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

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

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 ; Apoptosis* ; Biotin ; Fluorouracil* ; Injections, Intramuscular* ; Rats* ; Thymus Gland*

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 ; Streptavidin ; Biotin ; Luminescence ; Milk ; Antibodies, Monoclonal ; Goats ; Immunoassay/methods*

PURPOSE: We investigated the prognostic impacts of p53, c-erbB-2, nm23 and Ki-67 expression in patients with stage II and IIIA gastric carcinoma who underwent curative (RO) resections. MATERIALS AND METHODS: 261 paraffin-embedded gastric carcinoma tissues (stage II, 121; stage IIIA, 135) were stained with the monoclonal antibodies, p53, c-erbB-2, nm23 and Ki-67 using the labelled streptovidin biotin method. The positivity was determined by two pathologists who were kept blind for the patients outcome. RESULTS: The overexpression was seen in 51.7% for p53, 11.9% for c-erbB-2, and 70.1% for nm23. The mean Ki-67 labelling index was 25.5+ 16.7. The rates of overexpression between the stage II and stage IIIA were not significantly different in all these molecules. Overexpression of p53 was more likely to be associated with old age and lymph node metastasis. Overexpression of c-erbB-2 was more likely to be associated with Borrmann type I, II and well-differentiated tumor. However, nm23 was more frequently expressed in patients with older age and well-differentiated tumor. In survival analysis, the overexpressions of p53 and Ki-67 were significantly associated with poor prognosis of the patients (p<0.01), but c-erbB-2 and nm23 were not related to the patients outcome. In a multivariate analysis of prognostic factors, only. the lymph node metastasis was an independent prognostic factor. CONCLUSION: Although the values did not reach statistical significance in a multivariate analysis, the overexpression of p53 and Ki-67 tended to have adverse effects an the prognosis of patients with gastric cancer.
Antibodies, Monoclonal ; Biotin ; Humans ; Lymph Nodes ; Multivariate Analysis ; Neoplasm Metastasis ; Prognosis ; Stomach Neoplasms

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

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 ; Antibodies, Monoclonal ; Biotin ; Bowen's Disease ; Coloring Agents ; Diagnosis ; Melanoma ; Molecular Weight ; Skin ; Streptavidin

To evaluate prognostic significance of p53 gene mutation and p53 protein overexpression, 83 cases of primary gastric carcinoma (stage 1b, II, and III) were analyzed. Immunohistochemical assay using labeled streptavidine biotin method and mouse Anti-P53 was done on paraffin embedded tissues. The overall 5-year survival rate was 54.2%. Depth of invasion, lymph node metastasis, and stage were the prognostic factors (p<0.05). p53 was positive in 33 cases(39.8%). There were no significant association between p53 expression and other parameters such as age, sex, tumor location, size, gross finding, histologic type, depth of invasion, lymph node metastasis, and stage. The 5-year survival rates of p53 positive and negative groups were 62% and 42.4% respectively (p=0.0657). Survival distribution for p53 adjusted for location and depth of invasion showed significant difference. Five-year survival rates were 55%, 25%, 0% in p53 positive groups and 65.4%, 65%, 28% in negative groups at the lower, middle, and upper portion of the stomach, and 53.9%, 36.9%, 0% in p53 positive and 77.3%, 53%, 0% in negative groups in t2, t3 and t4. In comparison of 5-year survival rates of p53 positive and negative groups in the same stage, p53 positive groups showed worse prognosis than the negative, but they were not significant statistically. There were 57.1%, 92.9% in stage 1b (p=0.0678), 50%, 61.1% in stage II, 42.9%, 42.9% in stage IIIa, and 14.3%, 25% in stage IIIb respectively. So overexpression of p53 protein has little prognostic significance.
Animals ; Biotin ; Genes, p53 ; Lymph Nodes ; Mice ; Neoplasm Metastasis ; Paraffin ; Prognosis ; Stomach ; Streptavidin ; Survival Rate

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* ; Apoptosis ; Biopsy ; Biotin ; Epidermis ; Epithelium ; In Situ Nick-End Labeling ; Keratinocytes* ; Myeloid Cells ; Skin ; Transferases ; Zinc

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 ; Carcinogenesis ; Cell Cycle ; Genes, Retinoblastoma ; Melanoma* ; Nevus ; Nevus, Pigmented ; Peroxidase