Comparative analysis of glutamate transporter expression in rat brain using differential double in situ hybridization.
AuthorsBerger UV, Hediger MA
JournalAnat Embryol (Berl)
PubMed ID9683064
This study compares the mRNA expression pattern for the three glutamate transporters EAAC1, GLT1 and GLAST in rat brain, using a sensitive non-radioactive in situ hybridization technique. The results confirm the predominantly neuronal localization of EAAC1 mRNA, the astroglial and ependymal localization of GLAST mRNA and the astroglial and neuronal ... More
Co-localization of synaptophysin with different neuroendocrine hormones in the human gastrointestinal tract.
AuthorsPortela-Gomes GM, Stridsberg M, Johansson H, Grimelius L
JournalHistochem Cell Biol
PubMed ID9930883
Colocalisation of synaptophysin has been studied in different neuroendocrine cell types in histologically normal mucosa from human gastrointestinal tract (corpus, antrum, duodenum, ileum and colon) using double-immunofluorescence stainings. Numerous synaptophysin immunoreactive cells were seen in the antrum, while a smaller number were found in the intestinal tract. Synaptophysin immunoreactivity was ... More
Detection of CD5 antigen on B cell lymphomas in fixed, paraffin embedded tissues using signal amplification by catalyzed reporter deposition.
AuthorsLuo JH, Matsushima AY, Chen R, Szabolcs MJ
JournalEur J Histochem
PubMed ID9615189
CD5 surface antigen is expressed on some categories of B cell lymphomas. The detection of CD5 coexpression on malignant B cell infiltrates, particularly in small biopsy specimens, is useful in distinguishing between small lymphocytic lymphoma, mantle cell lymphoma, low grade marginal zone B cell lymphoma, and follicular small cleaved cell ... More
In situ identification of cyanobacteria with horseradish peroxidase-labeled, rRNA-targeted oligonucleotide probes.
AuthorsSchönhuber W, Zarda B, Eix S, Rippka R, Herdman M, Ludwig W, Amann R
JournalAppl Environ Microbiol
PubMed ID10049892
Individual cyanobacterial cells are normally identified in environmental samples only on the basis of their pigmentation and morphology. However, these criteria are often insufficient for the differentiation of species. Here, a whole-cell hybridization technique is presented that uses horseradish peroxidase (HRP)-labeled, rRNA-targeted oligonucleotides for in situ identification of cyanobacteria. This ... More
Rapid synthesis of biotin-, digoxigenin-, trinitrophenyl-, and fluorochrome-labeled tyramides and their application for In situ hybridization using CARD amplification.
AuthorsHopman AH, Ramaekers FC, Speel EJ
JournalJ Histochem Cytochem
PubMed ID9603790
A one-step procedure for the synthesis of different tyramide conjugates, which can be utilized in the catalyzed reporter deposition (CARD) amplification system, is described. Succinimidyl esters of biotin, digoxigenin, and of the fluorochromes fluorescein, rhodamine, aminomethylcoumarine acetic acid, and Cy3 were coupled to tyramine in dimethylformamide (DMF) adjusted to a ... More
Detection of oestrogen receptors in non-invasive and invasive transitional cell carcinomas of the urinary bladder using both conventional immunohistochemistry and the tyramide staining amplification (TSA) technique.
AuthorsKaufmann O, Baume H, Dietel M
JournalJ Pathol
PubMed ID9924432
In a retrospective immunohistochemical study, the expression of oestrogen receptors has been investigated in paraffin-embedded transitional cell carcinomas of the urinary bladder with antibody 6F11, using both a conventional immunohistochemical detection system and the highly sensitive tyramide staining amplification (TSA) technique. The study comprised 88 invasive and 97 non-invasive carcinomas ... More
Confocal microscopy assessment of lymphoid tissues with follicular hyperplasia from patients infected with human immunodeficiency virus type 1.
AuthorsSoontornniyomkij V, Wang G, Kapadia SB, Achim CL, Wiley CA
JournalArch Pathol Lab Med
PubMed ID9625421
OBJECTIVE: To characterize human immunodeficiency virus (HIV) infection of lymphoid tissues during follicular hyperplasia. METHODS: We examined 10 tonsil/adenoid, 3 parotid lymphoepithelial cyst, and 7 lymph node specimens that had been surgically removed from 13 patients infected with HIV-1. Characteristics of productive HIV-1 infection were assessed using immunocytochemistry for HIV-1 ... More
Advances in fluorescence in situ hybridization.
AuthorsRaap AK
JournalMutat Res
PubMed ID9685683
The techniques of in situ hybridization (ISH) are widely applied for analyzing the genetic make-up and RNA expression patterns of individual cells. This review focusses on a number of advances made over the last 5 years in the fluorescence ISH (FISH) field, i.e., Fiber-FISH, Multi-colour chromosome painting, Comparative Genomic Hybridization, ... More
Expression of brain-derived neurotrophic factor protein in activated microglia of human immunodeficiency virus type 1 encephalitis.
AuthorsSoontornniyomkij V, Wang G, Pittman CA, Wiley CA, Achim CL
JournalNeuropathol Appl Neurobiol
PubMed ID9888155
The role of neurotrophic factors and their therapeutic potential have been investigated in various neurodegenerative disorders. In neurodegeneration associated with human immunodeficiency virus (HIV) infection, neuronal function and survival may be affected by abnormal neurotrophic regulation involving HIV-infected microglia and reactive astrocytes. To characterize the cellular localization of brain-derived neurotrophic ... More
Mapping ESTs by fiber-FISH.
AuthorsHorelli-Kuitunen N, Aaltonen J, Yaspo ML, Eeva M, Wessman M, Peltonen L, Palotie A
JournalGenome Res
PubMed ID9927485
A visual transcript map of six genes was constructed on the chromosome 21q22.3 by high resolution fluorescence in situ hybridization (FISH). Expressed sequence tags (ESTs) from six genes-PWP2, KNP1, AIRE, C21orf3, SMT3A, and C21orf1-were successfully localized by fiber-FISH by use of sensitive tyramide-based detection. The sizes of the ESTs varied ... More
The EnVision++ system: a new immunohistochemical method for diagnostics and research. Critical comparison with the APAAP, ChemMate, CSA, LABC, and SABC techniques.
AuthorsSabattini E, Bisgaard K, Ascani S, Poggi S, Piccioli M, Ceccarelli C, Pieri F, Fraternali-Orcioni G, Pileri SA
JournalJ Clin Pathol
PubMed ID9797726
AIM: To assess a newly developed immunohistochemical detection system, the EnVision++. METHODS: A large series of differently processed normal and pathological samples and 53 relevant monoclonal antibodies were chosen. A chessboard titration assay was used to compare the results provided by the EnVision++ system with those of the APAAP, CSA, ... More
Comparison of an amplified enzyme-linked immunosorbent assay with procedures based on molecular biology for assessing human immunodeficiency virus type 1 viral load.
AuthorsGoldschmidt PL, Devillechabrolle A, Ait-Arkoub Z, Aubin JT
JournalClin Diagn Lab Immunol
PubMed ID9665959
The sensitivity of the enzyme-linked amplified sorbent test (ELAST) was compared with those of other classic enzyme-linked immunosorbent assays (ELISAs), with or without previous acidic immunocomplex dissociation (ICD), in a series of samples at different stages of human immunodeficiency virus type 1 (HIV-1) infection. The limit of viral detection of ... More
Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family.
Amino-acid transport across cellular plasma membranes depends on several parallel-functioning (co-)transporters and exchangers. The widespread transport system L accounts for a sodium-independent exchange of large, neutral amino acids, whereas the system y(+)L exchanges positively charged amino acids and/or neutral amino acids together with sodium. The molecular nature of these transporters ... More
Signal amplification at the ultrastructural level using biotinylated tyramides and immunogold detection.
AuthorsSchöfer C, Weipoltshammer K, Almeder M, Wachtler F
JournalHistochem Cell Biol
PubMed ID9387923
The tyramide amplification technique has recently been developed for signal enhancement in enzyme-linked immunosorbent assays and western blots. This method relies on using labelled tyramides as substrates for peroxidase, resulting in an immobilization of the labelled tyramide residues (tyramide reaction). We succeeded in establishing reliable protocols for the use of ... More
Sensitive in situ hybridization with catalyzed reporter deposition, streptavidin-Nanogold, and silver acetate autometallography: detection of single-copy human papillomavirus.
AuthorsZehbe I, Hacker GW, Su H, Hauser-Kronberger C, Hainfeld JF, Tubbs R
JournalAm J Pathol
PubMed ID9137082
The usefulness of standard in situ hybridization for viral nucleic acid detection is occasionally limited by its sensitivity limit of 10 to 50 copies per cell. A modified version of the recently described signal amplification method, catalyzed reporter deposition (CARD), and its application to formalin-fixed cells and tissue sections is ... More
Constant detection of CD2, CD3, CD4, and CD5 in fixed and paraffin-embedded tissue using the peroxidase-mediated deposition of biotin-tyramide.
AuthorsMalisius R, Merz H, Heinz B, Gafumbegete E, Koch BU, Feller AC
JournalJ Histochem Cytochem
PubMed ID9389770
Immunohistochemical methods are widely used for diagnostic purposes in histopathology. However, the use of most monoclonal anti-leukocyte antibodies is limited to frozen tissues. Initially, it was believed that formalin fixation in particular, which is the gold standard for morphological tissue preservation, destroys most of the antigen binding sites. In recent ... More
Immunohistochemical visualization of wild-type p53 protein in paraffin-embedded rat liver using tyramide amplification: zonal hepatic distribution of p53 protein after N-hydroxy-2-acetylaminofluorene administration.
Authorsvan Gijssel HE, van Gijlswijk RP, de Haas RR, Stark C, Mulder GJ, Meerman JH
JournalCarcinogenesis
PubMed ID9472715
P53 protein plays an important role in regulation of the cell cycle. Recently, a role in tumour genesis has also been suggested. The protein is induced after various forms of DNA damage. Immunohistochemical detection of p53 protein showed positive cells in human skin after UV-irradiation, in mouse skin after benzo[a]pyrene ... More
Simple monitoring of antiretroviral therapy with a signal-amplification-boosted HIV-1 p24 antigen assay with heat-denatured plasma.
AuthorsBöni J, Opravil M, Tomasik Z, Rothen M, Bisset L, Grob PJ, Lüthy R, Schüpbach J
JournalAIDS
PubMed ID9143600
OBJECTIVE: Virus load determination has become indispensable for the management of HIV patients, but depends on expensive assays of a low throughput. We evaluated whether a highly improved HIV-1 p24 antigen detection procedure which involves heat-mediated immune complex dissociation and signal-amplification-boosted enzyme-linked immunosorbent assay (ELISA) was suitable for antiretroviral treatment ... More
In situ polymerase chain reaction amplification of HIV-1 DNA in brain tissue.
AuthorsStrappe PM, Wang TH, McKenzie CA, Lowrie S, Simmonds P, Bell JE
JournalJ Virol Methods
PubMed ID9562406
A direct in situ polymerase chain reaction (IS-PCR) assay is described for the detection of HIV-1 proviral DNA in formalin fixed paraffin embedded brain tissue. Biotin-16-dUTP is incorporated during the PCR process and microwave pretreatment of tissue sections ensures that no non-specific incorporation into damaged or nicked genomic DNA occurs. ... More
Signal amplification of FISH for automated detection using image cytometry.
The purpose of this study was to improve the detection of FISH signals, in order that spot counting by a fully automated image cytometer be comparable to that obtained visually under the microscope. Two systems of spot scoring, visual and automated counting, were investigated in parallel on stimulated human lymphocytes ... More
High-performance liquid chromatographic separation of microcystins derivatized with a highly fluorescent dienophile.
AuthorsHarada K, Oshikata M, Shimada T, Nagata A, Ishikawa N, Suzuki M, Kondo F, Shimizu M, Yamada S
JournalNat Toxins
PubMed ID9496379
Microcystins are potent hepatotoxins produced by cyanobacteria, and are also tumor promoters as well as potent inhibitors of the catalytic subunits of protein phosphatases 1 and 2A. In order to establish a physicochemical method for individual detection and determination of trace amounts of microcystins, we developed a derivatization method for ... More
Target and signal amplification: approaches to increase the sensitivity of in situ hybridization.
AuthorsKomminoth P, Werner M
JournalHistochem Cell Biol
PubMed ID9387925
In situ hybridization (ISH) has proven to be a very important molecular tool in research and diagnosis. However, its applicability can be limited by its restricted detection sensitivity. During the last few years, several strategies have been developed to improve the threshold levels for ISH detection by amplification of either ... More
[New fluorogenic dienophile: synthesis, reaction with vitamin D, vitamin A and microcystins, and application to fluorometric assays]
AuthorsShimizu M, Iwasaki Y, Yamada S
JournalYakugaku Zasshi
PubMed ID7473056
We designed and synthesized a new sensitive and highly reactive fluorogenic reagent (1a, DMEQ-TAD) targeting a conjugated diene. DMEQ-TAD reacted quantitatively with major vitamin D metabolites and synthetic analogs under a variety of conditions to yield the corresponding 6,19-cycloadducts as a mixture of the C(6) epimers. The stereochemistry of the ... More
Ultra-sensitive FISH using peroxidase-mediated deposition of biotin- or fluorochrome tyramides.
AuthorsRaap AK, van de Corput MP, Vervenne RA, van Gijlswijk RP, Tanke HJ, Wiegant J
JournalHum Mol Genet
PubMed ID7633400
We describe a detection principle for indirect fluorescence in situ hybridization (FISH) methods that with only one or two antibody layers dramatically improves FISH signal intensities. The method uses as a first layer an anti-hapten immunoglobulin [or (strept)avidin] conjugated to peroxidase. The quintessence of the method is the use of ... More
Expression of tryptophan 5-hydroxylase gene during sea urchin neurogenesis and role of serotonergic nervous system in larval behavior.
AuthorsYaguchi S, Katow H
JournalJ Comp Neurol
PubMed ID14528449
Tryptophan 5-hydroxylase (TPH) is the rate-limiting enzyme in the biosynthesis of serotonin. cDNA cloning of TPH was carried out, and the occurrence of spatiotemporal transcription of TPH message was examined in larvae of the sea urchin, Hemicentrotus pulcherrimus (HpTPH), with in situ hybridization by using the tyramide signal amplification (TSA) ... More
Protein synthetic machinery in the dendrites of the magnocellular neurosecretory neurons of wild-type Long-Evans and homozygous Brattleboro rats.
AuthorsMa D, Morris JF
JournalJ Chem Neuroanat
PubMed ID11861124
There is growing evidence of local protein synthesis in neuronal dendrites, especially in relation to synaptic activity. The hypothalamic magnocellular system is a robust model for peptidergic neurons, especially for the study of dendrites. Quantitative electron microscopy, immunocytochemistry and non-radioactive in situ hybridization (with tyramide signal amplification) were used to ... More
Protein expression analysis using quantitative fluorescence image analysis on tissue microarray slides.
AuthorsRao J, Seligson D, Hemstreet GP
JournalBiotechniques
PubMed ID11962614
We have developed a tissue microarray (TMA)-based quantitative fluorescence image analysis (QFIA) method in which protein markers on TMA sections were labeled by immunofluorescence using tyramide signal amplification and a quantitative fluorescence detection system. Using this method, BRCA1 protein expression patterns were studied in the TMA sections of cell lines ... More
In situ analysis of the transcriptional activity of integrated viral DNA using tyramide-FISH.
Infection by the oncogenic human papillomavirus (HPV) types 16 and type 18 can progress to cancers. Two well studied cervical carcinoma cell lines, SiHa and CaSki, contain two to four copies, or several hundred copies of integrated HPV-16, respectively. To define the chromosomal loci from which HPV mRNAs are transcribed ... More
Strategies for signal amplification in nucleic acid detection.
AuthorsAndras SC, Power JB, Cocking EC, Davey MR
JournalMol Biotechnol
PubMed ID11697219
Many aspects of molecular genetics necessitate the detection of nucleic acid sequences. Current approaches involving target amplification (in situ PCR, Primed in situ Labeling, Self-Sustained Sequence Replication, Strand Displacement Amplification), probe amplification (Ligase Chain Reaction, Padlock Probes, Rolling Circle Amplification) and signal amplification (Tyramide Signal Amplification, Branched DNA Amplification) are ... More
Neuroendocrine differentiation in carcinoma of the breast. Tyramide signal amplification discloses chromogranin A-positive tumour cells in more breast tumours than previously realized.
AuthorsBofin AM, Qvigstad G, Waldum C, Waldum HL
JournalAPMIS
PubMed ID12529020
The aim of the study was to determine if, by means of tyramide signal amplification (TSA), the presence of chromogranin A (CgA)-positive tumour cells could be demonstrated in breast cancer cases found to be negative by conventional immunohistochemical staining. Sections from 44 cases of breast cancer (28 infiltrating ductal carcinomas, ... More
Recent developments in signal amplification methods for in situ hybridization.
AuthorsQian X, Lloyd RV
JournalDiagn Mol Pathol
PubMed ID12605030
In situ hybridization (ISH) allows for the histologic and cytologic localization of DNA and RNA targets. However, the application of ISH techniques can be limited by their inability to detect targets with low copies of DNA and RNA. During the last few years, several strategies have been developed to improve ... More
Closely related Prochlorococcus genotypes show remarkably different depth distributions in two oceanic regions as revealed by in situ hybridization using 16S rRNA-targeted oligonucleotides.
AuthorsWest NJ, Schönhuber WA, Fuller NJ, Amann RI, Rippka R, Post AF, Scanlan DJ
JournalMicrobiology
PubMed ID11429451
An in situ hybridization method was applied to the identification of marine cyanobacteria assignable to the genus Prochlorococcus using horseradish-peroxidase-labelled 16S rRNA-targeted oligonucleotide probes in combination with tyramide signal amplification (TSA). With this method very bright signals were obtained, in contrast to hybridizations with oligonucleotides monolabelled with fluorochromes, which failed ... More
Detection of gamma-globin mRNA in fetal nucleated red blood cells by PNA fluorescence in situ hybridization.
AuthorsLarsen RD, Schønau A, Thisted M, Petersen KH, Lohse J, Christensen B, Philip J, Pluzek KJ
JournalPrenat Diagn
PubMed ID12533814
OBJECTIVES: Fetal nucleated red blood cells (NRBC) that enter the peripheral blood of the mother are suitable for non-invasive prenatal diagnosis. The application of peptide nucleic acid (PNA) probes for tyramide amplified flow fluorescence in situ hybridization (FISH) detection of gamma-globin mRNA in fixed fetal NRBC is investigated. METHODS: Hemin-induced ... More
The expression of gastric H+-K+-ATPase mRNA and protein in developing rat fundic gland.
AuthorsYang DH, Tsuyama S, Murata F
JournalHistochem J
PubMed ID11508339
The proton pump H+-K+-ATPase is the final common pathway mediating the production and secretion of hydrochloric acid by gastric parietal cells. The present studies were undertaken to examine whether the expression of gastric H+-K+-ATPase mRNA and protein changes are associated with the development of H+-K+-ATPase activity in the rat fundic ... More
Tissue distribution of products of the mouse decay-accelerating factor (DAF) genes. Exploitation of a Daf1 knock-out mouse and site-specific monoclonal antibodies.
AuthorsLin F, Fukuoka Y, Spicer A, Ohta R, Okada N, Harris CL, Emancipator SN, Medof ME
JournalImmunology
PubMed ID11683962
Decay-accelerating factor (DAF) is a membrane regulator of C3 activation that protects self cells from autologous complement attack. In humans, DAF is uniformly expressed as a glycosylphosphatidylinositol (GPI)-anchored molecule. In mice, both GPI-anchored and transmembrane-anchored DAF proteins are produced, each of which can be derived from two different genes (Daf1 ... More
Quantitative assessment of picoeukaryotes in the natural environment by using taxon-specific oligonucleotide probes in association with tyramide signal amplification-fluorescence in situ hybridization and flow cytometry.
AuthorsBiegala IC, Not F, Vaulot D, Simon N
JournalAppl Environ Microbiol
PubMed ID12957941
Picoeukaryotes (cells of <3 micro m in diameter) contribute significantly to marine plankton biomass and productivity, and recently molecular studies have brought to light their wide diversity. Among the methods that have been used so far to quantify aquatic microorganisms, fluorescence in situ hybridization of oligonucleotide probes combined with flow ... More
Application of tyramide signal amplification for detection of N-glycolylneuraminic acid in human hepatocellular carcinoma.
AuthorsKoda T, Aosasa M, Asaoka H, Nakaba H, Matsuda H
JournalInt J Clin Oncol
PubMed ID14586758
BACKGROUND: N-Acetylneuraminic acid and N-glycolylneuraminic acid (NeuGc) are the most common sialic acids in mammals, and NeuGc has attracted attention as a tumor-associated antigen. METHODS: In frozen liver sections from patients with hepatocellular carcinoma, glycolipid-type NeuGc was detected on the surface of liver cancer cells in 9 of 17 samples ... More
A comparison of anti-biotin and biotinylated anti-avidin double-bridge and biotinylated tyramide immunohistochemical amplification.
AuthorsFreedman LJ, Maddox MT
JournalJ Neurosci Methods
PubMed ID11640956
Often it is difficult to detect very small amounts of antigen with conventional immunohistochemical techniques. We evaluate three amplification techniques involving anti-biotin or anti-avidin double-bridges or biotinylated tyramide amplification to enhance the sensitivity of serotonin transporter immunohistochemistry. For the anti-biotin double-bridge, after the secondary antibody, the sections were incubated in ... More
Fluorometric analysis of pectenotoxin-2 in microalgal samples by high performance liquid chromatography.
AuthorsSasaki K, Takizawa A, Tubaro A, Sidari L, Loggia RD, Yasumoto T
JournalNat Toxins
PubMed ID11122513
A rapid HPLC method with fluorescence detection of pectenotoxin-2 (PTX2), a polyether macrolide toxin, in microalgae is presented. A dienophile reagent, DMEQ-TAD, was used for precolumn fluorescence labeling. PTX2 could be quantitatively detected in the range 1-200 ng. This method confirmed the occurrence of PTX2 in net haul samples mostly ... More
Fluorescence in situ hybridization method for co-localization of mRNA and GEP.
AuthorsOliva AA, Swann JW
JournalBiotechniques
PubMed ID11464523
Co-localization studies using green fluorescent protein (GFP) and fluorescence immunohistochemistry have become commonplace. However, co-localization studies using GFP and mRNA in situ hybridization are rare, in large part because typical in situ hybridization reaction conditions often lead to the loss of GFP fluorescence. Here, we describe a new fluorescence mRNA ... More
Early pathogenesis of transmucosal feline immunodeficiency virus infection.
AuthorsObert LA, Hoover EA
JournalJ Virol
PubMed ID12021364
To identify the early target cells and tissues in transmucosal feline immunodeficiency virus (FIV) infection, cats were exposed to a clade C FIV isolate via the oral-nasal or vaginal mucosa and multiple tissues were examined by virus isolation coculture (VI), DNA PCR, catalyzed tyramide signal-amplified in situ hybridization (TSA-ISH), and ... More
Enhanced cyclooxygenase-2 expression in sporadic and familial adenomatous polyposis of the human colon.
AuthorsKhan KN, Masferrer JL, Woerner BM, Soslow R, Koki AT
JournalScand J Gastroenterol
PubMed ID11495083
BACKGROUND: The cyclooxygenase (COX) enzymes exist in two related but unique isoforms (COX-1 and COX-2) and catalyze the formation of prostaglandins (PGs). COX-1 is constitutively expressed, and is responsible for the synthesis of PGs necessary for gastroprotection and normal renal function. The COX-2 isoform is important in a variety of ... More
Visualization of transcription-dependent association of imprinted genes with the nuclear matrix.
AuthorsKagotani K, Nabeshima H, Kohda A, Nakao M, Taguchi H, Okumura K
JournalExp Cell Res
PubMed ID11900479
Genomic imprinting is characterized by allele-specific gene expression as a biological phenomenon. To analyze the participation of the nuclear matrix in the expression of imprinted genes, we first examined the allelic expression state of genes by simultaneously visualizing their primary transcripts and the gene sequences in individual cell nuclei using ... More
Vascular smooth muscle and nitric oxide synthase.
AuthorsBuchwalow IB, Podzuweit T, Bocker W, Samoilova VE, Thomas S, Wellner M, Baba HA, Robenek H, Schnekenburger J, Lerch MM
JournalFASEB J
PubMed ID11919152
The concept of endothelium-derived relaxing factor (EDRF) put forward in 1980 by Furchgott and Zawadzki implies that nitric oxide (NO) produced by NO synthase (NOS) in the endothelium diffuses to the underlying vascular smooth muscle, where it modulates vascular tone as well as vascular smooth muscle cell (VSMC) proliferation by ... More
Tyramide signal amplification enhances the detectable distribution of connexin-43 positive gap junctions across the ventricular wall of the rabbit heart.
AuthorsMcLachlan CS, Jusuf PR, Rummery N, Kummerfeld SK, Hambly B, McGuire MA, Turner V
JournalArch Histol Cytol
PubMed ID14692691
Previous mapping studies examinig the distribution and pattern of staining for connexin-43 expression (the major ventricular gap junction protein) across the ventricular wall have yielded variable findings. The aim of this study was to determine if variations in the distribution of connexin-43 were due to histochemical detection problems, i.e. cross-linking ... More
Immunocytochemical distribution of nitric oxide synthase in the human corpus cavernosum: an electron microscopical study using the tyramide signal amplification technique.
AuthorsStanarius A, Uckert S, Machtens SA, Stief CG, Wolf G, Jonas U
JournalUrol Res
PubMed ID11482440
Nitric oxide has proven to be an important mediator in the relaxation of human cavernosal smooth muscle. Nevertheless, there are many inconsistencies in the literature regarding the cellular and subcellular distribution of endothelial nitric oxide synthase in the human penis. The purpose of this study was to reexamine the localization ... More
Improved detection of differentially expressed genes in microarray experiments through multiple scanning and image integration.
AuthorsRomualdi C, Trevisan S, Celegato B, Costa G, Lanfranchi G
JournalNucleic Acids Res
PubMed ID14627839
The variability of results in microarray technology is in part due to the fact that independent scans of a single hybridised microarray give spot images that are not quite the same. To solve this problem and turn it to our advantage, we introduced the approach of multiple scanning and of ... More
DNA microarrays and beyond: completing the journey from tissue to cell.
AuthorsMills JC, Roth KA, Cagan RL, Gordon JI
JournalNat Cell Biol
PubMed ID11483971
For the cell biologist, identifying changes in gene expression using DNA microarrays is just the start of a long journey from tissue to cell. We discuss how chip users can first filter noise (false-positives) from daunting microarray datasets. Combining laser capture microdissection with real-time polymerase chain reaction and reverse transcription ... More
An improved protocol for quantification of freshwater Actinobacteria by fluorescence in situ hybridization.
AuthorsSekar R, Pernthaler A, Pernthaler J, Warnecke F, Posch T, Amann R
JournalAppl Environ Microbiol
PubMed ID12732568
We tested a previously described protocol for fluorescence in situ hybridization of marine bacterioplankton with horseradish peroxidase-labeled rRNA-targeted oligonucleotide probes and catalyzed reporter deposition (CARD-FISH) in plankton samples from different lakes. The fraction of Bacteria detected by CARD-FISH was significantly lower than after FISH with fluorescently monolabeled probes. In particular, ... More
Phosphorylation of period is influenced by cycling physical associations of double-time, period, and timeless in the Drosophila clock.
AuthorsKloss B, Rothenfluh A, Young MW, Saez L
JournalNeuron
PubMed ID11430804
The clock gene double-time (dbt) encodes an ortholog of casein kinase Iepsilon that promotes phosphorylation and turnover of the PERIOD protein. Whereas the period (per), timeless (tim), and dClock (dClk) genes of Drosophila each contribute cycling mRNA and protein to a circadian clock, dbt RNA and DBT protein are constitutively ... More
Detection in coal tar waste-contaminated groundwater of mRNA transcripts related to naphthalene dioxygenase by fluorescent in situ hybridization with tyramide signal amplification.
AuthorsBakermans C, Madsen EL
JournalJ Microbiol Methods
PubMed ID11943360
The ideal ecological metabolic activity assay would be applied to naturally occurring microbial populations immediately fixed in the field, and the assay would focus upon intracellular parameters indicative of a dynamic biogeochemical process. In this study, fluorescent in situ hybridization (FISH) with tyramide signal amplification (TSA) detected intracellular mRNA in ... More
Endothelin immunocytochemistry: indications of false-positive labeling patterns and non-detectable antigen concentrations.
AuthorsWolf WP, Weis M, von Scheidt W
JournalHistochem Cell Biol
PubMed ID11735005
Endothelin is an endothelium-derived peptide with potent vasoconstrictor and mitogenic properties. Since studies concerning the immunocytochemical localization of endothelin are often inconsistent we tried to clear up some of these discrepancies by comparing specificity and labeling patterns of different endothelin antibodies. Monoclonal and polyclonal endothelin antibodies ( n=7) were examined ... More
Detailed gene expression analysis but not microsatellite marker analysis of 9p21 reveals differential defects in the INK4a gene locus in the majority of head and neck cancers.
AuthorsGrüttgen A, Reichenzeller M, Jünger M, Schlien S, Affolter A, Bosch FX
JournalJ Pathol
PubMed ID11439363
The INK4a gene locus on chromosome 9p21 encodes two proteins, p16(INK4a) and p14(ARF), which influence cell cycle control regulated by pRb and p53. The objective of this study was to use different methods for the analysis of the incidence of changes at the INK4a locus in head and neck cancer ... More
Mild hypothermia reduces zinc translocation, neuronal cell death, and mortality after transient global ischemia in mice.
The authors sought to determine whether Zn translocation associated with neuronal cell death occurs after transient global ischemia (TGI) in mice, as has been previously shown in rats, and to determine the effect of mild hypothermia on this reaction. To validate the TGI model, carbon-black injection and laser-Doppler flowmetry were ... More
An improved protocol of biotinylated tyramine-based immunohistochemistry minimizing nonspecific background staining.
AuthorsKim SH, Shin YK, Lee KM, Lee JS, Yun JH, Lee SM
JournalJ Histochem Cytochem
PubMed ID12502763
An immunohistochemical method using biotinyl tyramine was recently introduced to amplify weak staining signals. Despite its high sensitivity, however, tyramine-based immunostaining has been limited by its increased background staining. In this study, to develop an improved protocol of biotinyl tyramine-based immunohistochemistry minimizing the background staining, we determined which staining steps ... More
The CCK-2 receptor is located on the ECL cell, but not on the parietal cell.
AuthorsBakke I, Qvigstad G, Sandvik AK, Waldum HL
JournalScand J Gastroenterol
PubMed ID11686210
BACKGROUND: The interrelationship between histamine and gastrin in the physiological regulation of gastric acid secretion is still a matter of dispute. CCK-2 receptors are located on enterochromaffin-like (ECL) cells in corpus mucosa and gastrin stimulates acid production by releasing histamine from the ECL cells, which in turn stimulates the parietal ... More
Ureaplasma in lung. 1. Localization by in situ hybridization in a mouse model.
Ureaplasma urealyticum is a common inhabitant of mucosal surfaces but is also associated with a higher incidence of pneumonia and bronchopulmonary dysplasia in preterm infants. Culture and polymerase chain reaction demonstrate high isolation rates of ureaplasma in clinical specimens documenting their presence but do not associate the organism directly with ... More
Immunohistochemical detection of SWC3, CD2, CD3, CD4 and CD8 antigens in paraformaldehyde fixed and paraffin embedded porcine lymphoid tissue.
AuthorsTingstedt JE, Tornehave D, Lind P, Nielsen J
JournalVet Immunol Immunopathol
PubMed ID12909409
Identification of the different cell types of the immune system is important for in situ studies on the pathogenesis of infectious diseases in various animals, including the pig. Unfortunately, many monoclonal anti-leukocyte antibodies are only useful for staining frozen tissue sections with inherent poor tissue morphology, and are not readily ... More
Involvement of macrophage mannose receptor in the binding and transmission of HIV by macrophages.
AuthorsNguyen DG, Hildreth JE
JournalEur J Immunol
PubMed ID12645947
The human immunodeficiency virus (HIV) is an enveloped virus whose surface glycoprotein gp120 binds CD4 on target cell membranes to initiate infection. About half of the carbohydrates on gp120 are terminally mannosylated, a pattern common to many pathogens. We have examined the ability of macrophage mannose receptor (MMR) on primary ... More
An evaluation of tyramide signal amplification and archived fixed and frozen tissue in microarray gene expression analysis.
Archival formalin-fixed, paraffin-embedded and ethanol-fixed tissues represent a potentially invaluable resource for gene expression analysis, as they are the most widely available material for studies of human disease. Little data are available evaluating whether RNA obtained from fixed (archival) tissues could produce reliable and reproducible microarray expression data. Here we ... More
Immunohistochemical detection of CD30 remains negative in nodular lymphocyte-predominant Hodgkin's disease using enhanced antigen retrieval.
AuthorsRoberts C, Jack F, Angus B, Reid A, Thompson WD
JournalHistopathology
PubMed ID11952861
AIMS: The aims of this study were to confirm that CD30 is reproducibly negative in cases of nodular lymphocyte-predominant Hodgkin's disease (nLPHD), and its relationship to further antibody targets for the distinction of L&H cells from classical Hodgkin's and Reed-Sternberg cells. METHODS AND RESULTS: We examined 16 cases of nLPHD ... More
Cytokine profile and supposed contribution to scarring in cicatricial pemphigoid.
AuthorsCaproni M, Calzolari A, Salvatore E, Giomi B, Volpi W, D'Agata A, Santucci M, Fabbri P
JournalJ Oral Pathol Med
PubMed ID12558956
BACKGROUND: The progressive scarring observed in cicatricial pemphigoid (CP) is still partially unexplained but recently the release of soluble fibrogenic factors by inflammatory infiltrating cells has been considered as pathogenically relevant. In the present study we evaluated the expression of mRNA for IL-4, IL-5, TGF-beta1, IFN-gamma in CP in comparison ... More
Detection of the integrated feline leukemia viruses in a cat lymphoid tumor cell line by fluorescence in situ hybridization.
AuthorsFujino Y, Satoh H, Hisasue M, Masuda K, Ohno K, Tsujimoto H
JournalJ Hered
PubMed ID12816966
Feline leukemia virus (FeLV) is a type-C retrovirus associated with lymphoid and hematopoietic malignancies in cats. The FeLV-induced tumors are thought to be caused, at least in part, by somatically acquired insertional mutagenesis in which the integrated provirus may activate a proto-oncogene or disrupt a tumor suppressor gene. This study ... More
Antral G cells in rats during dosing with a PPAR alpha agonist: a morphometric and immunocytochemical study.
Gastrin-producing G cells constitute one of the major populations of neuroendocrine cells in the antral mucosa of the stomach. The peroxisome proliferator-activated receptor (PPAR) alpha-agonist ciprofibrate is used as a lipid-lowering drug. Recently, ciprofibrate has been shown to induce hypergastrinemia in rats without reducing gastric acidity, which indicates a direct ... More
Immunohistochemical localization of feline immunodeficiency virus using native species antibodies.
AuthorsRogers AB, Mathiason CK, Hoover EA
JournalAm J Pathol
PubMed ID12368188
Feline immunodeficiency virus (FIV) is the feline analog of human immunodeficiency virus and a small animal model of human acquired immune deficiency syndrome (AIDS). We sought to identify early in vivo target cells in cats infected with clade B or C FIV. In tissues, however, neither mouse monoclonal nor rabbit ... More
Double-color fluorescence in situ hybridization with RNA probes.
AuthorsXi X, Roane DS, Zhou J, Ryan DH, Martin RJ
JournalBiotechniques
PubMed ID12765014
Validation of a more sensitive method for using spotted oligonucleotide DNA microarrays for functional genomics studies on bacterial communities.
AuthorsDenef VJ, Park J, Rodrigues JL, Tsoi TV, Hashsham SA, Tiedje JM
JournalEnviron Microbiol
PubMed ID14510847
Spotted oligonucleotide microarrays potentially offer a wide scope of applications for microbial ecology, especially as they improve the flexibility of design and the specificity of detection compared to PCR product based microarrays. Sensitivity, however, was expected to be problematic, as studies with the more sensitive PCR-based cDNA microarrays indicate that ... More
Immunocytochemical localization of acyl-CoA oxidase in the rat central nervous system.
AuthorsFarioli-Vecchioli S, Moreno S, Cerù MP
JournalJ Neurocytol
PubMed ID11577243
Peroxisomal beta-oxidation, consisting of four steps catalysed by an acyl-CoA oxidase, a multifunctional protein and a thiolase, is responsible for the shortening of a variety of lipid compounds. The first reaction of this pathway is catalysed by a FAD-containing acyl-CoA oxidase, three isotypes of which have been so far recognised. ... More
EnVision+, a new dextran polymer-based signal enhancement technique for in situ hybridization (ISH).
AuthorsWiedorn KH, Goldmann T, Henne C, Kühl H, Vollmer E
JournalJ Histochem Cytochem
PubMed ID11511676
Seventy paraffin-embedded cervical biopsy specimens and condylomata were tested for the presence of human papillomavirus (HPV) by conventional in situ hybridization (ISH) and ISH with subsequent signal amplification. Signal amplification was performed either by a commercial biotinyl-tyramide-based detection system [GenPoint (GP)] or by the novel two-layer dextran polymer visualization system ... More
Fluorescence in situ hybridization and catalyzed reporter deposition for the identification of marine bacteria.
AuthorsPernthaler A, Pernthaler J, Amann R
JournalAppl Environ Microbiol
PubMed ID12039771
Fluorescence in situ hybridization (FISH) with horseradish peroxidase (HRP)-labeled oligonucleotide probes and tyramide signal amplification, also known as catalyzed reporter deposition (CARD), is currently not generally applicable to heterotrophic bacteria in marine samples. Penetration of the HRP molecule into bacterial cells requires permeabilization procedures that cause high and most probably ... More
Mapping of the distribution of significant proteins and proteoglycans in small intestinal submucosa by fluorescence microscopy.
AuthorsHurst RE, Bonner RB
JournalJ Biomater Sci Polym Ed
PubMed ID11853391
Because small intestine submucosa (SIS) is a bioscaffold for tissue regeneration, we describe a method to analyze the material for growth peptides and for structural molecules. Immunofluorescence methods are described for relative quantification of abundant structural proteins. Additionally, a quantitative technique for comparison of the content of less abundant proteins ... More
Amplified in situ hybridization with peptide nucleic acid probes for differentiation of Mycobacterium tuberculosis complex and nontuberculous Mycobacterium species on formalin-fixed, paraffin-embedded archival biopsy and autopsy samples.
AuthorsZerbi P, Schønau A, Bonetto S, Gori A, Costanzi G, Duca P, Vago L
JournalAm J Clin Pathol
PubMed ID11710696
The aim of this study was to evaluate sensitivity and specificity of in situ hybridization (ISH) using peptide nucleic acid (PNA) probes and tyramide-based amplification for the differentiation between Mycobacterium tuberculosis (MTB) and mycobacteria other than tuberculosis (MOTT) on formalin-fixed, paraffin-embedded tissue samples. We performed ISH simultaneously with both probes ... More
Triple immunofluorescence staining with antibodies raised in the same species to study the complex innervation pattern of intrapulmonary chemoreceptors.
AuthorsBrouns I, Van Nassauw L, Van Genechten J, Majewski M, Scheuermann DW, Timmermans JP, Adriaensen D
JournalJ Histochem Cytochem
PubMed ID11897811
A general problem in immunocytochemistry is the development of a reliable multiple immunolabeling method when primary antibodies must be used that originate in the same species. We have developed a protocol for the immunodetection of three antigens in a single tissue preparation, using unconjugated primary antibodies raised in the same ... More
Enhancement of in situ hybridization signals in Caenorhabditis elegans by tyramide signal amplification.
AuthorsOgurusu T, Shingai R
JournalAnal Biochem
PubMed ID11969199
Age-associated changes in histology and gene-expression profile in the rat ventral prostate.
AuthorsLau KM, Tam NN, Thompson C, Cheng RY, Leung YK, Ho SM
JournalLab Invest
PubMed ID12746483
The incidence of prostate diseases rises dramatically with age in men, yet little is understood of the mechanisms underlying prostatic senescence and its contribution to disease development in the gland. In Noble rats, aging of the ventral prostate (VP) is characterized morphologically by widespread atrophy of acini, increased accumulation of ... More
FISHing for chick genes: Triple-label whole-mount fluorescence in situ hybridization detects simultaneous and overlapping gene expression in avian embryos.
AuthorsDenkers N, GarcÃa-Villalba P, Rodesch CK, Nielson KR, Mauch TJ
JournalDev Dyn
PubMed ID14991720
Multi-color whole-mount in situ hybridization is a powerful technique for comparing the spatial expression patterns of two or more genes in developing embryos. We have developed an amplified triple-label whole-mount fluorescence in situ hybridization (FISH) protocol that permits detection of three different mRNAs in a single embryo. Our protocol uses ... More
Direct detection of 16S rRNA using oligonucleotide microarrays assisted by base stacking hybridization and tyramide signal amplification.
AuthorsWang D, Zhu L, Jiang D, Ma X, Zhou Y, Cheng J
JournalJ Biochem Biophys Methods
PubMed ID15163522
A simple method has been developed and validated for direct, sensitive detection and specific identification of 16S rRNA. We first report our direct investigation of discrimination efficiency for sequence variations in RNA using oligonucleotide microarrays assisted by base stacking hybridization, and demonstrate that the sequence variations of double base substitution, ... More
Physical mapping of barley genes using an ultrasensitive fluorescence in situ hybridization technique.
AuthorsStephens JL, Brown SE, Lapitan NL, Knudson DL
JournalGenome
PubMed ID15060614
The primary objective of this study was to elucidate gene organization and to integrate the genetic linkage map for barley (Hordeum vulgare L.) with a physical map using ultrasensitive fluorescence in situ hybridization (FISH) techniques for detecting signals from restriction fragment length polymorphism (RFLP) clones. In the process, a single ... More
Viral RNA and p24 antigen as markers of HIV disease and antiretroviral treatment success.
AuthorsSchüpbach J
JournalInt Arch Allergy Immunol
PubMed ID14646380
HIV-1 RNA has become the standard for monitoring antiretroviral therapies. Dogma predicts, however, that a viral protein like p24 should be at least as good a marker of HIV disease activity, provided that it is measured with sufficient sensitivity and accuracy. Simple modifications including use of a more efficient virus ... More
Protein arrays for serodiagnosis of disease.
AuthorsBacarese-Hamilton T, Ardizzoni A, Gray J, Crisanti A
JournalMethods Mol Biol
PubMed ID15020797
Protein microarrays offer the possibility to circumvent most of the current limitations in the serodiagnosis of allergy, autoimmune, and infectious disease by allowing the simultaneous, multiparametric determination of specific subclasses of antibodies directed against many pathogenic antigens. Microarray immunoassays have been developed with these characteristics. A first-generation assay, for the ... More
Effects of one bout of endurance exercise on the expression of myogenin in human quadriceps muscle.
AuthorsKadi F, Johansson F, Johansson R, Sjöström M, Henriksson J
JournalHistochem Cell Biol
PubMed ID14997318
The objective of this study was to investigate the cellular localisation of MyoD and myogenin in human skeletal muscle fibres as well as the possible alterations in the expression of MyoD and myogenin in response to a single bout of endurance exercise at 40% and 75% of maximum oxygen uptake ... More
The c-SRC1 gene visualized by in situ hybridization on Xenopus laevis chromosomes.
AuthorsKrylov V, Mácha J, Tlapáková T, Takác M, Jonák J
JournalCytogenet Genome Res
PubMed ID15004482
Fluorescent in situ hybridization followed by tyramide signal amplification was used to map the site of the c-SRC1 gene on XENOPUS LAEVIS chromosomes. Positive results were obtained with a cDNA probe of about 1 kb. The c-SRC1 gene is located in the subcentromeric region in the long arm of one ... More
Simultaneous in situ detection of RNA, DNA, and protein using tyramide-coupled immunofluorescence.
AuthorsVan Tine BA, Broker TR, Chow LT
JournalMethods Mol Biol
PubMed ID15507711
The use of tyramide-coupled immunofluorescence at the single cell level provides expedient, clean, and sensitive signals for detection of DNA, RNA, or proteins. The principle is based on the ability of horseradish peroxidase (HRP) to cleave tyramides into a free radical species with a very short diffusion radius. The free ... More
A protein microarray ELISA for screening biological fluids.
AuthorsVarnum SM, Woodbury RL, Zangar RC
JournalMethods Mol Biol
PubMed ID15020788
Protein microarrays permit the simultaneous measurement of many proteins in a small sample volume and therefore provide an attractive approach for the quantitative measurement of proteins in biological fluids, including serum. This chapter describes a microarray enzyme-linked immunosorbent assay (ELISA). Capture antibodies are immobilized onto a glass surface; the covalently ... More
Detection of low copy numbers of HPV DNA by fluorescent in situ hybridization combined with confocal microscopy as an alternative to in situ polymerase chain reaction.
AuthorsLizard G, Chignol MC, Souchier C, Roignot P, Chardonnet Y, Schmitt D
JournalJ Virol Methods
PubMed ID9672129
'In genital lesions infected by human papillomavirus (HPV), histological criteria and HPV DNA typing are of prognostic value. Therefore, non-radioactive methods such as in situ hybridization are used extensively since they preserve the histological organization of the tissue, and allow the detection and characterization of HPV DNA. However, the sensitivity ... More
Attenuation of astroglial reactivity by interleukin-10.
AuthorsBalasingam V, Yong VW
JournalJ Neurosci
PubMed ID8622125
'Prominent responses that follow brain trauma include the activation of microglia, recruitment of blood-derived macrophages, and astroglial reactivity. Based on evidence that cytokines produced by macrophages/microglia may cause astrocytes to become reactive, the aim of this study was to determine whether astroglial reactivity could be attenuated by interleukin (IL)-10, a ... More
Biotinyl-tyramide: a novel approach for electron microscopic immunocytochemistry.
AuthorsMayer G, Bendayan M
JournalJ Histochem Cytochem
PubMed ID9358846
'The biotinyl-tyramide protocol recently introduced for sensitive light microscopic immunocytochemistry was applied to electron microscopy and revealed various tissue antigens with high resolution. The protocol consists of an indirect method in which thin tissue sections are incubated successively within a specific primary antibody, followed by a biotinylated secondary antibody, streptavidin-HRP, ... More
Galanin-5-hydroxytryptamine interactions: electrophysiological, immunohistochemical and in situ hybridization studies on rat dorsal raphe neurons with a note on galanin R1 and R2 receptors.
AuthorsXu ZQ, Zhang X, Pieribone VA, Grillner S, Hökfelt T
JournalNeuroscience
PubMed ID9722143
'Galaninergic mechanisms related to 5-hydroxytryptamine neurons in the dorsal raphe nucleus of the rat were analysed using electrophysiology, immunohistochemistry and in situ hybridization. Galanin caused a dose-dependent hyperpolarization accompanied by a decrease in membrane resistance in most 5-hydroxytryptamine-sensitive dorsal raphe neurons. The galanin-induced outward current reversed at about - 105 ... More
Thymocytes express the golli products of the myelin basic protein gene and levels of expression are stage dependent.
AuthorsFeng JM, Givogri IM, Bongarzone ER, Campagnoni C, Jacobs E, Handley VW, Schonmann V, Campagnoni AT
JournalJ Immunol
PubMed ID11067896
'The golli products of the myelin basic protein gene have been shown to be expressed in mouse thymus and brain. The full repertoire of thymic cell types expressing golli products has not yet been determined, although immunoreactivity has been found in some macrophages. We have analyzed the cellular expression of ... More
Fluorescein-labeled tyramide strongly enhances the detection of low bromodeoxyuridine incorporation levels.
AuthorsVan Heusden J, de Jong P, Ramaekers F, Bruwiere H, Borgers M, Smets G
JournalJ Histochem Cytochem
PubMed ID9016320
'Immunocytochemical detection of bromodeoxyuridine (BrdU) labeling can be hampered by low BrdU incorporation levels. We describe here an amplification method for weak BrdU immunosignals. The tyramide signal amplification method based on catalyzed reporter deposition (CARD) uses fluorescein-labeled tyramide as a substrate for horseradish peroxidase. The enzyme catalyzes the formation of ... More
Co-localization of the glial cell-line derived neurotrophic factor and its functional receptor c-RET in a subpopulation of rat dorsal root ganglion neurons.
AuthorsHonda T, Takahashi M, Sugiura Y
JournalNeurosci Lett
PubMed ID10554981
'Co-localization of the glial cell line-derived neurotrophic factor (GDNF) and its functional receptor c-RET was examined immunohistochemically in the dorsal root ganglion (DRG) of the 5th lumbar nerve in rats. The total 1728 DRG neurons were observed in three female rats (Wistar) with ca. 200 g body weight, 46.6% of ... More
Catalyzed reporter deposition, a novel method of signal amplification. II. Application to membrane immunoassays.
AuthorsBobrow MN, Shaughnessy KJ, Litt GJ
JournalJ Immunol Methods
PubMed ID1849153
'In a previous publication (Bobrow et al., J. Immunol. Methods (1989) 279-285), we described a novel signal amplification method, catalyzed reporter deposition (CARD), and its application to microplate immunoassays. The method utilizes the analyte-dependent reporter enzyme (ADRE) to catalyze the deposition of additional reporter onto the surface of a solid-phase ... More
Cryopreservation of organotypic multicellular spheroids from human gliomas.
AuthorsKaaijk P, van den Berg F, Van Amstel P, Troost D
JournalNeuropathol Appl Neurobiol
PubMed ID9004246
'Fresh human glioma tissue can be cultured on agarose to form organotypic multicellular spheroids (OMS). The major advantage of OMS is the preservation of the cellular heterogeneity and the tumour architecture, which is lost in conventional monolayer cultures. The present study was undertaken to assess the possibilities of storing frozen ... More
Expression patterns of transmembrane and released forms of neuregulin during spinal cord and neuromuscular synapse development.
'We mapped the distribution of neuregulin and its transmembrane precursor in developing, embryonic chick and mouse spinal cord. Neuregulin mRNA and protein were expressed in motor and sensory neurons shortly after their birth and levels steadily increased during development. Expression of the neuregulin precursor was highest in motor and sensory ... More
Sensitive multicolor fluorescence in situ hybridization using catalyzed reporter deposition (CARD) amplification.
AuthorsSpeel EJ, Ramaekers FC, Hopman AH
JournalJ Histochem Cytochem
PubMed ID9313806
'We describe the simultaneous localization of DNA sequences in cell and chromosome preparations by means of differently fluorochrome-labeled (AMCA, FITC, TRITC) tyramides using the catalyzed reporter deposition (CARD) procedure. For this purpose, repeated as well as single-copy DNA probes were labeled with biotin, digoxigenin, and FITC, hybridized, and visualized with ... More
'To gain a molecular understanding of tumor angiogenesis, we compared gene expression patterns of endothelial cells derived from blood vessels of normal and malignant colorectal tissues. Of over 170 transcripts predominantly expressed in the endothelium, 79 were differentially expressed, including 46 that were specifically elevated in tumor-associated endothelium. Several of ... More
Immunocytochemical localization of the NPY/PYY Y1 receptor in enteric neurons, endothelial cells, and endocrine-like cells of the rat intestinal tract.
AuthorsJackerott M, Larsson LI
JournalJ Histochem Cytochem
PubMed ID9389767
'Neuropeptide Y (NPY) and peptide YY (PYY) are structurally related peptides that are considered to mediate inhibitory actions on gastrointestinal motility, secretion, and blood flow. Several receptor subtypes for these peptides have been identified and the Y1, Y2, Y4/PP1, Y5, and Y5/PP2/Y2b receptors have been cloned. In this article we ... More
A protective role of the low density lipoprotein receptor-related protein against amyloid beta-protein toxicity.
AuthorsVan Uden E, Sagara Y, Van Uden J, Orlando R, Mallory M, Rockenstein E, Masliah E
JournalJ Biol Chem
PubMed ID10899157
'In order to delineate the neuroprotective role of the low density lipoprotein receptor-related protein (LRP) against amyloid beta-protein toxicity, studies were performed in C6 cells challenged with amyloid beta-protein in the presence or absence of activated alpha(2)-macroglobulin. Toxicity was assessed via two cell viability assays. We found that this endocytic ... More
Fluorescence in situ hybridization using horseradish peroxidase-labeled oligodeoxynucleotides and tyramide signal amplification for sensitive DNA and mRNA detection.
Authorsvan de Corput MP, Dirks RW, van Gijlswijk RP, van de Rijke FM, Raap AK
JournalHistochem Cell Biol
PubMed ID9792422
'We have used horseradish peroxidase-labeled 40 mer oligodeoxynucleotides (HRP-ODNs) specific for the human cytomegalovirus immediate early gene (HCMV-IE) and a novel dinitrophenol-tyramide signal amplification reagent (DNP-TSA plus) to evaluate their utility in fluorescence in situ hybridization (FISH). For DNA FISH, single or cocktails of HRP-ODNs were hybridized to metaphase chromosomes ... More
Enhancement of immunohistochemical detection of HIV-1 p24 antigen in brain by tyramide signal amplification.
AuthorsStrappe PM, Wang TH, McKenzie CA, Lowrie S, Simmonds P, Bell JE
JournalJ Virol Methods
PubMed ID9274823
'Human immunodeficiency virus type 1 (HIV-1) infection of the brain has been demonstrated in formalin fixed, paraffin embedded post-mortem brain tissue (PM) by chromogenic immunohistochemistry for the HIV p24 antigen. The sensitivity of antigen detection is increased significantly by tyramide signal amplification (TSA) compared to the conventional peroxidase labelled Avidin-Biotin ... More