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Method of Nitrate Nitrogen Determination for Plant, Soil and Water Analysis by E. coil Cells (E. coli 세포(細胞)를 이용한 식물(植物), 토양(土壤) 및 수질(水質)의 질산태(窒酸態) 질소(窒素) 분석방법(分析方法))

  • Sohn, Sang Mok;Kucke, Martin;Lee, Yoon Gun
    • Korean Journal of Soil Science and Fertilizer
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    • v.30 no.4
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    • pp.361-369
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    • 1997
  • A microbiological nitrate determination method by E. coli is modified in Korea, using K12 wildtype, KCTC 1116, for the quantitative reduction of $NO_3{^-}$ to $NO_2{^-}$. The nitrate in plant, soil or water sample is determined spectrophotometrically after being diazotized with sulfaniamide and N-(1-naphthl)-ethlenediamine. The modified E. coli cell method and principle for nitrate determination using Korean wildtype E. coli strain is described, and cell culture and preparation of stock suspension for E. coli as well. This modified E. coli cell method can be managed simply and fast, it is suitable for the investigation of the large serials, it can be also automated and has a high degree of sensitivity up to 0.01ppm $NO_3{^-}-N$ in the sample solution. The applicability of the modified E. coli cell method has been tested for plant, soil and water analysis on a wide range of different samples. Recovery rates of added nitrate have been determined and comparisons with other standard nitrate analytical procedures have been carried out. The results with the modified E. coli cell method show high correlation ($r^2=0.98$) with those gained by the standard analytical procedures. The advantages and disadvantages of the method are also discussed to other nitrate determination methods.

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

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

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Functional Analysis of Bacillus subtilis Isolates and Biological Control of Red Pepper Powdery Mildew Using Bacillus subtilis R2-1 (Bacillus subtilis 균주의 기능성 분석 및 이를 활용한 고추 흰가루병 방제)

  • Kim, Yong-Ki;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Choi, Eun-Jung;Lee, Min-Ho;Park, Jong-Ho;Han, Eun-Jung;An, Nan-Hee;Jee, Hyeong-Jin
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.201-209
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    • 2012
  • The multi-function of 18 Bacillus subtilis isolates collected from agricultural extension centers of local government and National Academy of Agricultural Science was investigated by measuring their antifungal activities against five plant pathogens, such as Rhizoctonia solani, Colletotrichum acutatum, Fusarium oxysporum, Magnaporthe oryzae and Phytophthora capsici, phosphorus solubilization ability, production of indole acetic acid (IAA) and siderophore, and nitrogen fixation. The B. subtilis isolates showed antifungal activity against several plant pathogens and nitrogen fixation activity, and produced siderophore and IAA. They could control pepper powdery mildew (Leveillula taurica), but there was no difference in control efficacy among the B. subtilis isolates. In fields, the control efficacy of B. subtilis R2-1 ($10^8$ cells/ml) was compared with two microbial fungicides, Q-pect and Topsid. In 2009, the control efficacy of B. subtilis R2-1 (37.7%) was lower than that of Topsid (47.6%), but higher than that of Q-pect (25.7%). In 2010, the control efficacy of B. subtilis R2-1 (83.3%) was higher than that of Topsid (67.9%). In order to elucidate mode of action of B. subtilis R2-1 for controlling pepper powdery mildew, spore germination rates of pepper powdery mildew pathogen collected on treated leaves was investigated when suspensions of B. subtilis R2-1 and two microbial fungicides (Q-pect and Topsid) were foliar-sprayed. They highly suppressed spore germination of the pathogen with inhibition values of 84.2% for B. subtilis R2-1, 97.9% for Q-pect and 94.7% for Topsid. Further study on the mass-culturing method and formulation is needed for development of a microbial fungicide.

Administration of Anti-Inhibin Serum and Bovine Follicular Fluid on Plasma Hormonal Level and Ovarian Follicular Development in Hanwoo (Anti-Inhibin Serum 및 Bovine Follicular Fluld 처리가 한우 난소의 난포 발달과 혈중 호르몬 변화에 미치는 영향)

  • Seong, H.H.;Yang, B.C.;Hwang, S.S.;Park, S.J.;Ko, Y.G.;Rho, W.K.;Yang, B.S.
    • Journal of Embryo Transfer
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    • v.21 no.4
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    • pp.315-322
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    • 2006
  • This study was performed to investigate the physiological effects of bovine follicular fluid (bFF) or anti-inhibin serum (AI) on follicular development in Hanwoo. Saline (0.95%), bFF or AI (total of 40 ml) were administered into the jugular vein in 9 Hanwoo cows. The plasma inhibin, estradiol-17 $\beta$ (E2), and progesterone (P4) levels were measured using RIA or ELISA kit and the number of ovarian follicles was observed by ultrasonography at 72 hr after ovulation. The plasma inhibin level in bFF treatment group was significantly increased and maintained higher level from 102 hr after ovulation compared to that of saline and AI groups (p<0.05). In plasma E2 level, AI treatment group showed significantly higher level from 36 hr to 108 hr after ovulation than that of saline and bFF groups (p<0.05). After that it showed decreasing tendency. The plasma P4 level was increased in control and AI treatment groups at 68 hr after ovulation. However, it was maintained significantly lower level in bFF group from 84 hr to 180 hr compared to that of saline and AI group (p<0.05). As a result of ultrasonography at 72 hr after ovulation, higher number of follicles was shown in AI treatment group compared to bFF groups, although the difference was not statistically significant. Taken together, it can be postulated that a treatment of synthesized AI inhibits the secretion of inhibin, stimulates FSH secretion inhibited by inhibin, and induces follicular development and estrogen secretion. According to these results, a development of ovarian follicle immediately after ovulation is associated closely with inhibin in Hanwoo heifers.

Effects of Phytic Acid on Viability of Vibrio vulnificus and on Septicemia-Induced Mice (피틴산이 비브리오균의 생존과 마우스의 패혈증에 미치는 영향)

  • Chung, Young-Ho;Park, Woo-Woung;Lee, Sang-Yong;Lee, Sun-Woo;Lim, Chi-Hwan;Yoon, Min-Ho
    • Applied Biological Chemistry
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    • v.49 no.1
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    • pp.15-20
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    • 2006
  • EDTA is known to have bacteriocidal effect on Vibrio vulnificus, pathogen of septicemia by osmotic shock in seafoods. Attempts were made to elucidate the bacteriocidal effect of phytic acid (PA) as a substitute for EDTA against V. vulnificus and its inhibition effect on the septicemia, which induces liver damage of the mice by the pathogen. Viable cells of V. vulnificus with the initial titre of $1.7{\times}10^6$ c.f.u. $ml^{-1}$ decreased by 90.6% after 1 min and 99.6% after 5 min in distilled water. The titre decreased by 65.9% and 94.5% in 2 mM solution of $Mg^{2+}$. In 0.1 mM solution of PA, the rate of decrease in titre was 97.4% after 1 min of incubation and 99.8% after 5 min, compared to 95.7% and 99.8% in 0.1 mM solution of EDTA. The bacteriocidal effect of PA solution at a concentration of 1 mM was marked: the rate of decrease in titre was 99.9% after 1 min. In relation to the bacteriocidal effect, PA was evaluated as a potential therapeutic agent for V. vulnificus septicemia in mouse. When the survival periods of mice were investigated by PA and EDTA treatment after the pathogen injection, the group of mice which infected by a low concentration of the strain survived longer than that inoculated at high concentration; also, the ratio of survival was 1.3 times higher in PA than in EDTA, showing that the fatal rate depended on the inoculation concentration. Although survival periods of mice induced with liver damage by carbon tetrachloride and then inoculated with the strain showed a similar trend, the fatal rate of mice was 2 times faster than those inoculated with only pathogen into normal liver, These results indicate that the infection by V. vulnificus was more fatal to those with liver disease. Also, symptoms of hemorrhage and inflammation on the mice with induced liver damage were reduced in case there was phytic acid treatment at each concentration.

Experimental Studies on Uterine Catecholamines (Catecholamines에 관(關)하여 -제5편(第五編) : 자궁(子宮) catecholamines에 관한 실험적(實驗的) 연구(硏究)-)

  • Lee, Woo-Choo
    • The Korean Journal of Pharmacology
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    • v.19 no.1
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    • pp.37-60
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    • 1983
  • The uterus receives adrenergic terminals from the mesenteric ganglia and considerably large amount of catecholamines have been shown to be contained in this organ. On the other hand, the activities of epinephrine, norepinephrine or adrenergic nerve on uterine motility is so complicated that many controversial results have been reporter. Recently, a large number of reports concerning the changes of uterine catecholamines content have appeared, but little is known about the role of uterine catecholamines in their activities on uterine motility. The present experiments were undertaken to determine the significance of the intrinsic uterine catecholamines in the physiology of uterus. Female albino rabbits weighing approximately 2 kg were employed in this experiment. uterine strip3 were prepared and suspended in a constant temperature $bath(38^{\circ}C)$ containing 100 ml of Locke's solution aerated with 95% oxygen and 5% carbon dioxide. Spontaneous motility was recorded on a smoked drum with an isotonic lever. The catecholamines concentration of the uterus was determined according to the Procedure described of Shore and Olin (1958). Human uterus obtained from patients was also used to determine the catecholam ines content of myometrium. Followings are summarized results. 1) On the non-pregnant rabbit uterine strips, epinephrine and norepinephrine significantly elevated the tonus and stimulated the spontaneous motility. Pretreatment with dichloroisoproterenol(DCI), an adrenergic beta-receptor blocker, enhanced the stimulatory activity of epinephrine or norepinephrine. On the other hand, pretreatment with dibenamine, an adrenergic alpha-receptor blocker, rendered the uterine muscle to exhibit inhibition after the administration of epinephrine or norepinephrine. Following the treatment with both DCI and dibenamine, epinephrine or norepinephrine produced no appreciable effects on the spontaneous motility of the uterus. These results suggest there exist both alpha and beta-adrenergic receptors in the uterine muscle and the response to epinephrine of the former is predominant over that of latter in the non-pregnant uterus of rabbits. The total catecholamines concentration of the non-pregnant uterus was $351\;m{\mu}g/g$ and the fractional concentrations of epinephrine and norepinephrine were $125\;m{\mu}g/g(35.7%)$ and $226\;m{\mu}g/g$ respectively. It is interesting to note that the catecholamines content of uterus was characterized by a high fractional corcentration of epinephrine relative to norepinephrine. 2) On the pregnant rabbit uterine strips, the effects of epinephrine and norepinephrine varied according to the period of pregnancy. The response to epinephrine of adrenergic beta receptor of uterus increased during pregnancy, and the effect of catecholamine was inhibitory in the early pregnancy but became stimulatory as the pregnancy progressed. This stimulating action on the uterine motility was found to occur through the action of norepinephrine. The uterine catecholamines concentration was markedly reduced during pregnancy. The catecholamines concentration was started to decrease in the early pregnancy, reached the lowest level in the mid-pregnancy and then started to increaae again in the late pregnancy when the total catecholamines content became the highest level of all. This increase of catefholamines in late pregnancy was chiefly due to the increase of norepinephrine. These results suggest that the uterine motility may be related to the catecholamines content, especially norepinephrine content in the uterus. 3) Bilateral oophorectomy of rabbits results in a marked shrink of the uterus in size. The spontaneous motility of the uterine segment of these animals was very weak and irregular. Norepinephrine produced inhibitory effect, whereas epinephrine was stimulatory or inhibitory effect on the uterine segment. The total catecholamines tontent in whole uterus was markedly reduced. The injection of estrogen into the oophorectornized rabbit increased the weight of uterus to approximately three times of that of oophorectornized animal. The apontaneous motility and the response to epinephrine and norepinephrine of the uterine segment were greatly enhanced. Both epinephrine and norepinephrine produced a marked stimulatory effects of the uterine motility. The uterine content of catecholamines, particularly epinephrine, was markedly increased. The injection of progesterone into the oophorectornized rabbit increaeed the weight of uterus to approximately 2.5 times of that of eophorectornized animal. The spontaneous motility of the uterine segment was weak and irregular. Epinephrine produced stimulatory effect at high concentrations but norepinephrine always prcdnced inhibitory effect on the uterine segment. The uterine content of catecholamines, particularly of norepinephrine, was markedly reduced. These results suggested that ovarian hormones play an important role not only on the growth and spontaneous norepinephrine of uterus but also on the catecholamines content and responee to epinephrine and norepinephrine of the uterus. 4) The intraperitoneal injection of reserpine(3 mg/kg) into the non-pregnant, pregnant and oophorectornieed rabbits markedly decreased the uterine content of catecholamines, particularly of the norepinephrine. The stimulatory response to epinephrine and. norepinephrine of the uterine segment of these reserpinized ratbits was markedly reduced whereas the inhibitory response to these catecholamines was enhanced. This finding further support the close relationship between the uterine catecholamines content and uterine response to epineptrire and norepinephrine. 5) In the human uterus, the concentration of epinephrine was actrally greater than that of norepinephrine and it was significantly greater during the proliferative phase of the menstrtal cycle. In the human pregnant uterus, the concentrations of toth epinephrine and ncrefinephrine were markedly reduced and showed about 45 percent rednction after 6-8 weeks of ectopic Pregnancy. At full term ana during labor, the concentrations of epinephrine and norepinephrine at placental sites were less than those found in the non-pregnant group. Of interest was the finding that the norepinephrine concentration of uterus from toxemic patients was two and half times higher than that of lower uterine segment of the nontoxemic pregnant individuals. Also the epinephrine concentraticn was slightly increaeed.

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Fusarium moniliforme Detected in Seeds of Corn and Its Pathological Significance (옥수수 종자(種子)에서 검출(檢出)된 Fusarium moniliforme와 그 병리학적(病理學的) 중요성(重要性))

  • Kim, Wan-Gyu;Oh, In-Seok;Yu, Seung-Hun;Park, Jong-Seong
    • The Korean Journal of Mycology
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    • v.12 no.3
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    • pp.105-110
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    • 1984
  • Seven seed samples of corn obtained from Kangweon Provincial Office of Rural Development, Kerea were tested for seed-borne fungi, and found that all the samples tested were infected with Fusarium moniliforme to an extent of $6.0{\sim}79.5%$. Severely infected seed samples showed poor germination on blotter. Seed component plating showed that the fungus present not only in tip caps, pericarps and endosperms, but also in embryos. Heavy infection of the fungus caused severe seed rot and seedling blight in soil, but the damage was not severe and many plants grew without any symptoms when the seeds with light infection were sown in soil. However the fungus was frequently detected from inside of the stems of healthy looking seedlings. The results indicate that the fungus transmit from seed to plant systemically. In inoculation experiments, the fungus produced stem rots on corn plants of 110 days old. The cultivar of Hwangok 3 was revealed more susceptible to the fungus than that of Suweon 19.

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EFFECTS OF ND:YAG LASER IRRADIATION AND FLUORIDE APPLICATION ON REMINERALIZATION OF THE ENAMEL (Nd:YAG 레이저 조사와 불화물 도포가 탈회법랑질의 재광화에 미치는 영향)

  • Cho, Hyun;Kim, Dae-Eop;Lee, Kwang-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.3
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    • pp.381-390
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    • 2004
  • The purpose of this in vitro study was to investigate the effects of Nd:YAG laser irradiation and fluorides on acid drink demineralized enamel. The materials were 30 freshly extracted permanent premolars with intact smooth enamel surfaces. They were demineralized with Coca-cola at $37^{\circ}C$ for 12hours and then irradiated by Nd:YAG laser with 6W power, $50mJ/cm^2$ energy density, and 20Hz pulse repetition. After laser irradiation, teeth were treated by three kinds of fluorides; (1)0.05% NaF fluoride solution (2)1.23% APF gel and (3)0.1%F fluoride varnish, microhardness(VHN) and Diagnodent scores were measured and the surfaces of each treated specimens were also observed with SEM under 1500 magnification. The results were as follows: 1. In the change of microhardness(VHN), it decreased to 34.68% from the initial micrhardness, increased to 78.37% after laser irradiation and increased to 82.62% after fluoride treatment, there were significant differences except when it was irradiated and treated with fluoride(P<0.05). 2. In the change of Diagnodent scores, it was decreased to 28.08% from the initial scores after demineralization, and then increased to 59.81% after laser irradiation, and increased to 82.17% after fluoride treatment. Scores were different significantly between the scores of initial, demineralization, laser irradiation and fluoride treatment(P<0.05). All the scores were not different significantly between fluoride types. 3. SEM observation showed that the lased enamel surfaces after demineralization were thermally degenerated and showed molten lava-like appearance and crater with cracks and many microholes.

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Serum Beta-2 Microglobulin: a Possible Marker for Disease Progression in Egyptian Patients with Chronic HCV Related Liver Diseases

  • Ouda, SM;Khairy, AM;Sorour, Ashraf E;Mikhail, Mikhail Nasr
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7825-7829
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    • 2015
  • Background: Egypt has the highest prevalence of HCV infection in the world (~14.7%). Around 10-15% of HCV-infected persons will advance to cirrhosis within the first 20 years. The incidence of HCC is expected to grow in the next two decades, largely due to HCV related cirrhosis, and detection of HCC at an early stage is critical for a favorable clinical outcome. No simple reliable non-invasive marker has been available till now. B2M, a non-glycosylated polypeptide composed of 99 amino acids, is one of the components of HLA class I molecules on the surfaces of all nucleated cells. It has been reported that the level of serum B2M is elevated in patients with chronic hepatitis C and HCV-related HCC when compared to HCV-negative patients or healthy donors. Determining the clinical utility of serum B2M as a marker for disease progression in Egyptian patients with HCV related chronic hepatitis, cirrhosis and hepatocellular carcinoma was the aim of the present study. Materials and Methods: In this analytical cross sectional study 92 participants were included in 4 equal groups: Group (1) non cirrhotic chronic HCV; Group (2) HCV related liver cirrhosis; Group (3) HCC on top of HCV,; and Group (4) healthy controls. History taking, clinical examination, routine labs and abdominal ultrasound were conducted for all patients, PCR and Metavir scores for group (1) patients, and triphasic CT abdomen and AFP for Group (3) patients. B2M levels were measured in serum with a fully-automated IMX system. Results: The mean serum B2M level of Group (1) was $4.25{\pm}1.48{\mu}g/ml$., Group (2) was $7.48{\pm}3.04$, Group (3) was $6.62{\pm}2.49$ and Group (4) was $1.62{\pm}0.63$. Serum B2M levels were significantly higher in diseased than control group (p<0.01) being significantly higher in cirrhosis ($7.48{\pm}3.04$) and HCC groups ($6.62{\pm}2.49$) than the HCV group ($4.25{\pm}1.48$) (p<0.01). There was a significant correlation between B2M Level and ALK, total and direct bilirubin and INR (p<0.05), and a significant inverse correlation between B2M level and albumin, total proteins, HB andWBCS values (p<0.05). There was no significant correlation between B2M level and viral load or Metavir score, largest tumour size or AFP (p>0.05). The best B2M cut-off for HCV diagnosis was 2.6 with a sensitivity of 100%, a specificity of 92%, a positive predictive value (PPV) of 97% and a negative predictive value (NPV) of 100%. The best B2M cut-off for HCC diagnosis was 4.55 which yielded sensitivity, specificity, positive predictive value, negative predictive values of 74%, 62%, 39.5, 87.8% respectively (p-value <0.01) while best cut-off for cirrhosis was 4.9, with sensitivity 74 % and specificity 74%.The sensitivity for HCC diagnosis increased upon B2M and AFP combined estimation to 91%, specificity to 79%, NPV to 95% and accuracy to 83%. Conclusions: Serum B2M level is elevated in HCV related chronic liver diseases and may be used as a marker for HCV disease progression towards cirrhosis and carcinoma.

Efficacy of Neoadjuvant Chemotherapy and Radiotherapy for the Histology-confirmed Intracranial Germinoma - Preliminary Report (조직학적으로 확진된 두개내 배아종의 전보조화학요법 후 방사선치료의 성적 - 예비적 결과)

  • Noh, Young-Ju;Kim, Hak-Jae;Heo, Dae-Seog;Shin, Hee-Yung;Kim, Il-Han
    • Radiation Oncology Journal
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    • v.20 no.2
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    • pp.93-99
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    • 2002
  • Purpose : We intended to decrease late CNS reaction after radical radiotherapy for an intracranial germinoma by using combined neoadjuvant chemotherapy and involved-field radiotherapy. The efficacy in terms of its acute toxicity and short-term relapse patterns was analyzed. Materials and Methods : Eighteen patients were treated with combined neoadjuvant chemotherapy and radiotherapy between 1995 and 2001. The chemotherapy regimen used was the Children's Cancer Group (CCG) 9921A (cisplatin, cyclophosphamide, VP-16, vincristine) for 5 patients younger than 16 years, BEP (bleomycin, VP-16, cisplatin) for 12 patients, and EP (VP-16, cisplatin) for 1 patient. The radiotherapy covered the whole craniospinal axis for 5 patients, the whole brain for 1, and the partial brain (involved field) for 12. the primary lesion received tumour doses between 3,960 and 5,400 cGy. Results : The male to female ratio was 16:2 and the median age was 16 years old. The tumors were located in the pineal gland in 12 patients, in the suprasellar region in 1, in the basal ganglia In 1, in the thalamus in 1. Three patients had multiple lesions and ventricular seedings were shown at MRI. In 3 patients, tumor cells were detected in the cerebrospinal fluid and MRI detected a spinal seeding in 2 patients. The response to neoadjuvant chemotherapy was complete remission in 5 patients, partial remission in 12, and no response in 1. However, after radiotherapy, all except 1 patient experienced complete remission. The toxicity during or after chemotherapy greater than or equal to grade III was remarkable; hematologic toxicity was observed in 11 patients, liver toxicity in none, kidney toxicity in none, and gastrointestinal toxicity in one. One patient suffered from bleomycin-induced pneumonitis. Radiotherapy was therefore stopped and the patient eventually died of respiratory failure. The other 17 are alive without any evidence of disease or relapse during an average of 20 months follow-up. Conclusion : A high response rate and disease control was experienced, which was the same as observed other studies and the morbidity from chemotherapy-induced toxicity was similar. With these results, the results from adjuvant chemotherapy and involved-field radiotherapy cannot be concluded to be equal to those from extended-field radiotherapy. The long term follow-up study on later complications are required in order to draw definite conclusions on the optimal management with minimum side effects.