• Journal of Periodontal and Implant Science
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• v.41 no.5
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• pp.218-226
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• 2011

Purpose: Micro-computed tomography (micro-CT) has been widely used in the evaluation of regenerated bone tissue but the reliability of micro-CT has not yet been established. This study evaluated the correlation between histomorphometric analysis and micro-CT analysis in performing new bone formation measurement. Methods: Critical-size calvarial defects were created using a 8 mm trephine bur in a total of 24 Sprague-Dawley rats, and collagen gel mixed with autogenous rat bone marrow stromal cells (BMSCs) or autogenous rat BMSCs transduced by adenovirus containing bone morphogenic protein-2 (BMP-2) genes was loaded into the defect site. In the control group, collagen gel alone was loaded into the defect. After 2 and 4 weeks, the animals were euthanized and calvaria containing defects were harvested. Micro-CT analysis and histomorphometric analysis of each sample were accomplished and the statistical evaluation about the correlation between both analyses was performed. Results: New bone formation of the BMP-2 group was greater than that of the other groups at 2 and 4 weeks in both histomorphometric analysis and micro-CT analysis (P=0.026, P=0.034). Histomorphometric analysis of representative sections showed similar results to histomorphometric analysis with a mean value of 3 sections. Measurement of new bone formation was highly correlated between histomorphometric analysis and micro-CT analysis, especially at the low lower threshold level at 2 weeks (adjusted $r^2=0.907$, P<0.001). New bone formation of the BMP-2 group analyzed by micro-CT tended to decline sharply with an increasing lower threshold level, and it was statistically significant (P<0.001). Conclusions: Both histomorphometric analysis and micro-CT analysis were valid methods for measurement of the new bone in rat calvarial defects and the ability to detect the new bone in micro-CT analysis was highly influenced by the threshold level in the BMP-2 group at early stage.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

• Parasites, Hosts and Diseases
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• v.55 no.5
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• pp.523-532
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• 2017

A field survey studying intestinal parasites in humans and microbial pathogen contamination at environment was performed in a Laotian rural village to identify potential risks for disease outbreaks. A parasitological investigation was conducted in Ban Lak Sip village, Luang Prabang, Lao PDR involving fecal samples from 305 inhabitants as well as water samples taken from 3 sites of the local stream. Water analysis indicated the presence of several enteric pathogens, i.e., Aeromonas spp., Vibrio spp., E. coli H7, E. coli O157: H7, verocytotoxin-producing E. coli (VTEC), Shigella spp., and enteric adenovirus. The level of microbial pathogens contamination was associated with human activity, with greater levels of contamination found at the downstream site compared to the site at the village and upstream, respectively. Regarding intestinal parasites, the prevalence of helminth and protozoan infections were 68.9% and 27.2%, respectively. Eight helminth taxa were identified in fecal samples, i.e., 2 tapeworm species (Taenia sp. and Hymenolepis diminuta), 1 trematode (Opisthorchis sp.), and 5 nematodes (Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, trichostrongylids, and hookworms). Six species of intestinal protists were identified, i.e., Blastocystis hominis, Cyclospora spp., Endolimax nana, Entamoeba histolytica/E. dispar, Entamoeba coli, and Giardia lamblia. Questionnaires and interviews were also conducted to determine risk factors of infection. These analyses together with a prevailing infection level suggested that most of villagers were exposed to parasites in a similar degree due to limited socio-economic differences and sharing of similar practices. Limited access to effective public health facilities is also a significant contributing factor.

• Pediatric Infection and Vaccine
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• v.17 no.1
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• pp.49-55
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• 2010

Purpose : The Purposes of this study are to identify the circulating etiologic viruses of acute lower respiratory tract infection in children and to understand the relation with clinical diagnosis. Methods : We obtained a total of 418 nasopharyngeal aspirates from children admitted for their acute lower respiratory tract infections at three tertiary hospitals in Seoul from September 2008 to March 2009. We performed multiplex RT-PCR to identify 14 etiologic viruses and analyzed their emerging patterns and clinical features. Results : Average age of patients was 16.4 months old and the ratio of male to female was 1.36. Viruses were detected in 56.2% of a total of 418 samples. Respiratory syncytial virus (35%) was the most frequently detected and followed by human rhinovirus (22%), human bocavirus, adenovirus, human metapneumovirus, parainfluenza virus, influenza virus and human coronavirus. Co-infection reached 21.9 % of positive patients. Conclusion : When we manage the patients with acute lower respiratory infectious diseases, we should remind the role of various viral pathogens, which might be circulating by seasons and by local areas.

• Journal of Life Science
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• v.18 no.3
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• pp.374-380
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• 2008

Rapid detection of enterovirus (EVs) is important in the management of aseptic meningitis. We examined the relative efficiency and specificity of the real-time nucleic acid sequence-based amplification (NASBA) comparing with the established reverse transcription polymerase chain reaction (RT-PCR) and viral culture method which were used for the detection of enterovirus RNA in clinical specimens. Of the total 292 samples, 145 were found to be positive to enterovirus RNA by real-time NASBA, 101 were positive by viral culture, and 86 were positive by RT-PCR. 147 samples and 46 samples were determined to be negative and positive by all methods respectively, but 4 samples were positive only by real-time NASBA. To compare the specificity of each method, various clinical samples which were diagnosed for herpes simplex virus (HSV)-1, HSV-2, adenovirus, mumps, and rhinovirus were applied. Except one rhinovirus sample which was false positive to enterovirus RNA by RT-PCR, the other different samples were negative to all three methods. The real-time NASBA procedure can be completed within 5 hours in contrast with 9 hours for the RT-PCR and 3-14 days for the viral culture. From this study, it was suggested that the real-time NASBA assay could be a standardized, rapid, specific, and sensitive procedure for the detection of enterovirus RNA.

• Molecules and Cells
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• v.37 no.11
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• pp.785-794
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• 2014

Mitophagy, a cellular process that selectively targets dysfunctional mitochondria for degradation, is currently a hot topic in research into the pathogenesis and treatment of many human diseases. Considering that hypoxia causes mitochondrial dysfunction, which results in cell death, we speculated that selective activation of mitophagy might promote cell survival under hypoxic conditions. In the present study, we introduced the Regulator of calcineurin 1-1L (Rcan1-1L) to initiate the mitophagy pathway and aimed to evaluate the effect of Rcan1-1L-induced mitophagy on cell survival under hypoxic conditions. Recombinant adenovirus vectors carrying Rcan1-1L were transfected into human umbilical vein endothelial cells and human adult cardiac myocytes. Using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay and Trypan blue exclusion assay, Rcan1-1L overexpression was found to markedly reverse cell growth inhibition induced by hypoxia. Additionally, Rcan1-1L overexpression inhibited cell apoptosis under hypoxic conditions, as detected by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay. Meanwhile, the mitochondria-mediated cell apoptotic pathway was inhibited by Rcan1-1L. In contrast, knockdown of Rcan1-1L accelerated hypoxia-induced cell apoptosis. Moreover, Rcan1-1L overexpression significantly reduced mitochondrial mass, decreased depolarized mitochondria, and downregulated ATP and reactive oxygen species production. We further delineated that the loss of mitochondrial mass was due to the activation of mitophagy induced by Rcan1-1L. Rcan1-1L overexpression activated autophagy flux and promoted translocation of the specific mitophagy receptor Parkin into mitochondria from the cytosol, whereas inhibition of autophagy flux resulted in the accumulation of Parkin-loaded mitochondria. Finally, we demonstrated that mitochondrial 1permeability transition pore opening was significantly increased by Rcan1-1L overexpression, which suggested that Rcan1-1L might evoke mitophagy through regulating mitochondrial permeability transition pores. Taken together, we provide evidence that Rcan1-1L overexpression induces mitophagy, which in turn contributes to cell survival under hypoxic conditions, revealing for the first time that Rcan1-1L-induced mitophagy may be used for cardioprotection.

• Korean Journal of Veterinary Service
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• v.43 no.2
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• pp.67-77
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• 2020

Canine infectious respiratory disease (CIRD), also known as infectious tracheobronchitis or kennel cough occurs in a multiple-dog environment such as a shelter. In this study, we were collected 300 of nasal swab samples from dogs and 145 of environmental samples from a shelter to investigate respiratory pathogens of dogs in the Gwangju metropolitan city animal shelter from February to October, 2019. Bacteria cultures for isolation of Bordetella (B.) bronchiseptica and polymerase chain reaction (PCR) tests were performed for detection of eleven canine respiratory pathogens, namely Mycoplasma (M.) cynos, canine distemper virus (CDV), canine influenza virus (CIV), canine parainfluenza virus (CPIV), canine respiratory coronavirus (CRCoV), alpha-coronavirus (CCoV), canine pneumovirus (CnPnV), canine hepacivirus (CHeV), canine adenovirus type 2 (CAdV-2), canine herpesvirus-1 (CHV-1) and canine bocavirus (CBoV). Among 300 nasal swab samples, 148 samples (49.3%) were positive for at least one pathogens. CHV-1 was the most common pathogen, found in 95/300 (31.7%) samples. Subsequently, M. cynos (22.0%), B. bronchiseptica (2.3%), CPIV (2.0%), CBoV (1.7%), CCoV (0.7%) were detected. The detection rates of M. cynos and CHV-1 according to the duration of stay in the shelter were statistically significant. Among environmental samples, M. cynos, CCoV, CBoV and CHV-1 were detected in 45/145 (31.0%). These results indicated the need for disease control and prevention systems in the shelter.

• Journal of Life Science
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• v.13 no.4
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• pp.529-534
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• 2003

This study was done to investigated the phytosterol compositions of safflower (Carthamus tinctorius L.) seed. The phytoestrogen activity was also determined using CAT-ELISA Kit in ethanol extract of safflower seed. The phytosterol of safflower seeds was identified using gas chromatography-mass spectrometry after saponification of the oils. The phytosterol content and composition of safflower seed oils were 4% and identified stigmast-5-en-3-ol (3$\beta$, 24S)-form, ${\gamma}$-sitosterol (clionasterol) with Wiley MS spectrum library. The synergistic effect of human estrogen receptor (hER) has been investigated using a minimal chimeric promoters composed of the TATA region of the adenovirus-2 major late promoter (A22MLP) and two consensus perfectly polindromic Xenopus vitellogenin A2 gene estrogen responsive elements (XVEREl19). Transient transfection experiments in tile human breast adenocarcinoma cell line MCF-7, which is known to express the estrogen receptor endogenously, revealed that phytoestrogen from Carthamus tinctorius L. acts as estrogen. We have observed the transcriptional activities stimulated methanol and ethanol extract of safflower seed in MCF-7, were 0.43 and 0.37 respectively, compared to that by $\beta$-estradiol as 1.0. Our data showed that safflower seeds have estrogenic activity methanol and ethanol extracts and ethanol lower than that of $\beta$-estradiol. This result provides the first evidence that the beneficial effect of safflower seeds may be mediated, at least in part, by the stimulating effect of phytoestrogen ell bone-protecting.

• Journal of the Korean Child Neurology Society
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• v.26 no.4
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• pp.233-239
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• 2018

Purpose: Viral gastroenteritis is a common disease in infants and children. Seizures can be associated with viral gastroenteritis as benign convulsions with mild gastroenteritis (CwG). After the development of the rotavirus vaccination, norovirus has become inreasingly significant in children. We retrospectively analyzed the clinical features in a pediatric population presenting with seizures and confirmed enteral viral infections, especially norovirus infections. Methods: We retrospectively reviewed the medical records of pediatric patients aged <15 years admitted due to convulsions and gastroenteritis to the Department of Pediatrics of Myongji Hospital between July 2014 and June 2016. Results: A total of 46 patients (24 male and 22 female) were included. Norovirus was detected in 21 (45.7%) patients, adenovirus in three (6.5%), rotavirus in two (4.3%), astrovirus in one (2.2%), and none of agents were detected in 19 (41.3%) patients. Patients in the norovirus gastroenteritis (NGE) group had a higher incidence of diarrhea than that among in the non-norovirus gastroenteritis (NNGE) group (61.9% vs 28.0%; P<0.05). Twelve patients experienced status epilepticus, including five (23.8%) in the NGE group and seven (33.0%) in the NNGE group (P=0.837). Seizures were effectively terminated by intravenous benzodiazepines in 8 (66.7%) of 12 patients. Additional long-acting antiepileptic drugs such as fosphenytoin or levetiracetam were required in three (25%) of 12 patients. Conclusion: Patients with CwG with and without noroviurs infection did not differ in terms of clinical features. However, status epilepticus was not uncommon among patients with CwG by definition.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.3
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• pp.329-335
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• 2019

Annually, millions of children die from respiratory virus infections. Human rhinovirus (HRV) is a causative agent of severe respiratory infections in young, elderly, and asthmatic patients with weak immunity. In this study, 9,010 respiratory virus specimens were collected from January 2012 to December 2018 at Dankook University Hospital, Cheonan and examined by real-time reverse transcription polymerase chain reaction. Twelve respiratory viruses were detected. The mean detection rate was 21.3% (N=1,920/9,010), and the mean age of HRV-positive patients was 6.5 years (median age: 1.6 years, range: 0.0~96.0). The detection rate was the highest in July (32.4%) and the lowest in February (8.3%). When the detection rate was analyzed by age group, the detection rate was the second highest in patients aged 10~19 years. The co-infection rate of HRV was 35.3%, and the most common combination was with Adenovirus. Respiratory virus infections are known to occur in children and elderly people with weak immunity. However, in this study, the detection rate was second highest in patients aged 10~19 years. Indeed, the detection rate in this age group was more than 15%, except in January and February. These results suggested that steady-state studies on the infection patterns of HRV are required.