• Title/Summary/Keyword: rapid screening

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Production of Colchicine Induced Tetraploids in Rye (Secale cereale L.) (콜히친 처리에 의한 4배체 호밀 육성)

  • Hwang, Jong-Jin;Kim, Dea-Wook;Kim, Chul-Woo;Son, Beum-Young;Baek, Seong-Beum;Park, Hyong-Ho;Ku, Ja-Hwan;Kim, Jung-Tae;Lee, Jin-Seok;Moon, Jung-Kyung;Kwon, Young-Up;Han, Ouk-Kyu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.4
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    • pp.343-352
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    • 2012
  • This experiment was carried out to report some data such as survival rate, tetraploid production efficiency, and agronomic characteristics of offspring from the induced tetraploid by the colchicine treatment in rye. The colchicine was soaked with 0.05%, 12 hours in dark condition and at two growth stages (green seed and 2nd leaf stage) in diploid ryes. Flow cytometry (FC) was proved to be efficient and rapid tool for screening ploidy levels in rye, showing around 40 to 60 in DNA amount (DAP1) corresponding to diploid and 80 to 110 tetraploid. There were 18.5% of survival rate at green seed treatment and 78% at 2nd leaf stage in average of two rye cultivars, Gogu and Jogreen, but in reverse 50.9% and 1.1% in the ratio of tetraploid to total tillers among the plants survived, respectively, resulting in 9.42% of tetraploid production rate in green seed treatment and 0.86% at 2nd leaf stage, respectively. In green seed treatment, there were 33% of survival rate in Gogu, 4% in Jogreen in 1st year, but 56% in Gogu, 21% in Jogreen and 49% in Charmgreen, respectively. The rate of tetraploid to total spikes among survived was 53.7% in Gogu, 32.4% in Jogreen, and 50.9% in average in 1st year, and 64.1% in Gogu, 51.5% in Jogreen, 60% in Charmgreen, and 60.5% in average in 2nd year. In green seed treatment, tetraploid production rate (survival rate ${\times}$ tetraploid ratio ${\times}$ 100) was 17.7% in Gogu and 1.3% in Jogreen and 9.42% in average in 1st year, and 35.9% in Gogu, 10.8% in Jogreen, 29.4% in Charmgreen, and 25.4% in average of three diploid rye cultivars. By the colchicine treatment with 0.05% for 12 hours in Gogu and Jogreen, 35 tetraploid plants were obtained and they produced 2,673 seeds with 148 spikes. There were 3.3-4.4 in the number of spikes per plant, 15.6-18.3 in grain number per spike, and 37.6 g in Gogu and 46.8 g in Jogreen in the 1,000-grain weight.

Comparison of the Standard Culture Method and Real-time PCR for the Detection of Vibrio parahaemolyticus in Seafoods and Vegetables (해산식품과 채소에서 Vibrio parahaemolyticus 검출을 위한 배지배양법과 real-time PCR의 비교검증)

  • Chon, Jung-Whan;Hyeon, Ji-Yeon;Hwang, In-Gyun;Kwak, Hyo-Sun;Han, Jeong-A;Chung, Yun-Hee;Song, Kwang-Young;Seo, Kun-Ho
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.355-360
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    • 2010
  • Vibrio parahaemolyticus (V. parahaemolyticus), which is commonly found in raw seafood, causes gastroenteritis in humans. Rapid and effective methods have been developed as culture methods require up to 5-7 days. In this study, real-time PCR was compared with the standard culture method for detecting V. parahaemolyticus in seafood and radish sprout samples. Five hundred grams of the samples were artificially contaminated with V. parahaemolyticus then divided into 20 samples. The samples were incubated in alkaline peptone water and then streaked onto thiosulfate-citrate-bile saltssucrose agar. Biochemical tests for suspicious colonies were performed using the API 20NE strip. In parallel, real-time PCR was performed targeting the toxR gene using the enrichment broth. The real-time PCR was sensitive in discriminating V. parahaemolyticus from other foodborne pathogens. The detection limit of the real-time PCR was $10^3\;CFU/mL$ in phosphate-buffered saline. Although the real-time PCR detected more positive samples (76 out of 180, 42%) than the culture method (66 out of 180, 37%), there was no significant statistical difference (p>0.05) between the two methods. In conclusion, real-time PCR assays could be an alternative to the standard culture method for detecting V. parahaemolyticus in seafood and radish sprouts, which has many advantages in terms of detection time, labor, and sensitivity.

Importance of Strain Improvement and Control of Fungal cells Morphology for Enhanced Production of Protein-bound Polysaccharides(β-D-glucan) in Suspended Cultures of Phellinus linteus Mycelia (Phellinus linteus의 균사체 액상배양에서 단백다당체(β-D-glucan)의 생산성 향상을 위한 균주 개량과 배양형태 조절의 중요성)

  • Shin, Woo-Shik;Kwon, Yong Jung;Jeong, Yong-Seob;Chun, Gie-Taek
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.220-229
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    • 2009
  • Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

Gene Expression of Metalloproteinases, Tissue Inhibitors of Metalloproteinases and Cytokines in Adriamycin-induced Cardiomyopathy (아드리아마이신으로 유도된 심근증에서 Metalloproteinase, Metalloproteinase 조직억제자, Cytokine 유전자 발현에 대한 연구)

  • Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.48 no.2
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    • pp.197-203
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    • 2005
  • Purpose : Changes in metalloproteinases(MMP) activity have been demonstrated in several disease states, including rheumatoid arthritis and tumor metastasis. More importantly, increased myocardial MMP activity has been reported to occur in both clinical and experimental forms of dilated cardiomyopathy. There was no report about MMP in adriamycin(ADR)-induced cardiomyopathy. The purpose of this study was to investigate gene expression of MMP and tissue inhibitor of metalloproteinases(TIMP) in ADR-induced cardiomyopathy and clarify the relationship between MMP and cytokines. Methods : Male Sprague-Dawley rats were divided into two groups. The first group was control. The second group was given intraperitoneal injections of ADR(5 mg/kg) twice a week over two weeks. Serum concentrations of MMP, TIMP, interleukin(IL)-6 and tumor necrosis factor(TNF)-${\alpha}$ were measured. RNA extraction was performed from frozen rat hearts. Reverse transcription polymerase chain reaction(RT-PCR) was employed. cDNA Microarray analysis was performed by using a set of 5,184 sequence-verified rat cDNA clones. Results : Serum MMP and TIMP levels were not significantly different between the two groups. IL-6 was $36.8{\pm}2.8pg/mL$ and TNF-${\alpha}$ $2.2{\pm}2.7pg/mL$ in the ADR group. They were significantly higher than in the control group. Serum MMP correlated significantly with TNF-${\alpha}$(r=0.41, P<0.05). There was no gene expression of MMP, IL-6 or TNF-${\alpha}$ in the hearts of both groups. Gene expression of TIMP was significantly depressed in the hearts of the ADR group. Conclusion : These results suggested a potential role for TNF-${\alpha}$ in the regulation of extracellular matrix remodeling in ADR induced cardiomyopathy. Rapid screening of multiple decreased gene expression by DNA chip may be a useful diagnostic test to detect early cardiac injury before developing ADR induced cardiomyopathy.

Frequency of Micronuclei in Lymphocytes Following Gamma and Fast-neutron Irradiations (방사선 조사량에 따른 인체 정상 림파구의 미세핵 발생빈도)

  • Kim Sung-Ho;Cho Chul-Koo;Kim Tae-Hwan;Chung In-Yong;Yoo Seong-Yul;Koh Kyoung-Hwan;Yun Hyong-Geun
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.35-42
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    • 1993
  • The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with $\gamma$-rays and neutrons in the 5 dose ranges was studied for a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-dependent increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model, frequencies per 1000 CB cells were ($0.31{\pm}0.049$)D+($0.0022{\pm}0.0002)D^2+(13.19{\pm}1.854) (r^2=1.000,\;X^2=0.7074,\;p=0.95$) following $\gamma$ irradiation, and ($0.99{\pm}0.528$)\;D+(0.0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309)\;(r^2=0.996,\;X^2=7.6834,\;p=0.11) following neutrons irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with $\gamma$-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was $2.37{\pm}0.17$. Since the MN assay is simple and rapid, it may be a good tool for evaluating the $\gamma$-ray and neutron response.

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Simultaneous estimation of fatty acids contents from soybean seeds using fourier transform infrared spectroscopy and gas chromatography by multivariate analysis (적외선 분광스펙트럼 및 기체크로마토그라피 분석 데이터의 다변량 통계분석을 이용한 대두 종자 지방산 함량예측)

  • Ahn, Myung Suk;Ji, Eun Yee;Song, Seung Yeob;Ahn, Joon Woo;Jeong, Won Joong;Min, Sung Ran;Kim, Suk Weon
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.60-70
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    • 2015
  • The aim of this study was to investigate whether fourier transform infrared (FT-IR) spectroscopy can be applied to simultaneous determination of fatty acids contents in different soybean cultivars. Total 153 lines of soybean (Glycine max Merrill) were examined by FT-IR spectroscopy. Quantification of fatty acids from the soybean lines was confirmed by quantitative gas chromatography (GC) analysis. The quantitative spectral variation among different soybean lines was observed in the amide bond region ($1,700{\sim}1,500cm^{-1}$), phosphodiester groups ($1,500{\sim}1,300cm^{-1}$) and sugar region ($1,200{\sim}1,000cm^{-1}$) of FT-IR spectra. The quantitative prediction modeling of 5 individual fatty acids contents (palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid) from soybean lines were established using partial least square regression algorithm from FT-IR spectra. In cross validation, there were high correlations ($R^2{\geq}0.97$) between predicted content of 5 individual fatty acids by PLS regression modeling from FT-IR spectra and measured content by GC. In external validation, palmitic acid ($R^2=0.8002$), oleic acid ($R^2=0.8909$) and linoleic acid ($R^2=0.815$) were predicted with good accuracy, while prediction for stearic acid ($R^2=0.4598$), linolenic acid ($R^2=0.6868$) had relatively lower accuracy. These results clearly show that FT-IR spectra combined with multivariate analysis can be used to accurately predict fatty acids contents in soybean lines. Therefore, we suggest that the PLS prediction system for fatty acid contents using FT-IR analysis could be applied as a rapid and high throughput screening tool for the breeding for modified Fatty acid composition in soybean and contribute to accelerating the conventional breeding.

Diagnosis of Pigs Producing PSE Meat using DNA Analysis (DNA검사기법을 이용한 PSE 돈육 생산 돼지 진단)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
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    • v.24 no.4
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    • pp.349-354
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    • 2004
  • Stress-susceptible pigs have been known as the porcine stress syndrome (PSS), swine PSS, also known as malignant hyperthermia (MH), is characterized as sudden death and production of poor meat quality such as PSE (pale, soft and exudative) meat after slaughtering. PSS and PSE meat cause major economic losses in the pig industry. A point mutation in the gene coding for the ryanodine receptor (RYR1) in porcine skeletal muscle, also known calcium (Ca$^{2+}$) release channel, has been associated with swine PSS and halothane sensitivity. We used the PCR-RFLP(restriction fragment length polymorphism) and PCR-SSCP (single strand conformation polymorphism) methods to detect the PSS gene mutation (C1843T) in the RYR1 gene and to estimate genotype frequencies of PSS gene in Korean pig breed populations. In PCR-RFLP and SSCP analyses, three genotypes of homozygous normal (N/M), heterozygous carrier (N/n) and homozygous recessive mutant (n/n) were detected using agarose or polyacrylamide gel electrophoresis, respectively. The proportions of normal, carrier and PSS pigs were 57.1, 35.7 and 7.1% for Landrace, 82.5, 15.8 and 1.7% far L. Yorkshire, 95.2, 4.8 and 0.0% for Duroc and 72.0, 22.7 and 5.3% for Crossbreed. Consequently, DNA-based diagnosis for the identification of stress-susceptible pigs of PSS and pigs producing PSE meat is a powerful technique. Especially, PCR-SSCP method may be useful as a rapid, sensitive and inexpensive test for the large-scale screening of PSS genotypes and pigs with PSE meat in the pork industry.y.

Ultrasonographic Evaluation in Patients with Inguinal Hernia (서혜부 탈장에서의 초음파 조영술)

  • Kwon, Oh-Kyoung;Jung, Jin-Hyang;Park, Jin-Young;Chang, Soo-Il
    • Advances in pediatric surgery
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    • v.8 no.1
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    • pp.16-22
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    • 2002
  • Between March 1999 and January 2000, 82 boys with the diagnosis of inguinal hernias (12 bilateral and 70 unilateral hernias), underwent Ultrasound (US) examination of both sides of the groin, a total of 164 inguinal imaging prior to surgery. The patients ages ranged from 3 days to 12 years with a mean of 32.6 months. Ninty four examinations were on the clinically symptomatic side and 70 were on the asymptomatic side. The US criteria for the diagnosis of an inguinal hernia were as follows: 1) visceral hernia, the presence of bowel loops, or omentum in the inguinal canal, 2) communicating hydrocele, the presence of fluid in the processus vaginalis, 3) widening of patent processus vaginalis at the level of internal inguinal ring. The width of patent processus vaginalis at the level of internal inguinal ring over 4 mm is considered an occult hernia. Among the 94 symptomatic groins, US findings showed 31 (33%) visceral hernias, 18 (19%) communicating hydroceles, and 38 (41%) widening of the internal inguinal ring, and 7 (7%) groins without abnormalities. In 70 asymptomatic groins, there were 4 (6%) visceral hernias, 5 (7%) communicating hydroceles, 11 (16%) widening of the internal inguinal ring, and 50 (70%) groins without abnormalities. Among the 70 asymptomatic groins there were US abnormalities in 20 (28%). One hundred and seven groins with positive US findings were surgically explored. Among 107 operated sites, the operative findings were compatible with the US diagnosis in 104, a sensitivity for US of 97.2%. In patients with US findings of widening of internal inguinal ring (>4 mm), there was patent processus vaginalis in 36 out of 38 symptomatic groins and 10 of 11 asymptomatic groins. The sensitivity of US to the operative findings in widening of internal inguinal ring was 93.8%. For visceral hernia and communicating hydrocele, the sensitivity of positive US findings was 100%. Ultrasonography for inguinal hernias appears to be a rapid, reliable, and noninvasive screening diagnostic tool with high positive specificity. Therefore, we recommend the use of US as a routine diagnostic tool in pediatric patients with inguinal hernias and hydroceles.

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The apoptotic fragment assay in rat peripheral lymphocytes and crypt cells with whole body irradiation with 60Co ϒ-rays and 50 MeV cyclotron fast neutrons (코발트-60 감마선과 50 MeV 싸이크로트론 고속 중성자선에 전신조사된 랫드의 말초 임파구와 음와 세포의 아포토시스 유도를 이용한 생물학적 선량 측정 모델 개발 연구)

  • Kim, Tae-hwan
    • Korean Journal of Veterinary Research
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    • v.41 no.2
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    • pp.203-210
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    • 2001
  • Here, we compared the effectiveness of 50 MeV($p{\to}RBe^+$) cyclotron fast neutrons versus $^{60}Co$ ${\gamma}$-rays by the apoptotic fragment frequency in both rat peripheral lymphocytes and crypt cells to check a radiobiological endpoint. The incidence of apoptotic cell death was increased in all irradiated groups, and radiation at all doses trigger rapid changes in both crypt cells and peripheral lymphocytes. These data suggest that apoptosis may play an important role in homeostasis of damaged radiosensitive target organ by removing damaged cells. The curve of dose-effect relationship for these data of apoptotic fragments frequencies was $y=0.3+(6.512{\pm}0.279)D(r^2=0.975)$ after neutrons, while $y=0.3+(4.435{\pm}0.473)D+(-1.300{\pm}0.551)D^2(r^2=0.988)$ after ${\gamma}$-rays. In addition, $y=3.5+(118.410{\pm}10.325)D+(-33.548{\pm}12.023)D^2(r^2=0.992)$ after ${\gamma}$-rays in rat lymphocytes. A significant dose-response relationship was found between the frequency of apoptotic cell and dose. These data show a trend towards increase of the numbers of apoptotic cells with increasing dose. Dose-response curves for high and low linear energy transfer (LET) radiation modalities in these studies were different. The relative biological effectiveness (RBE) value for crypt cells was 1.919. In addition, there were significant peaks on apoptosis induction at 4 and 6h after irradiation, and the morphological findings of the irradiated groups were typical apoptotic fragments in crypt cells that were hardly observed in the control group. Thus, apoptosis induction in both crypt cells and peripheral lymphocytes could be a useful endpoint of rat model for studying screening test and microdosimetic indicator to evaluate the biological effects of radiation-induced cell damage.

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Post-natal Outcome of Fetal Hydronephrosis Detected with Prenatal Ultrasonography (산전 초음파로 진단된 태아 수신증의 생후 경과 관찰)

  • Park, Youn Jin;Mun, Soon Jung;Bae, Chong Woo;Lee, Bum Ha;Kim, Jin Il
    • Clinical and Experimental Pediatrics
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    • v.45 no.10
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    • pp.1213-1218
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    • 2002
  • Purpose : Hydronephrosis constitutes a great portion of fetal anomalies screened by prenatal sonogram. The present authors made an attempt to access its natural courses through follow up neonatal hydronephrosis diagnosed by prenatal sonogram. Methods : The study was composed of 23 neonates(36 renal units) who were diagnosed with hydronephrosis through prenatal sonogram screening and confirmed 3-7 days after birth with sonographic evaluation at Kyung Hee University Hospital. The neonates were closely monitored for 12-24 months with renal sonogram, diuretic renogram, intravenous pyelography(IVP) and voiding cystoureterography(VCUG). Results : The underlying diseases were composed of 16 cases(44%) of functional abnormalities, 14 cases(39%) of ureteropelvic junction(UPJ) obstruction, three cases(8%) of vesicoureteral reflux (VUR) and on case each of multicystic dysplastic kidney, UPJ obstruction with ureteral stenosis and ureterovesical junction(UVJ) obstruction with VUR. The degree of hydronephrosis was divided into three classes according to its severity. In 30 renal units with UPJ obstruction and functional abnormalities, 26(87%) showed mild hydronephrosis, while four(13%) were moderate. During the follow up period, six cases(20%) showed natural resolution of hydronephrosis, 15 cases(50%) showed improvement while two cases(7%) were aggravated with improvement only after surgery of the underlying disease. The cases which showed natural resolution were all mild hydronephrosis at diagnosis and the cases which underwent surgery due to continuous aggravation were mild one case and moderate one case. Conclusion : Those with cases of mild hydronephrosis show rapid natural improvement. On the other hand, in some cases, follow up monitoring reveal aggravation of the situation, emphasizing the necessity for thorough follow up for a long period of time.