• Journal of fish pathology
• /
• v.6 no.2
• /
• pp.93-101
• /
• 1993

The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

• Clinical and Experimental Reproductive Medicine
• /
• v.19 no.1
• /
• pp.87-94
• /
• 1992

From December 1991 to March 1992, 34 anejacuratory patients with spinal cord injury underwent 90 of electric stimulations with Seager NRH model 12. The average patient age was 43.5 years with a range of 23 to 48 years. The level of cord injury was cervical in 7, thoracic in 6, lumbar in 11, lumbosacral in 7 and conus medullaris in 3. The average number of electric stimulation per a patient was 2.65 with a range of 1 to 4. The average voltage and amplitude per a stimulation were 17.72 volts and 309. 89 mAmp with ranges of 5 to 25 volts and 50 to 500 mAmp. The total and motile sperm number were evaluated microscopically and analyzed statistically by paired t-test according to the frequency of electroejaculation, level of cord injury and voiding pattern. The results were obtained as follows. 1. An overall success rate of electroejaculation was 85.3% among 34 patients and 82.2% among 90 electric stimulations. 2. The total and motile sperm number per a stimulation were not correlated the frequency of electric stimulation, level of cord injury and voiding pattern. 3. Complications occured in 10 cases; severe low abdominal pain in 5, hypertension in 2, sweating in 1, headache in 1 and neck stiffness in 1. All the copmlications subsided spontaneously within 5 to 10 minutes after transient interruption of the electric stimulation. In summary, rectal probe electroejaculation is an accepted safe means of procuring sperm from spinal cord injury patients with ejaculatory incompetence. However very poor sperm motility was found and it was not related with the frequency of electroejaculation, level of cord injury and voiding pattern. Further investigation would be needed to conclude and to identify the reasons for impaired sperm motility.

• Proceedings of the Materials Research Society of Korea Conference
• /
• 2009.11a
• /
• pp.41.1-41.1
• /
• 2009

Aerosol Deposition (AD) is anovel way to fabricate bioactive ceramic coatings in biomedical implants and prostheses applications. In the present work, silicon-substituted hydroxyapatite (HA) coatings on commercially pure titanium were prepared by aerosol deposition using Si-HA powders. The incorporation of silicon in the HA lattice is known to improve the bioactivity of the HA, makingsilicon-substitute HA an attractive alternative to pure HA in biomedical applications. Si-HA powders with the chemical formula $Ca_{10}(PO_4)_6-x(SiO_4)x(OH)_2-x$, having silicon contents up to x=0.5 (1.4 wt%), were synthesized by solid-state reaction of $Ca_2P_2O_7$, $CaCO_3$, and $SiO_2$. The Si-HA powders were characterized by X-ray diffraction (XRD), X-ray fluorescence spectrometry (XRF), and Fourier transform infrared spectroscopy(FT-IR). The corresponding coatings were also analyzed by XRD, scanning electron microscopy (SEM), and electron probe microanalyzer (EPMA). The results revealed that a single-phase Si-HA was obtained without any secondary phases such as $\alpha$- or $\beta$-tricalcium phosphate (TCP) for both the powders and the coatings.The Si-HA coating was about $5\;{\mu}m$ thick, had a densemicrostructure with no cracks or pores. In addition, the proliferation and alkaline phosphatase (ALP) activity of MC3T3-E1 preosteoblast cells grown on the Si-HA coatings were significantly higher than those on the bare Ti and pure HA coating. These results revealed the stimulatory effects induced by siliconsubstitution on the cellular response to the HA coating.

• Journal of Korean Society of Environmental Engineers
• /
• v.27 no.1
• /
• pp.59-66
• /
• 2005

[ $TiO_2$ ] was deposited by DC magnetron sputtering on glass surface under various sputtering parameters such as discharge power($0.6{\sim}5.2\;kW$, substrate temperature($R.T{\sim}350^{\circ}C$), Ar and $O_2$ flow ratio with $0{\sim}50\;sccm$($Ar+O_2$ 90 sccm) and about 1 mtorr of pressure. TiO-N thin film was prepared under same sputtering conditions for $TiO_2$ thin film except flow ratio($Ar+O_2+N_2$ 90 sccm). The sheet resistance of thin films deposited under these parameters was measured to analyze electronic characteristic and thin film's thickness(${\alpha}$-step), surface roughness(AFM) and formation construction(FE-SEM, XRD) were also measured to draw optimal sputtering parameters. In order to evaluate photo-activity of thin film($TiO_2$, TiO-N) made in optimal parameters for removal of pollutants, toluene among VOCs and Suncion Yellow among reactive dyes were chosen to probe organic compounds for photo-degradation. It was shown that the photo-catalytic thin films had a significant photo-activation for the chosen contaminants and especially TiO-N thin film showed maximum efficiency of 33% for toluene(5 ppm) removal in visible-light range.

• Proceedings of the Korean Radioactive Waste Society Conference
• /
• 2004.06a
• /
• pp.353-354
• /
• 2004

사용후 핵연료의 조성을 분석하거나 또는 반사전자상과 2차 전자상 등으로 시료를 관찰하기 위해서는 핫셀(Hot cell)에 증착기(coater)를 설치하여 시료표면을 전도성 물질인 탄소 등으로 증착시켜야 한다. 그러나 원격조정기를(manipulator)를 이용하여 수행되는 핫셀에서의 증착작업은 사용후 핵연료 시험의 선진분석기술을 갖고 있는 원자력 선진국에서도 핫셀내에 설치되어 있는 증착기의 탄소봉을 교체하는 작업과 진공장치의 성능 유지가 까다로워 시료표면에 균질하게 전도성 물질을 증착시키는 작업에 많은 어려움을 겪고 있다. 본 연구는 통상적으로 이용되는 증착기를 사용하지 않고 Silver Paint를 사용하여 사용후 핵연료를 분석할 수 있는 새로운 방법에 대한 연구를 수행하였다. 산화물 핵연료는 전기전도도가 매우 낮아($3{\times}10^{-1}~4{\times}10^{-8}/ohm{\cdot}cm$)입사된 전자의 이동이 원활하지 못해 일어나는 들뜸(Charging)현상이 발생한다. 그러나 Silver Paint 에 사용후 핵연료를 접착하면 모세관(capillary)현상에 의해 시료 주위와 핵연료의 결정립계로 Silver가 스며들어 입사된 전자의 이동이 원활해져 전도성이 극히 낮은 시료의 분석이 가능하게 된다. 본 시험에 사용된 EPMA는 (Electron Probe Micro Analyzer, SX-50R, CAMECA, Paris, France) 고 방사능을 띤 조사 핵연료의 시험을 수행할 수 있도록 기기의 적절한 부위에 납과 텅스텐으로 차폐되어 시편의 방사능 세기가 $3{\times}10^{10}Bq$까지 시험 가능한 기기이다. 그림 1은 JAERI 에 설치 운영중인 증착기 설비 사진이다. 그림에서 핫셀에 설치된 증착기의 진공을 유지하기 위해 핫셀 벽을 관통하여 증착기 본체까지 연결된 배출관의 형상과 복잡한 주변장치들을 볼 수 있다. 그림 2는 비조사 핵연료 시편을 Silver Pain떼 접착한 사진이다. 그림은 시료주위와 시료 표면까지 Silver Paint가 도포된 모습을 보여주고 있다. 상용발전소에서 연소도가 50,000 Mwd/tU인 사용후 핵연료를 상기와 같은 방법으로 만든 시편의 표면을 관찰한 사진을 그림 3~8에 나타내었다. 그림 3은 핵연료 중앙부위의 결정립을 나타낸 그림이다. Silver Paint만으로 접착한 시료의 표면관찰 및 정량분석이 그림에서 보듯이 가능함을 확인하였다. 그림 4는 사용후 핵연료시료를 중앙부위에서 가장자리까지를 다섯 부위로 나누어 그 중 중앙부위(1/5) 지점의 입계 및 형상을 관찰한 사진이다. 결정립의 크기가 다른 부위보다 상대적으로 크고, 결정립에 생성된 기공이 발달되어 있음을 볼 수 있다. 그림 5와 6과 7은 중심부위와 rim부위 사이 지점을 관찰한 사진으로서 결정립과 기공의 분포가 비슷한 형상을 나타내고 있음을 관찰할 수 있었다. 그림 8은 rim 부위 사진으로 전형적인 rim 영역 현상을 관찰할 수 있었다. 표 1은 그림 2와 같이 비조사 산화물 핵연료를 Silver Paint로 접착한 시편을 정량 분석한 결과이다. 시편의 조성은 33.6 at% U, 66.4 at% O의 결과를 얻었다. 산화물 핵연료의 표면 관찰 및 정량 분석 시험시 시편 표면을 전도성 물질로 증착시키지 않고, Silver Paint 에 시편을 접착하는 방법으로도 만족한 시험 결과를 얻을 수 있었다.

• Journal of Periodontal and Implant Science
• /
• v.26 no.3
• /
• pp.771-778
• /
• 1996

The assessment of alveolar bone changes on dental radiographs to indicate progression of periodontal diseases or healing response to therapy is routine procedure. However, the diagnostic accuracy in detecting small alveolar bone changes is very limited. Recently, guided bone regeneration therapy is popular, but the quantification of new bone is somewhat difficult with conventional evaluation method. To quantificate the amount of new bone, various evaluating methods have been introduced including histomorphometry, radiomorphometry, biochemical analysis, X-ray probe microanalysis, scanning electron microscope backscatter method. In this study, guided bone regeneration using resorbable membrane with & without PDGF-BB is quatificated through histomorphmetry to evaluate the efficacy of histomorphometric analysis. 4 beagle dogs and 8 Sprague-Dawley rats were selected as experimental animals. In beagle dog experiment, $4{\times}4mm$ Class II defects were created in maxillary both second premolars, and biodegradable membrane containing PDGF-BB(experimental group) were covered over one defect, and same membrane without PDGF-BB(control group) were covered over the other defect. At 2 weeks, 5 weeks after surgery, each beagle dogs were sacrificed, and the tissues were treated by undecalcified fixation. In Sprague-Dawley rat experiment, 5mm round defect were created in temporal bone, the same membranes were covered on the defects. At 1 week, 2 weeks after surgery, each rats were sacrificed, and undecalcified fixation were taken. After grinding tissue specimen, we analyse them histomorphometrically using image analysis system. In beagle dog 2 weeks specimens, new bone formation area were $0.03123mm^2$ in experimental group,and $0.03012mm^2$ in control group. At 5 weeks specimens, $0.15324mm^2$ in experimental group, and $0.09123mm^2$ in control group. In Sprague-Dawley rat specimens, new bone fomation area were $0.20448mm^2$ in 1 week experimental group, $0.03604mm^2$ in 1 week control group. At 2 weeks specimens, $0.46349mm^2$ in experimental group, $0.17741mm^2$ in control group. The results indicated that histomorphometric analysis of new bone formation using image analysis system is very effective quantification method to evaluate the efficacy of treatment modalities.

• Journal of Plant Biotechnology
• /
• v.33 no.1
• /
• pp.19-25
• /
• 2006

With the use of Agrobacterium and gene-gun, cold regulated gene (BN115) has been injected in Chrysanthemum leaf disc and transgenic plants have been produced successfully on the selection media containing phytohormone. To determine the presence of the transferred cold regulated gene (BN115) in the transgenic Chrysanthemum, PCR-amplification indicated the presence of that gene. Real-Time PCR for confirmation of the putative transgenic plants was established. The copy number of cold regulated gene (BN115) is extrapolated on the basis of a standard curve. Serial dilutions of known number of gene copies were in triplicates. In this diagram, PCR cycles are plotted against the fluorescence intensity. The cycle at which the fluorescence reaches a threshold cycle is inversely proportional to the starting amount of target DNA.

• Korean Journal of Materials Research
• /
• v.4 no.1
• /
• pp.81-89
• /
• 1994

Ti film of lOnm thickness and Co film of 18nm thickness were sequentially e-heam evaporated onto Si (100) substrates. Metal deposited samples were rapidly thermal-annt.aled(KTA) in thr N1 en vironment a t $900^{\circ}C$ for 20 sec. to induce the reversal of metal bilayer, so that $CoSi_{2}$ thin films could be formed. The sheet resistance measured by the 4-point probe was 3.9 $\Omega /\square$This valur was maintained with increase in annealing time upto 150 seconds, showing high thermal stab~lity. Thc XRII spectra idrn tified the silicide film formed on the Si substrate as a $CoSi_{2}$ epitaxial layer. The SKM microgr;iphs showed smooth surface, and the cross-sectional TKM pictures revealed that the layer formed on the Si substrate were composed of two Co-Ti-Si alloy layers and 70nm thick $CoSi_{2}$ epl-layer. The AES analysis indicated that the native oxide on Si subs~rate was removed by TI ar the beginning of the RTA, and Ihcn that Co diffused to clean surface of Si substrate so that epitaxial $CoSi_{2}$ film could bt, formed. In thc rasp of KTA at $700^{\circ}C$. 20sec. followed by $900^{\circ}C$, 20sec., the thin film showed lower sheet resistance, but rough surface and interface owing to $CoSi_{2}$ crystal growth. The application scheme of this $CoSi_{2}$ epilayer to VLSI devices and the thermodynarnic/kinetic mechan~sms of the $CoSi_{2}$ epi-layer formation through the reversal of Co/Ti bdayer were discussed.

• Journal of Animal Science and Technology
• /
• v.45 no.2
• /
• pp.169-182
• /
• 2003

To understand the structure and regulation of bovine ADRP (Adipocyte Differentiation Related Protein) gene, we have isolated the genomic clone of bovine ADRP and determined its sequence. A genomic Southern blot analysis confirmed that ADRP gene is present as a single copy in bovine genome and the ADRP gene spans 12 kb. Bovine ADRP genomic clone, HwADRPg-1, had 8 exons and 7 introns, and all splicing sites conformed to the GT/AG rule with the exon-intron boundaries located exactly. Analysis of the upstream 649 bp of the sequence of HwADRPg-1 showed that it does not contain any canonical TATAA boxes; however Sp1 binding sites and CAAT boxes are found. The promoter contained potential binding sites for AP-1, AP-2 and several putative transcription factor binding sites. The 5'-flanking region of HwADRPg-1 contained muscle specific transcription activator Myo G and C/EBP (CCAAT/ enhancer binding protein) recognizing site. These results suppose that the Myo G transcription activator regulate the transcription of bovine ADRP gene in muscular tissue and its transcriptional activity was triggered by degree of muscular development. Our results provide the necessary analysis for other flanking sequences are needed in addition to the proximal cis elements of this promoter to confer adipocyte differentiation-dependent or growth-dependent transcriptional control.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.12
• /
• pp.4915-4920
• /
• 2015

Background: : Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease mainly caused by mutations of the adenomatous polyposis coli (APC) gene with almost complete penetrance. These colorectal polyps are precancerous lesions that will inevitable develop into colorectal cancer at the median age of 40-year old if total proctocolectomy is not performed. So identification of APC germline mutations has great implications for genetic counseling and management of FAP patients. In this study, we screened APC germline mutations in Chinese FAP patients, in order to find novel mutations and the APC gene germline mutation characteristics of Chinese FAP patients. Materials and Methods: The FAP patients were diagnosed by clinical manifestations, family histories, endoscope and biopsy. Then patients peripheral blood samples were collected, afterwards, genomic DNA was extracted. The mutation analysis of the APC gene was conducted by direct polymerase chain reaction (PCR) sequencing for micromutations and multiplex ligation-dependent probe amplification (MLPA) for large duplications and/or deletions. Results: We found 6 micromutations out of 14 FAP pedigrees, while there were no large duplications and/or deletions found. These germline mutations are c.5432C>T(p. Ser1811Leu), two c.3926_3930delAAAAG (p.Glu1309AspfsX4), c.3921_3924delAAAA (p.Ile1307MetfsX13), c3184_3187delCAAA(p.Gln1061AspfsX59) and c4127_4126delAT (p.Tyr1376LysfsX9), respectively, and all deletion mutations resulted in a premature stop codon. At the same time, we found c.3921_3924delAAAA and two c.3926_3930delAAAAG are located in AAAAG short tandem repeats, c3184_3187delCAAA is located in the CAAA interrupted direct repeats, and c4127_4128 del AT is located in the 5'-CCTGAACA-3', 3'-ACAAGTCC-5 palindromes (inverted repeats) of the APC gene. Furthermore, deletion mutations are mostly located at condon 1309. Conclusions: Though there were no novel mutations found as the pathogenic gene of FAP in this study, we found nucleotide sequence containing short tandem repeats and palindromes (inverted repeats), especially the 5 bp base deletion at codon 1309, are mutations in high incidence area in APC gene,.