scholarly journals Investigation of Ruminant Encephalitic and Septicemic Listeriosis by the Immunofluorescence Method

2021 ◽  
Author(s):  
Emin KARAKURT ◽  
Fatih BÜYÜK ◽  
Serpil DAĞ ◽  
Enver BEYTUT ◽  
Özgür ÇELEBİ ◽  
...  
Keyword(s):  
Immunofluorescence Method

Express Analysis of Activity of Anti-Rabies Serum and Anti-Rabies Immunoglobulin in Cell Cultures by Immunofluorescence Method

2018 ◽  
Vol 34 (4) ◽  
pp. 83-88 ◽  
Author(s):  
Yu.K. Gavrilova ◽  
◽  
S.V. Generalov ◽  
E.G. Abramova ◽  
L.V. Savitskaya ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Immunofluorescence Method ◽  
Express Analysis

A Direct Immunoassay Assessment of Streptavidin- and N-Hydroxysuccinimide-Modified Biochips in Validation of Serological TNFα Responses in Hemophagocytic Lymphohistiocytosis

2008 ◽  
Vol 13 (6) ◽  
pp. 515-526 ◽  
Author(s):  
Weidong Du ◽  
Xueling Ma ◽  
E. Marion Schneider
Keyword(s):  
Hemophagocytic Lymphohistiocytosis ◽  
Correlation Coefficients ◽  
Molecular Complexes ◽  
Immunofluorescence Method ◽  
Receptor Complexes ◽  
Direct Immunoassay ◽  
Antibody Assays ◽  
Factor Α ◽  
Necrosis Factor ◽  
Biomarker Molecules

The authors report 2 biochip platforms on gold manufactured by either nanoscale biotinylated self-assembled architectures to streptavidin surface or proteins containing free NH 2 groups to N-hydroxysuccinimide (NHS)—activated surfaces and investigated the potential application of tumor necrosis factor—α (TNFα) serodiagnosis of hemophagocytic lymphohistiocytosis (HLH). Interactions of TNFα antigen and TNFα antibody on the biochips were optimized using an indirect immunofluorescence method. Variation coefficients were 1.87% to 4.56% on the streptavidin biochip and 5.03% to 8.64% on the NHS biochip. The correlation coefficients ( r) in TNFα and TNFα antibody assays in HLH patients between the 2 biochip formats were 0.9623 and 0.9386 and the concordance frequencies were 92.2% and 96.1%, respectively. To detect plasma TNFα-receptor complexes (TNFR1 and R2) in HLH, a biochip assay strategy was developed. Plasma levels of TNFα, TNFα antibody, and TNFα-receptor complexes (TNFR1 and R2) were detected in plasmas from 42 HLH cases using streptavidin biochips. Frequencies of the biomarkers in the plasmas were 40.5% (17/42) for TNFα, 30.9% (13/42) for TNFα antibody, 28.6% (12/42) for TNFα—receptor 1 complex, and 26.1% (11/42) for TNFα—receptor 2 complex, respectively. The streptavidin biochip format was more sensitive than the NHS surface and was demonstrated to be a valuable tool to identify individual biomarker molecules and molecular complexes in sera and cell lysates and to track therapeutic progress of patients. ( Journal of Biomolecular Screening 2008:515-526)

scholarly journals Development of a specific method to evaluate 8-hydroxy,2-deoxyguanosine in sperm nuclei: relationship with semen quality in a cohort of 94 subjects

Reproduction ◽  
2013 ◽  
Vol 145 (3) ◽  
pp. 227-235 ◽  
Author(s):  
M Cambi ◽  
L Tamburrino ◽  
S Marchiani ◽  
B Olivito ◽  
C Azzari ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Oxidative Damage ◽  
Oxidative Dna Damage ◽  
Semen Quality ◽  
Sperm Count ◽  
Specific Method ◽  
Sperm Dna Fragmentation ◽  
Immunofluorescence Method ◽  
Sperm Nuclei ◽  
The Relationship

Oxidative stress (OS) is involved in many disoders including male infertility. Human spermatozoa are very sensitive targets of reactive oxygen species (ROS) and most sperm functions are impaired in the case of OS. In addition unbalanced production of ROS is considered one of the most important causes of sperm DNA fragmentation, a semen trait of infertile men. The relationship between oxidative damage and semen quality is partially controversial, probably due to the different methods and/or targets used to reveal the OS. In this study, by fluorescence microscopy and flow cytometry, we compared two methods to reveal 8-hydroxy,2-deoxyguanosine (8-OHdG), the hallmark of oxidative DNA damage: an immunofluorescence method and the commercial OxyDNA kit. We found that although both methods localized the labelling in sperm nuclei they yielded different measures, and only with the immunofluorescence method was the labelling specific for sperm 8-OHdG. The immunofluorescence method, coupled to flow cytometry, was thus selected to analyse the 8-OHdG content in semen samples from 94 subfertile patients and to investigate the relationship with semen quality. We found that the percentages of spermatozoa with 8-OHdG (mean±s.d., 11.4±6.9%) were related to sperm count (Pearson's correlation coefficient (r)=−0.27, P=0.04 (ANOVA and student's t-test)), motility (progressive: r=−0.22, P=0.04; non-progressive: r=0.25, P=0.01), and normal morphology (r=−0.27, P=0.01). In conclusion, we demonstrate that immunofluorescence/flow cytometry is a reliable and specific method to detect 8-OHdG at single-cell level and show that oxidative damage only partially overlaps poor semen quality, suggesting that it could provide additional information on male fertility with respect to routine semen analysis.

scholarly journals Antibodies to platelets in patients with anti-phospholipid antibodies

Blood ◽  
1991 ◽  
Vol 77 (12) ◽  
pp. 2655-2659 ◽  
Author(s):  
HJ Out ◽  
PG de Groot ◽  
M van Vliet ◽  
GC de Gast ◽  
HK Nieuwenhuis ◽  
...  
Keyword(s):  
Platelet Activation ◽  
Lysosomal Membrane ◽  
Immunofluorescence Method ◽  
Exact Role ◽  
Flow Cytofluorometry ◽  
Healthy Donors ◽  
Lysosomal Membrane Proteins ◽  
Lysosomal Membrane Protein ◽  
Platelet Autoantibodies

Abstract Binding of anti-phospholipid antibodies to circulating platelets and its consequences on platelet activation and aggregation was investigated in 11 patients with anti-phospholipid antibodies. Seven patients had mild thrombocytopenia. Nine healthy donors served as controls. Binding to platelets was investigated by performing enzyme- linked immunosorbent assays (ELISAs) with phospholipids as antigen on platelet eluates. Platelet activation was measured by flow cytofluorometry using monoclonal antibodies to an activation-specific lysosomal membrane protein. Findings in ELISA were compared with results of a conventional immunofluorescence method to detect platelet autoantibodies. In seven patients antibodies to negatively charged phospholipids were present in platelet eluates. In all thrombocytopenic patients and controls the platelets were not activated and aggregation was not impaired. There was a positive concordance of 50% between the results of immunofluorescence and ELISA. No apparent relation was found between the results of ELISA or immunofluorescence and platelet counts. It is concluded that anti-phospholipid antibodies can bind to circulating platelets. This binding is not associated with measurable aggregation abnormalities nor with platelet activation characterized by exposure of lysosomal membrane proteins. More studies are necessary to determine the exact role of anti-phospholipid antibodies in the pathogenesis of thrombocytopenia and thrombosis.

scholarly journals Synthesis and assembly of the cytoskeleton of Naegleria gruberi flagellates.

1984 ◽  
Vol 98 (2) ◽  
pp. 449-456 ◽  
Author(s):  
C Walsh
Keyword(s):  
Monoclonal Antibody ◽  
Protein Synthesis ◽  
Indirect Immunofluorescence ◽  
Cell Protein ◽  
Basal Bodies ◽  
Immunofluorescence Method ◽  
Nonionic Detergent ◽  
Cytoplasmic Microtubules ◽  
Spherical Cells ◽  
Naegleria Gruberi

When Naegleria gruberi flagellates were extracted with nonionic detergent and stained by the indirect immunofluorescence method with AA-4.3 (a monoclonal antibody against Naegleria beta-tubulin), flagella and a network of cytoskeletal microtubules (CSMT) were seen. When Naegleria amebae were examined in the same way, no cytoplasmic tubulin-containing structures were seen. Formation of the flagellate cytoskeleton was followed during the differentiation of amebae into flagellates by staining cells with AA-4.3. The first tubulin containing structures were a few cytoplasmic microtubules that formed at the time amebae rounded up into spherical cells. The formation of these microtubules was followed by the appearance of basal bodies and flagella and then by the formation of the CSMT. The CSMT formed before the cells assumed the flagellate shape. In flagellate shaped cells the CSMT radiate from the base of the flagella and follow a curving path the full length of the cell. Protein synthetic requirements for the formation of CSMT were examined by transferring cells to cycloheximide at various times after initiation. One-half the population completed the protein synthesis essential for formation of CSMT 61 min after initiation of the differentiation. This is 10 min after the time when protein synthesis for formation of flagella is completed and 10-15 min before the time when the protein synthesis necessary for formation of the flagellate shape is completed.

scholarly journals LOCALIZATION OF α-FETOPROTEIN IN HUMAN AND RAT FETAL LIVERS AN IMMUNOHISTOCHEMICAL METHOD USING HORSERADISH PEROXIDASE

1974 ◽  
Vol 22 (6) ◽  
pp. 414-418 ◽  
Author(s):  
N. C. NAYAK ◽  
P. K. DAS ◽  
U. N. BHUYAN ◽  
ASHA MITTAL
Keyword(s):  
Connective Tissue ◽  
Horseradish Peroxidase ◽  
Immunohistochemical Method ◽  
Immunoperoxidase Technique ◽  
Serial Sections ◽  
Immunofluorescence Method ◽  
Immunohistochemical Technique

α-Fetoprotein (AFP) was successfully demonstrated in paraffin-embedded sections of human and rat fetal livers, by a multilayering immunohistochemical technique using the peroxidase-antiperoxidase system. Serial sections simultaneously subjected to immunofluorescence showed identical sites of localization in hepatocytic cytoplasm and sometimes in perivenous connective tissue. AFP-containing hepatocytes were located around efferent veins as well as randomly in the lobule. The immunoperoxidase technique has certain advantages over the immunofluorescence method which would justify the former's application in studies on the dynamics of AFP synthesis.

Immunofluorescence studies on deoxyribonuclease from mouse teratocarcinoma cells during cell differentiation

Development ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 37-46
Author(s):  
L. Soriano ◽  
D. Paulin
Keyword(s):  
Cell Differentiation ◽  
Embryonal Carcinoma ◽  
Cell Types ◽  
Carcinoma Cells ◽  
Nuclear Staining ◽  
Fluorescence Staining ◽  
Immunofluorescence Method ◽  
Teratocarcinoma Cells ◽  
Different Cell Types ◽  
Mouse Teratocarcinoma

Specific anti-DNase-I IgG have been used to detect deoxyribonuclease in teratocarcinoma cells by an indirect immunofluorescence method. All the cells studied show fluorescence staining. However, the patterns are quite different in embryonal carcinoma cells (amorphous cytoplasmic fluorescence and absence of nuclear staining) as compared to differentiated cell lines (diffuse, bright granular nuclear and fibrillar cytoplasmic fluorescence). It is possible by this method to distinguish different cell types derived from the same origin. Deoxyribonuclease from teratocarcinoma cells can therefore be considered as a marker of cell differentiation in this system.

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