• 제목/요약/키워드: infection experiments

검색결과 226건 처리시간 0.037초

적외선 촬영 영상 기반의 작물 병해 모니터링 가능성 타진을 위한 실내 감염 실험 (Preliminary growth chamber experiments using thermal infrared image to detect crop disease)

  • 정회정;정래동;류재현;오도혁;최선웅;조재일
    • 한국농림기상학회지
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    • 제21권2호
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    • pp.111-116
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    • 2019
  • 세균과 바이러스에 감염된 작물의 생물적 스트레스 탐지를 원격탐지 기술 기반의 열적외 센서를 이용하여 실내 생육 챔버에서 실험하였다. 감염으로 인한 엽온의 증가와 감염 농도에 따른 엽온 차이를 확인했다. 또한 엽온 기반으로 산출한 CWSI 값은 감염 잎에서 증가하였고, 그러한 현상은 육안으로 병징을 발견하기 하루 전에 시작되었다. 따라서 스마트팜 시스템의 작물 모니터링에 열적외 센서를 이용한다면, 병해의 탐지는 물론 피해 등급 평가, 조기 알람 등에 활용될 수 있을 것이다. 하지만 실제 스마트팜 적용을 위해서는 향후 엽온 측정 정확도 향상 기술, 자료 해석 방법, 생물 비생물적 스트레스 구별 알고리즘 연구 등이 추가로 필요할 것이다.

3D8 scFv 형질전환 돼지 개발 및 PRRS 저항성 평가 (Production of a transgenic pig expressing 3D8 single chain variable fragment (scFv) and its evaluation of PRRS resistant)

  • 이휘철;이건섭;김지윤;양현;이보람;박미령;황인설;이풍연;변승준;김원일;오건봉
    • 한국동물위생학회지
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    • 제43권4호
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    • pp.227-236
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    • 2020
  • In this study, we have developed 3D8 scFv transgenic pig (TG) by microinjection of fertilized one-cell pig zygotes (2.17%). The effect of 3D8 scFv TG on porcine reproductive and respiratory syndrome virus (PRRSV) resistance were evaluated through PRRSV VR2332 (1×103 TCID50/mL) challenge and transmission experiments. As a result, the average daily weight gain (ADWG) of TG increased compared to the wild type pigs (WT) in PRRSV challenge groups and the serum viremia levels of the TG was significantly lower than of WT on the 7 day and 21 day after infection, meaning that the viral shedding was suppressed by 3D8 scFv expression. These results suggest that the expression of 3D8 scFv in pig could suppress spreading of infected virus to pigs sharing a room.

Antiviral effects of Bovine antimicrobial peptide against TGEV in vivo and in vitro

  • Liang, Xiuli;Zhang, Xiaojun;Lian, Kaiqi;Tian, Xiuhua;Zhang, Mingliang;Wang, Shiqiong;Chen, Cheng;Nie, Cunxi;Pan, Yun;Han, Fangfang;Wei, Zhanyong;Zhang, Wenju
    • Journal of Veterinary Science
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    • 제21권5호
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    • pp.80.1-80.13
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    • 2020
  • Background: In suckling piglets, transmissible gastroenteritis virus (TGEV) causes lethal diarrhea accompanied by high infection and mortality rates, leading to considerable economic losses. This study explored methods of preventing or inhibiting their production. Bovine antimicrobial peptide-13 (APB-13) has antibacterial, antiviral, and immune functions. Objectives: This study analyzed the efficacy of APB-13 against TGEV through in vivo and in vitro experiments. Methods: The effects of APB-13 toxicity and virus inhibition rate on swine testicular (ST) cells were detected using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT). The impact of APB-13 on virus replication was examined through the 50% tissue culture infective dose (TCID50). The mRNA and protein levels were investigated by real-time quantitative polymerase chain reaction and western blot (WB). Tissue sections were used to detect intestinal morphological development. Results: The safe and effective concentration range of APB-13 on ST cells ranged from 0 to 62.5 ㎍/mL, and the highest viral inhibitory rate of APB-13 was 74.1%. The log10TCID50 of 62.5 ㎍/mL APB-13 was 3.63 lower than that of the virus control. The mRNA and protein expression at 62.5 ㎍/mL APB-13 was significantly lower than that of the virus control at 24 hpi. Piglets in the APB-13 group showed significantly lower viral shedding than that in the virus control group, and the pathological tissue sections of the jejunum morphology revealed significant differences between the groups. Conclusions: APB-13 exhibited good antiviral effects on TGEV in vivo and in vitro.

20 (S)-ginsenoside Rh2 inhibits colorectal cancer cell growth by suppressing the Axl signaling pathway in vitro and in vivo

  • Zhang, Haibo;Yi, Jun-Koo;Huang, Hai;Park, Sijun;Kwon, Wookbong;Kim, Eungyung;Jang, Soyoung;Kim, Si-Yong;Choi, Seong-kyoon;Yoon, Duhak;Kim, Sung-Hyun;Liu, Kangdong;Dong, Zigang;Ryoo, Zae Young;Kim, Myoung Ok
    • Journal of Ginseng Research
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    • 제46권3호
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    • pp.396-407
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    • 2022
  • Background: Colorectal cancer (CRC) has a high morbidity and mortality worldwide. 20 (S)-ginsenoside Rh2 (G-Rh2) is a natural compound extracted from ginseng, which exhibits anticancer effects in many cancer types. In this study, we demonstrated the effect and underlying molecular mechanism of G-Rh2 in CRC cells in vitro and in vivo. Methods: Cell proliferation, migration, invasion, apoptosis, cell cycle, and western blot assays were performed to evaluate the effect of G-Rh2 on CRC cells. In vitro pull-down assay was used to verify the interaction between G-Rh2 and Axl. Transfection and infection experiments were used to explore the function of Axl in CRC cells. CRC xenograft models were used to further investigate the effect of Axl knockdown and G-Rh2 on tumor growth in vivo. Results: G-Rh2 significantly inhibited proliferation, migration, and invasion, and induced apoptosis and G0/G1 phase cell cycle arrest in CRC cell lines. G-Rh2 directly binds to Axl and inhibits the Axl signaling pathway in CRC cells. Knockdown of Axl suppressed the growth, migration and invasion ability of CRC cells in vitro and xenograft tumor growth in vivo, whereas overexpression of Axl promoted the growth, migration, and invasion ability of CRC cells. Moreover, G-Rh2 significantly suppressed CRC xenograft tumor growth by inhibiting Axl signaling with no obvious toxicity to nude mice. Conclusion: Our results indicate that G-Rh2 exerts anticancer activity in vitro and in vivo by suppressing the Axl signaling pathway. G-Rh2 is a promising candidate for CRC prevention and treatment.

미세아교세포에서 GPR56 발현에 의한 이상 행동 (Abnormal Behavior Controlled via GPR56 Expression in Microglia)

  • 김현주
    • 생명과학회지
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    • 제33권6호
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    • pp.455-462
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    • 2023
  • 임신 중 감염에 의한 산모의 면역 활성화는 조현병과 자폐 스펙트럼 장애를 포함한 신경 발달 질환의 위험을 증가시킨다. 여러 연구에서 poly (I:C) 또는 LPS를 사용하여 모체 면역 활성화 유도한 자손에서 비정상적인 행동과 뇌 발달을 관찰하였다. 또한 최근 뇌에 상주하며 면역 세포로 기능하는 미세아교세포가 MIA 유발 자손의 행동 이상과 뇌 발달에 중요한 역할을 한다는 것이 보고되고 있으나 아직 메커니즘은 명확하지 않다. 본 연구에서는 GPCR의 구성원인 GPR56의 미세아교세포 특이적 억제가 행동 이상과 뇌 발달을 유발하는지 여부를 조사하였다. 먼저, MIA 유도는 발달 중인 뇌의 미세아교세포 집단에 영향을 미치지 않으나, 미세아교세포를 분리하여 GRP56의 발현을 조사한 결과, MIA 유도 태아에서 성별에 관계 없이 E14.5와 E18.5 사이에서 GPR56 발현이 억제됨을 관찰하였다. 그리고 미세아교세포 특이적 GPR56 억제는 MIA 유도 자손에게서 나타나는 사교성 결손, 반복적인 행동 패턴 및 증가된 불안 수준과 같은 비정상적인 행동을 관찰하였다. 미세아교세포 GPR56 억제 마우스에서는 MIA 유도 자손과 같은 비정상적인 피질 발달이 관찰되지 않았지만, c-fos 염색을 통해 뇌 활동이 관찰되었다. 따라서 본 연구는 미세아교세포 특이적 GPR56 결핍이 이상 행동을 유발함을 시사하며, 추후 연구를 통해 MIA 자손의 행동 결손 진단 및/ 치료 표적을 위한 바이오마커로 활용될 수 있음을 시사한다.

L-글루타민산 생산균 Brevibacterium lactofermentum의 Bacteriophag에 관한 연구 (Studies on the Bacteriophages of Brevibacterium lactofermentum)

  • 이태우
    • 미생물학회지
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    • 제17권3호
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    • pp.97-130
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    • 1979
  • Many industrial processes those employ bacteria are subjected to phage infestations. In L-glutamic acid fermentions using acetic acid, the phage infestations of the organisms have been recently recognized. In efforts to elucidate the sources of phage contamination involved in the abnormal fermentation, a series of study was conducted to isolate the phages both from the contents of abnormally fermented tanks and the soil or sewage samples from the surroundings of a fermentation factory, to define major charateristics of the phage isolates, and finally to determine the correlation between the phage isolates and temperate phages originating from the miscellaneous bacterial species isolated from the soil or sewage samples. The results are summarized as follows; 1) All phages were isolated from the irregular fermentation tanks and soil or sewage samples, and they were designated as phage PR-1, PR-2, PR-3, PR-4, PR-5, PR-6, and PR-7, in the order of isolation. These PR-series phages were proved to be highly specific for the variant strains of Br. lactofermentum only, namely, phage PR-1 and PR-2 for Br. lactofermentum No. 468-5 and phage PR-3~PR-7 for Br. lactofemrentum No. 2256. By cross-neutralization test, the 7 phagescould be subdivided into 3 groups, i. e., phage PR-I and PR-2 the first, phage PR-3, PR-4, PR-5, PR-6 the second, and the phage PR-7 the third. 2) The 7 phages were virulent under the experimental conditions. They produced plaques with clear and relatively sharp margins without distinct halo. The mean sizes of plaques were 1.5mm in diameter for phage PR-1 and PR-2, and 1. Omm for phages PR-3~PR-7. Double layer technique modified by Hongo and described by Adams, was applied to assay of the PR-series phages. The factors influencing the plaques were as follows;young age cells of host bacteria cultured for 3-6 hours represented the largest number and size, optimum was pH 7.0, incubation temperature was $30^{\circ}C$, and agar concentration and amount of overlayer medium were 0.6% and 0.2ml, respectively. 3) PR-series phages were stable in 0.05M tris buffer and 0.1M ammonium acetate buffer solution. The addition of $5{\times}10^{-3}M$ magnesium ion effectively increased the stability. Thermostability experiments indicated that PR-series phages were stable at the teinperture between $50^{\circ}{\sim}55^{\circ}C$ in nutrient medium, $45^{\circ}{\sim}50^{\circ}C$ in buffer solution. However, the phages mere completely inactivated at 603C and 65$^{\circ}$C within 10 minutes. The phages were stable at the range of pH6~9 in nutrient medium and of pH 8-9 in buffer solution, respectively. Exposure of the phages to UV for 25, 60 and 100 seconds resulted in the complete loss of infectivily, respectively. 4) Electron microscopy showed that PR-series phage particles exhibited rather similar morphology, differing in the size All of PR-series phages had a multilateral head and had a simple long tiil about three to five times long as compared with head. By the size, phage PR-1 and PR-2, PR-3, PR-4, PR-5, and PR-6 and PR-7 were classified into same groups, respectively. The head and tail size of phage PR-1, PR-5, PR-5(T) and PR-7 were 85nm, 74nm and 235nm and 350mm, and 72nm and 210nm, respectively. 5) Nucleic acids of PR-series phages were double stranded DNA. The G+C contents of phage PR-1, PR-5 and PR-7 were 56.1, 52.9 and 53.7, respectively. The values of G+C contents derived from the $T_m$ were in agreement with the chemically determined values. 6) PR-series phages effectively adsorbed on their host bacteria at the rate of more than 90% during 5 min. K value for phage PR-1, PR-5 and PR-7 were calculated to be $6{\times}10^9 ml$ per minute, respectiveky. The pH of the medium did effect adsorption rate, but both temperature and age of host cells did not. Generally, optimum adsorption condition of phages seemed to be almost same as optimum growth conditions of host bacteria. 7) In one-step growth experiments, the latent periods at $30^{\circ}C$ for PR-1, and PR-7 were about 70, 50 and 55 min, respectively. The corresponding average burst size was 200, 70 and 90, respectively. Lpsis period according to the multiplicity of infection and a phage series. In case of m. o. i. 100, strain No. 2256 (PR-5) and No. 468-5(PR-1) failed to grow and turbidity decreased after 50 and 70min, respectively. 8) In the lysate of a plaque purified phage PR-5 infected bacteria, there observed 2 types ofphage particles, i. e., phage PR-5 and PR-5 (T) of similar morphology but differing at the length of phage tail, and phage tail like particles. The phage taillike particles could be divided into 4 types by the length. Induction experiments of Br. lactofermentum with UV irradiation, mitomycin C or bacitracin treatment produced neither phage PR-5 (T) or phage tail-like particles. 9) No lysis occured when the growth of 7 strains of miscellaneous bacteria, isolated from soil and sewage samples, were inoculated with either phage PR-5 (T) or phage tail-like particles the inoculation of phage PR-5 pellet resulted in the growth inhibition of the orgainsms in the spot test. The lysates obtained from 3 miscellaneous soil derived bacteria following mitomycin C treatment the growth of Br. lactofermentum, but did not lyze the bacterium.

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한국산 양식김의 종류 (SPECIES OF CULTIVATED PORPHYRA IN KOREA)

  • 강제원
    • 한국수산과학회지
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    • 제3권2호
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    • pp.77-92
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    • 1970
  • Ueda, in the course of his systematic work on the lavers, Porphyra, in Japan and Korea in 1932, mentioned that most of the cultivated Porphyra belong to Porphyra tenera Kjellman. Then he, dividing the species into two forms, f. typica and f. kjellmani, put Korean cultivated Porphyra under the latter. From the 1930s to the early 1940s, Fujikawa, Kaneko and others worked on Physiological experiments or cultivational experiments of Porphyra in the culture-bed, but there was no mention about the cultivated Porphyra species. However, many fishermen generally recognize that the characteristics of cultivated Porphyra vary depending on their habitat or the picking season, and it is considered that these differences are due to the varieties of the species which are well adaptable to various environments. Recently, I have become aware of the predominant occurrence of P. yezoensis Ueda in most culture-beds of Korea as in the Tokyo Bay or other places in Japan. At present, since artificial seeding for the cultivation of Porphyra with Conchocelis has been carried out and peculiar species can be cultured, a study of the species of cultivated Porphyra has become an important subject. I collected the specimens from a number of culture-beds which are located in the legions shown in fig. 1 from January, 1968 to May, 1970 and found that there are five species, P. tenera Kjellman, P. yezoensis Ueda, P. kuniedai Kurogi, P. seriata Kjellman and P. suborbiculata Kjellman. Among them, P. kuniedai was treated as a round-type, a form of P. tenera, by Kunieda (1939) and Tanaka (1952) and the occurrence of this form is generally recognized by most fishermen. At present, as mentioned above, the most dominant species of cultivated Porphyra is P. yezoensis but the cultivation of P. tenera is restricted to certain culture-beds or the early half of the cultivation period. P. kunieda appears as a mixed species throughout most of the culture-beds, particulary in the later half of the period, while when it was picked in January it appeared dominantly in a place such as Gum-Dang where the 'Bal', splitted bamboo piece mat, was settled during the last of September. This is the first seeding process. The latter two species, P. suborbiculata and P. seriata appear frequently but in small amounts in the later half of the period particulary in the western region of the southern coast. However, it can not be ascertained when P. yezoensis becomes predominant, because specimens have not been available up until recent years but the process can be described as follows: We commonly recognize the ecological characteristics of P. tenera as follows; First, the conchospores of the species develop earlier and the period of its discharge is shorter than those of P. yezoensis; second, the microscopical buds discharge neutral spores which develop into new buds directly and buds develop repeatedly through a short period. Consequently, according to such above ecological characteristics, the species can grow thick on the 'Bal' exclusively. However, buds may disappear when they are harmed by disease such a 'infection by certain parasites or by other unusual environmental conditions. Thus P. yezoensis are enabled to grow on the 'Bal' instead of the former species since they not only develop later than the former but also macroscopical fronds discharge the neutral spore throughout the period from October to May. Likewise, if any disease appears in the culture-bed ill the later half of the period, the former is more severely damaged than the latter because the former have less resistance to the disease than the latter. Thus fewer frond survive and fewer carpospores which are the origin of the next generation can be discharged. However the latter by their nature can continue growing until early summer. In the case of the culture-bed where the above phenomenon occurs repeatedly P. yezoensis gradually may become the dominant species among cultivated Porphyra. In support of the validity of this process we find that according to the description and the plate of Wada (1941), P. tenera, P. yezoensis and P. kuniedai grow together in the culture-bed at the mouth of the Nakdong River where P. yezoensis occurs predominantly and mixed with P. kuniedai.

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하수처리수를 관개한 후 벼재배 시험구에 대한 미생물 위해성 평가 (Microbial Risk Assessment in Treated Wastewater Irrigation on Paddy Rice Plot)

  • 정광욱;안윤주;장재호;전지홍;윤춘경
    • 생태와환경
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    • 제38권2호통권112호
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    • pp.225-236
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    • 2005
  • 하수처리수는 이용할 수 있는 소중한 수자원이지만, 적절한 정책과 안전하고 효율적인 관리 없이는 이용할 수 없다. 본 연구에서는 국제적인 농업적 재이용수 수질기준을 고찰하고 하수처리수를 논에 관개한 후 E. coli의 농도를 모니터링 하였으며, Beta-Poisson 모형을 이용하여 두개의 시나리오를 바탕으로 미생물 위해성 평가를 실시하였다. 본 연구의 모니터링 결과 biofilter 유출수를 추가 처리 없이 관개한 처리구에서 높은 E. coli농도를 나타내었으며, 처리구와 모니터링 시기별로 큰 차이를 보였는데 이는 관개나 기타 영농작업시 저질이 교란되어 수체의 농도에 반영되고 처리구별 논의 생태 차이가 영향을 미친 것으로 판단된다. 하수처리수를 관개할 경우 Beta-poisson 모형을 이용하여 직접 영농활동에 참여하는 농부와 주변의 아이들에 미치는 위해성에 대해 평가하였다. 미생물 위해성 평가 결과 관개 직후보다 관개 후 24시간이 경과되면 위해도 값이 감소하는 것으로 나타났으며, 관개 후 1 ${\sim}$ 2일이 경과한 후에 작업에 임하는 것이 미생물 위험이 적은 것으로 나타났다. 2003년과 2004년 모두 관개 초기인 5월말과 6월초에 높은 위해도 값을 나타내었다. 본 연구에서 실시한 위해성 평가는 논에 관한 역학자료가 충분하지 않았으며,E. coli를 음용수로 섭취할 경우 발생할 수 있는 질병에 대한 평가 모형을 이용하였기 때문에 하수처리수의 논 관개시 위험을 평가하는데 한계를 갖고 있다. 하지만, 하수처리수를 관개용수로 사용할 경우 병원성 세균에 의한 감염의 개연성을 보여주었으며 하수처리수를 재이용하고자 할 때 추가적인 처리 등의 적절한 관리가 필요함을 시사한다.

한국에서 CMV에 항바이러스 효과를 나타내는 Serratia spp. Gsm01 균주의 분리 동정 및 효과 검정 (Isolation and Evaluation of an Antiviral Producing Serratia spp. Strain Gsm01 against Cucumber mosaic virus in Korea)

  • ;이선화;석정기;;서동욱;박덕환;조준모;박동식;허장현;임춘근
    • 농약과학회지
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    • 제10권4호
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    • pp.344-350
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    • 2006
  • 항바이러스 효과를 나타내는 세균은 강원도 홍천 인삼재배지의 수삼으로부터 분리하였다. 분리세균은 생리 생화학 테스트와 16s rRNA 유전자 분석을 통해 동정한 결과, Serratia 속으로 동정 되어져 본 세균을 Gsm01으로 명명하였다. Gsm01 균주를 MGY 액체배지에 증식시켜 배양 여액을 0.45 ${\mu}m$ filter에 통과시킨 Culture filtrate (CF)를 명아주 (Chenopodium amaranticolor)에 반엽법으로 처리한 결과, 오이모자이크 바이러스 (CMV)에 대한 억제율이 98%로 매우 높은 것을 확인하였다. 또한 전신유도저항성을 확인하기 위하여 담배 (Nicotiana tabacum cv. Xanthi-nc)의 하엽에 CF와 CMV-Y를 처리하였을 때, CF를 처리한 식물에서 접종15일 후까지 바이러스 증상이 관찰되지 않았다. 고추의 포장시험에서 무처리 식물과 비교해 보았을 때 바이러스 증상은 52.9% 감소한 것으로 보아 포장 시험에서 CF를 처리한 농작물의 산출량이 무처리 식물에 비해 14% 증가함을 확인 할 수 있었다. 한편, Gsm01 균주의 CF는 고추에 어떠한 약해도 나타내지 않아 매우 안전한 것으로 판단되었다.

설비공학 분야의 최근 연구 동향 : 2016년 학회지 논문에 대한 종합적 고찰 (Recent Progress in Air-Conditioning and Refrigeration Research : A Review of Papers Published in the Korean Journal of Air-Conditioning and Refrigeration Engineering in 2016)

  • 이대영;김사량;김현정;김동선;박준석;임병찬
    • 설비공학논문집
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    • 제29권6호
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    • pp.327-340
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    • 2017
  • This article reviews the papers published in the Korean Journal of Air-Conditioning and Refrigeration Engineering during 2016. It is intended to understand the status of current research in the areas of heating, cooling, ventilation, sanitation, and indoor environments of buildings and plant facilities. Conclusions are as follows. (1) The research works on the thermal and fluid engineering have been reviewed as groups of flow, heat and mass transfer, the reduction of pollutant exhaust gas, cooling and heating, the renewable energy system and the flow around buildings. CFD schemes were used more for all research areas. (2) Research works on heat transfer area have been reviewed in the categories of heat transfer characteristics, pool boiling and condensing heat transfer and industrial heat exchangers. Researches on heat transfer characteristics included the results of the long-term performance variation of the plate-type enthalpy exchange element made of paper, design optimization of an extruded-type cooling structure for reducing the weight of LED street lights, and hot plate welding of thermoplastic elastomer packing. In the area of pool boiling and condensing, the heat transfer characteristics of a finned-tube heat exchanger in a PCM (phase change material) thermal energy storage system, influence of flow boiling heat transfer on fouling phenomenon in nanofluids, and PCM at the simultaneous charging and discharging condition were studied. In the area of industrial heat exchangers, one-dimensional flow network model and porous-media model, and R245fa in a plate-shell heat exchanger were studied. (3) Various studies were published in the categories of refrigeration cycle, alternative refrigeration/energy system, system control. In the refrigeration cycle category, subjects include mobile cold storage heat exchanger, compressor reliability, indirect refrigeration system with $CO_2$ as secondary fluid, heat pump for fuel-cell vehicle, heat recovery from hybrid drier and heat exchangers with two-port and flat tubes. In the alternative refrigeration/energy system category, subjects include membrane module for dehumidification refrigeration, desiccant-assisted low-temperature drying, regenerative evaporative cooler and ejector-assisted multi-stage evaporation. In the system control category, subjects include multi-refrigeration system control, emergency cooling of data center and variable-speed compressor control. (4) In building mechanical system research fields, fifteenth studies were reported for achieving effective design of the mechanical systems, and also for maximizing the energy efficiency of buildings. The topics of the studies included energy performance, HVAC system, ventilation, renewable energies, etc. Proposed designs, performance tests using numerical methods and experiments provide useful information and key data which could be help for improving the energy efficiency of the buildings. (5) The field of architectural environment was mostly focused on indoor environment and building energy. The main researches of indoor environment were related to the analyses of indoor thermal environments controlled by portable cooler, the effects of outdoor wind pressure in airflow at high-rise buildings, window air tightness related to the filling piece shapes, stack effect in core type's office building and the development of a movable drawer-type light shelf with adjustable depth of the reflector. The subjects of building energy were worked on the energy consumption analysis in office building, the prediction of exit air temperature of horizontal geothermal heat exchanger, LS-SVM based modeling of hot water supply load for district heating system, the energy saving effect of ERV system using night purge control method and the effect of strengthened insulation level to the building heating and cooling load.