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Experience of Reticulocytes Measurement at 720 nm Using Spectrophotometer (분광광도계를 이용한 720 nm에서 망상적혈구 측정 경험)

  • Sung, Hyun-Ho;Seok, Dong-In;Jung, You-Hyun;Kim, Dae-Jung;Lee, Seok-Jae
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.382-389
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    • 2017
  • Currently, reticulocyte experimental calculation technology used in clinical laboratories are divided two types: manual and automated. Manual reticulocyte counting using a microscopy lacks accuracy due particularly to its low reproducibility, affecting the accuracy of manual reticulocyte count. Moreover, Automatic blood corpuscle analyzer flow cytometry is difficult to be used in underdeveloped countries and small scale laboratories due to relatively high cost. Therefore, this study tried to find a new method to complement these drawbacks. The aim of this study was to compare the stained reticulocytes count by spectrophotometer and also to analyze the statistics of spectrophotometer and flow cytometer. The same 8 EDTA samples were repeated 36 times to compare the agreement between spectrophotometer and flow cytometer. This study measured the specimen diluted 600 times at 700~780 nm by 10 differences. Wavelength between 710 to 730 by absorbance showed a positive correlation between standard data and test data (r=0.967, p<0.01), presenting a correlation between variables. Statistical analyses of regression for test and standard parametric data, the optimal dilution factor was 600 times. Therefore, this study tried to technical utilizes such as contributing economical for the reticulocyte absorbance apply from the auto spectrophotometer, a monitoring system for the reticulocyte relation anemia, etc. Therefore, more extensive studies, including an auto chemical analyzer application, will be needed.

Clonal Dissemination of Multidrug Resistant Acinetobacter baumannii Isolates Harboring blaOXA-23 at One University Hospital in Daejeon, Korea (대전지역 소재 대학병원에 blaOXA-23 유전자를 가지고 있는 다제내성 Acinetobacter baumannii의 확산)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.94-101
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    • 2016
  • Acinetobacter species isolates are important opportunistic pathogens and commonly implicated in nosocomial infections. The therapeutic options for treatment of the bacterial infections are limited because the bacteria isolates are usually multidrug resistant (MDR). In the current study, we investigated various carbapenemase genes in 68 Acinetobacter species isolates. Antimicrobial susceptibilities were tested using the disk diffusion method. Screening of carbapenemase genes was performed via multiplex PCR. In addition, PCR and DNA sequencing were used to identify the carbapenemase genes. Repetitive extragenic palindromic-PCR (REP-PCR) was also performed to assess the clonality of isolates. In our study, A. baumannii isolates were highly resistant to all agents tested while all non-A. baumannii isolates were susceptible to all agents tested, with the exception of aztreonam and cefotaxime. All 51 A. baumannii isolates contained the $bla_{OXA-51}$ gene and 37 (72.5%) isolates also harbored the $bla_{OXA-23}$ gene. In addition, 39 MDR A. baumannii isolates were identified in our study and 37 isolates contained the $bla_{OXA-23}$ gene. The 37 MDR strains harboring $bla_{OXA-23}$ showed type I (n=22) or type II (n=15) banding patterns on their REP-PCR profiles. Our results suggest clonal relation and horizontal spreading of MDR A. baumannii isolates containing the $bla_{OXA-23}$ gene at the hospital located in Daejeon. Continuous investigation of antimicrobial resistant determinants and monitoring emergence and dissemination of MDR isolates is required to prevent and control infection and colonization of MDR A. baumannii isolates.

Evaluation of Analyzer and Measurement Conditions of Blood Ammonia (혈중 암모니아의 측정조건과 분석기기의 평가)

  • Kim, Sang-Su;Kim, Myong Soo;Lee, Seung Mo;Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.68-73
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    • 2016
  • Ammonia is very toxic, and causes neuronal damage via excitotoxicity, oxidative stress, and inflammation. Because the liver is the primary organ for ammonia metabolism, compromised liver function can result from inborn errors of metabolism. Measurement of blood ammonia has some limitations. Recently, several laboratories examined possible concurrent increases in plasma ammonia. However, the collection, handling, storage, and analysis of blood samples are all potential sources of error. For evaluation of rapidity and reliability of measurement of blood ammonia, the DRI-CHEM 100 (Fuji Film Co., Japan) and COBAS 8000 (Roche Diagnostic Ltd., Switzerland) analyzer were used for analysis of ammonia level values. The results of this study detected a high correlation between analyzer. Therefore, one-step measurement was suitable for ammonia analysis. After sampling of the ammonia in the time slot for measurement an increase to 46.5, 57.4, and 79.0 (${\mu}g/dL$) was observed at 30, 90, and 180 minutes. In addition, specific capacity of the ammonia, 7, 10, and 13 (${\mu}L$), was measured as 39, 46, and 43 (${\mu}g/dL$), respectively, and the FDC-100 analyzer was more effective in $10{\mu}L$ (p<0.001). In conclusion, the evaluated analysis may offer useful information for clinical application.

Usefulness of the Neutrophil Gelatinase-Associated Lipocalin (NGAL) Kit for Acute Kidney Injury Patients at the Emergency Medical Center in Daegu (대구지역 응급의료센터에 내원한 급성 콩팥손상 환자의 진단을 위한 호중구 젤라티나제 관련 리포칼린 키트의 유용성)

  • Lee, Seung-Jin;Park, Sangwook
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.49-53
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    • 2016
  • Acute kidney injury (AKI) is a common syndrome resulting in kidney damage and malfunction within a few days or even a few hours. The diagnosis of AKI depends on routine biochemical tests, including serum creatinine, aspartate aminotransferase (AST), alanine aminotransaminase (ALT), blood urea nitrogen (BUN), and electrolytes. Plasma neutrophil gelatinase-associated lipocalin (NGAL) is a biomarker that shows correlation with the severity of acute infections and kidney injuries. The predictive value in other conventional assays for kidney functions has been reported to cause distraction for AKI syndrome. The aim of this study is to verify the predictive value of plasma NGAL in patients with established AKI. The NGAL kit for checkup demonstrates sensitivity of ${\geq}300$ (92.2%), ${\geq}200$ (95.6%), ${\geq}100$ (99.6%), specificity of ${\geq}300$ (95.1%), ${\geq}200$ (97.3%), ${\geq}100$ (99.4%), positive predictability of ${\geq}300$ (93.3%), ${\geq}200$ (93.4%), ${\geq}100$ (99.2%), and negative predictability of ${\geq}300$ (96.7%), ${\geq}200$ (97.7%), ${\geq}100$ (98.1%), respectively. The plasma NGAL compared with the enzyme-linked immunosorbent assay (ELISA) has been shown to be an early predictive biomarker of AKI. The NGAL kit, recently developed for point-of-care of plasma specimens, is thought to be a useful and reliable biomarker for the early diagnosis of decreased kidney functions.

Correlation Between p53 and p21 Proteins Expression and Prognostic Factors Related with Colon Cancer

  • Kim, Tai-Jeon;Kim, Tae-Geun
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.2
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    • pp.128-135
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    • 2007
  • This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins by immunohistochemical staining and tumor prognostic factors including the tumor size, histological differentiation and Dukes' stage of tumor prognostic factors in colon cancer, and to acquire necessary data for the presumption of diagnosis, treatment and prognosis of colon cancer patients. From January 2000 to January 2003 at Hanyang University Guri Hospital, the paraffin blocks of 35 patients diagnosed with colon cancer whose pathologic reports were possible to review were selected. Harris hematoxylin & eosin (H&E) staining and immunohistochemical staining by ABC (Avidin Biotin Conjugate) method were performed. The histological differentiation grade and stage were classified according to the classification of the World Health Organization (WHO) and modified Dukes's stage from H&E staining. The expression rate of p53 and p21 proteins were analyzed by immunohistochemical staining. The results was analyzed statistically by SPSS (Windows version 8.0). As a result, the expression rate of p53 protein was 11.4% (4 cases) in clear differentiation, 48.6% (17 cases) in moderate differentiation, and 17.1% (6 cases) in poor differentiation. In other words, the poorer the differentiation, the higher the expression rate of p53 protein (p<0.05). The expression rate of p21 was 17.1% (6 cases) in clear differentiation, 40.0%(14 cases) in moderate differentiation, and 8.6% (3 cases) in poor differentiation, According to the progression of histological malignant degeneration, the expression rate of p21 protein decreased distinctively (p<0.05). However, the correlation between the two above mentioned proteins and the tumor-size and Dukes' stage was not of statistical significance. In the comparison of the expression rate of p53 protein with that of p21 protein, in 10 cases, p53 protein expression was positive while p21 protein expression was negative, and in 6 cases, p53 protein expression was negative whereas p21 protein expression was positive. Consequently a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed (p<0.05). In conclusion, we found a significant correlation between histological differentiation and the expression rate of p53 and p21 proteins (p<0.05), and a significant inverse correlation between the expression rate of p53 protein and that of p21 protein (p<0.05). Also, it could be confirmed that the over expression of p53 and p21 proteins is closely associated with the occurrence of colon cancer and its progress. Therefore, it is thought that this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

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The Development of Diagnosis for Atopic Dermatitis by Evaluating the Expression of Skin Barrier Proteins Using a Non-Invasive Method (비침습적 피부장벽단백질 측정을 통한 아토피 피부염의 진단 및 유용성)

  • Kim, In Sik;Lee, Ji-Sook
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.395-400
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    • 2017
  • Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.

Distribution of Antimicrobial Resistant Genes in Acinetobacter calcoaceticus-baumannii Complex Isolated from Clinical Specimens in Chungcheong, Korea (충청지역의 임상검체로부터 분리된 Acinetobacter calcoaceticus-baumannii Complex를 대상으로 항균제 내성 유전자 비교분석)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.4
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    • pp.427-434
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    • 2017
  • Species that belong to the Acinetobacter calcoaceticus-baumannii (Acb) complex are major causes of hospital-acquired infections. They are important opportunistic pathogens. These species are usually multidrug resistant (MDR), and the therapeutic options to treat the infections caused by these species are limited. In the present study, we investigated fluoroquinolone resistance mechanisms in 53 ciprofloxacin resistant Acinetobacter species isolates in Chungcheong, Korea. Antimicrobial susceptibilities were determined using the disk-diffusion method. Detections of genes and identification of mutations associated with fluoroquinolone resistance were carried out using PCR and DNA sequencing. In our study, 47 out of 53 ciprofloxacin resistant Acinetobacter isolates harbored sense mutations at the 83rd residue (serine to leucine) in the gyrA gene as well as at the 80th residue (serine to leucine) in the parC gene. Among the 47 isolates harboring sense mutations in gyrA and parC gene, 44 isolates were A. baumannii and 3 isolates were A. pittii. Plasmid-mediated quinolone resistance (PMQR) determinants were detected in isolates in our study. Among the 46 ciprofloxacin resistant A. baumannii isolates, 41 showed type A, B, or F banding patterns on their REP-PCR profiles. This result suggests that clonal relation and horizontal spreading of the bacterial isolates have been around hospitals in Chungcheong area. To prevent colonization and disseminations of fluoroquinolone resistance Acb complex isolates, continuous investigation and monitoring of antimicrobial resistant determinants of MDR isolates are needed.

Changes of Plasma and Urinary $TNF-{\alpha}$ in Children with Minimal Change Nephrotic Syndrome and Its Role in Albumin Permeability (미세변화신증후군 환아에서 Tumor Necrosis Factor-${\alpha}$의 혈중 및 요중 변화와 알부민 투과성에 미치는 영향)

  • Cho Min-Hyun;Lee Hwan-Seok;Oh Hyun-Hee;Chung Ki-Young;Koo Ja-Hoon;Ko Cheol-Woo
    • Childhood Kidney Diseases
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    • v.7 no.1
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    • pp.16-22
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    • 2003
  • Purpose : Minimal Change Disease(MCD) is the most common primary nephrotic syndrome in children. Some suggested that tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$) are involved in the pathogenesis of MCD. This study was done to see the changes of plasma and urinary $TNF-{\alpha}$, and their effects on the permeability of glomerular basement membrane. Methods : Study patients consisted of 19 biopsy-proven MCD children aged 2-15 years old. Both plasma and urinary $TNF-{\alpha}$ were measured. Employing the Millicell system, $TNF-{\alpha}$ were screened for the permeability factors. Results : Urinary $TNF-{\alpha}$ during relapse was significantly increased(P<0.01). No significant change was seen in the plasma $TNF-{\alpha}$ during relapse when compared to those in remission and the healthy controls. Furthermore, in the in vitro Millicell system, $TNF-{\alpha}$ did not produce a significant change in albumin permeability. Conclusion : Therefore, it seems that $TNF-{\alpha}$ may not play a disease-specific role in the pathogenesis of MCD.

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Causes of Bacterial Growth in Gels and Gel Containers Used for Ultrasonography (초음파 검사용 젤과 젤 용기의 세균증식 원인)

  • Kim, A-Young;Cho, Pyoung-Kon;Song, Do-Young;Kim, Su-Jung
    • Journal of radiological science and technology
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    • v.43 no.5
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    • pp.359-365
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    • 2020
  • This study aimed to investigate the causes of bacterial growth to prevent infection caused by ultrasound gel and gel containers in contact with patients during ultrasonography. To investigate bacterial contamination during manufacturing or storage, we cultured ultrasound gels originally supplied from three manufacturers. To analyze bacterial growth according to the lapse of time and frequency of use of the ultrasound gel container, the gel and container were cultured at regular intervals every week for 4 weeks. In addition, to determine the source of infection, the examiner's hand was inspected with hand plate and the degree of bacterial contamination was measured before the test. As a result of the study, bacteria were not detected in the gel provided at the initial supply, and in the gel and gel container used repeatedly for 4 weeks, the same bacteria residing on the skin were identified in the examiner's hand, such as Staphylococcus epidermidis, Micrococcus luteus, Leuconostoc mesenteroid spp cremoris, Kocuria rhizophila, and etc. Separated strains were classified as those of the low- or non-pathogenicity; however, most of these strains may render fatal consequences to patients of lower level of immunity due to acquired tolerance to antibiotics. At week 1, when the number of tests was the highest, 44 colonies were identified, and at week 4, when the number of tests was the lowest, 4 colonies were identified. As r=0.994, it was found that the number of colonies increased as the number of tests increased. In conclusion, it was confirmed that the cause of the infection was not the ultrasound gel, but the examiner's hand. The ultrasound gel or gel container may be contaminated by skin flora of examiner's hands, which can cause opportunistic infection in patients with low immunity. The ultrasound gel or gel container may be contaminated by skin flora of examiner's hands, which can cause opportunistic infection in patients with low immunity. Therefore, it was confirmed that thorough hand disinfection was necessary to block healthcare-associated infections.

A Retrospective Sectional Study about the Effect of the Interaction of Herbal Medicines and Warfarin on Prothrombin Time(INR) in Stroke Patients (뇌경색 입원환자대상으로 한약과 Warfarin 의 복합 투여시 미치는 Prothrombin Time (INR) 의 변화에 대한 후향적 단면연구)

  • Lee, Sang-Hun;Kim, Young-Seok;Kang, Chul-Ho;Song, Mun-Gu;Doo, Ho-Kyung;Ahn, Se-Young;Ahn, Young-Min;Lee, Byung-Cheol
    • The Journal of Internal Korean Medicine
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    • v.28 no.3
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    • pp.464-472
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    • 2007
  • Objectives : Nowadays the combined use of oriental herbal medicines and western biomedical medicines has been prevalent but controversial. Warfarin has been much reported to interact with some herbal medicines so that it influences prothrombin time(PT) & international normalized ratio(INR). This study was aimed to examine how much warfarin interacts with herbal medicines during treatment of stroke patients Methods : This was a retrospective case control study of 53 patients whowere treated with concomitant treatment of herbal medicines & warfarin. They were within normal limit in liver function, renal function, hematocrit, hemoglobin, and platelet count at first admission lab. We classified them into 2 classes: study group (taking herbal medicines including Panax ginseng, Angelica sinensis, Zingiber officinale, Salvia miltiorrhiza that were reported to interact with warfarin to impact PT (INR) and control group (taking other herbal medicines). We followed up PT (INR) at 5-10 days interval with AST, ALT, BUN, creatinine, hematocrit, hemoglobin, and platelet count. Results : AST, BUN, creatinine, hemoglobin, hematocrit, and platelet count were not changed significantly between first and final tests during the admission period. Only ALT decreased significantly in the control group. Neither baseline nor peak PT (INR) was significantly different between the groups. However, only warfarin dose was significantly correlated with PT and INR (r=0.810, r=0.798, p<0.01). Conclusions : It was concluded that PT(INR) was not influenced with herbal medicines and warfarin but by far dependent on warfarin dose in stroke patients restricted with normal liver function, renal function, and hematocrit, hemoglobin, and platelet count. Further prospective study is needed on larger samples to conclude that the combined therapy of herbal medicines and warfarin is safe.

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