scholarly journals Methodological improvement of fluorescein isothiocyanate peanut agglutinin (FITC-PNA) acrosomal integrity staining for frozen-thawed Japanese Black bull spermatozoa

2019 ◽  
Vol 81 (5) ◽  
pp. 694-702 ◽  
Author(s):  
Reza RAJABI-TOUSTANI ◽  
Quzi Sharmin AKTER ◽  
Essam A. ALMADALY ◽  
Yoichiro HOSHINO ◽  
Hiromichi ADACHI ◽  
...  
Keyword(s):  
Fluorescein Isothiocyanate ◽  
Peanut Agglutinin ◽  
Methodological Improvement ◽  
Acrosomal Integrity

Peanut agglutinin receptors in the epididymai spermatozoa of the palm squirrel (Funambulus penanti)

1993 ◽  
Vol 51 ◽  
pp. 324-325
Author(s):  
S. R. Bawa ◽  
R. Bawa ◽  
H. K. Bains
Keyword(s):  
Fluorescence Microscopy ◽  
Colloidal Gold ◽  
Fluorescein Isothiocyanate ◽  
Maturation Process ◽  
Peanut Agglutinin ◽  
Epididymal Spermatozoa ◽  
Phosphate Buffered Saline ◽  
Epididymal Duct ◽  
Mammalian Spermatozoa

Physiological ripening of the mammalian spermatozoa during their sojourn in the epididymal duct is also mirrored on the morphological plane by lectin-labeling studies. In this report peanut agglutinin (PNA), which specifically labels methyl α-D-galactopyranoside, has been employed to monitor the maturation process of the epididymal spermatozoa of the Indian palm squirrel (Funambulus penanti) employing fluorescence microscopy and transmission electror microscopy (TEM in toto).Spermatozoa were released from the epididymis by gently mincing the tissues in phosphate-buffered saline (pH 7.4). For fluorescence microscopy, sperm suspensions were smeared onto poly-L-lysine coated coverslips and labeled with 10 ¼l of fluorescein isothiocyanate-linked PNA (100 ¼l/L). For TEM in toto sperm suspensions were adsorbed onto poly-L-lysine and Formvar coated grids and subsequently labeled with colloidal gold-linked PNA. After washing in buffer the grids were examined with a JEOL 1200 EX transmission electror microscope. Controls were prepared by incubating sperm samples in 0.2 M α-D galactopyranoside.

scholarly journals Discrete Distribution of binding sites for Dolichos biflorus agglutinin (DBA) and for peanut agglutinin (PNA) in mouse organ tissues.

1981 ◽  
Vol 29 (7) ◽  
pp. 779-780 ◽  
Author(s):  
M Watanabe ◽  
T Muramatsu ◽  
H Shirane ◽  
K Ugai
Keyword(s):  
Binding Sites ◽  
Cell Lineage ◽  
Discrete Distribution ◽  
Fluorescein Isothiocyanate ◽  
Superficial Layer ◽  
Pancreatic Ducts ◽  
Peanut Agglutinin ◽  
Dolichos Biflorus ◽  
Dolichos Biflorus Agglutinin ◽  
Collecting Tubules

The distribution of binding sites for two lectins with different specificities was studied in adult mice by staining paraffin sections with lectins labeled either with fluorescein isothiocyanate or horseradish peroxidase. Binding sites for Dolichos biflorus agglutinin, a lectin specific to terminal alpha-N-acetylgalactosamine residue, were detected only in several restricted regions, such as collecting tubules and Bowman's capsules of the kidney, bile ducts, pancreatic ducts, sperms, oocytes, some secreting cells, and the secreted mucin itself. Binding sites for peanut agglutinin (PNA), a lectin specific to terminal beta-galactosyl residue, were distributed more widely. However, differentiation-dependent alterations in the expression of PNA binding sites have been observed in two types of cell lineage. During the course of spermatogenesis, the binding sites were expressed at the stage of spermatocytes. In the epithelium of esophagus, the binding sites were present in cells of the superficial layer.

scholarly journals COMPARATIVE STUDY OF DIFFERENT METHODS FOR DOG SEMEN CRYOPRESERVATION AND TESTING UNDER CLINICAL CONDITIONS

2000 ◽  
Vol 48 (3) ◽  
pp. 325-333 ◽  
Author(s):  
F. Szász ◽  
G. Gábor ◽  
László Solti
Keyword(s):  
Clinical Practice ◽  
Sperm Quality ◽  
Intrauterine Insemination ◽  
Fluorescein Isothiocyanate ◽  
Final Conclusion ◽  
Peanut Agglutinin ◽  
Semen Cryopreservation ◽  
Average Litter Size ◽  
Acrosome Integrity ◽  
Clinical Conditions

The extenders and freezing rates from three different freezing protocols were combined and compared to each other in order to study the post-thawing acrosome integrity and fertility of frozen dog sperm. A commercial bovine TRIS-base extender (TRILADYL) and two self-made canine semen extenders (Norwegian and Dutch) were combined with a conventional bovine and two canine freezing regimes, and acrosome integrity of frozen/thawed spermatozoa was assessed by fluorescein isothiocyanate conjugated peanut agglutinin staining (FITC-PNA). Differences between freezing/thawing protocols were reflected in the proportion of cells with acrosomal damage and not based on motility results. It was concluded that during dog semen cryopreservation extenders had less influence on the post-thawing sperm quality than did the freezing rates. The optimal extender/freezing rate combination (TRILADYL/Norwegian) was used in the clinical practice to evaluate the fertility of frozen sperm administered by intrauterine insemination using a surgical approach. The pregnancy rate was 57% (4/7), but the average litter size was low (2.8). This may have been due to the insufficient sperm numbers contained in an insemination dose and/or to the incorrect timing of artificial insemination (AI). The final conclusion is that the commercial bovine extender is useful for freezing dog semen, and the TRILADYL/Norwegian freezing protocol is recommended as the most advantageous combination for the freezing of canine semen in the clinical practice.

Physiological Effects of Nonimmune Platelet Associated Immunoglobulin G

1981 ◽  
Vol 45 (01) ◽  
pp. 027-033 ◽  
Author(s):  
K Sugiura ◽  
M Steiner ◽  
M Baldini
Keyword(s):  
Shape Change ◽  
Fluorescein Isothiocyanate ◽  
Recovery Phase ◽  
Human Platelets ◽  
Serotonin Release ◽  
Platelet Volume ◽  
Igg Antibody ◽  
Heterologous Antiserum ◽  
Igg Binding ◽  
Series Of Experiments

SummaryThe function of nonimmune IgG associated with platelets is unknown. In a series of experiments we have investigated this problem, relating amount of platelet-associated IgG (PAIgG) to platelet volume, serotonin release, adherence of platelets to monocytes and platelet senescence. Most of these studies were performed with human platelets. Platelets freed of preexisting PAIgG by incubation at 22° C were incubated with IgG in a series of concentrations ranging from 0.4 — 27.0 X10-6 M. The IgG preparations used were demonstrably free of aggregated forms of the protein. The amount of PAIgG bound to platelets was determined by the use of fluorescein isothiocyanate-conjugated anti-IgG antibody (F-anti-IgG antibody) which was quantified in a fluorospectrophotometer. Newly bound IgG was assayed similarly by the use of F-IgG. A dose-dependent increase in platelet volume was associated with the binding of nonimmune IgG by platelets. The process which leveled off at an IgG concentration of 1.2 —1.5 X10-5 M was almost fully reversible and was not due to platelet shape change or aggregation. Release of serotonin from IgG-treated platelets was relatively small but to the extent that it occurred was positively related to the IgG concentration to which platelets were exposed. Adherence to autologous monocytes studied quantitatively by the use of formaldehyde-fixed cells was also positively related to the amount of IgG on the platelets. Normal or IgG-defident serum had a potent inhibitory (noncompetitive) action on the binding of F-IgG and F-anti-human IgG antibody to human platelets. Cohorts of platelets prepared in rabbits during the recovery phase of immunological thrombocytopenia induced by injection of heterologous antiserum, showed an age-dependent increase of PAIgG and of IgG binding. These results suggest that PAIgG plays a role in the clearance of senescent platelets.

Further Studies on Aggregation of Platelet-Type von Willebrand’s Disease Platelets by Human von Willebrand Factor

1986 ◽  
Vol 55 (03) ◽  
pp. 338-341 ◽  
Author(s):  
H Takahashi ◽  
W Tatewaki ◽  
M Hanano ◽  
R Nagayama ◽  
A Shibata
Keyword(s):  
Von Willebrand Factor ◽  
Ricinus Communis ◽  
Divalent Cations ◽  
Peanut Agglutinin ◽  
Von Willebrand's Disease ◽  
Von Willebrand’S Disease ◽  
Ricinus Communis Agglutinin ◽  
Von Willebrand ◽  
Induced Aggregation ◽  
Willebrand Factor

SummaryPlatelet-type von Willebrand’s disease (vWD) is a bleeding disorder characterized by a heightened interaction between platelets and von Willebrand factor (vWF) as the result of an intrinsic platelet abnormality (probably in GPIb). Platelet aggregability was nearly normal in response to thrombin, wheat germ agglutinin and Ricinus communis agglutinin in this disorder. Unmodified platelets showed no aggregation upon the addition of peanut agglutinin. Partially purified human vWF induced little aggregation of washed patient platelets, but the aggregation was greatly enhanced in the presence of plasma devoid of vWF. Monoclonal antibodies directed against GPIb and GPIIb/IIIa as well as EDTA completely inhibited vWF-induced aggregation. These results indicate that human vWF induces aggregation of platelet-type vWD platelets in the presence of divalent cations and some plasma cofactor(s), and that both GPIb and GPIIb/IIIa are involved in this aggregation.

Interspecific variation of zona pellucida glycoconjugates in several species of marsupial

Reproduction ◽  
2000 ◽  
pp. 111-120 ◽  
Author(s):  
JA Chapman ◽  
OW Wiebkin ◽  
WG Breed
Keyword(s):  
Glycine Max ◽  
Sialic Acid ◽  
Zona Pellucida ◽  
Fluorescein Isothiocyanate ◽  
Interspecific Variation ◽  
Lectin Histochemistry ◽  
Sialic Acids ◽  
Arachis Hypogea ◽  
Marsupial Species ◽  
Similar Intensity

The zona pellucida glycoconjugate content of several marsupial species was investigated using differential lectin histochemistry. Ovaries from fat-tailed dunnarts, a southern brown bandicoot, grey short-tailed opossums, brushtail possums, ringtail possums, koalas and eastern grey kangaroos were fixed, embedded in paraffin wax, sectioned and stained with ten fluorescein isothiocyanate-conjugated lectins. Sections were also incubated with either neuraminidase or saponified, respectively, before incubation with the lectins to identify saccharide residues masked by sialic acids or O-acetyl groups on sialic acids. The zonae pellucidae surrounding the oocytes of the marsupials demonstrated interspecific variation in glycoconjugate content, with mannose-containing glycoconjugates exhibiting the greatest variation. Some of the zona pellucida glycoconjugates of all species, except those of the opossums, were masked by sialic acid with an increase in fluorescence with lectins from Arachis hypogea (PNA), and Glycine max (SBA), after desialylation. The disaccharide beta-galactose(1-4)N-acetyl-D-glucosamine appeared to be conformationally masked by O-acetyl groups of sialic acids in the zonae pellucidae of all species, with an increase in fluorescence with the lectin from Erythrina cristagalli (ECA), after saponification. Similar intensity and localization of beta-(1-4)-N-acetyl-D-glucosamine, as shown by staining of the lectin from Triticum vulgaris (WGA), to the inner and outer regions of the zona pellucida, were found to those reported in eutherian species. WGA fluorescence became uniform throughout the zonae pellucidae after saponification, indicating differential O-acetylation of sialic acids on the internal compartment of the zonae pellucidae.

scholarly journals Specific and rapid reverse assaying protocol for detection and antimicrobial susceptibility testing of Pseudomonas aeruginosa based on dual molecular recognition

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yong He ◽  
Hang Zhao ◽  
Yuanwen Liu ◽  
He Zhou
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antimicrobial Susceptibility ◽  
Magnetic Particles ◽  
Susceptibility Testing ◽  
Fluorescein Isothiocyanate ◽  
High Specificity ◽  
Antimicrobial Susceptibility Testing ◽  
Tail Fiber ◽  
Isolation And Identification ◽  
Magainin Ii

AbstractThe worldwide emergence and spread of antimicrobial resistance is accelerated by irrational administration and use of empiric antibiotics. A key point to the crisis is a lack of rapid diagnostic protocols for antimicrobial susceptibility testing (AST), which is crucial for a timely and rational antibiotic prescription. Here, a recombinant bacteriophage tail fiber protein (TFP) was functionalized on magnetic particles to specifically capture Pseudomonas aeruginosa, while fluorescein isothiocyanate-labeled-magainin II was utilized as the indicator. For solving the magnetic particles’ blocking effects, a reverse assaying protocol based on TFP recognition was developed to investigate the feasibility of detection and AST of P. aeruginosa. P. aeruginosa can be rapidly, sensitively and specifically detected within 1.5 h with a linear range of 1.0 × 102 to 1.0 × 106 colony forming units (CFU)⋅mL−1 and a detection limit of 3.3 × 10 CFU⋅mL−1. Subsequently, AST results, which were consistent with broth dilution results, can be obtained within 3.5 h. Due to the high specificity of the TFP, AST can actually be conducted without the need for bacterial isolation and identification. Based on the proof-of-principle work, the detection and AST of other pathogens can be extended by expressing the TFPs of their bacteriophages.

scholarly journals Silver Nanoparticles Functionalized by Fluorescein Isothiocyanate or Rhodamine B Isothiocyanate: Fluorescent and Plasmonic Materials

2021 ◽  
Vol 11 (6) ◽  
pp. 2472
Author(s):  
Ilaria Fratoddi ◽  
Chiara Battocchio ◽  
Giovanna Iucci ◽  
Daniele Catone ◽  
Antonella Cartoni ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Rhodamine B ◽  
Photoelectron Spectroscopy ◽  
Transient Absorption ◽  
Fluorescein Isothiocyanate ◽  
Dye Molecules ◽  
Chemical Surface ◽  
Vis Spectroscopy ◽  
Rhodamine B Isothiocyanate ◽  
Fluorescent Molecules

This paper presents the synthesis of silver nanoparticles (AgNPs) functionalized with fluorescent molecules, in particular with xanthene-based dyes, i.e., fluorescein isothiocyanate (FITC, λmax = 485 nm) and rhodamine B isothiocyanate (RITC, λmax = 555 nm). An in-depth characterization of the particle–dye systems, i.e., AgNPs–RITC and AgNPs–FITC, is presented to evaluate their chemical structure and optical properties due to the interaction between their plasmonic and absorption properties. UV–Vis spectroscopy and the dynamic light scattering (DLS) measurements confirmed the nanosize of the AgNPs–RITC and AgNPs–FITC. Synchrotron radiation X-ray photoelectron spectroscopy (SR-XPS) was used to study the chemical surface functionalization by structural characterization, confirming/examining the isothiocyanate–metal interaction. For AgNPs–RITC, in which the plasmonic and fluorescence peak are not superimposed, the transient dynamics of the dye fluorescence were also studied. Transient absorption measurements showed that by exciting the AgNPs–RITC sample at a wavelength corresponding to the AgNP plasmon resonance, it was possible to preferentially excite the RITC dye molecules attached to the surface of the NPs with respect to the free dye molecules in the solution. These results demonstrate how, by combining plasmonics and fluorescence, these AgNPs can be used as promising systems in biosensing and imaging applications.

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