Primitive adult hematopoietic stem cells can function as osteoblast precursors.
AuthorsOlmsted-Davis EA, Gugala Z, Camargo F, Gannon FH, Jackson K, Kienstra KA, Shine HD, Lindsey RW, Hirschi KK, Goodell MA, Brenner MK, Davis AR
JournalProc Natl Acad Sci U S A
PubMed ID14673088
'Osteoblasts are continually recruited from stem cell pools to maintain bone. Although their immediate precursor is a plastic-adherent mesenchymal stem cell able to generate tissues other than bone, increasing evidence suggests the existence of a more primitive cell that can differentiate to both hematopoietic and mesenchymal cells. We show here ... More
Qualification of embryonal carcinoma 2102Ep as a reference for human embryonic stem cell research.
AuthorsJosephson R, Ording CJ, Liu Y, Shin S, Lakshmipathy U, Toumadje A, Love B, Chesnut JD, Andrews PW, Rao MS, Auerbach JM
JournalStem Cells
PubMed ID17284651
'As the number of human embryonic stem cell (hESC) lines increases, so does the need for systematic evaluation of each line''s characteristics and potential. Comparisons between lines are complicated by variations in culture conditions, feeders, spontaneous differentiation, and the absence of standardized assays. These difficulties, combined with the inability of ... More
Osteoclast responses to lipopolysaccharide, parathyroid hormone and bisphosphonates in neonatal murine calvaria analyzed by laser scanning confocal microscopy.
'Because the development and activity of osteoclasts in bone remodeling is critically dependent on cell-cell and cell-matrix interactions, we used laser confocal microscopy to study the response of osteoclasts to lipopolysaccharide (LPS; 10 microg/ml), parathyroid hormone (PTH; 10(-8) M), and bisphosphonates (BPs; 1-25 microM clodronate or 0.1-2.5 microM risedronate) in ... More
Fluorescence-based staining for tartrate-resistant acidic phosphatase (TRAP) in osteoclasts combined with other fluorescent dyes and protocols.
AuthorsFilgueira L
JournalJ Histochem Cytochem
PubMed ID14966208
Osteoclasts are the only bone-resorbing cells. In addition to other specific properties, osteoclasts are characterized by their expression of tartrate-resistant acidic phosphatase (TRAP), which is usually detected using a histochemical method for light microscopy. Using ELF97 phosphatase substrate, this study describes a new fluorescence-based method for TRAP detection. The fluorescence-based ... More
Novel method for screening bacterial colonies for phosphatase activity.
Current methods for screening large numbers of bacterial colonies for phosphatase activity, rely heavily on the use of colorimetric assays. While such methods have been applied extensively in the laboratory, they are not without their drawbacks. We here describe a precipitating fluorescent probe that can be used to screen phosphatase ... More
Functional dissociation of the basolateral transcytotic compartment from the apical phago-lysosomal compartment in human osteoclasts.
AuthorsMeagher J, Zellweger R, Filgueira L,
JournalJ Histochem Cytochem
PubMed ID15872059
Tartrate-resistant acid phosphatase (TRAP) is essential for elimination of Staphylococcus aureus, the main infectious agent responsible for osteomyelitis. This in vitro study investigated uptake and processing of fluorescence-labeled S. aureus by human osteoclasts and dendritic cells. The cells were stained for TRAP and the acidic compartment using a fluorescence-based protocol. ... More
A phosphoenol pyruvate phosphatase transcript is induced in the root nodule cortex of Phaseolus vulgaris under conditions of phosphorus deficiency.
AuthorsBargaz A, Ghoulam C, Amenc L, Lazali M, Faghire M, Abadie J, Drevon JJ,
JournalJ Exp Bot
PubMed ID22771853
Although previous studies on N2-fixing legumes have demonstrated the contribution of acid phosphatases to their phosphorus (P) use efficiency under P-deficient growth conditions, localization of these enzymes in bean nodules has not been demonstrated. In this study, phosphoenol pyruvate phosphatase (PEPase) gene transcripts were localized within the nodule tissues of ... More
Modulation of symbiont lipid A signaling by host alkaline phosphatases in the squid-vibrio symbiosis.
AuthorsRader BA, Kremer N, Apicella MA, Goldman WE, McFall-Ngai MJ,
JournalMBio
PubMed ID22550038
The synergistic activity of Vibrio fischeri lipid A and the peptidoglycan monomer (tracheal cytotoxin [TCT]) induces apoptosis in the superficial cells of the juvenile Euprymna scolopes light organ during the onset of the squid-vibrio symbiosis. Once the association is established in the epithelium-lined crypts of the light organ, the host ... More
Osteoclast size is controlled by Fra-2 through LIF/LIF-receptor signalling and hypoxia.
AuthorsBozec A, Bakiri L, Hoebertz A, Eferl R, Schilling AF, Komnenovic V, Scheuch H, Priemel M, Stewart CL, Amling M, Wagner EF,
JournalNature
PubMed ID18548006
Osteoclasts are multinucleated haematopoietic cells that resorb bone. Increased osteoclast activity causes osteoporosis, a disorder resulting in a low bone mass and a high risk of fractures. Increased osteoclast size and numbers are also a hallmark of other disorders, such as Paget's disease and multiple myeloma. The protein c-Fos, a ... More
Validation and quantitation of an in vitro M-cell model.
AuthorsTyrer P, Ruth Foxwell A, Kyd J, Harvey M, Sizer P, Cripps A
JournalBiochem Biophys Res Commun
PubMed ID12445810
This study has evaluated an in vitro model of the follicle-associated epithelia that overlie Peyer's patches of the small intestine. The model shares many phenotypic characteristics of M cells in vivo. Co-cultures of the human adenocarcinoma cell line Caco-2 and freshly isolated Peyer's patch cells were established. Fluorescence microscopy and ... More
AuthorsMcBeath R, Pirone DM, Nelson CM, Bhadriraju K, Chen CS
JournalDev Cell
PubMed ID15068789
Commitment of stem cells to different lineages is regulated by many cues in the local tissue microenvironment. Here we demonstrate that cell shape regulates commitment of human mesenchymal stem cells (hMSCs) to adipocyte or osteoblast fate. hMSCs allowed to adhere, flatten, and spread underwent osteogenesis, while unspread, round cells became ... More
Use of a new fluorogenic phosphatase substrate in immunohistochemical applications.
AuthorsLarison KD, BreMiller R, Wells KS, Clements I, Haugland RP
JournalJ Histochem Cytochem
PubMed ID7822768
We used the phosphatase substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6- chloro-4-[3H]-quinazolinone, with standard alkaline phosphatase-mediated immunohistochemical techniques, to visualize a number of antibodies that bind to adult zebrafish retinal tissue. This compound, known as the ELF (enzyme-labeled-fluorescence) phosphatase substrate, produces a precipitate that fluoresces at approximately 500-580 nm (bright yellow-green). We show that the ... More
Semi-automated positional analysis using laser scanning microscopy of cells transfected in a regenerating newt limb.
AuthorsPecorino LT, Brockes JP, Entwistle A
JournalJ Histochem Cytochem
PubMed ID8666741
Limb regeneration in urodele amphibians such as the newt is a key system for investigating the positional identity of cells. The regenerate arises locally from blastemal cells, mesenchymal progenitors that normally give rise to structures distal to the amputation plane but which can be respecified (proximalized) by treatment with retinoic ... More
HGF inhibits BMP-induced osteoblastogenesis: possible implications for the bone disease of multiple myeloma.
AuthorsStandal T, Abildgaard N, Fagerli UM, Stordal B, Hjertner O, Borset M, Sundan A
JournalBlood
PubMed ID17138824
The bone disease in multiple myeloma is caused by an uncoupling of bone formation from bone resorption. A key difference between patients with and patients without osteolytic lesion is that the latter have fewer and less active osteoblasts. Hepatocyte growth factor (HGF) is often produced by myeloma cells and is ... More
A high-resolution, fluorescence-based method for localization of endogenous alkaline phosphatase activity.
AuthorsCox WG, Singer VL
JournalJ Histochem Cytochem
PubMed ID10544217
We describe a high-resolution, fluorescence-based method for localizing endogenous alkaline phosphatase in tissues and cultured cells. This method utilizes ELF (Enzyme-Labeled Fluorescence)-97 phosphate, which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. We compared zebrafish intestine, ovary, and kidney cryosections stained for endogenous alkaline phosphatase using ... More
Extracellular phosphatase activity of natural plankton studied with ELF97 phosphate: fluorescence quantification and labelling kinetics.
AuthorsNedoma J, Strojsová A, Vrba J, Komárková J, Simek K
JournalEnviron Microbiol
PubMed ID12755713
ELF(R)97 phosphate (ELFP) is a phosphatase substrate which produces ELF(R)97 alcohol (ELFA), a fluorescent water-insoluble product, upon hydrolysis. We studied the kinetics of ELFA precipitation in freshwater samples at levels of total plankton and single phytoplankton cells, and tested the suitability of ELFP for measurement of surface-bound algal extracellular phosphatases. ... More
Fluid shear stress induces less calcium response in a single primary osteocyte than in a single osteoblast: implication of different focal adhesion formation.
AuthorsKamioka H, Sugawara Y, Murshid SA, Ishihara Y, Honjo T, Takano-Yamamoto T
JournalJ Bone Miner Res
PubMed ID16813522
The immediate calcium response to fluid shear stress was compared between osteocytes and osteoblasts on glass using real-time calcium imaging. The osteoblasts were responsive to fluid shear stress of up to 2.4 Pa, whereas the osteocytes were not. The difference in flow-induced calcium may be related to differences in focal ... More
Detection of endogenous and antibody-conjugated alkaline phosphatase with ELF-97 phosphate in multicolor flow cytometry applications.
AuthorsTelford W, Cox W, Singer V
JournalCytometry
PubMed ID11169576
BACKGROUND: The fluorogenic alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (ELF(R)-97 phosphate, for Enzyme-Labeled Fluorescence) has been used primarily in microscope-based imaging applications to detect endogenous AP activity, antigens and various ligands in cells and tissues, and nucleic acid hybridization. In a previous study, we demonstrated the applicability of ELF-97 phosphate for ... More
Spatial patterns of alkaline phosphatase expression within bacterial colonies and biofilms in response to phosphate starvation.
AuthorsHuang CT, Xu KD, McFeters GA, Stewart PS
JournalAppl Environ Microbiol
PubMed ID9546188
The expression of alkaline phosphatase in response to phosphate starvation was shown to be spatially and temporally heterogeneous in bacterial biofilms and colonies. A commercial alkaline phosphatase substrate that generates a fluorescent, insoluble product was used in conjunction with frozen sectioning techniques to visualize spatial patterns of enzyme expression in ... More
Phosphate stress in cultures and field populations of the dinoflagellate prorocentrum minimum detected by a single-cell alkaline phosphatase assay
AuthorsDyhrman ST, Palenik B
JournalAppl Environ Microbiol
PubMed ID10388722
Alkaline phosphatase activity is a common marker of phosphate stress in many phytoplankton, but it has been difficult to attribute alkaline phosphatase activity to specific organisms or groups of phytoplankton in the field with traditional biochemical procedures. A new alkaline phosphatase substrate, ELF-97 (enzyme-labeled fluorescence), shows promise in this regard. ... More
Detection of endogenous alkaline phosphatase activity in intact cells by flow cytometry using the fluorogenic ELF-97 phosphatase substrate.
BACKGROUND: The alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (ELF((R))-97 for enzyme-labeled fluorescence) has been found useful for the histochemical detection of endogenous AP activity and AP-tagged proteins and oligonucleotide probes. In this study, we evaluated its effectiveness at detecting endogenous AP activity by flow cytometry. METHODS: The ELF-97 phosphatase substrate was ... More
Seasonal variations in extracellular enzymatic activity in marine snow-associated microbial communities and their impact on the surrounding water.
Authors
JournalFEMS Microbiol Ecol
PubMed ID30299466
Detection of extracellular phosphatase activity at the single-cell level by enzyme-labeled fluorescence and flow cytometry: the importance of time kinetics in ELFA labeling.
Authors
JournalCytometry A
PubMed ID19051328
Enslavement in the water body by toxic Aphanizomenon ovalisporum, inducing alkaline phosphatase in phytoplanktons.
Authors
JournalCurr Biol
PubMed ID20705465
Synergistic combination treatment to break cross talk between cancer cells and bone cells to inhibit progression of bone metastasis.
Authors
JournalBiomaterials
PubMed ID31654872
Revealing the influence of electron beam melted Ti-6Al-4V scaffolds on osteogenesis of human bone marrow-derived mesenchymal stromal cells.
Authors
JournalJ Mater Sci Mater Med
PubMed ID34406475
Stromal-Initiated Changes in the Bone Promote Metastatic Niche Development.
Authors
JournalCell Rep
PubMed ID26725121
Morpho-physiological adaptations of Leptocylindrus aporus and L. hargravesii to phosphate limitation in the northern Adriatic.
Authors
JournalSci Rep
PubMed ID35177689
High performance magnesium anode in paper-based microfluidic battery, powering on-chip fluorescence assay.
Authors
JournalBiomicrofluidics
PubMed ID25332741
Insights into the life strategy of the common marine diatom Chaetoceros peruvianus Brightwell.
Authors
JournalPLoS One
PubMed ID30208090
Cell-based RNAi screening and high-content analysis in primary calvarian osteoblasts applied to identification of osteoblast differentiation regulators.