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Utilizing cell-free DNA to validate targeted disruption of MYO7A in rhesus macaque pre-implantation embryos

  • Junghyun Ryu;Fernanda C. Burch;Emily Mishler;Martha Neuringer;Jon D. Hennebold;Carol Hanna
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.4
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    • pp.292-297
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    • 2022
  • Direct injection of CRISPR/Cas9 into zygotes enables the production of genetically modified nonhuman primates (NHPs) essential for modeling specific human diseases, such as Usher syndrome, and for developing novel therapeutic strategies. Usher syndrome is a rare genetic disease that causes loss of hearing, retinal degeneration, and problems with balance, and is attributed to a mutation in MYO7A, a gene that encodes an uncommon myosin motor protein expressed in the inner ear and retinal photoreceptors. To produce an Usher syndrome type 1B (USH1B) rhesus macaque model, we disrupted the MYO7A gene in developing zygotes. Identification of appropriately edited MYO7A embryos for knockout embryo transfer requires sequence analysis of material recovered from a trophectoderm (TE) cell biopsy. However, the TE biopsy procedure is labor intensive and could adversely impact embryo development. Recent studies have reported using cell-free DNA (cfDNA) from embryo culture media to detect aneuploid embryos in human in vitro fertilization (IVF) clinics. The cfDNA is released from the embryo during cell division or cell death, suggesting that cfDNA may be a viable resource for sequence analysis. Moreover, cfDNA collection is not invasive to the embryo and does not require special tools or expertise. We hypothesized that selection of appropriate edited embryos could be performed by analyzing cfDNA for MYO7A editing in embryo culture medium, and that this method would be advantageous for the subsequent generation of genetically modified NHPs. The purpose of this experiment is to determine whether cfDNA can be used to identify the target gene mutation of CRISPR/Cas9 injected embryos. In this study, we were able to obtain and utilize cfDNA to confirm the mutagenesis of MYO7A, but the method will require further optimization to obtain better accuracy before it can replace the TE biopsy approach.

The relationship between the prevalence of anemia and dietary intake among adults according to household types based on data from the 7th (2016-2018) Korea National Health and Nutrition Examination Survey (국민건강영양조사 제7기 (2016-2018년)에서의 가구 유형에 따른 성인의 빈혈 유병율과 식이 섭취)

  • Hye Won Kim;Ji-Myung Kim
    • Journal of Nutrition and Health
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    • v.56 no.5
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    • pp.510-522
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    • 2023
  • Purpose: In this study, data from the 7th Korea National Health and Nutrition Examination Survey (2016-2018) were used to examine the relationship between the prevalence of anemia and dietary intake among adults according to household types. Methods: Using data from a total of 10,646 subjects (4,428 men and 6,218 women), the general information, body measurements, results of biochemical examination, food and nutrient intake, and meal quality evaluation were analyzed according to the type of household. Results: The prevalence of anemia was higher in men belonging to single-person households (SPH) than in those from multi-person households (MPH), while anemia prevalence was higher among the women in the MPH than in the SPH. The men in SPH had a lower total food intake of nuts, vegetables, fruits, fish, and seaweed than the men in MPH, and consumed higher quantities of milk, oil, and processed foods. The women from SPH had a lower intake of seaweed and a higher intake of milk than those belonging to the MPH. In addition, the men in SPH had a lower iron intake and iron intake per 1,000kcal than the men in MPH, lower iron intake through plant-based foods, and a lower iron intake ratio compared to the reference nutrient intake. The total Korean Healthy Eating Index (KHEI) score was lower in both men and women in SPH than in those from the MPH. When analyzing the relationship between household type and anemia risk after correcting for the confusion variable, the risk of anemia in men in SPH increased compared to those belonging to the MPH. However, women showed no such significant correlation. There was no relationship between the total KHEI score and the risk of anemia by gender and household type. Conclusion: In conclusion, since anemia in men belonging to SPH is a matter of concern, it is essential to develop guidelines for anemia-related nutrition education for men living alone.

An Analytical Study on Stem Growth of Chamaecyparis obtusa (편백(扁栢)의 수간성장(樹幹成長)에 관(關)한 해석적(解析的) 연구(硏究))

  • An, Jong Man;Lee, Kwang Nam
    • Journal of Korean Society of Forest Science
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    • v.77 no.4
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    • pp.429-444
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    • 1988
  • Considering the recent trent toward the development of multiple-use of forest trees, investigations for comprehensive information on these young stands of Hinoki cypress are necessary for rational forest management. From this point of view, 83 sample trees were selected and cut down from 23-ear old stands of Hinoki cypress at Changsung-gun, Chonnam-do. Various stem growth factors of felled trees were measured and canonical correlaton analysis, principal component analysis and factor analysis were applied to investigate the stem growth characteristics, relationships among stem growth factors, and to get potential information and comprehensive information. The results are as follows ; Canonical correlation coefficient between stem volume and quality growth factor was 0.9877. Coefficient of canonical variates showed that DBH among diameter growth factors and height among height growth factors had important effects on stem volume. From the analysis of relationship between stem-volume and canonical variates, which were linearly combined DBH with height as one set, DBH had greater influence on volume growth than height. The 1st-2nd principal components here adopted to fit the effective value of 85% from the pincipal component analysis for 12 stem growth factors. The result showed that the 1st-2nd principal component had cumulative contribution rate of 88.10%. The 1st and the 2nd principal components were interpreted as "size factor" and "shape factor", respectively. From summed proportion of the efficient principal component fur each variate, information of variates except crown diameter, clear length and form height explained more than 87%. Two common factors were set by the eigen value obtained from SMC (squared multiple correlation) of diagonal elements of canonical matrix. There were 2 latent factors, $f_1$ and $f_2$. The former way interpreted as nature of diameter growth system. In inherent phenomenon of 12 growth factor, communalities except clear length and crown diameter had great explanatory poorer of 78.62-98.30%. Eighty three sample trees could he classified into 5 stem types as follows ; medium type within a radius of ${\pm}1$ standard deviation of factor scores, uniformity type in diameter and height growth in the 1st quadrant, slim type in the 2nd quadrant, dwarfish type in the 3rd quadrant, and fall-holed type in the 4 th quadrant.

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Comparison of Health Status and Nutrient Intakes of Elders Who Participated in MOW and Free Congregate Meal Services (가정배달급식과 무료 회합급식 이용 노인의 건강 및 영양섭취상태 비교)

  • Chung, Eun-Jung;Shim, Eu-Gene
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.11
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    • pp.1399-1408
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    • 2007
  • This study was conducted to compare health and nutritional status of 45 home-living elderly people receiving free Meals on Wheels (MOW) (13 men, 32 women) and 81 low income class elderly people receiving free congregate meals (CM) (10 men, 71 women) served in Seoul. Data were obtained from questionnaires, anthropometry and interviews for the 24-hour dietary recall methods. There were no significant differences between the two groups in age and body mass index. Education level, type of housing, family type and income of the two groups also were not significantly different. In MOW, frequencies of exercise were lower while the prevalence of stroke, respiratory disease and loneliness were higher, compared with the CM. The scores of ADL, IADL and food habit of MOW were lower than those of CM. The average daily nutritional intake of both MOW and CM were as a whole under the DRI for Koreans. Energy and macro-nutrient intakes of MOW were tended to be lower than CM (except protein intakes for female). Ca, K, vitamin A, vitamin $B_1$, vitamin $B_2$, vitamin C and folate intakes of MOW were less than 50% of DRI. Percentages of subjects consuming energy less than 75% of EER and 4 nutrients intakes less than EAR were higher in MOW (42.2%) than in CM (1.2%). Our results indicated that dietary nutritional status of MOW was very poor, especially in the case of female elderly groups. For the welfare of the home-living elderly people receiving free MOW, meal service programs should be improved in quality of diet by national supports.

Effect of Tillage System and Livestock Manures on the Silage corn Production and NO3-N Concentration in Leaching Water (경운방법과 가축분뇨 시용이 옥수수의 생산성 및 질소의 용탈에 미치는 영향)

  • Jung, Min-Woong;Jo, Nam-Chul;Kim, Jong-Geun;Lim, Young-Chul;Choi, Ki-Choon;Yoon, Sei-Hyung;Lee, Ki-Won;Yook, Wan-Bang
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.29 no.3
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    • pp.211-216
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    • 2009
  • Approximately 43 million tons of livestock manure (LM) are produced each year on Korean farms. LM can be utilized as a valuable resource and/or it can contaminate water by runoff and leaching through the soil, when LM has been thoughtlessly applied to the land and directly discharged into the water. This experiment was carried out to investigate the effect of no-till system and LM application on dry matter (DM) yield of silage corn and $NO_3$-N concentration in leaching water of lysimeter installed in the experimental field. The treatments were replicated three times in split plot design. Main plots consisted of tillage systems, such as conventional tillage (CT) and no-tillage (NT). Sub plots consisted of the type of LM, such as chemical fertilizer (CF), composted cattle manure (CCM) and composted swine manure (CSM). The control plots were fertilized as commercial chemical fertilizer. DM yields of corn increased significantly in order to CF > CCM > CSM (p<0.05). DM yield of corn in CT increased as comparing with that of corn in NT. Plant height, ear height and stem diameter also increased in order to CF > CCM > CSM. In addition, the root weight in CT was increased as comparing with that of corn in NT. However, there was no interaction effects of between type of LM and tillage system. $NO_3$-N concentration in leaching water of LM application was less than 10 ppm, but $NO_3$-N concentration in CF exceeded 10 ppm which is safety level of drinking water during summer time (rainfall season).

A Clinical Study on Binaural Hearing Aid (양이 보청효과에 관한 연구)

  • 김기령;김영명;심윤주
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1978.06a
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    • pp.9.2-9
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    • 1978
  • Monaural and binaural hearing aid performance under quiet and noisy conditions were compared in regard to (1) the degree of hearing impairment, (2) the symmetry of pure tone audiogram, (3) the automatic gain control of the hearing aid. (4) hearing impairement with recruitment and, word discrimination ability. Performance using binaural hearing aids was consistently superior to that using monaural hearing aids. The results were as follows. 1. Speech detection thresholds were enhanced by a mean of 4.25dB when tested with danavox 747 PP stereo type hearing aid and by a mean of 4.12 dB when tested hearing aids connected seperately to the right and left ears. 2. Binaurally tested speech reception thresholds were superior to monaurally tested thresholds by a mean of 3.56dB when tested in quiet and by a mean of 5.56dB when tested in noise. 3. Binaurally tested word discrimination scores were also superior by a mean of 17.09% in quiet and by a mean 19.63% in noise. 4. Both SRT and word discrimination scores were performed best by subjects with moderately-severe impairement. The performance by one mildly impaired subject was the poorest of all performances. The levels of performance order were; moderately-severe loss, severe loss. moderate loss and mild loss. 5. The data obtained using AGC aids when compaired with that of linear amplification show that when AGC aids were worn in both ears. the results were very poor but when one AGC aid was worn in one ear and linear amplification in the other. the results were good. 6. The advantages of binaural hearing aids were obvious even in cases 1) with great diferences in hearing thresholds between right and left ears, 2) when the subject was unable to discriminate words without vision and. 3) when the subject had extreme recruitme t phenomenon.

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A Literature Study about Comparison of Eastern-Western Medicine on the Acne (여드름의 동(東)·서의학적(西醫學的) 문헌(文獻) 고찰(考察))

  • Joo, Hyun-A;Bae, Hyeon-Jin;Hwang, Chung-Yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.25 no.2
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    • pp.1-19
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    • 2012
  • Objective : The purpose of this study is to investigate about comparison of Eastern-Western medicine on the acne. Methods : We searched Eastern and Western medicine books for acne. We analyzed these books and examined category, definition, etiology, classification, internal and external methods of treatment of acne. Results : The results were as follows. 1. In Eastern medicine, Acne belongs to the category of the Bunja(粉刺), Jwachang(痤瘡), Pyepungbunja(肺風粉刺). In Western medicine, the other name of Acne is acne vulgaris. 2. In Eastern medicine, the definition of Acne includes manual extraction of comedones and skin appearance. In Western medicine, Acne is a common skin disease during adolescence and a chronic inflammatory disease of pilosebaceous unit of self localization. It is characterized by noninflammatory, open or closed comedones and by inflammatory papules, pustules, and nodules and it affects the areas of skin with the densest population of sebaceous follicles, these areas include the face, neck, back, and the upper part of the chest. 3. In Eastern medicine, the cause and mechanism of Acne arose from the state of internal dampness-heat and spleen-stomach internal qi deficiency due to dietary irregularities and then invaded external pathogen such as wind-dampness-heat-cold-fire in lung meridian lead to qi and blood heat depression stagnation. So it appears in skin. In Western medicine, the etiology and pathogenesis of Acne is clearly not identified, but there are most significant pathogenic factors of blood heat depression stagnation. So it appears in skin. In Western medicine, the etiology and pathogenesis of Acne is clearly not identified, but there are most significant pathogenic factors of Acne; Androgen-stimulated production of sebum, hyperkeratinization and obstruction of sebaceous follicles, proliferation of Propionibacterium acnes and inflammation, abnormaility of skin barrier function, genetic aspects, environmental factors etc. 4. In Eastern medicine, differentiation of syndromes classifies clinical aspects, and cause and mechanism of disease; the former is papular, pustular, cystic, nodular, atrophic, comprehensive type; the latter is lung blood heat, intestine-stomach dampness-heat, phlegm-stasis depression, thoroughfare-conception disharmony, heat toxin type. In Western medicine, it divides into an etiology and invasion period, and clinical aspects; Acne neonatorum, Acne infantum, Acne in puberty and adulthood, Acne venenata; Acne vulgaris, Acne conglobata, Acne fulminans, Acne keloidalis. 5. In Eastern medicine, Internal methods of treatment of Acne are divided into five treatments; general treatments, the treatments of single-medicine and experiential description, the treatments depending on the cause and mechanism of disease, and clinical differentiation of syndromes, dietary treatments. In Western medicine, it is a basic principles that regulation on production of sebum, correction on hyperkeratinization of sebaceous follicles, decrease of Propionibacterium acnes colony and control of inflammation reaction. Internal methods of treatment of Acne are antibiotics, retinoids, hormone preparations etc. 6. In Eastern medicine, external methods of treatment of Acne are wet compress method, paste preparation method, powder preparation method, pill preparation method, acupuncture and moxibustion therapy, ear acupuncture therapy, prevention and notice, and so on. In Western medicine, external method of treatments of Acne are divided into topical therapy and other surgical therapies. Topical therapy is used such as antibiotics, sebum regulators, topical vitamin A medicines etc and other surgical therapies are used such as surgical treatments, intralesional injection of corticosteroids, skin dermabrasion, phototherapy, photodynamic therapy, and so on. Conclusions : Until now, there is no perfect, effective single treatment. We think that Eastern medicine approach and treatment can be helpful to overcome the limitations of acne cure.

Mechanical and Operational Factors Affecting the Efficiency of Rice Polishing Machines (정미기의 능률에 미치는 기계적 요인및 작동조건에 관한 연구)

  • No, Sang-Ha
    • Journal of Biosystems Engineering
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    • v.1 no.1
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    • pp.15-15
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    • 1976
  • In analyzing the operational characteristics of a rice whitening machine, the internal radial pressure of the machine was measured using strain gage equipment. Changes in cylinder and feed screw configurations, screen type, cylinder speed and counter-pressure levels were examined to determine their impact on the quality and quantity of milled rice and the performance of the machine. The results are summarized as follows: 1. The internal radial pressure in the whitening chamber varied with the surface condition of the grain being processed. During the first or second pass through the machine, pressure was relatively low, reached a maximum after two to three passes with combinations I and II, three to six with combination III and then began to fall. 2. The pitch of the feed screw and the size of the feed gate opening which determine the rate of entry of grain into the whitening chamber, appeared to be the most important factor aff-::cting the degree of radial pressure, quality and quantity of milled rice and the efficiency of the machine. Using a feed screw with a wide pitch (4.8cm), radial pressure was relatively high and head rice recovery ratio \vere quite low. In this case capacity and machine effic?iency were much higher than obtained when using a feed screw with a narrow pitch (2.3cm). Very significant responses in radial pressure, head rice recovery rates and machine capacity were observed with changes in cylinder speed and counter-pressure levels when using the wide pitch feed screw. 3. The characteristics of the screen which surrounds the whitening chamber had an important effect on whitening efficiency. The existence of small protuberances on the original screen resulted in significant increases in both machine capacity and efficiency but without a significant decrease in head rice recovery or development of excessive radial pressure. Further work is required to determine the effects of screen surface conditions and the shape of the cylinderical steel roller on the rate of bran removal, machine efficiency and recovery rates. The size of the slotted perforations 0:1 the screen affects total milled rice recovery. The opening size on the original screen was fabricated to accommodate the round shape of Japonica rice varieties but was not suitable for the more slender Indica type. Milling Indica varieties with this screen resulted in a reduction in total milled rice recovery. 4. An increase in cylinder speed from 380 to 820 rpm produced a positive effect on head rice recovery for all machine combinations at every level of counter-pressure used in the tests. Head rice recovery was considerably lower at 380rpm using a wide screw pitch when compared to the results obtained at speeds from 600 to 820 r.p.m. The effects of cylinder speed On radial pressure, capacity and machine efficiency showed contrasting results, depending on the width of the feed screw pitch. With a narrow feed screw pitch (2.3cm), a direct proportional relationship was observed bet?ween cylinder speed and both radial pressure and machine efficiency. In contrast, using a 4.8 centimeter pitch feed roller produced a series of inverse relationships between the above variables. Based on the results of this study it is recommended when milling Indica type long grain rice varieties that the cylinder speed of the original machine be increased from 500-600 rmp up to a minimum of 800 rpm to obtain a greater abrasive effect between the grain and the screen. The pitch of the feed screw should be also reduced to decr?ease the level of internal radial pressure and to obtain higher machine efficiency and increased quality of milled rice with increased cylinder speeds. Further study on the interaction between cylinder speed and feed screw pitch is recommended. 5. An increase in the counter pressure level produced a negative effect On the head rice recovery with an increase in radial pressure, capacity, and machine efficiency over all combinations and at every level of cylinder speed. 6. Head rice recovery rates were conditioned primarily by the pressure inside the whitening chamber. According to the empirical cha racteristics curve developed in this study, the relationships of head rice recovery ($Y_h$) and machine capacity ($Y_c$/TEX>) to internal radial pressure ($X_p$) followed an inverse quadratic function and a linear function respectively: $$Y_h^\Delta=\frac{1}{{1.4383-0.2951X_p^\ast+0.1425X_p^{\ast\ast}}^2} , (R^2=0.98)$$$$Y_c^\Delta=-305.83+374.37X_p^{\ast\ast}, (R^2=0.88)$$The correlation between capacity and power consumption per unit of brown rice expressed in the following exponential function: $$Y_c^\Delta=1.63Y_c^{-0.7786^\{\ast\ast}, (R^2=0.94)$$These relationships indicate that when radial pressure increases above a certain range (1. 6 to 2.0 kg/$cm^2$ based On the results of the experiment) head ricerecovery decrea?ses in a quadratic relation with a inear increase in capacity but without any decrease in power consump tion per unit of brown rice. On the other hand, if radial pressure is below the range shown above, power consumption increases dramatically with a lin?ear decrease in capacity but without significant increases in head rice recovery. During the operation of a given whitening machine, the optimum radial pressure range or the correct capacity range should be selected by controlling the feed rate and/or counter-pressure keeping in mind the condition of the grain, particulary the hardness. It was observed that the total number of passes is related to radial pessure level, feed rate and counter-pressure level. The higher theradial pressure the fewer num?ber of pass required but with decreased head rice recovery. In particular, when using high feed rates, the total number of passes should be increased to more than three by reducing the counter-pressure level to avoid decreaseases in head rice recovery (less than 65 percent head rice recovery on the basis of brown rice) at every cylinder speed. 7. A rapid rise in grain temperature seemed to have a close relationship with the pressure generated inside the whitening chamber and, subsequently with head rice reco?very rates. The higher the rate of increase, the lower were the resulting head rice recoveries.

Serotypes and Antimicrobial Susceptibility of Streptococcus pneumoniae (폐구균의 혈청형 분포와 항생제 감수성에 관한 연구)

  • Choi, Kyoung-Min;Kim, Jeong-Hyun;Shin, Kyoung-Mi;Yeon, Soo-In;Shin, Jeon-Soo;Yong, Dong-Eun;Lee, Kyoung-Won;Kim, Dong-Soo
    • Pediatric Infection and Vaccine
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    • v.10 no.2
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    • pp.159-166
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    • 2003
  • Purpose : Streptococcus pneumoniae is part of the normal flora but is also responsible for causing many invasive diseases such as pneumonia, meningitis, and sepsis in addition to noninvasive diseases such as otitis in children. Multi-drug resistant strains has raised a lot of concern worldwide and thus the importance of prevention has been emphasized. We have analyzed the current serotypes and antibiotic sensitivity of each serotype as a baseline study to estimate the efficacy of the pneumococcal vaccine in Korean children. Methods : One hundred sixteen cases of pneumococcus cultured at Yonsei Medical Center from September 2001 to January 2003 were analyzed. The serotyping was done with the Quellung reaction and penicillin resistance was tested using the oxacillin disc diffusion method. Results : Pneumococcus were cultured from the sputum in 76 cases(65.5%), from the blood in 13 cases(11.2%), from the ear discharge in 12 cases(10.3%), from the throat in 7 cases(6.0%), from the nasal cavity in 2 cases(1.7%), and one case(0.9%) each from the cerebrospinal fluid, eye discharge, peritoneal fluid, post-operational wound, brain abscess, and catheter tip. Serotyping was possible with 98 cases and the following serotypes were found; 15 cases of type 19F(15.3%), 11 cases of 19A(11.2%), 8 cases of 11A(8.2%), 7 cases each of 6A, 14 and 3(7.1%), 6 cases each of 35, 6B and 23F(6.1%). Eighty two cases(70.7%) out of 116 cases were penicillin resistant and serotypes 19F, 19A, 11A, 23F, 6A, 9V constituted the majority, 48 cases(59.8%). These serotypes showed resistance to cotrimoxazole (74.4%), tetracycline(69.5%), and erythromycin(90.3%) as well. In the 22 cases cultured from children, 19A and 19F were found in 25.0%, 6A, 6B, and 23F in 10.0%, 11A, 14, 19, and 29 in 5.0%. Fifty percent(10/20) of the clinical isolates were represented in the current 7-valent pneumococcal protein conjugate vaccine, and 85%(17/20) when the cross-reacting serotypes were included. Penicillin resistance was found in 86.4%(19/22). Conclusion : The percentage of serotypes included in the 7 valent pneumococcal protein conjugate vaccine found in our study was 40.8% which was less than other prior studies. In anticipation of a change of pneumococcal serotypes, a nationwide multicenter study is needed before the initiation of pneumococcal vaccines in Korea.

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Evaluation of Nutritional Health Camp in Obese Elementary Students (일부 비만 초등학생을 위한 건강영양캠프의 효과 평가)

  • Jun, Ye-Sook;Lee, Ji-Eun;Lee, Yong-Sook;Bae, Yun-Jung;Kim, Mi-Hyun;Lee, Yoon-Shin;Kim, Ae-Jung;Sung, Chung-Ja;Choi, Mi-Kyeong
    • The Korean Journal of Food And Nutrition
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    • v.20 no.1
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    • pp.79-87
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    • 2007
  • An evaluation of the influence of nutritional health camp for in obese elementary students was assessed. Anthropometric measurement, 24-hour recall of dietary intake, MMTIC(Murphy-Meisgeier Type Indicator for Children) and questionnaire response(nutritional knowledge, nutritional attitude, and self-satisfaction) was documented in 43 obese elementary students(29 boys and 14 girls). The mean age of the elementary students was 11.2 years. The mean height, body weight and obesity index was 144.7 cm, 55.4 kg and 41.0% in boys, and 136.1 cm, 45.2 kg, 40.0% in girls. The mean energy intake was 1,522.4 kcal in boys and 1,494.9 kcal in girls. The subjects taking less than estimated average requirement(EAR) for vitamin ${B_2}$, folate, vitamin C and calcium was over 70%. A considerable number of students(65.1%) showed U band(undetermined character). The students demonstrated a propensity towards increased average nutritional knowledge and self-satisfaction after attending the nutritional health camp. In addition, a significant increase in average nutritional attitude was observed(p<0.001). The results of this study suggest that nutritional health camp positively influenced the nutritional attitude of elementary students.