• Title/Summary/Keyword: Plasma Diagnosis

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Noninvasive fetal RHD genotyping using cell-free fetal DNA incorporating fetal RASSF1A marker in RhD-negative pregnant women in Korea

  • Han, Sung-Hee;Yang, Young-Ho;Ryu, Jae-Song;Kim, Young-Jin;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.100-108
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    • 2015
  • Purpose: Conventional methods for the prenatal detection of fetal RhD status involve invasive procedures such as fetal blood sampling and amniocentesis. The identification of cell-free fetal DNA (cffDNA) in maternal plasma creates the possibility of determining fetal RhD status by analyzing maternal plasma DNA. However, some technical problems still exist, especially the lack of a positive control marker for the presence of fetal DNA. Therefore, we assessed the feasibility and accuracy of fetal RHD genotyping incorporating the RASSF1A epigenetic fetal DNA marker from cffDNA in the maternal plasma of RhD-negative pregnant women in Korea. Materials and Methods: We analyzed maternal plasma from 41 pregnant women identified as RhD-negative by serological testing. Multiplex real-time PCR was performed by amplifying RHD exons 5 and 7 and the SRY gene, with RASSF1A being used as a gender-independent fetal epigenetic marker. The results were compared with those obtained by postnatal serological analysis of cord blood and gender identification. Results: Among the 41 fetuses, 37 were RhD-positive and 4 were RhD-negative according to the serological analysis of cord blood. There was 100% concordance between fetal RHD genotyping and serological cord blood results. Detection of the RASSF1A gene verified the presence of cffDNA, and the fetal SRY status was correctly detected in all 41 cases. Conclusion: Noninvasive fetal RHD genotyping with cffDNA incorporating RASSF1A is a feasible, reliable, and accurate method of determining fetal RhD status. It is an alternative to amniocentesis for the management of RhD-negative women and reduces the need for unnecessary RhIG prophylaxis.

The effect of Westergren tube angle, temperature and osmobility with the different percent of NaCl solution on erythrocyte sedimentation rate (적혈구 침강속도에 대한 Westergren tube 각도, 온도 및 NaCl 농도가 미치는 영향)

  • Kim, Do-kyun;Kim, Young-hong;Yu, Chang-jun
    • Korean Journal of Veterinary Research
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    • v.36 no.1
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    • pp.75-82
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    • 1996
  • Erythrocyte sedimentation rate is influenced by plasma protein, red cell itself and physiological conditions and it is clear that the measurement of erythrocyte sedimentation rate varies with technique and various environmental factors. The effect of temperature, angle of test tube, and osmobility with different percent of NaCl solution on erythrocyte sedimentation rate on sheep was determined by modified Westergren method. In sheep, as the angle of Westergren tube was decreased from $90^{\circ}$ to $45^{\circ}$, erythrocyte sedimentation rate was increased in both diluted plasma and NaCl solutions. As temperature was increased from $4^{\circ}C$ to $20^{\circ}C$ at $45^{\circ}$ angle. erythrocyte sedimentation rate was increased. In ruminants, this modified Westergren methods, erythrocyte 1 : plasma 9 instead of erythrocyte 4 : plasma 6(whole blood), enable them to have meanings as dog therefor, using this method, clinical can determine the erythrocyte sedimentation rate of ruminants for diagnosis. Because erythrocyte sedimentation rate was changed according to the angle of Westergren tube, temperature as erythrocyte diluted with NaCl, this study detected that the change of temperature, the angle of Westergren tube could change erythrocyte sedimentation rate by effecting red cell itself. The increase of osmobility owing to change of NaCl percent resulted in the decrease of rapid erythrocyte sedimentation rate. So this fact indicate that deformibility and the change of red cell volume have meaning in the change of erythrocyte sedimentation rate.

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Plasma Peptidome as a Source of Biomarkers for Diagnosis of Cholangiocarcinoma

  • Kotawong, Kanawut;Thitapakorn, Veerachai;Roytrakul, Sittiruk;Phaonakrop, Narumon;Viyanant, Vithoon;Na-Bangchang, Kesara
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1163-1168
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    • 2016
  • Cholangiocarcinoma (CCA) is the bile duct cancer which constitutes one of the important public health problems in Thailand with high mortality rate, especially in the Opisthorchis viverrini (a parasite risk factor for CCA) endemic area of the northeastern region of the country. This study aimed to identify potential biomarkers from the plasma peptidome by CCA patients. Peptides were isolated using 10 kDa cut-off filter column and the flow-through was then used as a peptidome for LC-MS/MS analysis. A total of 209 peptides were obtained. Among these, 15 peptides were concerned with signaling pathways and 12 related to metabolic, regulatory, and biosynthesis of secondary metabolite pathways. Five exclusive peptides were identified as potential biomarkers, i.e. ETS domain-containing transcription factor ERF (P50548), KIAA0220 (Q92617), phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit beta isoform isoform 1 (P42338), LP2209 (Q6XYC0), and casein kinase II subunit alpha (P19784). Three of these biomarkers are signaling related molecules. A combination of these biomarkers for CCA diagnosis is proposed.

Diagnosis of Alzheimer's disease through measurement of anti-beta amyloid antibody in patient's serum

  • Sohn, Ji-Hoon;Kim, Hee;So, Jung-On;Huh, Ji-Yeon;Kim, Jong-Won;Kim, Man-Ho;MookJung, In-Hee;Kim, Young-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2006.04a
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    • pp.139-149
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    • 2006
  • Amyloid beta (A$\beta$) is believed one of the major pathogens of Alzheimer's disease (AD), and the reduction of A$\beta$ is considered a primary therapeutic target. Immunization with A$\beta$ can reduce A$\beta$ burden and pathological features in transgenic AD model mouse. This means anti-A$\beta$ autoantibodies may have a role in AD pathology. Recent findings suggest anti-A$\beta$ autoantibodies level decrease in AD patients. The early detection of AD is important for treatment of this disease. However, diagnosis on AD has only been possible through limited methods such as neuropsychological examination or MRI. To investigate whether it was possible to detect the presence and different levels of naturally occurring anti-A$\beta$ autoantibodies in the plasma of patients with AD and age-matched controls. An advanced ELISA was performed to detect levels of naturally occurring anti-A$\beta$ autoantibodies in the plasma. The level of anti-A$\beta$ auto-antibodies from patients with idiopathic Parkinson's disease or stroke and from normal controls were compared to that of AD patients. Our results showed a significantly lower anti-A$\beta$ autoantibodies level in AD compared to those with other neurological diseases or control. The level of anti-A$\beta$ autoantibodies in the serum may be used to diagnose the presence of AD.

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Diagnosis of Alzheimer's disease through measurement of anti-beta amyloid antibody in patient's serum

  • Sohn, Ji-Hoon;Kim, Hee;So, Jung-On;Huh, Ji-Yeon;Kim, Jong-Won;Kim, Man-Ho;MookJung, In-Hee;Kim, Young-Ho
    • 한국약용작물학회:학술대회논문집
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    • 2006.04a
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    • pp.137-149
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    • 2006
  • Amyloid beta ($A{\beta}$) is believed one of the major pathogens of Alzheimer's disease (AD), and the reduction of $A{\beta}$ is considered a primary therapeutic target. Immunization with $A{\beta}$ can reduce $A{\beta}$ burden and pathological features in transgenic AD model mouse. This means $anti-A{\beta}$ autoantibodies may have a role in AD pathology. Recent findings suggest $anti-A{\beta}$ autoantibodies level decrease in AD patients. The early detection of AD is important for treatment of this disease. However, diagnosis on AD has only been possible through limited methods such as neuropsychological examination or MRI. To investigate whether it was possible to detect the presence and different levels of naturally occurring $anti-A{\beta}$ autoantibodies in the plasma of patients with AD and age-matched controls. An advanced ELISA was performed to detect levels of naturally occurring $anti-A{\beta}$ autoantibodies in the plasma. The level of $anti-A{\beta}$ auto-antibodies from patients with idiopathic Parkinson's disease or stroke and from normal controls were compared to that of AD patients. Our results showed a significantly lower $anti-A{\beta}$ autoantibodies level in AD compared to those with other neurological diseases or control. The level of $anti-A{\beta}$ autoantibodies in the serum may be used to diagnose the presence of AD.

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Microsurgical re-treatment of an endodontically treated tooth with an apically located incomplete vertical root fracture: a clinical case report

  • Taschieri, Silvio;Fabbro, Massimo Del;Kabbaney, Ahmed El;Tsesis, Igor;Rosen, Eyal;Corbella, Stefano
    • Restorative Dentistry and Endodontics
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    • v.41 no.4
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    • pp.316-321
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    • 2016
  • Although it is challenging, the early diagnosis of a vertical root fracture (VRF) is crucial in order to ensure tooth preservation. The purpose of this clinical case report was to describe reparative surgery performed to treat a tooth affected by an incomplete VRF. A 26 year old male patient was suspected to have a VRF in a maxillary left central incisor, and an exploratory flap was performed in order to confirm the diagnosis. After detecting the fracture, the lesion was surgically treated, the fracture and the infected root-end were removed, and a platelet-rich plasma membrane was used to cover the defect in order to prevent bacterial migration. A 24 month clinical and radiological follow-up examination showed that the tooth was asymptomatic and that the healing process was in progress. The surgical approach described here may be considered an effective treatment for a combined endodontic-periodontal lesion originating from an incomplete VRF and a recurrent periapical lesion.

Comparison of the bovine blood gas parameters produced with three types of portable blood gas analyzers

  • Ro, Younghye;Choi, Woojae;Hong, Leegon;Kim, Eunkyung;Choe, Eunhui;Kim, Danil
    • Journal of Veterinary Science
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    • v.23 no.4
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    • pp.60.1-60.6
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    • 2022
  • Background: A definite diagnosis should be made in the bovine practice field, however, it was difficult to perform laboratory analysis immediately. Currently, three types of portable blood gas analyzers are available in Korea. Objectives: This study aimed to evaluate the correlations among these three analyzers. Methods: Seventy-two plasma samples from Holstein-Friesian cows were used for blood gas analysis, and three instruments (EDAN i15 Vet, VETSCAN i-STAT, and EPOC) were operated simultaneously. Moreover, plasma calcium levels were compared between these portable analyzers and blood chemistry device, which is usually used in a laboratory environment. Pearson analysis was performed to confirm the correlation of each parameter produced with the three instruments and blood chemistry analyzer. Results: As results, high correlation was observed in parameters of pH, pO2, potassium ion, ionized calcium, and glucose (p < 0.001, r > 0.7). In addition, pCO2 showed a moderate correlation among the three analyzers (p < 0.001, r > 0.5), and there was no correlation among all instruments for sodium ions. There was also a high correlation between ionized calcium from the three portable devices and total calcium from the biochemistry analyzer (p < 0.001, r > 0.9). Conclusions: In conclusion, there was a high correlation between results from the three different blood gas analyzers used in the bovine clinical field in Korea. Thus, a consistent diagnosis can be made even with different equipment if the operator is aware of the strengths and weaknesses of each piece of equipment and operates it properly.

Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease (패브리병(Fabry) 진단을 위한 혈장 중 Globotriaosylceramide (Gb3)의 탠덤매스 분석법 개발과 임상 응용)

  • Yoon, Hye-Ran;Cho, Kyung-Hee;Yoo, Han-Wook;Choi, Jin-Ho;Lee, Dong-Hwan;Zhang, Kate;Keutzer, Joan
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.45-52
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    • 2007
  • Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

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Milk Transfer and Toxicokinetics of Valproic Acid in Lactating Cynomolgus Monkeys

  • Lee, Jong-Hwa;Yu, Wook-Joon;Jeong, Eun Ju;Chung, Moon-Koo
    • Toxicological Research
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    • v.29 no.1
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    • pp.53-60
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    • 2013
  • Studies on milk transfer of drugs in non-human primates (NHPs) are among the crucial components in the assessment of peri- and postnatal toxicity because of the similarity between NHPs and humans. To evaluate the milk transfer of valproic acid (VPA) in NHPs, the toxicokinetics of VPA, an antiepileptic drug, were studied in pregnant cynomolgus monkeys. VPA was administered once daily to pregnant cynomolgus monkeys at doses of 0, 30, 90, and 270 mg/kg by oral gavage from Day 100 of gestation (GD 100) to Day 31 of lactation (LD 31). Concentrations of VPA and its metabolite, 4-ene-VPA, in the maternal plasma on GD 100, GD 140, and LD 30, and concentrations of VPA and 4-ene-VPA in the offspring plasma and milk on LDs 30 and 31, respectively, were quantified using liquid chromatography tandem mass spectrometry (LC/MS/MS). After administration of a single oral dose of VPA to pregnant monkeys on GD 100, the concentrations of VPA and 4-ene-VPA were generally quantifiable in the plasma of all treatment groups up to 24 hr after administration, which showed that VPA was absorbed and that the monkeys were systemically exposed to VPA and 4-ene-VPA. After administration of multiple doses of VPA to the monkeys, VPA was detected in the pup's plasma and in milk taken on LD 30 and LD 31, respectively, which showed that VPA was transferred via milk, and the pup was exposed to VPA. Further, the concentration of VPA in the milk increased with an increase in the dose. Extremely low concentrations of 4-ene VPA were detected in the milk and in the pup plasma. In conclusion, pregnant monkeys were exposed to VPA and 4-ene-VPA after oral administration of VPA at doses of 30, 90, and 270 mg/kg/day from GD 100 to LD 31. VPA was transferred via milk, and the VPA exposure to the pup increased with an increase in the dose of VPA. The metabolite, 4-ene VPA, was present in extremely low concentrations (< 0.5 ${\mu}g/ml$) in the milk and in the pup plasma. In this study, we established methods to confirm milk transfer in NHPs, such as mating and diagnosis of pregnancy by examining gestational sac with ultrasonography, collection of milk and pup plasma and determination of toxicokinetics, using cynomolgus monkeys.

A Comparative Evaluation of Three Rapid Tests of Syphilis and ARCHITECT Syphilis TP

  • Kim, Won-Shik
    • Korean Journal of Clinical Laboratory Science
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    • v.43 no.1
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    • pp.1-5
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    • 2011
  • The infection rate of syphilis is still increasing in the world especially in developing countries and the infection is often seen in large amounts of clinical specimens. For the diagnosis of this disease, Rapid Plasma Reagin (RPR)/Venereal Disease Research Laboratory (VDRL) has still been used as one of major primary methods to diagnose syphilis even though the test readings are somewhat subjective with high false positive rates. Recently, the automatic ARCHITECT Syphilis TP, which is based on the detection of the TP-specific antibodies, has been introduced in many laboratories. Therefore, the clinical assessment of the method is needed to provide primary diagnosis of syphilis at the moment. We evaluated 3 different manual rapid kits and ARCHITECT Syphilis TP comparing with RPR/FTA-ABS and analysed their diagnostic properties. From February 2006 to April 2008, 203 positive and 250 negative specimens, obtained from Chungbuk National University Hospital were used for the evaluation. In the evaluation between manual rapid kits, their specificities were as high as 99.2 ~ 99.6% while their sensitivities were observed with little differences; 98.0% (199/203) for Kit A, 96.6% (196/203) for Kit B, and 97.4% (197/203) for Kit S. In the case of ARCHITECT Syphilis TP test, it showed 100% specificity (250/250) and 98.5% sensitivity (249/250). Kappa values comparing with RPR/FTA-ABS were 0.978 for Kit A, 0.964 for Kit B and Kit S, and 0.987 for ARCHITECT Syphilis TP. From our evaluation, we found out that manual rapid tests and ARCHITECT Syphilis TP have very good clinical accuracies and high kappa agreements with RPR/FTA-ABS. Due to its automation and quick simultaneous diagnosis with another serological markers, we suggest that the ARCHITECT Syphilis TP is one of best suitable method for the primary diagnosis of syphilis and that it might be able to replace RPR method in the laboratories.

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