• Childhood Kidney Diseases
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• 제24권2호
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• pp.83-90
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• 2020

Purpose: To investigate the association between urinary neutrophil gelatinase-associated lipocalin (uNGAL) and leukocyte differential count in children with urinary tract infections (UTIs). Methods: A retrospective chart review was performed in children undergoing uNGAL measurements between June 2018 and September 2019. Patients with suspected or diagnosed UTIs were included. The relationship between uNGAL and blood leukocyte differential count was investigated in children. Results: A total of 197 children were included in this study, 119 of whom (60%) had UTIs. The non-UTI patients (n=78) were diagnosed with pneumonia, acute gastroenteritis, viral upper respiratory infection, and others. After adjusting for age, gender, and fever duration, the leukocyte count, monocyte count, and uNGAL levels were higher in the UTI group than in the non-UTI group (P<0.05). uNGAL showed positive correlations with neutrophil counts, monocyte counts, the neutrophil-to-lymphocyte ratio, and the monocyte-to-lymphocyte ratio in the UTI group (P<0.05). uNGAL levels were only associated with the neutrophil-to-lymphocyte ratio in the non-UTI group (P<0.05). In a multivariable logistic regression analysis, only uNGAL was associated with the presence of UTI (P<0.05). The area under the receiver operating characteristic curves for uNGAL and monocyte counts to identify UTI were 0.89 (95% confidence interval (CI): 0.824-0.939; P=0.025) and 0.7 (95% CI: 0.627-0.774; P=0.038), respectively. Conclusions: In children with UTIs, uNGAL levels may be associated with blood leukocyte differential counts. uNGAL measurements and monocyte counts can be helpful in children with suspected UTIs.

• BMB Reports
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• 제32권1호
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• pp.60-66
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• 1999

Matrix metalloproteinase-2 (MMP-2; 72-kDa gelatinase; 72-kDa type IV collagenase; gelatinase A) plays an important role in normal physiological processes and in many pathologic processes such as arthritis and metastasis of cancer. Tissue inhibitor of metalloproteinases-2 (TIMP-2) binds to proMMP-2 or mature MMP-2 at a 1:1 ratio and inhibits the catalytic activity of MMP-2. We demonstrated that the baculovirus/insect cell system does not have TIMP-2 activity. The human proMMP-2 free of TIMP-2 was expressed in the expression system and purified by one-step affinity chromatography using gelatin-Sepharose. The free proMMP-2 was autoactivated to the mature MMP-2 and autodegraded into smaller molecular weight forms in the absence of external activator. The activation and autodegradation of the proMMP-2 was much more rapid in the presence of 4-aminophenylmercuric acetate (APMA). Addition of TIMP-2 inhibits both APMA-induced activation and autodegradation of the free proMMP-2. However, an increasing concentration of TIMP-2 more readily inhibited activation of the free proMMP-2 than autodegradation. These results demonstrate that TIMP-2 plays roles in inhibition of both activation and autodegradation of the free proMMP-2 in addition to inhibition of the catalytic activity of MMP-2.

• 한국수정란이식학회지
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• 제27권2호
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• pp.93-100
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• 2012

To investigate the biochemical nature of changes in vaginal physiology during estrus and pregnancy, we examined the cytology and viscosity, and monitored the protein expression profile in vaginal mucus during estrus and pregnancy. The viscosity progressively decreased from estrus to pregnancy. Cell type analysis revealed that white blood cells progressively increased from estrus to pregnancy, while red blood cells progressively decreased during pregnancy. The cornification index (CI) was higher in estrus than in pregnancy. Protein mass spectrumetry identified the presence of ribosome-binding protein 1, GRIP 1 (Glutamate receptor-interacting protein 1)-associated protein 1, DUF729 (Domain of unknown function729) domain-containing protein 1, prolactin precursor, dihydrofolatereductase, and MMP (Matrix metalloprotease)-9 in vaginal mucus. MMP-2 and MMP-9 proteins in the vaginal mucus were active throughout estrus and gestation, as measured by a gelatinase assay, but most abundant in the vaginal mucus on day 0 of estrus. Results from ELISA of MMP-2 and MMP-9 were in accordance with the gelatinase assay. In light of the crucial role of metalloproteinases in extracellular matrix remodeling, the level of MMP-9 in vaginal mucus might be useful as an indicator of estrus and pregnancy to increase the efficiency of reproduction.

• Clinical and Experimental Pediatrics
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• 제61권1호
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• pp.24-29
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• 2018

Purpose: The aim of this study was to evaluate the diagnostic accuracy of urinary biomarkers, such as neutrophil gelatinase-associated lipocalin (uNGAL) and ${\beta}-2$ microglobulin (uB2MG), in early detection of urinary tract infection (UTI) in infants aged <3 months with fever. Methods: A total of 422 infants aged <3 months (male:female=267:155; mean age, 56.4 days), who were admitted for fever, were retrospectively included in this study. We compared uNGAL and uB2MG between the UTI and non-UTI groups at the time of admission. The sensitivity, specificity, accuracy, and area under the curve (AUC) of uNGAL and uB2MG for use in diagnosing UTI were assessed. Results: Among 422 patients, 102 (24.2%) were diagnosed with UTI. Levels of uNGAL were higher in the UTI group than in the non-UTI group (366.6 ng/mL vs. 26.9 ng/mL, P<0.001). Levels of uB2MG were not different between the 2 groups. Multivariate analysis revealed that uNGAL was an independent predictive factor for UTI (P=0.033). The sensitivity, specificity, and accuracy were 90.2%, 92.5%, and 91.9% for uNGAL, and 48.0%, 43.8%, and 44.8% for uB2MG, respectively. AUC of uNGAL was 0.942 and that of uB2MG was 0.407. Conclusion: Accuracy of uNGAL in the diagnosis of UTI is high in febrile infants aged <3 months. uNGAL can help in the early diagnosis and treatment of UTI in infants.

• Childhood Kidney Diseases
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• 제25권2호
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• pp.84-91
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• 2021

Purpose: We aimed to study the association of plasma neutrophil gelatinase-associated lipocalin (pNGAL) and leukocyte differential count in children with febrile urinary tract infection (UTI). Methods: Medical records of 154 children aged 1 month to 13 years with febrile UTI who were hospitalized were retrospectively reviewed. Associations between pNGAL levels and blood leukocyte differential count at admission and after 48 hours of treatment were investigated in children with or without acute pyelonephritis (APN). Results: The APN group (n=82) showed higher pNGAL levels, neutrophil count, monocyte count, and neutrophil-to-lymphocyte ratio (NLR), compared to the non-APN group (n=72) (all P<0.05). After adjustment for age and sex, pNGAL showed positive correlations with neutrophil count and NLR in both groups (all P<0.05). Additionally, it was correlated with the monocyte-to-lymphocyte ratio (MLR) only in the APN group (P<0.05). Before and after treatment, pNGAL was positively correlated with neutrophil count, NLR, and MLR in patients with APN while it was related with neutrophil count and NLR in those without APN (all P<0.05). Areas under the receiver operating curve of pNGAL, neutrophil count, NLR, and MLR for predicting APN were 0.804, 0.760, 0.730, and 0.636, respectively (all P<0.05). Only pNGAL was independently associated with the presence of APN in a multivariable logistic regression analysis (P<0.05). Conclusion: In children with febrile UTIs, pNGAL might be associated with leukocyte differential count and the presence of APN.

• 동국한의학연구소논문집
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• 제9권
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• pp.179-191
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• 2000

한방에서 건강골(建强骨)시키는 효능이 있는 5가지 약물(골쇄보(骨碎補), 상기생(桑寄生), 김모구척(金毛狗脊), 별갑(別甲), 법파고지(法破古紙))로 구성된 처방(CEDR로 약칭)으로 실험을 하였다. 생쥐의 calvarial osteoblast를 분리하고 배양한 후 실험을 행하였는데, 골 재흡수 인자인 PTH, $1,25(OH)_2D_3$, $TNF-\alpha$, IL-1에 자극받은 osteoblasts는 활동적인 gelatinase를 생산하므로서 collagen 용해를 증가시켰다. 암컷 생쥐의 long bone organ을 culture하여 골 재흡수를 자극하는 IL-1를 실험한 결과, IL-1은 골의 재흡수를 자극하였으며 동시에 존재해있을 때에는 뚜렷한 재흡수를 보였다. 더욱이, indomethacin과 dexamethasone이 $IL-1{\alpha}$의 투여량 증가에 대한 영향을 관찰하였을 때에는 indomethacin과 dexamethasone은 IL-1의 투여량에 대한 곡선 그래프를 오른쪽으로 이동시켰다. 시험관내에서 세포 독성에 대하여 관찰하였을 때, CEDR추출물은 $1-60{\mu}g/ml$의 농도에서 아무런 세포 독성이 나타나지 않았으며, 뿐만 아니라 생쥐의 두개골 세포에서는 $120{\mu}g/ml$ 농도에서 전혀 세포독성이 관찰되지 않았다. CEDR 추출물은 생쥐의 calvarial cells에서 PTH (5 units/ml), $IL-1{\alpha}$ (1 ng/ml), $TNF-\alpha$, $1,25(OH)_2D_3$ (20 ng/ml) 및 $IL-1{\alpha}$, $IL-1{\beta}$에 유도되는 collagen 용해를 보호하는 효과가 있었다. CEDR 추출물을 1시간동안 전처리하였을 때, 그 자체로는 세포 생존에 영향이 없었으며, collagen 용해를 증가시키지도 않았으며 전처치로 collagen 용해를 유의성있게 감소시키지도 않았다. 게다가, 추출물은 $IL-1{\alpha}$와 $IL-1{\beta}$에 의해 유도되는 collagen 용해를 방지하는 효과가 있었다. 1시간 동안 전처리로 한 결과, collagen 용해를 유의성있게 감소시켰다. 흥미로운 것은 CEDR 추출물이 gelatinase 활성과 PTH, $1,25(OH)_2D_3$, $TNF-\alpha$, $IL-1{\beta}$ 및 $IL-1{\alpha}$의 재흡수 인자에 유도되어 진행되는 활성을 억제하였으며, CEDR 전처리시에는 강력한 보호 효과를 나타내었다. CEDR 추출물은 $IL-1{\alpha}$와 $IL-1{\beta}$에 자극되는 골 재흡수를 억제하는 효과를 나타내였으며, 또한 농도를 다양하게 한 CEDR의 전처리시에는 유의성이 있었다. Indomethacin과 dexamethasone의 비 스테로이드성 항 염증 인자에 의한 IL-1에 자극받은 골 재흡수를 억제하는 정도와 현상은 CEDR 추출물을 생쥐의 두개골 배양 시스템에 적용시켜 얻은 결과와 유사하였다. 이러한 결과들로 보건데, CEDR 추출물은 임상적으로 골다공증의 치료에 매우 안정적으로 적용할 수 있음을 제시하고 있다.

• 식물병연구
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• 제20권3호
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• pp.216-218
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• 2014

Lysobacter antibiocus HS124 is a chitinase-producing rhizobacterium with proven capacities to suppress plant diseases. Bacterial cultures of L. antibioticus HS124 showed strong biocontrol efficacies against various plant diseases compared to those of bacterial cultures of Bacillus subtilis QST713 which is an active ingredient of a commercial biopesticide, Serenade. Here, we report the draft genome sequence and automated annotation of strain HS124. This draft genome sequence indicates the novelty of L. antibiocus HS124 and a subset of gene functions that may be related to its biocontrol activities.

• 한국동물위생학회지
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• 제41권3호
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• pp.157-163
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• 2018

In order for monitoring of pathogenic bacterial contamination in the freshly slaughtered poultry meats produced in Gyeong-Nam province, we first isolated 4 strains of Salmonella spp. and 32 strains of Enterococcus faecalis from the total 164 samples, then we analyzed potential virulence gene profiles of the bacterial isolates by PCR using species-specific primer. The potential virulence genes we selected in this study were stn, invA, fimA, spvR, and spvC for the isolates of Salmonella spp. and those of esp, cylM, cylA, cylB, gelE, fsrA, fsrB, and fsrC were for the isolates of E. faecalis. The PCR results showed that all 5 virulence genes were detected simultaneously in the all isolates of Salmonella spp. However, there was a diverse occurrence pattern of the virulence genes in the case of E. faecalis. The gene for enterococcal surface protein (esp) was not detected among the isolates (0/32), and the haemolysin gene prevalence rate of cylA, cylB, and cylM were 3.1% (1/32), 9.3% (3/32), and 9.3% (3/32), respectively. Moreover, the genes of gelE, fsrA, fsrB, and fsrC that associated with gelatinase activity were detected in the rate of 53.1% (17/32), 53.1% (17/32), 53.1% (17/32), and 53.1% (17/32), respectively. In conclusion, in the isolates of Salmonella spp., all possessed 5 virulence genes tested, suggesting that they are all related with each other clonally. However, in the case of E. faecalis isolates, the occurrence of the haemolysin genes (cylM, cylA, cylB) and the gelatinase genes (gelE, fsrABC) was highly variable among the isolates.

• 한국토양비료학회지
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• 제43권4호
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• pp.490-497
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• 2010

근권토양으로부터 고추역병균을 포함한 다양한 식물 병원성 곰팡이에 대하여 항균활성이 강한 세균을 분리하였다. 이 세균은 16S rRNA gene서열 분석 결과 Lysobacter enzymogens로 동정되었고 LE429로 명명 하였다. LE429는 chitinase, ${\beta}-1$, 3-glucanase, protease, gelatinase, lipase 및 항생물질과 같은 다양한 이차대사산물을 분비하였다. 항생물질은 diaon HP-20 및 sephadex LH-20 컬럼크로마토그래피 및 HPLC로 정제하여, GC-EI 및 GC-CI분석을 통하여 phenylacetic acid로 동정되었다. Field 실험에서 LE429의 고추 병해 억제 효과를 조사하기 위해 LE429배양액(CB), Neem oil 용액 (NO), LE429배양액과 Neem oil 용액을 섞은 혼합액(CB+NO), 그리고 대조구로서 물(CON)을 각각 고추에 처리하였다. 고추의 수량구성요소는 일반적으로 CB 처리구가 가장 높았고, CB+NO, CON 그리고 NO 순서로 나타났다. CB 처리구에서 병원성 곰팡이는 강하게 억제 되었지만, 몇몇 해충이 발견되었다. NO 처리구에서는 해충은 발견 되지 않았지만, 병원성 곰팡이가 발견 되었다. 하지만, CB+NO 처리구에서 병원성 곰팡이 및 해충이 전혀 발견 되지 않았다. 결론적으로, 2차 대 사산물을 생산하는 LE429와 Neem oil의 혼합액은 고추에 발생하는 병원성 곰팡이와 해충에 대한 좋은 생물학적 방제제가 될 수 있다고 사료된다.

• Journal of Periodontal and Implant Science
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• 제29권2호
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• pp.311-326
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• 1999

본 연구에서는 치주질환과 관련이 깊은 것으로 알려진 구강내 spirochetes 균 중 Treponema denticola(TDC)와 가장 최근에 분리 배양된 Treponema lecithinolyticum(TLC)이 치주인대세포에 미치는 영향을 알아보기 위하여 두 spirochtes 균주를 배양한후 MTT test를 이용한 치주인대 세포의 증식 억제효과를 알아보았다. 또한, 위상차 현미경을 이용한 세포형태의 변화를 관찰 하였으며, LDH(lactate dehydrogenase) test를 이용한 세포독성 실험과, gelatin zymography를 시행하여 교원질 분해효소의 하나인 gelatinase의 활성화 여부를 측정한 결과 다음과 같은 결론을 얻었다. 1. 일정한 반응시간에서 농도에 따른 세포증식 억제효과에서는 TLC의 경우 높은 농도$(150{\mu}g/well)$에서부터 세포증식 억제 효과가 나타났으며, TDC에서는 낮은 농도$(9.4{\mu}g/well)$에서도 억제 효과가 나타났다. 2. 일정한 농도에서 시간에 따른 세포증식 억제효과에서는 TLC의 경우 2일째 $150{\mu}g/well$ 농도에서부터 세포증식 억제효과가 나타났으며, TDC에서는 $9.4{\mu}g/well$ 농도에서는 2일째에 세포증식 억제효과가 나타났다. 3. 열처리한 세균의 세포증식 억제 효과에서는 TDC의 경우에서 열처리 시킨 경우와 그렇지 않은 경우에 차이가 나타났으나, TLC의 경우에는 차이가 없었다. 4. 위상차현미경으로 관찰한 치주인대세포의 형태변화는 대조군에 비해 실험군에서 세포 형태의 손상으로 방추형이 소실되었고 세포증식이 억제되었으며, 세포끼리의 연결 또한 끊어져 분리되어 있었다. 5. 세포독성을 알아보기 위한 LDH test에서는 대조군, 실험군 모두 큰 차이가 없었다. 6. Zymography를 통한 교원질 분해에 미치는 영향에서는 TDC와 TLC에 의한 분자량 72kDa의 progelatinase A가 활성형으로 발현되었다. 이상의 결과를 보아 TLC와 TDC는 세포증식 억제효과를 통해 치주인대세포에 영향을 미치며, 교원질 분해 효소 Type IV의 하나인 progelatinase A를 활성형으로 발현시킴을 확인하였다.