Localization of Single Avidin�Biotin Interactions Using Simultaneous Topography and Molecular for new users. Using well-characterized, high-affinity biolog- ical binding partners, such as avidin�biotin. The robust and well-described avidin�biotin interaction offers an ideal pair of binding partners. ...

Here we report an approach to the design and production of antibody/ligand pairs, to achieve functional affinity far greater than avidin/biotin. Using fundamental chemical principles, we have developed antibody/ligand pairs that retain the binding specificity of the antibody, but do not dissociate. Choosing a structurally characterized ...

The molecular host cucurbit[7]uril forms an extremely stable inclusion complex with the dicationic ferrocene derivative bis(trimethylammoniomethyl)ferrocene in aqueous solution. The equilibrium association constant for this host-guest pair is 3 � 10¹⁵ M^?1 (K_d = 3 � 10^?16 M), equivalent to that exhibited ...

chemical (Cu-catalyzed azide�alkyne cycloaddition) and biochemical (avidin�biotin binding) interactions of the affinity mass spectrometry theme, enabled by the use of cleavable covalent linkages and highly active with concomitant loss of DIOS activity. We have recently discovered that the lost DIOS function of oxidized p

A two site electrochemical enzyme immunoassay for thyrotropin (TSH) has been developed. This assay is based on the use of an immobilized capture antibody and a biotinylated second antibody. Detection is achieved via avidin labelled with alkaline phosphatase. The substrate 1-naphthyl phosphate was used and the product 1-naphthol was detected at disposable screen-printed carbon ...

Polymersomes are vesicles whose membranes are comprised of self-assembled amphiphilic block co-polymers. Synthetic control of block co-polymer chemistry provides an advantageous diversity of polymersome functions, ranging from tunable materials strength, superior encaspulation of hydrophobic and hydrophilic drugs and optical dyes, and facile functionalization. We have exploited polymersome ...

Advances in immunohistochemical technologies have greatly improved the ability to visualize antigens in formalin-fixed paraffin-embedded tissues. Nonetheless, there are occasions in which there may be no alternative to the use of cryostat-sectioned frozen tissues. While the basic chemistries and techniques used in staining cryostat-sectioned tissues are identical to those employed for staining ...

Our second system employed avidin-biotin chemistry. ... With carbohydrate- selectin or avidin-biotin interactions, large NA/NB leads to the formation of ...

... Both peroxidase-antiperoxidase and avidin biotin complex techniques -e used to identify intermediate filament proteins keratin, desmin, vimentin ...

Longitudinally bisected hypothalami were sectioned 20 >m thick and stained with a conventional avidin-biotin-immunoperoxidase procedure. ...

... were incubated for 3 hours in a 1% avidine-biotin complex (ABC: Streptavidin Biotin Complex Peroxidase kit, ... ...

To expand the electron microscopist's options in localization and visualization, a new and general staining technique has been tested. The avidin-biotin complex serves as a coupling between the electron-dense marker, ferritin, and points of interest in biological samples. When specific cellular components are tagged with biotin, those components may be visualized with ferritin-linked avidin. ...

A non-isotopic receptor assay for benzodiazepine drugs was developed using a biotin-labeled ligand, biotin-1012S. Biotinylated bovine serum albumin (biotin-BSA) was immobilized onto the wall of microtiter plate wells by simple adsorption. Avidin peroxidase conjugate could be extracted from solution owing to its strong interaction with biotin. The amount of avidin peroxidase taken up on the wall ...

The application of avidin-biotin technology, originally designed to facilitate and improve purification and localization procedures for biologically active macromolecules, has led to major advances in medical diagnostics. Among other new applications are ...

The testes of 8 specimens of Triturus marmoratus were collected during each month of 1987 and processed for electron microscopy and light microscopy demonstration of testosterone (T) following the ABC (avidin-biotin peroxidase complex) method. According to their staining affinity for anti-T antibodies, the glandular tissue cells were classified as T-, T+, ...

PurposeFor periodontal tissue engineering, it is a primary requisite and a challenge to select the optimum types of cells, properties of scaffold, and growth factor combination to reconstruct a specific tissue in its natural form and with the appropriate function. Owing to fundamental disadvantages associated with using a two-dimensional substrate, several methods of seeding cells into ...

Recently, a correlation has been suggested between a loss of E-cadherin (E-CD) and increased invasiveness of neoplastic cells. In this study, E-CD expression in breast cancer was investigated using an affinity-purified antibody (ECCD-2) in an immunoenzymatic (avidin-biotin-alkaline phosphatase) test. Intensity and extension of E-CD immunoreactivity were ...

Nanocrystals of the fluorogenic precursor fluorescein diacetate (FDA) were applied as labels in order to improve on the assay sensitivity achieved in our previous studies. Each FDA nanocrystal can be converted into approximately 2.6x10(6) fluorescein molecules, which is useful for improving immunoassay sensitivity and limits of detection. NeutrAvidin was simply adsorbed onto ...

Eight commercially available monoclonal antibodies directed against respiratory syncytial virus antigens were tested for ability to detect bovine respiratory syncytial virus (BRSV) antigen in formalin fixed, paraffin embedded bovine lung using avidin-biotin complex immunohistochemical staining. Monoclonal antibodies from clone 18B2 purchased from Biosoft, Paris, France and ...

A conventional enzyme-linked immunosorbent assay (ELISA) for alpha-fetoprotein was modified to incorporate the avidine-biotin complex, in order to enhance sensitivity. Five modifications of this assay were compared, of which the 2-step noncompetitive avidine-biotin assay (NABA) was found to be the most rapid and sensitive (detection limit 0.05 ng/ml). Levels of AFP in human ...

Metalloenzymes and electron transfer proteins influence the electrochemical properties of metal cofactors by controlling the second-sphere environment of the protein active site. Properties that tune this environment include the dielectric constant, templated charge structure, van der Waals interactions, and hydrogen bonds. By systematically varying the binding of a redox-active ligand with a ...

Metalloenzymes and electron transfer proteins influence the electrochemical properties of metal cofactors by controlling the second-sphere environment of the protein active site. Properties that tune this environment include the dielectric constant, templated charge structure, van der Waals interactions, and hydrogen bonds. By systematically varying the binding of a redox-active ligand with a ...

as an attractive and exclusive site for attaching cysteine-containing biotin. Using these novel biotinylation of using biotin over other tags (e.g., GST, His-tag, etc.) as the method of choice in protein purification, there is an urgent need to develop alternative methodologies that address these issues. Since the 1980s, avidin-biotin

-peptide sequence of L-3,4-dihydrox- yphenylalanine (DOPA3-PEG3400-biotin; DPB) were incorporated within a DOPA3-PEG avidin. The resulting biotin-avidin-biotin linkages were stable for prolonged periods under continuous perfusion, even in the presence of excess free biotin. Hematopoietic M07e cells bound to immobilized peptide

Exploiting self-assembly systems with biological building blocks is of significant interest in the fabrication of advanced biomaterials. We assessed the potential use of site-specific ligand labeling of protein building blocks in designing functional protein self-assemblies by combining site-specifically biotinylated bacterial alkaline phosphatase (as a bidentate or tetradentate ligand unit) and ...

1 FORCED UNBINDING OF THE AVIDIN�BIOTIN COMPLEX Vincent T. Moy*, Ernst�Ludwig Florin, Matthias Rief to develop a model for the dissociation of the avidin�biotin complex. Atomic force microscope (AFM) measurements of the unbinding force of five avidin�biotin homologues were found to be proportional

1 FORCED UNBINDING OF THE AVIDIN-BIOTIN COMPLEX Vincent T. Moy*, Ernst-Ludwig Florin, Matthias Rief to develop a model for the dissociation of the avidin-biotin complex. Atomic force microscope (AFM) measurements of the unbinding force of five avidin-biotin homologues were found to be proportional

In this study, biomaterials are patterned at the micro- and nanometer scale for biosensor applications and cell-based research. Two enabling technologies were photoactivatable biotin and the polymer lift-off technique. Avidin-biotin complexes have been im...

Med] Biotin is a protein found in mammalian cells and is considered to be a vitamin because it has constant (Kd) of 4 � 10-14 M (1,2). The avidin� biotin interaction has been described in detail by Wilchek to break the avidin/streptavidin� biotin bond, and the released proteins cannot be reused for any other

Elevated concerns of ecological safety and environmental pollution have facilitated massive research in the area of chemical and biological agent detection. Large numbers of anthropogenic chemicals were introduced in the environment within last 50 years, which posed a serious threat to the natural ecological balance. The series of unfortunate events in the past could have been avoided or minimized ...

We have pursued the use of biological crosslinking molecules of several types to make colloidal materials at relatively low volume fraction of colloidal particles. The objective is to make binary alloys of colloidal particles, made of two different colloidal particles coated with complementary biological lock-and-key binding molecules, which assemble due to the biological specificity. The ...

In this review, the preparation and properties of protein architectures constructed by layer-by-layer (LbL) deposition through avidin-biotin and concanavalin A (Con A)-sugar interactions are discussed in relation to their use for optical and electrochemical biosensors. LbL films can be constructed through the alternate deposition of avidin and biotin-labeled enzymes on the ...

Avidin-biotin-peroxidase complex immunohistochemistry was used on formalin fixed, paraffin embedded, trypsin digested, skin biopsies to detect immunoglobulin deposition in dogs with autoimmune skin disease. Immunostaining by the avidin-biotin-peroxidase complex technique revealed intercellular and/or basement membrane immunoglobulin deposits in 27 of 28 dogs considered to have ...

Resonant microcavities have recently become popular for research in optical biosensor applications. Whispering Gallery Mode (WGM) microresonators are preferred as active sensing surfaces due to their high-Q values which provide measurable shifts in mode frequencies as surface characteristics change. In this study, the active sensing surface is a microring resonator vertically coupled to waveguides ...

The distribution of different carbohydrates in dog major and minor salivary glands was investigated using a peroxidase-labelled avidin-biotin method to demonstrate binding of six lectins (Canavalia ensiformis agglutinin [Con A], Dolichos biflorus agglutinin [DBA], Arachis hypogaea (peanut) agglutinin [PNA], Glycine max agglutinin [SBA], Tetragonolobus purpurea agglutinin [TGP] ...

A biomimetic, microscale system using the mechanics of swimming bacteria has been fabricated and controlled in a low Reynolds number fluidic environment. The microswimmer consists of a polystyrene microbead conjugated to a magnetic nanoparticle via a flagellar filament using avidin-biotin linkages. The flagellar filaments were isolated from the bacterium, Salmonella ...

Spatiotemporal control of gene delivery, particularly signaling gradients, via biomaterials poses significant challenges because of the lack of efficient delivery systems for therapeutic proteins and genes. This challenge was addressed by using chemical vapor deposition (CVD) polymerization in a counterflow set-up to deposit copolymers bearing two reactive chemical gradients. FTIR spectroscopy ...

The products of biotechnology, recombinant proteins, monoclonal antibodies, antisense, RNA interference, or non-viral gene transfer, cannot be developed as pharmaceuticals for the brain, unless these molecules are re-formulated to enable transport across the blood-brain barrier (BBB). Large molecule drugs, and plasmid DNA, can be delivered across the BBB with receptor-specific molecular Trojan ...

Nanoparticles have emerged as key materials for biomedical applications because of their unique and tunable physical properties, multivalent targeting capability, and high cargo capacity. Motivated by these properties and by current clinical needs, numerous diagnostic and therapeutic nanomaterials have recently emerged. Here we describe a novel nanoparticle targeting platform that uses a rapid, ...

Nanoparticles have emerged as key materials for biomedical applications because of their unique and tunable physical properties, multivalent targeting capability, and high cargo capacity1,2. Motivated by these properties and by current clinical needs, numerous diagnostic3�10 and therapeutic11�13 nanomaterials have recently emerged. Here we describe a novel nanoparticle targeting platform that ...

Nanoparticles have emerged as key materials for biomedical applications because of their unique and tunable physical properties, multivalent targeting capability, and high cargo capacity. Motivated by these properties and by current clinical needs, numerous diagnostic and therapeutic nanomaterials have recently emerged. Here we describe a novel nanoparticle targeting platform that uses a rapid, ...

Various methods have been applied for serum ferritin detection, however, these methods still have some limitations. Over the last few years, quantum dots (QDs) have become very attractive for immunoassays because of their enormous potentials in ultrasensitive analysis. In this study, a Western blotting method combined with QDs-labeled avidin-biotin system for detecting human ...

We demonstrate simultaneous transverse dynamic force microscopy and molecular recognition imaging using tuning forks as piezoelectric sensors. Tapered aluminum-coated glass fibers were chemically functionalized with biotin and anti-lysozyme molecules and attached to one of the prongs of a 32kHz tuning fork. The lateral oscillation amplitude of the tuning fork was used as feedback signal for ...

A compositional analysis of the antibody response in rainbow trout was conducted using an affinity-based immunopartitioning assay. Trout were immunized with TNP-keyhole limpet hemocyanin (TNP-KLH) and individual serum titers and their affinity distributions analyzed over a period of 27 weeks. The kinetics of antibody affinity ...

One approach to prepare protein binding ligands is to join two low-affinity ligands that bind different sites on the target protein to create a high-affinity bivalent ligand. This typically requires some knowledge of the ligand binding site and requires exquisite orientation of the ligands in order to achieve maximum binding ...

The discovery of maraviroc 17 is described with particular reference to the generation of high selectivity over affinity for the HERG potassium channel. This was achieved through the use of a high throughput binding assay for the HERG channel that is known to show an excellent correlation with functional effects. PMID:16782336

The purpose of this study was to investigate the morphology of nerve endings in spinal ligaments of scoliotic and normal patients. Nervous elements were investigated by means of gold chloride (AuCl2) impregnation and an indirect avidine-biotine peroxydase immunohistochemical method. Antiserum to a neurofilament protein (NFP) and S-100 protein were used as specific markers of ...

A 6-week-old female BALB/c mouse was immunized with a preparation of partially purified sieve tubes of periwinkle (Catharantus roseus) plants infected with the apple proliferation mycoplasma-like organism. After fusion of the immunized spleen cells with myeloma cells, the supernatant fluids of the resulting hybridoma cultures were screened for the presence of antibodies in a differential ...

A rat model of lumbar root constriction with an additional sympathectomy in some animals was used to assess whether the sympathetic nerves influenced radicular pain. Behavioural tests were undertaken before and after the operation. On the 28th post-operative day, both dorsal root ganglia and the spinal roots of L4 and L5 were removed, frozen and sectioned on a cryostat (8 microm to 10 microm). ...

We aim to create a new class of materials that self-assemble and self-replicate. Biotin-terminated DNA strands are attached to neutravidin coated polystyrene particles via the well established avidin-biotin coupling mechanism. The DNA is composed of a 61-base strand and a 50-base complementary strand, leaving a single sticky end of 11 bases to interact with its complement ...

STD (sexually transmitted disease, Gonorrhoea) sensor based on nucleic acid probe (from Opa, a multi-copy gene of Neisseria gonorrhoeae) functionalized nanostructured-polyaniline coated onto indium-tin-oxide-coated glass plate has been fabricated using avidin-biotin as cross-linking agent. This DNA functionalized electrode can specifically detect upto 0.5 x 10(-15)M of ...

It has been reported that the immune response due to ?-Gal epitopes is an important factor in tissue valve failure. The elimination of the interaction between the natural anti-Gal antibodies and ?-gal epitopes on the xenografts is a prerequisite to the success of xenografts in humans. Previously, we reported that the green coffee bean ?-galactosidase could remove all ?-Gal epitopes from cell ...

Recombinant TNF-related apoptosis-inducing ligand (TRAIL) is considered a powerful and selective inducer of tumor cell death. We hypothesize that TRAIL�s potential as anticancer agent can be enhanced further by promoting its accumulation in tumor tissue. For this purpose, we developed TRAIL complexes that bind to angiogenic endothelial cells. We employed an avidin�biotin pretargeting approach, ...

The right bronchus of the short-tailed shrew, Blarina brevicauda, terminates in a nonrespiratory pulmonary appendix (PA) containing two bronchial extensions. The experimentally demonstrated ability of these structures to collect and peristaltically expel aspirated material was initially assumed to be a sufficient reason for their developmental persistence, but as bronchus associated lymphoid ...

Cross-linking analysis of protein complexes and structures by tandem mass spectrometry (MS/MS) has advantages in speed, sensitivity, specificity, and the capability of handling complicated protein assemblies. However, detection and accurate assignment of the cross-linked peptides are often challenging due to their low abundance and complicated fragmentation behavior in collision-induced ...

We attempted to develop an in situ hybridization procedure with maize meiotic pachytene chromosomes for single copy genes using non-radioactive probes. Protocols were used incorporating biotin into probe DNA using an avidin-biotin-fluorescein molecule as the detector system. We also tried avidin-biotin-horseradish peroxidase that leads to a precipitate at the site of ...

We investigated carbonic anhydrase IV (CA IV) in rat and human heart with immunohistochemical methods by both light and electron microscopy. In cryosections that were incubated with anti-CA IV/FITC, the capillaries showed a strong reaction for CA IV. In paraffin and semithin sections treated with anti-CA IV/ABC (avidin-biotin-peroxidase complex) blood vessels, capillaries, and ...

We have demonstrated label-free THz sensing of living body-related molecular binding using a thin metallic mesh and a polyvinylidene difluoride (PVDF) membrane. Metallic meshes in the THz region are designed for anomalous transmission phenomena derived from a resonant excitation of surface waves. Additionally, they are designed to have a sharp dip in transmittance. The metallic mesh is very ...

In this work, the kinetics and dissociation constant for the binding of a biotin-modified oligonucleotide to microparticle-immobilized avidin were measured. Avidin has been immobilized by both covalent coupling and bioaffinity capture to a surface prefunctionalized with biotin. The measured rate and equilibrium dissociation constants of avidin immobilized by these different methods have been ...

By using immunocytochemistry method of Strept Avidin-Biotin-Complex, four kinds of antisera raised against rabbits were applied to observe the immunoreactive neurons and neuropils of serotonin (5-HT), neuropeptide Y (NPY), substance P (SP) and ?-Endorphin (?-Ep) in optic ganglia and brain of Metapenaeus ensis. The results showed that, the 5-HT-immunoreactive cells were located ...

Human placental lactogen is one of the major hormones secreted by the placental syncytiotrophoblast and detected in the maternal circulation. Other sources of this hormone in intrauterine tissues at term have been sought by means of immunohistochemistry and northern analysis. Avidin-biotin immunoperoxidase staining with a specific polyclonal antibody to human placental ...

A significant increase in cutaneous leishmaniasis (CL) and its geographic expansion has motivated the development of techniques to help with diagnosis of the disease. Here we describe immunocytochemical (ICC) and immunohistochemical (IHC) techniques for the diagnosis of CL in the laboratory. Polyclonal antibodies and a modified avidin-biotin complex (Ultra Streptavidin(�)) ...

From January 1987 to August 1988, cytomorphologic criteria of both herpes simplex virus (HSV) and radiation effects were observed in Papanicolaou smears from 3 of 1,340 patients who had received radiotherapy for squamous cell carcinoma of the cervix. Avidin-biotin immunoperoxidase staining, using a rabbit IgG polyclonal HSV antibody, confirmed the presence of HSV antigen in ...

Enkephalin-like immunoreactive amacrine cells were visualized using the highly sensitive avidin-biotin method. The somas of these cells were situated in the inner nuclear and ganglion cell layers. Enkephalin-stained processes were observed in layers 1, 3, and 5 of the inner plexiform layer. The biosynthesis of sulfur-containing compounds in the goldfish retina was studied by ...

Microarray performance depends upon the ability to screen samples against a vast array of probes with the appropriate sensitivity and selectivity. While these factors are significantly influenced by probe design, they are also subject to the particular detection methodology and reagents employed. Herein we describe the incorporation of super avidin-biotin system (SABS) and ...

A method is described for indirect electron microscopic visualization and mapping of tRNA and other short transcripts hybridized to DNA. This method depends upon the attachment of the electron-dense protein ferritin to the RNA, the binding being mediated by the remarkably strong association of the egg white protein avidin with biotin. Biotin is covalently attached to the 3' end of tRNA using an ...

Effective Schwann cells (SCs) attachment is a prerequisite for the successful construction of tissue-engineered nerve. The present study aimed to investigate the role of an avidin-biotin binding system (ABBS) for neural tissue engineering. The attachment, proliferation, and morphology of biotinylated SCs on avidin-treated scaffolds were examined, and the effects of avidin, ...

The goal of brain drug targeting technology is the delivery of therapeutics across the blood-brain barrier (BBB), including the human BBB. This is accomplished by re-engineering pharmaceuticals to cross the BBB via specific endogenous transporters localized within the brain capillary endothelium. Certain endogenous peptides, such as insulin or transferrin, undergo receptor-mediated transport (RMT) ...

Flagella have been employed as nanoactuators for biomimetic microswimmers in low Reynolds number fluidic environments. The microswimmers utilize flagellar filaments isolated from Salmonella typhimurium to mimic the spiral-type propulsion mechanism of flagellated bacteria. The microswimmer included a polystyrene microbead conjugated to one or multiple magnetic nanobeads via flagellar filaments ...

In the last years, multiple studies indicated the potential effectiveness of the polymerase chain reaction (PCR) for detection of Giardia in pathological specimens. By using the difference at the triose phosphate isomerase (tpi) gene locus, we investigated the genotype groups of Giardia lamblia isolated directly from fecal specimens collected from humans living in Dakahlia Governorate, Egypt. ...

The basic recognition schemes underlying the principles of standard enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) protocols are increasingly being adapted for use with new detection devices. A direct comparison was made using a fiber optic biosensor that employs evanescent wave detection and an ELISA using avidin-biotin. The assays were developed for the ...

Click chemistry is explored as a potential cost-effective and selective immobilization method for the production of an enzyme-linked immunosorbent assay (ELISA). Coatings were formulated containing either a terminal alkyne or a bicyclo[6.1.0]non-4-yne (BCN) chemical handle and a diagnostic peptide was subsequently immobilized onto these coatings by the copper catalyzed azide-alkyne 1,3-dipolar ...

A solid phase immunoassay utilizing avidin-biotin binding has been developed for measuring anticapsular polysaccharide antibodies. Capsular polysaccharides of Escherichia coli K1, Haemophilus influenzae type b, Staphylococcus aureus types 5 and 8, and levan from Aerobacter levanicum have been biotinylated through -OH or COOH groups with retention of antigenicity. ...

The distributions of serotonin- (5HT-), substance P- (SP-), small cardioactive peptideb- (SCPb-), and histamine- (HA-) like immunoreactivities were examined in the adult lobster supraesophageal ganglion. Vibratome sections were labeled using avidin-biotin-peroxidase immunocytochemical methods. The localization patterns for each substance were assessed in 21 regions within the ...

Purification of alpha-galactosidase from the roots of Verbascum thapsus L. was difficult to achieve using conventional methods due to the presence of coloured contaminants. A newly developed procedure, hybrid affinity chromatography, which was based on a mixed matrix separation procedure, using a substrate analogue and an immobilized metal ...

Synthetic polymer nanoparticles with antibody-like affinity for a hydrophilic peptide have been prepared by inverse microemulsion polymerization. Peptide affinity was achieved in part by incorporating the target (imprint) peptide in the polymerization reaction mixture. Incorporation of the imprint peptide assists in the creation of ...

Nitrilotriacetic acid (NTA) has moderate affinity (10 �M) for hexahistidine (His6) and is widely used in the purification of His6-tagged proteins. The affinity can be increased significantly (10 nM) through multivalency such as using a tris-NTA. We show that the binding affinity of tris-NTA is dependent on the flexibility and length ...

The use of genetically encoded small peptide tags such as polyhistidine and tetracysteine tags has become important for protein purification and enrichment. An improved affinity purification of tetracysteine (CCXXCC) tagged proteins has been achieved using a nonfluorescent, photochemically stable bisarsenical affinity ligand SplAsH. ...

The population dynamics theory of B cells in a typical germinal center could play an important role in revealing how affinity maturation is achieved. However, the existing models encountered some conflicts with experiments. To resolve these conflicts, we present a coarse-grained model to calculate the B cell population development in ...

The study of the activation processes for achieving negative electron affinity (NEA) in Ge-doped GaAs layers grown by liquid phase epitaxy is described. Apart from the III-V compounds, NEA has been reported in silicon. The literature suggests that the det...

Methods for the isolation of antithrombin III using affinity chromatography on heparin sepharose are being developed. Best results are achieved when heparin is linked to sepharose thru a hydroxyl group. Preliminary animal studies show that an effect on th...

BackgroundThere is a significant need for affinity reagents with high target affinity/specificity that can be developed rapidly and inexpensively. Existing affinity reagent development approaches, including protein mutagenesis, directed evolution, and fragment-based design utilize large libraries and/or require structural information ...

We propose a new method of quantitative measurement of cell surface blood group antigen (BGA) in superficial transitional cell carcinoma (TCC) of the urinary tract by using flow cytometry. Flow cytometric analysis was performed on 22 cases of superficial urinary tract TCC registered at the department of Urology, Okayama University Hospital during April, 1992-Feb. 1993. Fresh samples were divided ...

Uroplakins (UPs) Ia, Ib, II, and III, transmembrane proteins constituting the asymmetrical unit membrane of urothelial umbrella cells, are the first specific urothelial differentiation markers described. We investigated the presence and localization patterns of UPs in various human carcinomas by applying immunohistochemistry (avidin-biotin-peroxidase complex method), using ...

OBJECTIVE:: The objective of the study was to evaluate hypoxia-inducible factor 1 (HIF-1), which plays a major role in the stimulation of angiogenesis in placental tissues, by using immunohistochemical staining in preeclampsia model of rats, developed by N-nitro-l-arginine methyl ester (l-NAME) METHODS:: Thirty pregnant rats were randomized into 2 groups (n = 15 in each group) on day 10 of ...

We have developed surfaces for the selective presentation of biotinylated peptides and proteins in a background that resists nonspecific protein adsorption; controlled amounts of biotinylated polyethylene glycol (MW 3400; PEG3400) anchored to titanium-dioxide-coated surfaces via an adhesive tri-peptide sequence of l-3,4-dihydroxyphenylalanine (DOPA₃-PEG3400-biotin; DPB) were ...

The surface plasmon resonance (SPR) technique is a well-established method for the measurement of molecules binding to surfaces and the quantification of binding constants between surface-immobilized proteins and proteins in solution. In this paper we describe an extension of the methodology to study bacteriophage-bacterium interactions. A two-channel microfluidic SPR sensor device was used to ...

Skin and superficial lymph nodes from the 65 juvenile (< 1 year old) and adult free-ranging American badgers (Taxidea taxus) of both sexes that were killed from late July to late October 1991 as part of the recovery program for the endangered black-footed ferret (Mustela nigripes) were examined for evidence of Filaria taxideae. Fifty-one badgers (51/64, 80%) were infected. Both adult badgers ...

Extrusion of hydrated lipid suspensions is frequently employed to produce vesicles of uniform size, and the resulting vesicles are often reported to be unilamellar. We describe a method for the quantitative fluorescence image analysis of individual vesicles to obtain information on the size, lamellarity, and structure of vesicles produced by extrusion. In contrast to methods for bulk analysis, ...

AIMS--To study the immunohistochemical expression of carcinoembryonic antigen (CEA) in ductal hyperplasia of the breast and to investigate its putative relation with atypia and co-existing infiltrating ductal carcinoma. METHODS--Paraffin wax embedded tissue from 37 cases of isolated ductal hyperplasia (five with atypia and 32 without atypia) and 25 cases of ductal hyperplasia associated ...

Thanks to Dynamic Force Spectroscopy (DFS) and developments of massive data analysis tools, such as YieldFinder, Atomic Force Microscopy (AFM) becomes a powerful method for analyzing long lifetime ligand-receptor interactions. We have chosen the well-known system, (strept)avidin-biotin complex, as an experimental model due to the lack of consensus on interpretations of the ...

Natural variation in neuroendocrine traits is poorly understood, despite the importance of variation in brain function and evolution. Most rodents in the temperate zones inhibit reproduction and other nonessential functions in short winter photoperiods, but some have little or no reproductive response. We tested whether genetic variability in reproductive seasonality is related to individual ...

In this study, a ToF-SIMS operating in the event-by-event bombardment/detection mode was used to characterize avidin-biotin assemblies on silane-modified glass substrates. SIMS was used to analyze several variants of the biointerface, including avidin physically adsorbed on a monofunctional acryl silane surface and covalently attached on a mono-functional (amine terminated) ...

Receptor-mediated targeting of nanometric contrast agents or drug carriers holds great potential for treating cardiovascular and vascular-associated diseases. However, predicting the ability of these vectors to adhere to diseased cells under dynamic conditions is complex due to the interplay of transport, hydrodynamic force, and multivalent bond formation dynamics. Therefore, we sought to ...

The adhesive plaques of Mytilus byssus are investigated increasingly to determine the molecular requirements for wet adhesion. Mfp-2 is the most abundant protein in the plaques, but little is known about its function. Analysis of Mfp-2 films using the surface forces apparatus detected no interaction between films or between a film and bare mica; however, addition of Ca²⁺ and ...

The objective of the present study was to determine the prevalence of porcine reproductive and respiratory syndrome virus (PRRSV) antigen-positive uterine tissue in gilts culled due to reproductive disturbance in relation to age at culling, reasons for culling, herds, and PRRSV vaccination. Uterine tissues of 100 gilts from six swine herds in Thailand were collected. The immunohistochemistry was ...

In a previous study we applied a three-step avidin-biotin pretargeting approach to target 90Y-biotin to the tumour in patients with recurrent high grade glioma. The encouraging results obtained in this phase I-II study prompted us to apply the same approach in an adjuvant setting, to evaluate (i) time to relapse and (ii) overall survival. We enrolled 37 high grade glioma ...

In a previous study we applied a three-step avidin�biotin pretargeting approach to target ⁹⁰Y-biotin to the tumour in patients with recurrent high grade glioma. The encouraging results obtained in this phase I�II study prompted us to apply the same approach in an adjuvant setting, to evaluate (i) time to relapse and (ii) overall survival. We enrolled ...

We have performed nine CD34 selection procedures on peripheral blood stem cells harvested from eight patients with myeloma using the Cellpro avidin-biotin immunoaffinity column (Ceprate). They all received CVAMP chemotherapy to maximum response prior to mobilisation. Six of the patients have been transplanted using these cells, one receiving successive autografts. Median ...

The evolution of mammalian brain function depends in part on levels of natural, heritable variation in numbers, location, and function of neurons. However, the nature and amount of natural genetic variation in neural traits and their physiological link to variation in function or evolutionary change are unknown. We estimated the level of within-population heritable variation in the number of ...

Mouse monoclonal antibodies (OV-TL 3) were raised against human ovarian tumor-associated antigens for diagnostic purposes. A cloned hybridoma cell line was obtained by fusion of murine myeloma cells with spleen lymphocytes from BALB/c mice immunized with a tumor cell suspension prepared from an ovarian endometrioid carcinoma. The antibodies were initially screened for their ability to bind on ...

Recent publications suggest the utility of temozolomide (TMZ) in the management of aggressive pituitary adenomas and carcinomas, resistant to conventional treatments. The response to TMZ is inversely correlated with tumoral expression of O-6 methylguanine DNA methyl transferase (MGMT). Therefore, we aimed to assess MGMT immunoexpression in pure GH-secreting pituitary adenomas, in an effort to ...

Angiogenic factors, such as vascular endothelial growth factor (VEGF), its receptors and epidermal growth factor receptor (EGF-R), are involved in increased progression in many carcinomas. The aim of this study was to investigate the role of angiogenesis and immunolocalization of VEGF, its receptors, EGF-R and Ki 67 in leiomyomas and leiomyosarcomas using an indirect immunohistochemical method. ...

Proliferating cells have been immunophenotypically characterized in lymph node and bronchoalveolar lavage (BAL) samples obtained from patients with active and inactive sarcoidosis with the cell-cycle-related antigen Ki67. Ki67 monoclonal antibody was used by combined immunohistochemical methods together with antibodies recognizing macrophage- and T-cell-subset-related antigens using ...

F. Schmitt, B. Cochand-Priollet, M. Toetsch, B. Davidson, A. Bondi and P. Vielh Immunocytochemistry in Europe: results of the European Federation of Cytology Societies (EFCS) inquiry Objective:? This report describes the results of the first chosen topic of the European Federation of Cytology Societies (EFCS) scientific committee, which concerns the application of immunocytochemistry (ICC) to ...

Commercially-available monoclonal antibodies to B lymphocytes were evaluated for immunohistochemical staining of feline B lymphocytes in frozen and formalin-fixed, paraffin-embedded tissues using an avidin biotin complex immunoperoxidase immunohistochemical technique. Three monoclonal antibodies: F46A and F72A raised to "carnivore" B lymphocytes and ...

Samples of intestinal content from thirty fattened pigs of six farms slaughtered at an abattoir in North-Western Germany, and faecal samples of four pigs kept as laboratory animals at the Federal Institute for Risk Assessment (BfR, Berlin, Germany) were investigated for the occurrence of microsporidia by light microscopy, PCR and sequencing. A modified Webers trichrome staining and the ...

This study was designed to investigate the harmful effects of toluene inhalation in the liver of rats and possible protective effects of melatonin on these detrimental effects. For this purpose, 21 adult male Wistar-albino rats were randomly divided into three equal groups. Animals in group I were used as control. The rats in group II were exposed to toluene (3000 ppm/1 hour/day) for 4 weeks, ...

PurposeTo better understand the histogenesis of ocular hemangioblastomas associated with von Hippel-Lindau (VHL) disease.MethodsWe found that co-expression of Epo and EpoR may mediate developmental stagnation and induce proliferation of hemangioblastoma. All lesions were frozen and/or fixed in formalin and embedded in paraffin. The specimens were sectioned and subjected to routine histology, ...

Aim�To study p53 expression in relation to proliferative status in normal and nondysplastic, dysplastic and malignant lesions of the oral mucosa.Method�The standard avidin-biotin complex (ABC) immunohistochemical staining method was used to study the expression of p53 and Ki67 on frozen sections of oral leukoplakias and carcinomas.Results�Of the leukoplakia and carcinoma ...

Multicellular heterospheroids including two or more cell types have some tissue/organ properties and can be used in cell-to-cell interaction studies. However, the spheroid formation is difficult to control because the adhesion efficacy is different in each cell type. To solve this, we applied a rapid cell-to-cell adhesion method, avidin-biotin (AB) binding, to spheroid ...

Abstract Background: Malaria infection with Plasmodium falciparum is an important basic health problem in the tropical and sub-tropical countries. The standard diagnostic method is blood film examination to visualize parasite morphology. However, in cases of low parasitemia or mixed infection with very low cryptic species, microscopy is not sensitive enough. Therefore, molecular techniques have ...

Monocyte procoagulant activity is mainly due to tissue factor (TF) expression, but functional assays may not be sufficiently accurate in clinical use, making useful a determination of TF antigen level. The aim of this study was to compare the results of one functional and three immunological TF assays (ELISA, immunocytochemical staining on slides and flow immuno cytometric analysis), in normal ...

The following papers are included: "Consortia and Collaborative Research: Getting Started" (Hansen); "Coordination of the Health Care System in the State of Michigan" (Burian, Boyden, Herbert); "Health Promotion and Disease Prevention in Allied Health" (Doiron, Douglas); "Interprofessional Collaboration in the Analysis of Public Policy" (Dunn); "Surveying Research Interests of Allied Health ...

In order to evaluate the correlation between immunohistochemical and morphometric data on the same histological sections, we have developed a flexible color image analyzer (Microcomputer-Assisted Picture Processing System type II, MAPPS-II), and established an effective method to analyze the immunostained colorectal neoplasms based on the color recognition theory of human visual system. Colorectal ...

Electrochemically fabricated nano-composite film of chitosan (CH)-iron oxide (Fe(3)O(4)) has been used to detect gonorrhoea, a sexually transmitted disease (STD) via immobilization of biotinylated probe DNA (BDNA) using avidin-biotin coupling for rapid and specific (mismatch-discriminating) DNA hybridization. The presence of Fe(3)O(4) nanoparticles (?18nm) increases the ...

A microdroplet can act as a high quality factor optical cavity that supports Morphology Dependent Resonances(MDRs). Enhanced radiative energy transfer through these optical resonances can also be utilized as a transduction mechanism for chemical and biological sensing. Enhancement in radiative energy transfer is observed when a donor/acceptor pair is present in the resonant medium of a ...

Background: There has recently been controversy regarding whether clone My10 is superior to clone QBEND-10 for labeling cells of tricholemmal lineage. Moreover, there have been no previous reports on the CD34 expression in human vellus hair follicles. Methods: We performed a comprehensive study of the CD34 expression in human terminal and vellus hair follicles and in 10 tricholemmomas using both ...

Background and purposePrimary and dynamically maintained periprosthetic bone formation is essential for osseointegration of hip implants to host bone. Bone morphogenetic proteins (BMPs) play a role in osteoinductive bone formation. We hypothesized that there is an increased local synthesis of BMPs in the synovial membrane-like interface around aseptically loosened total hip replacement (THR) ...

Implantation of sheet-like liver tissues is a promising method in hepatocyte-based therapies, because angiogenesis is expected to occur upon implantation from the surrounding tissues. In this context, we introduce here a new methodology for the formation of a functional thick hepatic tissue usable for cell sheet technology. First, we report the formation of composite tissue elements in suspension ...

We describe a method for the preparation of novel long (hundreds of nanometers), uniform, inter-molecular G4-DNA molecules composed of four parallel G-strands. The only long continuous G4-DNA reported so far are intra-molecular structures made of a single G-strand. To enable a tetra-molecular assembly of the G-strands we developed a novel approach based on avidin�biotin biological recognition. ...

Antisense radiopharmaceuticals could be used to image gene expression in the brain in vivo, should these polar molecules be made transportable through the blood-brain barrier. The present studies describe an antisense imaging agent comprised of an iodinated peptide nucleic acid (PNA) conjugated to a monoclonal antibody to the rat transferrin receptor by using avidin-biotin ...

Antisense radiopharmaceuticals could be used to image gene expression in the brain in vivo, should these polar molecules be made transportable through the blood�brain barrier. The present studies describe an antisense imaging agent comprised of an iodinated peptide nucleic acid (PNA) conjugated to a monoclonal antibody to the rat transferrin receptor by using ...

Mycobacterium leprae antigens could be detected in the cerebrospinal fluid (CSF) of patients with leprosy, using a monoclonal-antibody-based sandwich immunoradiometric assay (SIRMA). Antigens of 12 kD, 35 kD and 30-40 kD were detected using ML06, ML04, and ML34 monoclonal antibodies, respectively. The 30-40-kD polysaccharide antigen, although present in larger amounts in M. leprae than the 12-kD ...

The occurrence of neoplastic and nonneoplastic dural based masses that mimic meningiomas is infrequent and may not be considered during radiological and intraoperative analysis. We describe single institute study of 20 such rare cases. This study included total of 20 cases of meningioma mimics. The clinical, radiological and histopathological findings were evaluated. Tissue fixed in 10% formalin ...

ObjectivesActively targeting ultrasound contrast agents to tumor vasculature improves contrast-enhanced sonography of tumor angiogenesis. This report summarizes an evaluation of multitargeted microbubbles, comparing single-, dual-, and triple-targeted motifs.MethodsMicrobubbles were avidin-biotin linked to antibodies against mouse ?_V?-integrin, ...

Extremely high affinity and selectivity are two of the most sought-after properties of drug molecules. Selectivity has been difficult to achieve, especially for targets that belong to large families of structurally and functionally related proteins. There are essentially two ways by which selectivity can be improved during lead optimization: a chemical ...

Affinity purification coupled to mass spectrometry provides a reliable method for identifying proteins and their binding partners. In this study we have used Drosophila melanogaster proteins triple tagged with Flag, Strep II, and Yellow fluorescent protein in vivo within affinity pull-down experiments and isolated these proteins in their native complexes ...

Affinity purification coupled to mass spectrometry provides a reliable method for identifying proteins and their binding partners. In this study we have used Drosophila melanogaster proteins triple tagged with Flag, Strep II, and Yellow fluorescent protein in vivo within affinity pull-down experiments and isolated these proteins in their native complexes ...

Protein engineering is becoming increasingly important for pharmaceutical applications where controlling the specificity and affinity of engineered proteins is required to create targeted protein therapeutics. Affinity increases of several thousand-fold are now routine for a variety of protein engineering approaches, and the structural and energetic bases ...

Affinity maturation is an important part of the therapeutic antibody development process as in vivo activity often requires high binding affinity. Here, we describe a targeted approach for affinity improvement of therapeutic antibodies. Sets of CDR residues that are solvent accessible and relatively diverse in natural antibodies are ...

An androgen affinity label, 17 beta-[(bromoacetyl)-oxy]-5 alpha-androstan-3-one, has been synthesized in both radioactive and nonradioactive forms. The affinity label (170 Ci/mmol) was characterized and found to have a high degree of purity. Affinity labeling of the androgen receptor from rat ventral prostate was androgen specific and ...

We report on recent progress toward development of secondary emission ''amplifiers'' for photocathodes. Secondary emission gain of over 300 has been achieved in transmission mode and emission mode for a variety of diamond samples. Techniques of sample preparation, including hydrogenation to achieve negative electron ...

Semisynthetic aminoglycoside derivatives may provide a means to selectively target viral RNA sites, including the HIV-1 Rev response element (RRE). The design, synthesis, and evaluation of derivatives based upon neomycin B, kanamycin A, and tobramycin conjugates of 9-aminoacridine are presented. To evaluate the importance of the acridine moiety, a series of dimeric aminoglycosides as well as ...

Primary cultures of rabbit articular chondrocytes have been maintained for 10 days and labeled with [35S]sulfate, [3H]leucine, and [35S]cysteine in pulse-chase protocols to study the structure and hyaluronate binding properties of newly synthesized proteoglycan monomers. Radiolabeled monomers were purified from medium and cell-layer fractions by dissociative CsCl gradient centrifugation with ...