The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of micromethodologies and increasing availability of immunological and molecular biological reagents. The technological advances led to consider the diagnostic use of saliva for an array of analytes and DNA source. The purpose of the present study was to compare DNA from saliva with those from blood and buccal swab, to evaluate diagnostic and forensic application of saliva, to investigate the changes of genomic DNA in saliva according to the storage temperature and period of saliva samples, and to evaluate the integrity of the DNA from saliva stored under various storage conditions by PCR analysis. Peripheral venous blood, unstimulated whole saliva, stimulated whole saliva, and buccal swab were obtained from healthy 10 subjects (mean age: $29.9{\pm}9.8$ years) and genomic DNA was extracted using commercial kit. For the study of effects of various storage conditions on genomic DNA from saliva, stimulated whole saliva were obtained from healthy 20 subjects (mean age: $32.3{\pm}6.6$ years). After making aliquots from fresh saliva, they were stored at room temperature, $4^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$. Saliva samples after lyophilization and dry-out procedure were stored at room temperature. After 1, 3, and 5 months, the same experiment was performed to investigate the changes in genomic DNA in saliva samples. In case of saliva aliquots stored at room temperature and dry-out samples, the results in 2 weeks were also included. Integrity of DNA from saliva stored under various storage conditions was also evaluated by PCR amplification analysis of $\beta$-globin gene fragments (989-bp). The results were as follows: 1. Concentration of genomic DNA extracted from saliva was lower than that from blood (p<0.05), but there were no significant differences among various types of saliva samples. Purities of genomic DNA extracted from stimulated whole saliva and lyophilized one were significantly higher than that from blood (p<0.05). Purity of genomic DNA extracted from buccal swab was lower than those from various types of saliva samples (p<0.05). 2. Concentration of genomic DNA from saliva stored at room temperature showed gradual reduction after 1 month, and decreased significantly in 3 and 5 months (p<0.05, p<0.01, respectively). Purities of DNA from saliva stored for 3 and 5 months showed significant differences with those of fresh saliva and stored saliva for 1 month (p<0.05). 3. In the case of saliva stored at $4^{\circ}C$ and $-20^{\circ}C$, there were no significant changes of concentration of genomic DNA in 3 months. Concentration of DNA decreased significantly in 5 months (p<0.05). 4. There were no significant differences of concentration of genomic DNA from saliva stored at $-70^{\circ}C$ and from lyophilized one according to storage period. Concentration of DNA showed decreasing tendency in 5 months. 5. Concentration of genomic DNA immediately extracted from saliva dried on Petri dish were 60% compared with that of fresh saliva. Concentration of DNA from saliva stored at room temperature after dry-out showed rapid reduction within 2 weeks (p<0.05). 6. Amplification of $\beta$-globin gene using PCR was successful in all lyophilized saliva stored for 5 months. At the time of 1 month, $\beta$-globin gene was successfully amplified in all saliva samples stored at $-20^{\circ}C$ and $-70^{\circ}C$, and in some saliva samples stored at $4^{\circ}C$. $\beta$-globin gene was failed to amplify in saliva stored at room temperature and dry-out saliva.
Purpose: Acute febrile illness in children frequently accompanies with abdominal symptoms such as vomiting, diarrhea, and abdominal pain, even if its etiology is not occured from the gastrointestinal tract. If the etiology of fever was unknown and the fever was accompanied with abdominal symptoms, we should be concerned about whether the etiology of fever was originated from the gastrointestinal tract or interpretated from the abnormality in the gastrointestinal tract. This study was performed to evaluate the diagnostic value of abdominal ultrasonography in patients with fever and abdominal symptoms. Methods: We reviewed retrospectively the medical records of abdominal ultrasonographic (US) findings of 60 cases of acute febrile illness with abdominal symptoms at department of Pediatrics, Ulsan Dongang General Hospital during the period from January 1994 to June 1995. Results: The abnormal abdominal US findings obtained were as follows. 1) The abnormal US findings were seen in 56 cases (93.3%). 2) The most common abnormal US finding was the enlarged mesenteric lymph nodes in 52 cases (86.7%) 3) The enlarged mesenteric lymph nodes with or without intra-abdominal fluid were seen in 30 cases (50.0%) of a wide variety of illnesses, so their diagnostic values were absent. 4) The enlarged mesenteric lymph nodes and splenomegaly with or without ileocecitis were seen in 7 cases. Among them, 4 cases (6.7%) were confirmed as typhoid fever. 5) The US findings in 6 cases of typhoid fever were the enlarged mesenteric lymph nodes in all cases (100%), splenomegaly in 4 cases (66.7%), ileocecitis in 1 case (16.7%), enlarged mesenteric lymph nodes and splenomegaly in 4 cases (66.7%), enlarged mesenteric lymph nodes, splenomegaly and ileocecitis in 1 cases (16.7%). 6) The enlarged mesenteric lymph nodes and the abnormalities around the appendix were seen in 7 cases (11.7%), which were confirmed as appendicitis all. 7) The thickening of wall in urinary bladder was seen in 2 cases (3.3%) of acute cystitis and acute hemorrhagic cystitis. 8) The subtle thickening of wall in colon was seen in 1 cases (1.7%) of shigellosis. Conclusions: The enlarged mesenteric lymph nodes, splenomegaly and ileocolitis on the abdominal ultrasonography in patients with fever and abdominal symptoms aree suggestive findings of typhoid fever. The enlarged mesenteric lymph nodes and the abnormalities around the appendix on abdominal ultrasonography make the rapid diagnosis of acute appendicitis and its complications, when physical examination is difficult in small children and diagnosis of their illnesses is obscure in patients with fever and abdominal pain.
Shin, Kook-Sik;Lim, Woo-Jin;Lee, Sang Eun;Lee, Jae Seon;Cha, Geun Sig
Korean Journal of Soil Science and Fertilizer
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v.42
no.6
/
pp.513-521
/
2009
The primary goal of this research was to develop an optimized analytical procedure for soil analysis based on ion-selective microelectrodes for agricultural purposes, which can perform on-site measurement of various ions in soil easily and rapidly. For the simple and rapid on-site diagnosis, an analysis of soil chemicals was performed employing a multicomponent-in-situ-extractant and an evaluation of ionselective microelectrodes were conducted through the regressive correlation method with a standard analytical approach widely employed in this area. Examination of sensor responses between various soil nutrient extractants revealed that 0.01M HCl and 1M LiCl provided the most ideal Nernstian response. However, 1M LiCl deteriorated the selective response for analytes due to high concentration (1M) of lithium cation. Thus, employing either 0.1M HCl as an extractant followed by 10 times dilution, or 0.01M HCl as an extractant without further dilution was chosen as the optimal extractant composition. A study of regressive correlation between results from ion-selective microelectrodes and those from the standard analytical procedure showed that analyses of $K^+$, $Na^+$, $Ca^{2+}$, and $NO_3{^-}$ showed the excellent consistency between two methods. However, the response for $NH_4{^+}$ suffered the severe interference from $K^+$. In addition, the selectivity for $Mg^{2+}$ over $Ca^{2+}$ was not sufficient enough since available ionophores developed so far do not provide such a high selectivity for $Mg^{2+}$. Therefore, as an agricultural on-site diagnostic instrument, the device in development requires further research on $NH_4{^+}$ analysis in the soil sample, development of $Mg^{2+}$-selective ionophore, and more detailed study focused on potassium, one of the most important plant nutrients.
Purpose: The Helicobacter pylori stool antigen (HpSA) enzyme immunoassay is a non-invasive test for the diagnosis and monitoring of H. pylori infection. But, there are few validation studies on the HpSA test after eradication in children. The aim of this study was to assess the diagnostic accuracy of HpSA enzyme immunoassay for the detection of H. pylori to confirm eradication in children. Methods: From January 2001 to October 2003, 164 tests were performed in 146 children aged 1 to 17.5 years (mean $9.3{\pm}4.3$ years). H. pylori infection was confirmed by endoscopy-based tests (rapid urease test, histology, and culture). All H. pylori infected children were treated with quadruple regimens (Omeprazole, amoxicillin, metronidazole and bismuth subcitrate for 7 days). Stool specimens were collected from all patients for the HpSA enzyme immunoassay (Primier platinum HpSA). The results of HpSA tests were interpreted as positive for $OD{\geq}0.160$, unresolved for $$0.140{\leq_-}OD$$<0.160, and negative for OD<0.140 at 450 nm on spectrophotometer. Results: 1) One hundred thirty-one HpSA tests were performed before treatment. The result of HpSA enzyme immunoassay showed three false positive cases and one false negative case. The sensitivity, specificity, positive predictive value, and negative predictive value of HpSA enzyme immunoassay before treatment were 96.4%, 97.1%, 90%, and 99%, respectively. 2) Thirty-three HpSA enzyme immunoassay were performed at least 4 weeks after eradication therapy. The results of HpSA enzyme immunoassay showed two false positive cases and one false negative case. The sensitivity, specificity, positive predictive value, and negative predictive value after treatment were 88.9%, 91.7%, 80%, and 95.7%, respectively. Conclusion: Diagnostic accuracy of the HpSA enzyme immunoassay after eradication therapy was as high as that of the HpSA test before eradication therapy. The HpSA enzyme immunoassay was found to be a useful non-invasive method to confirm H. pylori eradication in children.
The Journal of the Korean bone and joint tumor society
/
v.20
no.1
/
pp.14-21
/
2014
Purpose: To analyze the clinical features and treatment outcome of Langerhans' cell histocytosis. Materials and Methods: From August 1996 to June 2013, 28 patients who histologically proven with LCH were analyzed of medical records, radiography, pathologic character retrospectively. Results: A total of 28 cases of LCH including 22 child has been reported. Onset age was 0.6 to 51 years old, occurred in the average age was 14.8 years. Follow-up period was 6 months to 134 months average was 44.6 months. The M:F ratio was 2.5:1. The initial symptoms was pain in 18 cases, 5 cases of pathologic fracture, 3 case of palpable mass, 1 case of discovered by accident in radiography, 1 case of torticollis. In radiological examination osteolysis was seen all cases, 7 cases showed a periosteal reaction, 1 case showed soft tissue extension. Clinical type of all cases were eosinophilic granuloma. 25 cases were classified as unifocal disease and 3 cases were multifocal single systemic diseases. In all cases, incisional biopsy was performed. After histologic confirmed, 14 cases was treated with curettage or surgical excision of the lesion and the other 14 cases were followed up without treatment. There is no death during follow up period. 11 cases has no radiological improvement after 3-6 months observation, intralesional steroid injection was performed. Conclusion: Patients with LCH who has rapid systemic onset is very rare, so if you meet the young children who suspected LCH, you shoulder avoid the examination which cause excessive radiation exposure to the young patient. In order to confirm the diagnosis of disease, biopsy is needed. Close observation after confirmed by histological method will bring the satisfactory results. But the patients who had pathologic fracture or wide bone destruction already may need curettage and bone grafting to lesion or internal fixation. The lesion which has no radiological improvement after 3-6 months observation or appear with pain interferes daily life may need local steroid injection as a good treatment.
Purpose: The C-14 urea breath test (C-14 UBT) is the most specific noninvasive method to detect Helicobacter (H) pylori infection. We investigated if the C-14 UBT can reflect the presence and degree of H. pylori detected by gastroduodenoscopic biopsies (GBx). Materials and methods: One hundred fifty patients (M:F=83:67, age $48.6{\pm}11.2$ yrs) underwent C-14 UBT, rapid urease test (CLO test) and GBx on the same day. For the C-14 UBT, a single breath sample was collected at 10 minutes after ingestion of C-14 urea (137 KBq) capsule and counting was done in a liquid scintillation counter for 1 minute, and the results were classified as positive (${\geq}200dpm$), Intermediate ($50{\sim}199dpm$) or negative (<50 dpm). The results of CLO tests were classified as positive or negative according to color change. The results of GBx on giemsa stain were graded 0 (normal) to 4 (diffuse) according to the distribution of H. pylori by the Wyatt method. We compared C-14 UBT results with GBx grade as a gold standard. Results: In the assessment of the presence of H. pylori infection, the C-14 UBT global performance yielded sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of 92.5%, 88.4%, 97.1%, 88.4% and 91.3%, respectively. However, the CLO test had sensitivity, specificity, PPV, NPV and accuracy of 83.2%, 81.4%, 91.8%, 81.4% and 82.7%, respectively. The quantitative values of the C-14 UBT were $45{\pm}27$ dpm in grade 0, $707{\pm}584dpm$ in grade 1, $1558{\pm}584dpm$ in grade 2, $1851{\pm}604dpm$ in grade 3, and $2719{\pm}892dpm$ in grade 4. A significant correlation (r=0.848, p<0.01) was found between C-14 UBT and the grade of distribution of H. pylori infection on GBx with giemsa stain. Conclusion: We conclude that the C-14 UBT is a highly accurate, simple and noninvasive method for the diagnosis of ongoing H. pylori infection and reflects the degree of bacterial distribution.
Cho Hee-Yeon;Chung Dae-Lim;Kang Ju-Hyung;Ha Il-Soo;Choi Yong;Cheong Hae-Il
Childhood Kidney Diseases
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v.8
no.2
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pp.176-185
/
2004
Purpose: Rapidly progressive glomerulonephritis (RPGN) is a clinicopathologic entity characterized by extensive crescent formation and rapid deterioration of renal function within few months. For better understanding of its clinical course and designing better treatment strategies, a clinicopathological study of childhood RPGN was performed. Methods: The clinical manifestations and pathological findings were reviewed retrospectively in 12 children who were diagnosed as having RPGN by clinical manifestations and renal biopsy during a period from 1991 to 2003. Several clinicopathological parameters were analyzed as prognostic factors. Results: Among a total of 12 patients, 4 were male and 8 were female. The median onset age was 11.5 years(range 5.5-14.6 years), and the median period of follow-up was 25 months(range 7 months-6.6 years). According to the pathological classification, 10 patients (83%) were type II RPGN(immune-complex mediated glomerulonephritis), 2 patients were type III RPGN(pauci-immune glomerulonephritis), and none was type I RPGN(anti-glomerular basement membrane nephritis). All patients were treated with oral steroid in various combinations with methylprednisolone pulse therapy(10 patients, 83%), cyclophosphamide(8 patients, 67%), or plasmapheresis(4 patients, 33%). Clinical outcomes of 12 patients were complete remission in 1(8%), end-stage renal disease in 2(17%), chronic renal insufficiency with persistent proteinuria in 2(17%), and normal renal function with persistent proteinuria in 7(58%) at the last follow-up. Poor prognosis is associated with increased serum creatinine level, severe anemia and younger age at the time of diagnosis. Conclusion: Immune-complex mediated glomerulonephritis is the major cause RPGN in children and most cases showed improvement of renal function with aggressive management. For better understanding of this rare disease, a prospective multicenter study should be done.
Purpose: Aortic Dissection is very dangerous, prognostic disease, which the bloodstream flow out of the true lumen of the aorta by the bursting of aortic intima resulting in a rapid dissociation of inner and outer layer from the media. It is difficult to diagnose aortic dissection clinically by normal X-ray. This study was to investigate the occurrence frequency by age and number of patients who are identified to be aortic dissection by CT (Computed Tomography) scan. Materials and methods: We investigated the trend of yearly fluctuation, gender, age, and department of clinical research of the 112 patients who conducted CT scan in C- University Hospital for two years from January 2005 to December 2006. The MIP and SSD which reconstructed CT image and the VRT image were obtained for the accurate observation. The result was investigated by comparing normal X-ray and CT scan. Results and Conclusion: 1. The yearly check of 112 patients conducted CT scan showed 37 people (41.9%) in 2005, and it was increased to 65 (58.1%) in 2006 by 1.4 times. 2. The gender distribution of patients given a CT scan showed 45 males (40.1%), and female 67 (59.9 %). The aortic dissection patients were 9 (20%) out of 45 males, 21 (31.3%) out of 67 females and women were 1.6 times more than men. Women are also 1.5 times more than men in the number of examinee. 3. The age distribution of patient's who conducted CT scan revealed that there was no patient under 30 years old while 88.3% of all patients were through 41 to 80 years old. The higher the age was, the higher the occurrence of aortic dissection was. The difference in the occurrence frequency of age was statistically significant (p<0.01). 4. The departments that requested CT scan were the emergency department 46 (41.1%), circulatory internal medicine 37 (33.0%), chest surgery 13 (11.6%), and others 6 (14.3%). The combined ratio of emergency medicine and circulatory internal medicine was 74.1% of all. The results show that the aortic dissection is a very dangerous disease whose patients visit mainly via the emergency room. 5. The aortic dissection patients had normal X-ray readings in 22 (73.3%) out of 30, and only 8 (26.7 percent) are abnormal in the X-ray diagnosis. Therefore, the CT scan needs to be enforced in order to assess accurately the disease of aortic dissection.
Jo, Eun-Kyeong;Song, Chang-Hwa;Park, Jeong-Kyu;Baek, Young-Jong;Rhu, Hye-Young;Lee, Jae-Ho;Hwang, Tai-Ju;Kook, Hoon
Clinical and Experimental Pediatrics
/
v.45
no.2
/
pp.183-191
/
2002
Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied the cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells from two siblings and one cousin with XLA, as well as additional family members. Methods : Btk protein expression was analyzed by flow cytometry. Isolation of the coding sequence of the Btk gene was performed by amplification using the reverse transcription-polymerase chain reaction(RT-PCR) technique. Sequence alterations were screened by the single-stranded conformation polymorphism(SSCP) method and characterized by standard sequencing protocols. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in three patients with XLA. In addition, Btk protein analysis clearly showed cellular mosaicism in monocytes from four obligate carriers, findings further supported by SSCP. A single base pair mutation(T to C) in Btk-exon three, which encodes the PH domain, was identified in four XLA patients. A diagnostic sequencing analysis was established to detect heterozygotic pattern in 4 carrier females. Furthermore, we found significant clinical heterogeneity in individuals with the same gene mutation. Conclusion : The implicating genetic alteration provided valuable clues to the pathogenesis of XLA in Korea and the flow cytometric analysis was suggested as a useful tool for rapid detection of XLA patients and carriers. The present study has identified a genetic mutation in the Btk coding region and demonstrated heterogeneity in clinical manifestations among patients with the same mutation. A flow cytometric analysis was found to be informative in establishing a deficiency of Btk protein in both patients and carriers and is recommended as a frontline procedure in the molecular diagnosis and work-up of XLA.
Purpose : To identify clinical characteristics of severe respiratory syncytial virus (RSV) in neonatal period and early infancy and provide information in clinical practice. Methods : Twelve neonates and young infants (<6 months) who were infected by respiratory syncytial virus and required mechanical ventilation between March 2005 and July 2007 were enrolled. Diagnosis of RSV infection was made based on the positive results by rapid antigen immunoassay or polymerase chain reaction. Results : There were four premature infants, of whom three were near-term. Birth weight of subject patients was $2.8{\pm}0.6kg$, gestational age was $37{\pm}2weeks$ and the age at the time of admission was $35{\pm}15days$. Nine of them showed apnea and in five patients, apnea itself was an indication for mechanical ventilation. In seven of the apneic patients, apnea was the first manifestation of RSV infection. In three of these seven apneic patients, apnea preceded definite respiratory distress signs or typical stethoscopic findings by 1-3 days. Mean duration of mechanical ventilation was $3{\pm}2days$, and mean duration of stay in intensive care unit was $6{\pm}2days$. Conclusion : RSV is a major cause of severe respiratory tract infection in term or near-term infant younger than 2 months. For apnea could be the first manifestation of the RSV infection, high level of suspicion is required in practice of neonates or young infants who show any upper respiratory infection symptoms during RSV season.
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