Simultaneous measurement of RBC velocity, flux, hematocrit and shear rate in vascular networks.
AuthorsKamoun WS, Chae SS, Lacorre DA, Tyrrell JA, Mitre M, Gillissen MA, Fukumura D, Jain RK, Munn LL,
JournalNat Methods
PubMed ID20581828
Not all tumor vessels are equal. Tumor-associated vasculature includes immature vessels, regressing vessels, transport vessels undergoing arteriogenesis and peritumor vessels influenced by tumor growth factors. Current techniques for analyzing tumor blood flow do not discriminate between vessel subtypes and only measure average changes from a population of dissimilar vessels. We ... More
Fluorescein and tetramethyl rhodamine as haptens in enzyme immunohistochemistry.
AuthorsHaaijman JJ, Coolen J, Kröse CJ, Pronk GJ, Ming ZF
JournalHistochemistry
PubMed ID3522496
'Fluorescein (Fl) and tetramethyl rhodamine (Rh) were evaluated as possible candidates for a double hapten sandwich system in enzyme immunohistology. Monoclonal antibodies were raised against Fl and Rh. Their fine-specificity was tested with a competition-like assay. A pair of Mab''s was selected for immunohistology in which they functioned as a ... More
Disruption of nuclear lamin organization blocks the elongation phase of DNA replication.
AuthorsMoir RD, Spann TP, Herrmann H, Goldman RD
JournalJ Cell Biol
PubMed ID10851016
'The role of nuclear lamins in DNA replication is unclear. To address this, nuclei were assembled in Xenopus extracts containing AraC, a reversible inhibitor that blocks near the onset of the elongation phase of replication. Dominant-negative lamin mutants lacking their NH(2)-terminal domains were added to assembled nuclei to disrupt lamin ... More
Resolution of fluorescence correlation measurements.
AuthorsMeseth U, Wohland T, Rigler R, Vogel H
JournalBiophys J
PubMed ID10049342
'The resolution limit of fluorescence correlation spectroscopy for two-component solutions is investigated theoretically and experimentally. The autocorrelation function for two different particles in solution were computed, statistical noise was added, and the resulting curve was fitted with a least squares fit. These simulations show that the ability to distinguish between ... More
Signal- and energy-dependent nuclear transport of SV40 Vp3 by isolated nuclei. Establishment of a filtration assay for nuclear protein import.
AuthorsDean DA, Kasamatsu H
JournalJ Biol Chem
PubMed ID8106464
'Nuclear transport signal (NTS)-containing proteins are transported into the nucleus through the nuclear pore complex by a mechanism that is not well understood. To better characterize the mechanisms of transport, we have established an homologous in vitro system using an NTS-containing structural protein of simian virus 40 (SV40) and isolated ... More
Calmodulin stabilization of kinetochore microtubule structure to the effect of nocodazole.
AuthorsSweet SC, Rogers CM, Welsh MJ
JournalJ Cell Biol
PubMed ID3198685
'To investigate the function of calmodulin (CaM) in the mitotic apparatus, the effect of microinjected CaM and chemically modified CaMs on nocodazole-induced depolymerization of spindle microtubules was examined. When metaphase PtK1 cells were microinjected with CaM or a CaM-TRITC conjugate, kinetochore microtubules (kMTs) were protected from the effect of nocodazole. ... More
Optimized conditions to couple two water-soluble biomolecules through alkylamine thiolation and thioetherification.
AuthorsMeunier L, Bourgerie S, Mayer R, Roche AC, Monsigny M
JournalBioconjug Chem
PubMed ID10077469
'A simple method for introducing, in buffered saline, a reactive sulfhydryl group on water-soluble molecules bearing an alkyl-amino group is described. This method is based on the use of two water-soluble reagents: 2-iminothiolane and 6,6''-dithiodinicotinic acid. The first one is open upon reaction with an amino group, and the generated ... More
Trafficking of Shigella lipopolysaccharide in polarized intestinal epithelial cells.
'Bacterial lipopolysaccharide (LPS) at the apical surface of polarized intestinal epithelial cells was previously shown to be transported from the apical to the basolateral pole of the epithelium (Beatty, W.L., and P.J. Sansonetti. 1997. Infect. Immun. 65:4395-4404). The present study was designed to elucidate the transcytotic pathway of LPS and ... More
Molecular motors are differentially distributed on Golgi membranes from polarized epithelial cells.
AuthorsFath KR, Trimbur GM, Burgess DR
JournalJ Cell Biol
PubMed ID8045931
'Microtubules (MT) are required for the efficient transport of membranes from the trans-Golgi and for transcytosis of vesicles from the basolateral membrane to the apical cytoplasm in polarized epithelia. MTs in these cells are primarily oriented with their plus ends basally near the Golgi and their minus-ends in the apical ... More
Passive and facilitated transport in nuclear pore complexes is largely uncoupled.
'Nuclear pore complexes provide the sole gateway for the exchange of material between nucleus and cytoplasm of interphase eukaryotic cells. They support two modes of transport: passive diffusion of ions, metabolites, and intermediate-sized macromolecules and facilitated, receptor-mediated translocation of proteins, RNA, and ribonucleoprotein complexes. It is generally assumed that both ... More
Quantitation of the volume of liquid injected into cells by means of pressure.
AuthorsMinaschek G, Bereiter-Hahn J, Bertholdt G
JournalExp Cell Res
PubMed ID2767158
'The amount of TRITC-labeled bovine serum albumin (TRITC-BSA) released from the tip of standardized microcapillaries at different injection pressures and times was investigated. Three test systems were used: (a) formation of droplets of the test solution (TRITC-BSA) in paraffin oil, (b) injection of the test solution into buffer droplets suspended ... More
Cellular remodeling of HCO3(-)-secreting cells in rabbit renal collecting duct in response to an acidic environment.
AuthorsSatlin LM, Schwartz GJ
JournalJ Cell Biol
PubMed ID2549077
'The renal cortical collecting duct (CCD) consists of principal and intercalated cells. Two forms of intercalated cells, those cells involved in H+/HCO3- transport, have recently been described. H+-secreting cells are capable of apical endocytosis and have H+ATPase on the apical membrane and a basolateral Cl-/HCO3- exchanger. HCO3(-)-secreting cells bind peanut ... More
Spatial and temporal imaging of gas phase protein and DNA produced by matrix-assisted laser desorption.
AuthorsHeise TW, Yeung ES
JournalAnal Chem
PubMed ID8135376
'The sensitivity of a transient imaging technique based on laser-excited fluorescence is shown to be within the levels required for studying matrix-assisted laser-desorption (MALD) of large biomolecules. Results show that film morphology, particularly film thickness, has a major influence on plume dynamics. Fluorescent labeling of protein and of DNA is ... More
What do measures of flux tell us about vascular wall biology?
AuthorsHuxley VH
JournalMicrocirculation
PubMed ID9789252
A novel flow cytometric method for measuring protein digestion within the phagocytic vacuole of polymorphonuclear neutrophils.
AuthorsHaynes AP, Fletcher J, Garnett M, Robins A
JournalJ Immunol Methods
PubMed ID2273255
When rhodamine is attached to albumin at a high molar ratio its fluorescence is quenched but fluorescence is released when the protein is digested and the dye released. Using this observation it is possible to measure protein digestion within the phagocytic vacuole of neutrophils. The assay is simple, rapid and ... More
Adhesive and mammalian transglutaminase substrate properties of Candida albicans Hwp1.
AuthorsStaab JF, Bradway SD, Fidel PL, Sundstrom P
JournalScience
PubMed ID10066176
The pathogenesis of candidiasis involves invasion of host tissues by filamentous forms of the opportunistic yeast Candida albicans. Morphology-specific gene products may confer proinvasive properties. A hypha-specific surface protein, Hwp1, with similarities to mammalian small proline-rich proteins was shown to serve as a substrate for mammalian transglutaminases. Candida albicans strains ... More
A photoactivatable GFP for selective photolabeling of proteins and cells.
AuthorsPatterson GH, Lippincott-Schwartz J
JournalScience
PubMed ID12228718
We report a photoactivatable variant of the Aequorea victoria green fluorescent protein (GFP) that, after intense irradiation with 413-nanometer light, increases fluorescence 100 times when excited by 488-nanometer light and remains stable for days under aerobic conditions. These characteristics offer a new tool for exploring intracellular protein dynamics by tracking ... More
Early endosome membrane dynamics characterized by flow cytometry.
AuthorsChavrier P, van der Sluijs P, Mishal Z, Nagelkerken B, Gorvel JP
JournalCytometry
PubMed ID9298810
Early endosomes are very dynamic intracellular membrane organelles that undergo multiple fusion and fission events. In this study, we developed a novel assay based on multiparametric flow cytometric analyses and early endosome sorting to characterize better the mechanisms of early endosome membrane dynamics in vitro. In particular, we have investigated ... More
A bioassay based on the ultrafast response of a reporter molecule.
AuthorsQuentmeier CC, Wehling A, Walla PJ,
JournalJ Biomol Screen
PubMed ID17307887
The capability of using ultrafast detection technologies for a fast analysis of biomolecular reactions has been explored. As an example, the ultrafast response of tetramethylrhodamine (TMR)-labeled bovine serum albumin (BSA) as a function of different extents in proteolytic cleavage was investigated. The authors compared 4 samples of masses differing over ... More
Transient diffusion of albumin in aortic walls: effects of binding to medial elastin layers.
AuthorsGoriely AR, Baldwin AL, Secomb TW,
JournalAm J Physiol Heart Circ Physiol
PubMed ID17208986
The goal of this study was to measure diffusive transport of albumin through artery walls experimentally and to analyze the results theoretically, taking into account the binding of albumin to elastic lamellae. Segments of rabbit aorta were placed in solutions of fluorescently labeled albumin for periods of 30, 60, 90, ... More
Basal and adenosine-mediated protein flux from isolated coronary arterioles.
AuthorsHuxley VH, Williams DA
JournalAm J Physiol
PubMed ID8853347
We have developed a new method to quantify solute flux per unit surface area and concentration gradient (JS/S delta C) from arterioles isolated from pig hearts. The apparent permeability (Ps) was assessed from measurements of JS/S delta C for two proteins, alpha-lactalbumin (alpha-lactalb) and porcine serum albumin (PSA), labeled with ... More
Low-efficiency (macro-)pinocytic internalization of non-pathogenic Escherichia coli into HEp-2 cells.
AuthorsSinai AP, Hayes SF, Small PL, Bavoil PM
JournalRes Microbiol
PubMed ID8584786
HEp-2 cells internalize non-pathogenic Escherichia coli bacteria by a low-efficiency internalization mechanism which is upregulated in Pho-derepressed strains (as shown by Sinai and Bavoil in 1993), and is independent of microfilament integrity but requires functional microtubules. Here, we further characterize the microtubule requirement of this pathway using various effectors of ... More
Differential phototoxicity of fluorescent dye-labeled albumin conjugates.
AuthorsRumbaut RE, Sial AJ
JournalMicrocirculation
PubMed ID10501094
OBJECTIVE: Fluorescent dyes are commonly used as probes for assessment of macromolecular permeability. Despite numerous examples of light-dye induced toxicity in the microvasculature, little is known regarding the relative phototoxicity of commonly used fluorescent conjugates. We, therefore, compared the phototoxicity of four fluorescent conjugates of bovine serum albumin (BSA) available ... More
Monolithic triglyceride matrices: a controlled-release system for proteins.
AuthorsVogelhuber W, Magni E, Mouro M, Spruss T, Guse C, Gazzaniga A, Göpferich A
JournalPharm Dev Technol
PubMed ID12665199
Matrices made of glyceryl trimyristate as a bioerodible and biocompatible material were manufactured by compression in dimensions that would still allow an application via injection. Pyranine, as a low molecular hydrophilic compound with a low detection limit, and tetramethylrhodamine labeled bovine serum albumin (TAMRA-BSA), as a high molecular weight (66 ... More
Measurement of material extravasation in microvascular networks using fluorescence video-microscopy.
We have developed a new method using fluorescence videomicroscopy to quantitate the extravasation of intravenously injected materials. This method can measure the relative plasma concentration of, and the vascular permeability to, these materials in microcirculatory preparations which contain multiple blood vessels in a field of view. The image of a ... More
A multienzyme network functions in intestinal protein digestion by a platyhelminth parasite.
AuthorsDelcroix M, Sajid M, Caffrey CR, Lim KC, Dvorák J, Hsieh I, Bahgat M, Dissous C, McKerrow JH
JournalJ Biol Chem
PubMed ID17028179
Proteases frequently function not only as individual enzymes but also in cascades or networks. A notable evolutionary switch occurred in one such protease network that is involved in protein digestion in the intestine. In vertebrates, this is largely the work of trypsin family serine proteases, whereas in invertebrates, cysteine proteases ... More
Contact sensitizers modulate mechanisms of receptor-mediated endocytosis but not fluid-phase endocytosis in murine epidermal Langerhans cells.
AuthorsBecker D, Lempertz U, Enk A, Saloga J, Knop J
JournalExp Dermatol
PubMed ID8535616
In order to define the influence of contact allergens on the fluid-phase endocytosis (FPE) of soluble molecules of murine epidermal Langerhans cells (LC), we studied the internalization of FITC-labeled bovine serum albumin (FITC-BSA), TRITC-labeled dextrane (TRITC-DEX) as well as horseradish peroxidase by LC. A 3-parameter flow-cytometric technique was performed for ... More
Fluorescent characterization of collecting duct cells: a second H+-secreting type.
AuthorsSchwartz GJ, Satlin LM, Bergmann JE
JournalAm J Physiol
PubMed ID2461095
We have used three fluorescent probes to label acid-base transporting cells with specific physiological properties in the rabbit collecting duct. Rhodamine albumin identified cells active in luminal endocytosis; rhodamine peanut agglutinin (PNA) identified cells with apical surface PNA ligands; and 6-carboxyfluorescein (6-CF) diacetate identified cells with alkaline pH or acetazolamide-sensitive ... More
The stiffness of rabbit skeletal actomyosin cross-bridges determined with an optical tweezers transducer.
AuthorsVeigel C, Bartoo ML, White DC, Sparrow JC, Molloy JE
JournalBiophys J
PubMed ID9726944
Muscle contraction is brought about by the cyclical interaction of myosin with actin coupled to the breakdown of ATP. The current view of the mechanism is that the bound actomyosin complex (or "cross-bridge") produces force and movement by a change in conformation. This process is known as the "working stroke." ... More
Dynamic intravital fluorescence microscopy--a novel method for the assessment of microvascular permeability in acute pancreatitis.
Authorsvon Dobschuetz E, Pahernik S, Hoffmann T, Kiefmann R, Heckel K, Messmer K, Mueller-Hoecker J, Dellian M
JournalMicrovasc Res
PubMed ID14709403
Edema formation is the first manifestation of acute pancreatitis. Microcirculatory derangements like leukocyte-endothelial cell interaction and perfusion failure result in enhancement of microvascular permeability to large molecules playing a pivotal role in the progression of the acutely altered pancreatic tissue. Due to the lack of suitable methods the crucial mechanisms ... More
Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion.
Authors
JournalAutophagy
PubMed ID29940786
In utero estrogenic endocrine disruption alters the stroma to increase extracellular matrix density and mammary gland stiffness.
Authors
JournalBreast Cancer Res
PubMed ID32370801
Universal autofocus for quantitative volumetric microscopy of whole mouse brains.
Authors
JournalNat Methods
PubMed ID34312564
The intracellular fate of zonula occludens 2 is regulated by the phosphorylation of SR repeats and the phosphorylation/O-GlcNAcylation of S257.
Authors
JournalMol Biol Cell
PubMed ID23804652
Proteolytically released Lasso/teneurin-2 induces axonal attraction by interacting with latrophilin-1 on axonal growth cones.
Authors
JournalElife
PubMed ID30457553
In vivo label-free measurement of lymph flow velocity and volumetric flow rates using Doppler optical coherence tomography.
Authors
JournalSci Rep
PubMed ID27377852
Distinct forms of the actin cross-linking protein α-actinin support macropinosome internalization and trafficking.
Authors
JournalMol Biol Cell
PubMed ID34010028
A Surface Plasmon Resonance Plastic Optical Fiber Biosensor for the Detection of Pancreatic Amylase in Surgically-Placed Drain Effluent.
Authors
JournalSensors (Basel)
PubMed ID34063347
A cerebellopontine angle mouse model for the investigation of tumor biology, hearing, and neurological function in NF2-related vestibular schwannoma.
Authors
JournalNat Protoc
PubMed ID30617350
Combined Rehabilitation Promotes the Recovery of Structural and Functional Features of Healthy Neuronal Networks after Stroke.