited the [Ca2+]i spikes in the absence or presence of DHPG. The IP3 receptor antagonist 2-aminoethoxydiphenyl borate or the ryanodine receptor antagonist 8-(diethylamino)octyl 3,4,5-trimethoxybenzoate also significantly inhibited the [Ca2+]i spikes in the absence or presence of DHPG. The TRPC channel inhibitors SKF96365 and flufenamic acid significantly inhibited the [Ca2+]i spikes in the absence or presence of DHPG. The mGluR5 antagonist MPEP significantly increased the neuronal cell survival, but mGluR1 antagonist LY367385 did not. These results suggest a possibility that mGluR5 is involved in synaptically-induced [Ca2+]i spikes and neuronal cell death in cultured rat hippocampal neurons by releasing Ca2+ from IP3 and ryanodine-sensitive intracellular stores and activating TRPC channels.