1.The use of an in-house biotin-avidin linked immunosorbent assay to detect Aspergillus antigens in sera of immunocompromised patients.
S Abdul Samad ; H Yusoff ; S A Fadilah
The Medical journal of Malaysia 2001;56(1):32-8
A biotin-avidin-linked immunosorbent assay was developed to detect Aspergillus antigens in sera of immunocompromised patients. The assay was based on a double antibody sandwich ELISA using polyclonal antibodies raised against water-soluble antigens of Aspergillus fumigatus. Aspergillus antigens were positive in sera of 9 of 16 (56%) patients who were studied prospectively and in 13 of 73 (19%) patients studied retrospectively. The 9 prospectively studied patients who were antigen positive were febrile neutropenic hematological malignancy patients who exhibited a high risk of acquiring invasive aspergillosis.
Antigens
;
assay
;
Aspergillus
;
Immunocompromised Host
;
Biotin
2.Urea Synthesis in the Intact and in the Isolated Perfused Liver of the Biotin-Deficient Rats.
Je Hyun KIM ; Moo Youn CHO ; Byung Woo KIM ; Chug Suk SONG
Yonsei Medical Journal 1971;12(1):13-16
Biotin-deficient rats were raised on a purified ration containing raw egg white plus avidin. Urea synthesis and excretion were compared between the biotin-deficient and the pair-fed control rats. 24hrurinary urea excretion and the specific activities of carbamylphosphate synthetase, ornithine transcarbamylase, and arginase in the liver mitochondria fraction were no different between these two groups. The net urea production in the liver slice and in the isolated perfused liver of the biotin-deficient rats was similar to that of the pair-fed control. Thus the conclusion must be that biotin is not in urea in mea biosynthesis in the rat.
Animal
;
Avitaminosis/metabolism
;
Biotin*
;
Liver/metabolism*
;
Rats
;
Urea/biosynthesis*
;
MH -
;
Substances:
;
Urea
;
Biotin
3.Apoptosis in Rat Thymus after Bolus Intramuscular Injection of 5-Fluorouracil.
Kyung Hee KIM ; Hae Joung SUL ; Dae Young KANG
Korean Journal of Pathology 2000;34(6):413-418
We induced apoptosis in normal rats by intramuscular injection of 5-fluorouracil (5-FU) and immunohistochemically evaluated the thymus for the TdT-mediated dUTP biotin nick end labelling on the 1st, 3rd, 6th, 9th, 15th and 21st days following the bolus intramuscular injection. The injections of 5-FU induced a greater extent of apoptosis in the thymus. In the thymus, a mild increase in apoptosis was observed 24 hours after injection. The greatest number of apoptotic cells were seen at 72 hours. The size of the thymus decreased and the cortex thinned with hypocellularity. The injection of 5-FU caused massive cell loss in the thymus. Most apoptotic cells were scattered in the cortex and lower levels of apoptosis were also observed in the medulla. After 72 hours, the level of apoptosis returned to the control level. Considering the above results, we think that 5-FU induced toxicity may be related to 5-FU induced apoptosis in normal tissue, especially the thymus.
Animals
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Apoptosis*
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Biotin
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Fluorouracil*
;
Injections, Intramuscular*
;
Rats*
;
Thymus Gland*
4.Comparison of two luminescence detection methods for staphylococcal enterotoxin C content in simulated milk samples.
Yuling ZHENG ; Ye WANG ; Qingyu LYU
Chinese Journal of Cellular and Molecular Immunology 2023;39(12):1089-1093
Objective To compare the sensitivity and accuracy of amplified luminescent proximity homogeneous assay linked immunosorbent assay (AlphaLISA) and magnetic particles-based chemiluminescence immunoassay (MP-CLIA) for detection of staphylococcal enterotoxin C (SEC) in the simulated milk samples. Methods The AlphaLISA was constructed using goat anti-SEC polyclonal antibody-coupled receptor microspheres, biotin-labeled SEC monoclonal antibody and streptavidin-coupled donor microspheres. The MP-CLIA was constructed using goat anti-SEC polyclonal antibody conjugated alkaline phosphatase, biotin-labeled anti-SEC monoclonal antibody and streptavidin conjugated magnetic beads. Results The sensitivity of AlphaLISA to detect SEC content in simulated milk samples was 4.04 ng/L, and the coefficient of variation (CV) was 1.98%~9.82%. The sensitivity of MP-CLIA was 108.19 ng/L and CV was 4.63%~20.40%. Conclusion Compared with MP-CLIA, AlphaLISA is more sensitive and accurate to detecting SEC.
Animals
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Streptavidin
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Biotin
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Luminescence
;
Milk
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Antibodies, Monoclonal
;
Goats
;
Immunoassay/methods*
5.An Immunohistochemical Stain in Paget's Disease.
Hong Tak LEE ; Seung Ho CHANG ; Tae Young YOON
Korean Journal of Dermatology 1998;36(5):796-803
BACKGROUND: Generally, there are few problems in the diagnosis of Paget's disease(PD) using the H&E stain. However, the differentiation of PD from the clonal type of Bowens disease and superficial spreading melanoma in situ that shows pagetoid spreading of tumor cells, may present diagnostic difficulties. In addition, the specia1 stains used for demonstrating the presenee of Pagets cells, such as PAS and mucicarmim, are non-specific and not always sensitive. So, inenunohistochemical stains with monoclonal antibodies against various antigens may be helpful for differentiating PD from ather morphologically similar skin lesions. OBJECTIVE: The purpose of this study was to investigate the diagnostic accuracy of immunohisto- chemical staining for diagnostic use in PD. METHODS: Immunohistochemical stains used in the biotin streptavidin amplificxl technique with monoclonal antibodies to several low rnolecular weight cytokeratin(CK)s, EMA and CEA, were performed on formalin-fixed, paraffin-embedded tissue. Twelve cases of PD(10 cases of extranmmmary PD and 2 cases of mammary PD), five cases of superficial spreading melanoma in situ and five cases of Bowens disease were investigated. RESULTS: The results were as follows. 1. Positive reactions with variable intensity using CK7, CKS, CK19 were seen in all cases(100%) of PD and the. staining intensity tor CK7 or CK19 was stronger than that of CKS. 2. Of the 12 cases of PD, both CK18 and CAM5.2 staining showed positivity in 11 cases(92%). 3. EMA and CEA staining showed positivity in 10(83%) and 9(75%) of 12 cases, respectively. 4. Some Pagets cells were negative for CK8, CK18 and EMA, although other positive cells were observed in the same sections. 5. All antigens were consistently negative in all cases of Bowens disease and superficial spreading melanoma in situ. CONCLUSION: The results show that moaoclonal antibodies to low molecular weight CKs are more sensitive than EMA or CEA in the demonstration of Pagets cells. Moreover, among the low molecular weight CK series, CK7 and CK19 are most useful for their high sensitivity and intensity.
Antibodies
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Antibodies, Monoclonal
;
Biotin
;
Bowen's Disease
;
Coloring Agents
;
Diagnosis
;
Melanoma
;
Molecular Weight
;
Skin
;
Streptavidin
6.Molecular Links between Caloric Restriction and Sir2/SIRT1 Activation.
Diabetes & Metabolism Journal 2014;38(5):321-329
Ageing is the most significant risk factor for a range of prevalent diseases, including cancer, cardiovascular disease, and diabetes. Accordingly, interventions are needed for delaying or preventing disorders associated with the ageing process, i.e., promotion of healthy ageing. Calorie restriction is the only nongenetic and the most robust approach to slow the process of ageing in evolutionarily divergent species, ranging from yeasts, worms, and flies to mammals. Although it has been known for more than 80 years that calorie restriction increases lifespan, a mechanistic understanding of this phenomenon remains elusive. Yeast silent information regulator 2 (Sir2), the founding member of the sirtuin family of protein deacetylases, and its mammalian homologue Sir2-like protein 1 (SIRT1), have been suggested to promote survival and longevity of organisms. SIRT1 exerts protective effects against a number of age-associated disorders. Caloric restriction increases both Sir2 and SIRT1 activity. This review focuses on the mechanistic insights between caloric restriction and Sir2/SIRT1 activation. A number of molecular links, including nicotinamide adenine dinucleotide, nicotinamide, biotin, and related metabolites, are suggested to be the most important conduits mediating caloric restriction-induced Sir2/SIRT1 activation and lifespan extension.
Biotin
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Caloric Restriction*
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Cardiovascular Diseases
;
Diptera
;
Humans
;
Longevity
;
Mammals
;
NAD
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Negotiating
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Niacinamide
;
Risk Factors
;
Sirtuins
;
Yeasts
7.Apoptotic Keratinocytes in Acrodermatitis Enteropathica.
Young Lip PARK ; Kyu Uang WHANG ; Sung Won BANG ; Young Keun KIM ; Hee Joon YU ; Sook Ja SON
Korean Journal of Dermatology 1996;34(5):753-756
BACKGROUND: The cause of acrodermatitis enteropathica(AE) is closely related to zinc deficiency. Zinc is a potent inhibitor of endonuclease. Acute rises in the apoptosis in lymphoid and myeloid cell lines during zinc deficiency has recently been reported. The method of terminal transferase mediated dUTP biotin nick end labeling(TUNEL) is used in situ labelling of apoptotic nuclei in routine tissue sections. OBJECTIVE: The purpose of this study is to clarify our hypothesis that apoptosis resulted from zinc deficiency might cause keratinocytes damages in AE. METHOD: We stained 6 AE biopsy specimen with TUNEL technique. RESULTS: In acroderrratitis enteropathica, apoptotic keratinocytes were shown in the entire epidermis as compared to normal, controlled skin, in which it was found only at the uppermost layer of this stratified epithelium. CONCLUSION: This result suggests that apoptosis resulting from zinc deficiency might play a role in keratinocyte death in AE.
Acrodermatitis*
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Apoptosis
;
Biopsy
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Biotin
;
Epidermis
;
Epithelium
;
In Situ Nick-End Labeling
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Keratinocytes*
;
Myeloid Cells
;
Skin
;
Transferases
;
Zinc
8.An Immunohistochemical Study of pRb, p16, p53 and p21 Protein Expression in Malignant Melanoma.
Sung Eun CHANG ; Hyun Soo KIM ; Il Jung PARK ; Jee Ho CHOI ; Kyung Jeh SUNG ; Kee Chan MOON ; Jai Kyoung KOH
Korean Journal of Dermatology 2001;39(3):274-279
BACKGROUND: The proteins regulating the cell growth cycle such as the products of rb, p53, p16 and p21 genes are recently believed to have a role in various tumorigenesis. OBJECTIVE: The purpose of this study was to investigate an expression pattern and a role of p53, pRb, p16, and p21 in the pathogenesis of non-metastatic malignant melanomas. METHODS: Immunohistochemical study of p53, pRb, p16, and p21 was done in 10 invasive non-metastatic malignant melanomas compared to 5 benign melanocytic nevi. Standard streptavidin- biotin peroxidase method using the monoclonal antibodies was used. RESULTS: pRb was positive in all malignant melanomas tested. p16 was positive in 100% of benign nevi and only 40% of the malignant melanomas showed loss of p16 protein. p53 was positive in only 30% of malignant melanomas. p21 expression was seen diffusely in 70% of the malignant melanomas. In benign nevi, significant reactivity of p53, pRb, and p21 was not shown. CONCLUSION: In non-metastatic malignant melanomas, rb gene is not likely to be deleted and the overexpression of pRb might from activation of pRb protein facilitating cell cycle progression. In non-metastatic malignant melanomas, neither loss of p16 nor p53 overexpression was not frequent as previous reports of metastatic malignant melanomas. p21 overexpression in malignant melanomas might be related to invasiveness of malignant melanoma.
Antibodies, Monoclonal
;
Biotin
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Carcinogenesis
;
Cell Cycle
;
Genes, Retinoblastoma
;
Melanoma*
;
Nevus
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Nevus, Pigmented
;
Peroxidase
9.A Case of Lennox-Gastaut Syndrome due to 3-Methylcrotonyl CoA Carboxylase Deficiency.
Yu Sok HAN ; Hoon Chul KANG ; Hong Jin LEE ; Heung Dong KIM
Journal of the Korean Child Neurology Society 2004;12(1):92-98
3-Methylcrotonyl-CoA carboxylase(MCC) is a biotin-dependent enzyme involved in the leucine metabolism. We describe a patient with MCC deficiency who manifested with Reye syndrome-like illness with status epilepticus, metabolic acidosis, hypoglycemia, hyperammonemia, elevated liver enzymes and neurologic impairments after a viral gastroenteritis and then suffered from Lennox-Gastaut syndrome. Urinary organic acid analysis revealed increased excretions of 3-hydroxyisovaleric acid and 3-methylcrotonylglycine. This patient was managed with a leucine restriction diet and supplementation of biotin and carnitine, which was not so effective. He suffered from neurologic sequelae such as Lennox-Gastaut syndrome, motor and cognitive impairements.
Acidosis
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Biotin
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Carnitine
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Diet
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Gastroenteritis
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Humans
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Hyperammonemia
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Hypoglycemia
;
Leucine
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Liver
;
Metabolism
;
Status Epilepticus
10.Immunohistochemical Staining in the Differential Diagnosis of Trichoblastoma and Basal cell Carcinoma.
Kwang Hyun CHO ; Kwang Ho HAN ; Sang Eun MOON ; Jai Il YOUN ; Hee Chul EUN ; Chul Woo KIM
Korean Journal of Dermatology 1999;37(10):1423-1429
BACKGROUND: Trichoblastoma is a benign skin tumor with follicular differentiation which sometimes is difficult to distinguish histologically from basal cell carcinoma (BCC). OBJECTIVE: Our purpose is to investigate the usefulness of the immunostains in the differential diagnosis of trichoblastoma and BCC. METHODS: The authors analyzed 11 trichoblastoma, 8 keratotic BCCs, 9 nodular BCCs, 6 borderline cases. The monoclonal antibodies to bcl-2, CD34, Ki-67, p53 were applied to formalin-fixed paraffin sections in all cases, using a labelled streptoavidin biotin peroxidase complex method. RESULTS: Most nodular BCCs demonstrated diffuse cytoplasm labelling for bcl-2. In contrast, 'peripheral' bcl-2 staining of trichoblastomas was noted in 9 of 11. Keratotic BCCs demonstrated mixed staining pattern for bcl-2. CD34 staining failed to discriminate between trichoblastomas and BCCs. In BCC, Ki-67 positive cells were mainly distributed in the perphery of tumors. CONCLUSION: Immunostain using monoclonal antibodies to bcl-2, CD34, Ki-67, p53 can not differentiate between trichoblastomas and BCCs. But it could be an adjunctive method to establish the definite diagnosis of trichoblastomas and BCCs.
Antibodies, Monoclonal
;
Biotin
;
Carcinoma, Basal Cell*
;
Cytoplasm
;
Diagnosis
;
Diagnosis, Differential*
;
Immunohistochemistry
;
Paraffin
;
Peroxidase
;
Skin