Herein, we report a case in which cholestasis caused discrepancy in high-density lipoprotein (HDL)-cholesterol levels measured using various methods. The discrepancy in HDL-cholesterol level originated from the abnormal increase in the level of an unusual lipoprotein, apo E-rich HDL, in the patient's serum. An abnormal slow alpha-migrating lipoprotein was observed on agarose gel electrophoresis, and an abnormal large-sized HDL was observed in a lipoprotein subfraction study. The level of apolipoprotein E was elevated.
Apolipoproteins ; Cholestasis ; Electrophoresis, Agar Gel ; Lipoproteins

Herein, we report a case in which cholestasis caused discrepancy in high-density lipoprotein (HDL)-cholesterol levels measured using various methods. The discrepancy in HDL-cholesterol level originated from the abnormal increase in the level of an unusual lipoprotein, apo E-rich HDL, in the patient's serum. An abnormal slow alpha-migrating lipoprotein was observed on agarose gel electrophoresis, and an abnormal large-sized HDL was observed in a lipoprotein subfraction study. The level of apolipoprotein E was elevated.
Apolipoproteins ; Cholestasis ; Electrophoresis, Agar Gel ; Lipoproteins

BACKGROUND: Recently biphasic agarose gel electrophoresis method using Chol/Trig ComboTM for simultaneous detection of cholesterol and triglyceride on lipoprotein fractions has been developed to facilitate the classification and interpretation of abnormal lipoprotein patterns of patients with hyperlipidemia. The author analysed serum lipid profiles using Chol/Trig ComboTM in the patients of OPD in Department of Internal Medicine. METHODS: Measurement of serum cholesterol and triglyceride using enzymatic method was performed in the sera of 415 patients from April, 2006 to July, 2006. Simultaneously, we electrophoresed serum cholesterol and triglyceride using Chol/Trig ComboTM with analysis software (ED BANK, Helena Laboratories, Saitama, Japan). RESULTS: According to ATP III guideline, we set up standard cholesterol as 200 mg/dL and triglyceride as 150 mg/dL, respectively, and the patients were classified into control, hypercholesterolemic, hypertriglyceridemic and hypercholesterolemic/hypertriglyceridemic groups, respectively. Hypercholesterolemic group had higher HDL and LDL, hypertriglyceridemic group had higher HDL, VLDL and the rates of positive modified LDL, hypercholesterolemic/hypertriglyceridemic group have higher VLDL and the rates of positive modified LDL than the control. Otherwise, the rates of positive modified LDL were 19.8% in the control, 23.6% in the hypercholesterolemic group, 50.9% in hypertriglyceridemic group, and 39.3% in hypercholesterolemic/hypertriglyceridemic group. CONCLUSIONS: The frequency of the rates of positive modified LDL and VLDL was higher in the hypertriglyceridemic group than those in the control.
Adenosine Triphosphate ; Cholesterol ; Electrophoresis ; Electrophoresis, Agar Gel ; Humans ; Hyperlipidemias ; Lipoproteins

BACKGROUND: Recently biphasic agarose gel electrophoresis method using Chol/Trig ComboTM for simultaneous detection of cholesterol and triglyceride on lipoprotein fractions has been developed to facilitate the classification and interpretation of abnormal lipoprotein patterns of patients with hyperlipidemia. The author analysed serum lipid profiles using Chol/Trig ComboTM in the patients of OPD in Department of Internal Medicine. METHODS: Measurement of serum cholesterol and triglyceride using enzymatic method was performed in the sera of 415 patients from April, 2006 to July, 2006. Simultaneously, we electrophoresed serum cholesterol and triglyceride using Chol/Trig ComboTM with analysis software (ED BANK, Helena Laboratories, Saitama, Japan). RESULTS: According to ATP III guideline, we set up standard cholesterol as 200 mg/dL and triglyceride as 150 mg/dL, respectively, and the patients were classified into control, hypercholesterolemic, hypertriglyceridemic and hypercholesterolemic/hypertriglyceridemic groups, respectively. Hypercholesterolemic group had higher HDL and LDL, hypertriglyceridemic group had higher HDL, VLDL and the rates of positive modified LDL, hypercholesterolemic/hypertriglyceridemic group have higher VLDL and the rates of positive modified LDL than the control. Otherwise, the rates of positive modified LDL were 19.8% in the control, 23.6% in the hypercholesterolemic group, 50.9% in hypertriglyceridemic group, and 39.3% in hypercholesterolemic/hypertriglyceridemic group. CONCLUSIONS: The frequency of the rates of positive modified LDL and VLDL was higher in the hypertriglyceridemic group than those in the control.
Adenosine Triphosphate ; Cholesterol ; Electrophoresis ; Electrophoresis, Agar Gel ; Humans ; Hyperlipidemias ; Lipoproteins

BACKGROUND: Polymerase chain reacation (PCR)-based methods have been described for rapid detection and identification of Candida spp. Multiplex PCR assay was developed using internal transcribed spacers and topoisomerase II gene for the accurate identification of Candida species. METHODS: We designed Dual Specificity Oligo (DSO) primers for multiplex PCR. Multiplex PCR was followed by agarose gel electrophoresis to test 8 type strains (C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis) and 96 clinical isolates (C. albicans 51 isolates, C. parapsilosis 10 isolates, C. glabrata 10 isolates, C. tropicalis 9 isolates, C. krusei 6 isolates, C. guilliermondii 5 isolates, C. lusitaniae 5 isolates) of Candida spp. RESULTS: With multiplex PCR using DSO primers, the eight Candida type strains each could be easily differentiated and all 96 clinical isolates were identified as the same species as were identified by the conventional method. CONCLUSION: Multiplex PCR followed by electrophoresis can be useful for the simple and rapid identification of Candida species in routine laboratories.
Candida* ; DNA Topoisomerases, Type II ; Electrophoresis ; Electrophoresis, Agar Gel ; Multiplex Polymerase Chain Reaction* ; Sensitivity and Specificity

OBJECTIVETo analyse clinical feasibility of two electrophoresis procedures of seminal plasma proteins, agarose gel electrophoresis and SDS-agarose gel electrophoresis.

METHODSSixty-nine semen samples were examined and classified into three groups: the asthenozoospermia (n = 22), the asthenoteratozoospermia (n = 19), and the relative normal group (n = 28) with normal routine and special test results, according to WHO routine and special test criterion. Then, the seminal plasma protenis were separated by two different electrophoresis, with SDS-agarose and agarose support medium, the buffer pH 7.0 and 9.2 respectively. The agarose gel electrophoresis was done under various sample loading time, motion power and staining modules. The completed gels were scanned and compared the each other statistically.

RESULTSSeminal plasma proteins can be separated into 4 strips by SDS-agarose gel electrophoresis with acid crystal violet, and the strips were diffusion and with dark background. However, 6 clear strips named A, B, C, D, E, and F can be obtained by agarose gel electrophoresis with 6 min. After samples were loaded and stained by amidoblack, there showed appropriate spaces among strips, and it was very easy to scan the drying gel by a densitometer. Using agarose gel electrophoresis, the statistical difference in strip C and E was shown between the asthenozoospermia and the relative normal group, and between the asthenozoospermia and the asthenoteratozoospermia, however, not between the relative normal and the asthenoteratozoospermia group. Moreover, the samples in the relative normal group with normal routine and special test results were in different electrophoresis maps.

CONCLUSIONAgarose gel electrophoresis of seminal plasma proteins with buffer pH 9.2, 6 min. sample loading and amidoblack stain was a simple, fast and fit technique for clinic.

Adult ; Electrophoresis, Agar Gel ; methods ; Electrophoresis, Polyacrylamide Gel ; methods ; Humans ; Male ; Proteins ; analysis ; Semen ; chemistry ; Staining and Labeling

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) has a heteroresistant nature, so methicillin resistance is influenced by various culture conditions, such as temperature, incubation time, and NaCl content in the medium. Mueller Hinton (MH) agar containing 2% NaCl and mannitol salt agar (MSA) with oxacillin disk were evaluated for the detection of methicillin resistance. METHODS: Disk diffusion test on plain Mueller- Hinton (MH) agar, 2% NaCl MH agar, and MSA with 1 microgram oxacillin disk was performed in 70 Stap hylococcus aureus isolates. Oxacillin MIC was determined by E-test. As a gold standard of methicillin resistance, mecA gene was amplified by PCR and detected by agarose gel electrophoresis. RESULTS: Plain MH agar could not detect heterogeneous resistance in 12 S. aureus isolates (18%), but 2% NaCl MH agar and MSA could correctly detect homogeneous and heterogeneous resistance. S. aureus isolates from stool have as much as 48% heterogeneous resistance, while those from non-stool specimen have 5%. CONCLUSION: 2% NaCl and MSA can be used reliably for accurate susceptibility testing of methicillin resistance in routine laboratory.
Agar* ; Diffusion ; Electrophoresis, Agar Gel ; Mannitol* ; Methicillin Resistance* ; Methicillin* ; Methicillin-Resistant Staphylococcus aureus ; Oxacillin ; Polymerase Chain Reaction ; Staphylococcus aureus* ; Staphylococcus*

The principal purpose of this study is to set up efficient purification techniques of small DNAs which are suitable for isolation of from tens to three hundred bases of genes. On the bases of the technique, purification methods for big DNA fragments are established. In the experiment, the DNA bands were cut after agarose gel electrophoresis and put into 0.5 mL of tubes with silica wool, glass wood, absorbent cotton and cotton at the bottom. And then 10 000 r/min for 2 min, the liquid was collected. The results indicated that silica wool was the best of the materials. The recovery rate for DNAs below 200bp was over 90%, 85% to approximately 90% for 300bp. And the technique can be applied to purify bigger DNA fragments. The kits for DNA purification hardly recovered DNA below 150bp. The recovery rate for 150bp of DNA was 5%, 60% even for 300bp. The efficiencies of enzymic digestion and enzymic connection for the DNAs purified by the technique were the same as those for the DNAs isolated by the kits. So, the technique is obviously superior to kit purification methods.
DNA ; genetics ; isolation & purification ; Electrophoresis, Agar Gel ; methods ; Silicon Dioxide ; chemistry

BACKGROUND: The recently developed nucleic acid amplification methods may provide us with very sensitive, specific and rapid tests for the detection of M. tuberculosis. So the aim of this study was to compare the commercial Amplicor M. tuberculosis kit and our in-house polymerase chain reaction (PCR) with the conventional culture and direct AFB staining method. Materials and METHODS: Among the total of 2,340 clinical specimens, 1,314 sputum samples were tested for the presence of M. tuberculosis by Amplicor PCR and 1,026 sputum samples were tested by in-house PCR performing with resin matrix preparation and DNA extraction, synthesized primer pair, detection using agarose gel electrophoresis. RESULTS: One hundred-seventeen specimens were positive by Amplicor PCR, 105 were positive by in-house PCR, 185 were positive by culture. The sensitivity of the Amplicor PCR for all of the specimens and for smear-positive and smear-negative specimens was 92.9%, 97.9% and 88.2%, respectively after discrepant analysis. The sensitivity of the in-house PCR for all of the specimens and for smear-positive and smear-negative specimens was 80.0%, 93.6% and 65.5%, respectively after discrepant analysis. The specificity of the Amplicor PCR and in-house PCR for all of the specimens was 97.9% and 99.0%, respectively. CONCLUSIONS: Amplicor PCR was more sensitive than in-house PCR, but there was another problems such as high false positive rate and high cost. So PCR may certainly become very useful in microbiological laboratories if PCR method is selected according to the laboratory conditions.
DNA ; Electrophoresis, Agar Gel ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymerase Chain Reaction* ; Sensitivity and Specificity ; Sputum* ; Tuberculosis

PURPOSE: We compared the extent of prostatic involution after treatment with finasteride, a potent 5alpha-reductase inhibitor, with those caused by castration. MATERIALS AND METHODS: Rats were divided into two groups: finasteride-treated and castrated ones. Finasteride-treated rats were given 1mg/kg a day orally. The rats were sacrificed immediately, and on days 1, 3, 5, 7, 10, 14 and 21 after treatment. The prostate was immediately removed, weighed and either prepared for histological examination detecting apoptotic bodies by hematoxylin and rosin staining or frozen in liquid nitrogen for the determination of intraprostatic DHT concentration and DNA content, for detection of 180-bp DNA ladder by agarose gel electrophoresis. RESULTS: Either finasteride or castration decreased prostate weight and DNA content, which the degree of decrease was more pronounced in castrated group than in finasteride-treated group. Both finasteride and castration caused a decrease in intraprostatic DHT concentration, which these processes occurred to a more rapid and greater extent in finasteride-treated group. By 3 days of finasteride treatment, the intraprostatic DHT concentration decreased to a greater extent with no further significant change thereafter, while castration decreased gradually in intraprostatic DHT concentration up to day 10, with no further significant decrease thereafter. In the pattern of cell loss determined by hematoxylin and eosin staining, apoptotlc bodies were observed typically in castration group but not in finasteride-treated group. In agarose gel electrophoresis, 180-bp DNA ladder was found in finasteride-treated group through experimental days but in castrated after 3 days of experiment this ladder was not found. CONCLUSIONS: We concluded that castration caused a more profound involution of rat ventral prostate than finasteride. But the extent of prostatic involution was not correlated with intraprostatic DHT concentration. Although we could not found the apoptotic body in finasteride-treated group, the 180-bp DNA ladder suggesting apoptosis was detected.
Animals ; Apoptosis ; Castration* ; DNA ; Electrophoresis, Agar Gel ; Eosine Yellowish-(YS) ; Finasteride* ; Hematoxylin ; Nitrogen ; Prostate* ; Rats*