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Development and Expression of Porcine Embryos by Direct Injection of Sperm Treated with Exogenous DNA (외래유전자 도입정자를 이용한 돼지 체외성숙 난포란의 Intracytoplasmic Sperm Injection (ICSI) 후 후기 배로의 발달율과 외래유전자의 발현에 관한 연구)

  • 정기화;조성근
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.45-53
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    • 2002
  • The main goal of this study was to produce transgenic porcine embryos by direct injection of sperm-mediated exogenous DNA. Spermatozoa (6$\times$10$^{6}$ sperms of final concentration) were mixed with pcDNA LAC Z (20 ng/$\mu$l) and subjected into electroporation (300~750 volts, 25 $\mu$F, 0.4 cm electrode). After sperm injection, the oocytes were activated electrically (1.7 KV/cm, 30$\mu$sec, single pulse) in 0.3 M mannitol solution or not. The sperm injected eggs were cultured in NCSU 23 medium (0.4% BSA) at 39$^{\circ}C$, 5% $CO_2$ in air fur 144 h. The rates of cleavage and development into blastocyst stage in activation group were significantly higher than those of non-activation group (79.6% and 24.1% vs. 46.3% and 14.4%, respectively, p<0.05). Control oocytes and shame injection were developed to blastocysts low (2.5%). Sixty five (27.1%) out of 240 embryos observed in activation and non-activation groups were showed positive by X-gal staining. However, all embryos in both groups were expressed partial or mosaic pattern. These results suggested that electrical stimulation far oocytes activation after sperm injection enhances the incidence of both fertilization and development fellowing sperm injection in the pig. Our study also suggested that sperm-mediated transfer of exogenous DNA by ICSI would be used as a valuable tool for the production of transgenic porcine embryos.

A STUDY ON THE DISTRIBUTION OF CALCITONIN GENE-RELATED PEPTIDE CONTAINING NERVE FIBERS IN RAT PULP FOLLOWING DENTINAL INJURY (상아질 손상 후 흰쥐 대구치 치수의 calcitonin gene-related peptide(CGRP) 함유 신경섬유 분포에 관한 연구)

  • Moon, Joo-Hoon;Park, Sang-Jin;Min, Byung-Soon;Choi, Ho-Young;Cho, Gi-Woon
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.100-115
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    • 1999
  • The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.

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Temporal and Spatial Variability of the Middle and Lower Tropospheric Temperatures from MSU and ECMWF (MSU와 ECMWF에서 유도된 중간 및 하부 대류권 온도의 시 ${\cdot}$ 공간 변동)

  • Yoo, Jung-Moon;Lee, Eun-Joo
    • Journal of the Korean earth science society
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    • v.21 no.5
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    • pp.503-524
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    • 2000
  • Intercomparisons between four kinds of data have been done to estimate the accuracy of satellite observations and model reanalysis for middle and lower tropospheric thermal state over regional oceans. The data include the Microwave Sounding Units (MSU) Channel 2 (Ch2) brightness temperatures of NOAA satellites and the vertically weighted corresponding temperature of ECMWF GCM (1980-93). The satellite data for midtropospheric temperatures are MSU2 (1980-98) in nadir direction and SC2 (1980-97) in multiple scans, and for lower tropospheric temperature SC2R (1980-97). MSU2 was derived in this study while SC2 and SC2R were described in Spencer and Christy (1992a, 1992b). Temporal correlations between the above data were high (r${\ge}$0.90) in the middle and high latitudes, but low(r${\sim}$0.65) over the low latitude and more convective regions. Their values with SC2R which included the noises due to hydrometeors and surface emission were conspicuously low. The reanalysis shows higher correlation with SC2 than with MSU2 partially because of the hydrometeors screening. SC2R in monthly climatological anomalies was more sensitive to surface thermal condition in northern hemisphere than MSU2 or SC2. The first EOF mode for the monthly mean data of MSU and ECMWF shows annual cycle over most regions except the tropics. The mode in MSU2 over the Pacific suggests the east-west dipole due to the Walker circulation, but this tendency is not clear in other data. In the first and second modes for the Ch2 anomalies over most regions, the MSU and ECMWF data commonly indicate interannual variability due to El Ni${\tilde{n}$o and La Ni${\tilde{n}$a. The substantial disagreement between observations and model reanalysis occurs over the equatorial upwelling region of the western Pacific, suggesting uncertainties in the model parameterization of atmosphere-ocean interaction.

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A study of Conjunctival Cellular Changes in Dry Eye Patients by Impression Cytology (Impression cytology를 이용한 건성안의 결막 세포변화에 관한 연구)

  • Kim, Jai-Min;Kho, Eun-Gyung;Chae, Soo-Chul;Kim, Soon-Ae
    • Journal of Korean Ophthalmic Optics Society
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    • v.9 no.2
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    • pp.333-343
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    • 2004
  • Impression cytology refers to application of cellulose acetate filter material to the ocular surface to remove the superficial layers of the conjunctival epithelium. The technique is non-invasive, is easy to perform, causes minimal discomfort to the patient, and can be used to follow changes in the conjunctival ocular surface over time. With this method, the morphology of the conjunctival ocular surface can be studied and the degree of squmaous metaplasia assessed. This study was performed to evaluate the conjunctival surface by impression cytology in dry eye patients. A total of 70 students with no contact lens wearing history were recruited. Subjects were required to fill in a McMonnies dry eye symptom questionnaire. The non-invasive tear thinning time(TIT) test of each subject was measured, followed by Schirmer tear test(STI), tear film break-up time(TBUT) tests and Rose-bengal staining were performed as a baseline. Conjunctival epithelial cells from the inferior bulbar conjunctiva were harvested by the impression cytology technique. The specimens collected were labelled and stained with PAS(Periodic Acid Schift)-haematoxylin. The goblet cells and conjunctival epithelial cells were observed under a light microscope of 400x magnification. The specimens were classified according to the Nelson Grading scale which was based on the degree of squamous metaplasia such as changes of goblet cells density, size/form, N:C(nucleus : cytoplasm) ratio. Dry eye patients were observed morphological changes of the epithelial cells, different nuclear alterations, decrease of the goblet cells density. The degree of cytological changes was related to severity of dry eye conditions. When the epithelial cell morphology was graded according to the system described by Nelson, specimens from the control group revealed 91.43% of the eyes to be grade 0 and 8.57% to be grade 1, whereas of the dry eye patients, 20% were grade 0, 42.86% grade 1, 34.29% grade 2 and 2,86% grade 3. Impression cytology represents a non- or minimally invasive biopsy of the ocular surface epithelium with no side effects or contraindications. It has demonstrated to be a useful diagnostic aid for a wide variety of processes involving the ocular surface. This technique is a safe, simple method and may help increase understanding of various ocular surface alterations in dry eye patients.

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Two Dimensional Size Effect on the Compressive Strength of Composite Plates Considering Influence of an Anti-buckling Device (좌굴방지장치 영향을 고려한 복합재 적층판의 압축강도에 대한 이차원 크기 효과)

  • ;;C. Soutis
    • Composites Research
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    • v.15 no.4
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    • pp.23-31
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    • 2002
  • The two dimensional size effect of specimen gauge section ($length{\;}{\times}{\;}width$) was investigated on the compressive behavior of a T300/924 $\textrm{[}45/-45/0/90\textrm{]}_{3s}$, carbon fiber-epoxy laminate. A modified ICSTM compression test fixture was used together with an anti-buckling device to test 3mm thick specimens with a $30mm{\;}{\times}{\;}30mm,{\;}50mm{\;}{\times}{\;}50mm,{\;}70mm{\;}{\times}{\;}70mm{\;}and{\;}90mm{\;}{\times}{\;}90mm$ gauge length by width section. In all cases failure was sudden and occurred mainly within the gauge length. Post failure examination suggests that $0^{\circ}$ fiber microbuckling is the critical damage mechanism that causes final failure. This is the matrix dominated failure mode and its triggering depends very much on initial fiber waviness. It is suggested that manufacturing process and quality may play a significant role in determining the compressive strength. When the anti-buckling device was used on specimens, it was showed that the compressive strength with the device was slightly greater than that without the device due to surface friction between the specimen and the device by pretoque in bolts of the device. In the analysis result on influence of the anti-buckling device using the finite element method, it was found that the compressive strength with the anti-buckling device by loaded bolts was about 7% higher than actual compressive strength. Additionally, compressive tests on specimen with an open hole were performed. The local stress concentration arising from the hole dominates the strength of the laminate rather than the stresses in the bulk of the material. It is observed that the remote failure stress decreases with increasing hole size and specimen width but is generally well above the value one might predict from the elastic stress concentration factor. This suggests that the material is not ideally brittle and some stress relief occurs around the hole. X-ray radiography reveals that damage in the form of fiber microbuckling and delamination initiates at the edge of the hole at approximately 80% of the failure load and extends stably under increasing load before becoming unstable at a critical length of 2-3mm (depends on specimen geometry). This damage growth and failure are analysed by a linear cohesive zone model. Using the independently measured laminate parameters of unnotched compressive strength and in-plane fracture toughness the model predicts successfully the notched strength as a function of hole size and width.

Viral Patterns and Clincal Analysis of Acute Respiratory Tract Infections of Children in Korea(September, 1998~August, 2002) (바이러스에 의한 소아 급성 하기도 감염의 유행 및 임상양상(1998년 9월~2002년 8월))

  • Lee, Su-Jin;Park, Eun-Young;Oh, Phil-Soo;Lee, Kon-Hee;Kim, Kwang-Nam;Lee, Kyu-Man
    • Pediatric Infection and Vaccine
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    • v.10 no.1
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    • pp.102-113
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    • 2003
  • Purpose : Acute respiratory tract infections are the most common illnesses in children. The great majority of these infections involving lower respiratory tracts infections(LRTI) are caused by respiratory viruses such as respiratory syncytial virus(RSV), parainfluenza virus (PIV), influenza virus(INF), adenovirus(ADV), etc. Our purpose of this study was to examine the seasonal occurrence, clinical and laboratory features of each viral infections. Methods : Nasopharyngeal aspirates(NPA) were collected from 913 hospitalized children on the first day of admission. They were all admitted to Hangang Sacred Heart hospital and diagnosed as LRTIs. The study period was from September 1998 to August 2002. Specimens were inoculated onto HEp-2 cell, LLC-MK2 cell and MDCK cell. Viruses were detected by immunofluorescent method performed at day 10 postinoculation. Respiratory viruses were detected in 251(27%) cases. Medical records of 208 cases diagnosed as acute LRTIs were all taken reviewed and analyzed. Results : The identified pathogens were RSV 122 cases(58%), PIV 30 cases(14%), INFA 20 cases(10%), INFB 21 cases(11%) and ADV 15 cases(7%) during the study period. Outbreaks of RSV infections occurred every year but mostly on December, 1998 and November, 1999. LRTIs by PIV and ADV occurred during all seasons of the year. INFA was isolated moslty on March, 1999 and January, 2000. INFB infections occurred largely on April, 2002. The clinical diagnoses were bronchiolitis 88 cases(43%), pneumonia 87 cases(42%), croup 19 cases(9%) and tracheobronchitis 14 cases(6%). Common symptoms and signs were cough, rhinorrhea, sputum and fever. WBC counts in peripheral blood showed normal findings in 123 cases(59%). Each viral infections also showed some differences in auscultatory findings and chest X-ray findings. Conclusion : We learned that viruses are one of the major and important etiologic agents of acute LRTIs of children in Korea. However, we still need to find out more about its characteristic clinical features and continue studying on their seasonal occurrence to focus on their management and also prevention.

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Studies on the Generation-to-Generation Transmission of Cytoplasmic Polyhedrosis Virus and the Effect of Their Activation on the Induction in the Next Generation in the Silkworm, Bombyx mori L. (Virus의 경란전염이 차대의 잠작에 미치는 영향에 관한 연구)

  • 임종성;김근영
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.85-92
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    • 1974
  • Many of studies on the transovarial transmission of occult virus and their activation due to various stresses such as cold or heat treatment, chemical feeding, and nutritional deficiency, etc., in the silkworm, Bombyx mori L. have been made, but any attempts have been not made to control virus diseases by detection of the occult virus-carried moths in the production of silkworm egg of hybrids, because of difficulty to detect occult virus in any stage. Therefore, it may be worth while to disclose whether a sublethal infection of the moths from which active virus are detectable, has the same level of induction rate as that of occult virus activation, thus to apply its results for the reduction of the occurence of virus diseases in silkworm rearing. For these purposes, the following experiment was conducted as one of preliminary steps. In this study, investigations on the generation-to-generation transmission of occult virus and a sublethal infection, and the role of chromosomal gene of the host, Jam 103 and Jam 104 in the Previous generation, and Jam 103 x Jam 103 and Jam 104 f Jam 104 in the next generation were made for the induction of virus diseases due to the transmitted virus. The frequency of cytoplasmic polyhedrosis due to the induction in the F$_1$ generation was markedly higher in the cross-batches, male$\times$female and male$\times$female in which inoculated individuals were used as fem ale parents than in the cross-batches, male$\times$female and male$\times$female in which virus has been not inoculated or inoculated only to male in the previous generation. The tendency of increasing rate was observed in any treatments; such as the inoculations of cytoplasmic polyhedrosis virus (10$\^$5/, 10$\^$6/ 10$\^$7, and 10$\^$8//ml ill different concentration of inocula) , cold-treatment (5$^{\circ}C$, 12hrs or 24hrs), and formalin-feeding treatment (2% or 3%). The shape of polyhedra (tetragonal in outline) examined in the F, larvae was identified as that of the inoculated polyhedra with partial application of immunofluorescent techniques. These results suggests that the cytoplasmic polyhedrosis virus in B. meri L. are transmitted to the next generation through the egg, apparently in the occult state. And the experimental results of various cross-batches revealed the egg cytoplasm plays an important part i the transmission of the occult virus of the cytoplasmic polyhedrosis virus,

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Effect of Dietary Fish Oil on Lipid Peroxidation in Rats Liver and Brain During Postnatal Development (어유섭취가 출생후 발달과정의 흰 쥐의 간과 뇌조직의 지질과산화와 그 관련기능에 미치는 영향)

  • 박명희
    • Journal of Nutrition and Health
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    • v.20 no.2
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    • pp.111-121
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    • 1987
  • Lipie peroxide formation, antiperoxidative s system and body adaptability for handling lipid p peroxide were examined in the first and second g generations of rats fed fish oil. Mackerel oil(MO) was used and four other dietary oils and fat, i.e. soybean oil(SO), perilla oil(PO), rapeseed oil(RO) and beef tallow(BT) were also employed to compare the effect of fish oil. Synthetic diets containing these five dietary fats at the level of 1O%(w/w), were given to the correspond­m ing groups of male and female rats weighing about 70 grams. After 34 days of feeding, male a and female rats were mated and their offsprings were raised throughout suckling (17, 26 days) and weanling (39 days) periods. Liver lipid pero­x xide level was highest in MO group of both first (mother rats after lactation) and second genera­t tions of 17 and 26 days old, but not of 39 days old. During suckling period, liver lipid peroxide level was well matched to total unsaturation of dietary fat. Brain lipid peroxide levels were not different among five groups. Liver $alpha$-tocopherol a and reduced glutathione (GSH) levels were lowest in MO fed first generation. In second generation, $alpha$-tocopherol level was also low in MO group, although the effect was less pronoun­c ced, but GSH level was not different from other groups. Oxidized glutathione (GSSG) level did not consistently vary by change in dietary fat. Glutathione peroxidase activity increased as young rats grew up to 39 days. Superoxide d dismutase activity change was insignificant by a age, but was shown as lowest in MO group. At the age of 26 and 39 days, liver glutatione peroxidase activity was increased as was level of lipid peroxide, suggesting that this is the one of the mechanisms responsible for body adapta­b bility for protection against the accumulation of lipid peroxide.

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P53 Overexpression and Outcome of Radiation Therapy in Head & Neck Cancers (두경부종양 환자에서 p53의 과발현과 방사선치료결과)

  • Kim In Ah;Choi Ihl Bhong;Kang Ki Mun;Jang Ji Young;Kim Kyung Mi;Park Kyung Shin;Young Shin Kim;Kang Chang Suk;Cho Seung Ho;Kim Hyung Tae
    • Radiation Oncology Journal
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    • v.17 no.1
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    • pp.1-8
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    • 1999
  • Purpose : Experimental studies have implicated the wild type p53 In cellular response to radiation. Whether altered p53 function can lead to changes in clinical radiocurability remains an area of ongoing study. This study was performed to investigate whether any correlation between change of p53 and outcome of curative radiation therapy in patients with head and neck cancels. Methods : Immunohistochemical analysis with a mouse monoclonal antibody (DO-7) specific for human p53 was used to detect to overexpression of protein in formalin fixed, paraffin-embedded tumor sample from 55 head and neck cancer patients treated with curative radiation therapy (median dose of 7020 cGy) from February 1988 to March 1996 at 51. Mary's Hospital. Overexpression of p53 was correlated with locoregional control and survival using Kaplan-Meier method. A Cox regression multi-variate analysis was peformed that included all clinical variables and status of p53 expression. Results : Thirty-seven (67.2$\%$) patients showed overexpression of p53 by immunohistochemical staining in their tumor. One hundred percent of oral cavity, 70$\%$ of laryngeal, 66.7$\%$ of oropharyngeal, 66.7$\%$ of hypopharyngeal cancer showed p53 overexpression (P=0.05). The status of p53 had significant relationship with stage of disease (P=0.03) and history of smoking (P=0.001). The overexpression of p53 was not predictive of response rate to radiation therapy. The locoregional control was not significantly affected by p53 status. Overexpression of p53 didn't have any prognostic implication for disease free survival and overall survival. Primary site and stage of disease were significant prognostic factors for survival. Conclusions : The p53 overexpression as detected by immunohistochemical staining had significant correlation with stage, primary site of disease and smoking habit of patients. The p53 overexpression didn't have any predictive value for outcome of curative radiation therapy in a group of head and neck cancers.

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A Study on the Development of an Instrument for Evaluating the Quality of Nursing Care (간호의 질 평가도구 개발에 관한 일 연구)

  • 유지수
    • Journal of Korean Academy of Nursing
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    • v.7 no.2
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    • pp.11-21
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    • 1977
  • Many in nursing look back on Nursing Research history and proudly point to the fact that emphasis in nursing research has changed from studying the nurse to studying nursing practice. In recent years, much emphasis has been placed on seeking a method of evaluating the quality of nursing care. In spite of these attempts, however, an instrument for evaluating the quality of nursing care that is actually applicable in the clinical area has not been found. The Purposes of this study are as follows: 1) To develop the instrument to be used in evaluating the quality of nursing care provided in the Neuro - Surgery Constant Care Unit of Severance Hospital 2) To evaluate the quality of nursing care in the clinical area. 3) To provide the necessary information for improvement of quality of nursing care. The instrument for evaluating the quality of nursing care, developed by the investigator, was composed of 7 nursing goals and divided into 65 standards of nursing performance. The 7 nursing goal are as follows : 1) Maintenance of airway 2) Maintenance of fluid at electrolyte balance 3) Maintenance of elimination 4) Personal hygiene 5) Optimum activity 6) Prevention of accidents 7) Emotional care The study population defined was composed of all the case (51) who were admitted in the Neuro- Surgery Constant Care Unit of Severance Hospital from May 7-13, 1976. The observation method was used and the data was subjected to the %, X²-test, T-test, F-test and Correlation. The results of tile study were as follows : 1. Levels of nursing performance regarding nursing goals. Seven different nursing care indices were constructed in terms of nursing goals. The index scores were grouped arbitrarily into ,j categories such as "excellent", "good", "moderate", "incomplete", and "poor"based upon the investigator′s personal judgement. a. The nursing index of maintaining airway showed that 78% of the patients fell within the "excellent" and 22% of the patients, fell within the "good" category. b. The nursing index of maintaining fluid & electrolyte balance showed that 95% of the patients fell within the "excellent" and 5 % of the patients fell within the "good" category. c. The nursing index of maintaining elimination showed that 100% of the patients fell within the "excellent" category. d. The nursing index of personal hygiene revealed that 49% of the patients fell within the "excellent" and 51% of the patients fell within the "good" category. e. The nursing index of optimum activity showed that 63% of the patients fell within the "excellent" and 32% of the patients fell within tile "good" and 5% of patients fell within the "moderate" category. f. The nursing index of prevention of accidents showed that 100% of the patients foil within the "excellent" category. g. The nursing index of emotional cart revealed that 27% of the patients fell within the "excellent", 24 % of the patients fell within tile "good", 29 % of the patients fell within the "incomplete" category. From these findings it is disclosed that the quality of nursing care provided in the Neuro- Surgery Constant Care Unit of Severance Hospital was excellent. h. There were statistically significant differences between the nursing index of physical care and emotional care. (t=8.73, D. F. =100. p<0.01) It is revealed that more physical care then emotional care was carried out by nurses. 2. Levels of nursing performance regarding general characteristics of the patients. No significant differences were observed statistically with the nursing indices of nursing goals according to the sex (t=0.084, D. F. =12, p>0.05). Age (F=0.1251, D. F. : 3.18. p 〉0.05), absence or presence of operating experiences (t=0.6032, D. F. =12, p〉0.05, levels of consciousness (F=0.31, D. F. :3. 18, p >0.05) 3. Relationship between the levels of consciousness and the nursing index of each nursing goal. There was negative correlation between the levels of consciousness and the nursing index of maintaining airway (r=-0. 5449, p<0.01) and personal hygiene (r= -0.4075, p<0.01) There was positive correlation between the levels of consciousness and the nursing index of optimum activity (r=0.3936, p <0.01) and emotional care (r=0.7819, p〈0.01). There was slight correlation between the levels of consciousness and the nursing index of maintaining fluid & electrolyte balance (r=-0.3418, 0.010.05) and preventing accidents (r=0.1441, p>0.05.

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