Journal of Histochemistry and Cytochemistry This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.

New Automated Single-Cell Technique for Segmentation and Quantitation of Lipid Droplets
We describe here a novel three-stage hybrid method for automated segmentation and quantitation of lipid droplets. In this method, objects are initially identified by thresholding. They are then tested for circularity to distinguish single lipid droplets from clusters. Clusters are subjected to a secondary watershed segmentation. We provide a characterization of this method in simulated images. Additionally, we apply this method to images of fixed cells containing stained lipid droplets and GFP-tagged proteins to provide a proof-of-principle that this method can be used for colocalization studies. The circularity measure ca...
Source: Journal of Histochemistry and Cytochemistry - November 25, 2014 Category: Biochemistry Authors: Dejgaard, S. Y., Presley, J. F. Tags: Articles Source Type: research

The Adaptor Protein p62 Is Involved in RANKL-induced Autophagy and Osteoclastogenesis
Previous studies have implicated autophagy in osteoclast differentiation. The aim of this study was to investigate the potential role of p62, a characterized adaptor protein for autophagy, in RANKL-induced osteoclastogenesis. Real-time quantitative PCR and western blot analyses were used to evaluate the expression levels of autophagy-related markers during RANKL-induced osteoclastogenesis in mouse macrophage-like RAW264.7 cells. Meanwhile, the potential relationship between p62/LC3 localization and F-actin ring formation was tested using double-labeling immunofluorescence. Then, the expression of p62 in RAW264.7 cells was ...
Source: Journal of Histochemistry and Cytochemistry - November 25, 2014 Category: Biochemistry Authors: Li, R.-F., Chen, G., Ren, J.-G., Zhang, W., Wu, Z.-X., Liu, B., Zhao, Y., Zhao, Y.-F. Tags: Articles Source Type: research

Expression of ORAI1, a Plasma Membrane Resident Subunit of the CRAC Channel, in Rodent and Non-rodent Species
We determined the expression of ORAI1 protein in rodent and non-rodent tissues using a monoclonal antibody directed against an extracellular loop of the protein. Previous reports using antibodies directed at the C-terminus of ORAI1 have not detected central nervous system (CNS) expression. Our results demonstrate broad tissue expression that includes the CNS using a unique monoclonal antibody specific to an extracellular loop of ORAI1. In addition, we present in situ hybridization (ISH) results using a probe within the middle of the mouse coding region showing CNS expression of Orai1 RNA. We contrast the patterns of rodent...
Source: Journal of Histochemistry and Cytochemistry - November 25, 2014 Category: Biochemistry Authors: Guzman, R., Valente, E. G., Pretorius, J., Pacheco, E., Qi, M., Bennett, B. D., Fong, D. H., Lin, F.-F., Bi, V., McBride, H. J. Tags: Articles Source Type: research

Brightfield Proximity Ligation Assay Reveals Both Canonical and Mixed Transforming Growth Factor-{beta}/Bone Morphogenetic Protein Smad Signaling Complexes in Tissue Sections
Transforming growth factor-β (TGF-β) is an important regulator of cellular homeostasis and disease pathogenesis. Canonical TGF-β signaling occurs through Smad2/3–Smad4 complexes; however, recent in vitro studies suggest that elevated levels of TGF-β may activate a novel mixed Smad complex (Smad2/3-Smad1/5/9), which is required for some of the pro-oncogenic activities of TGF-β. To determine if mixed Smad complexes are evident in vivo, we developed antibodies that can be used with a proximity ligation assay to detect either canonical or mixed Smad complexes in formalin-fixed paraffin-embedded s...
Source: Journal of Histochemistry and Cytochemistry - November 25, 2014 Category: Biochemistry Authors: Flanders, K. C., Heger, C. D., Conway, C., Tang, B., Sato, M., Dengler, S. L., Goldsmith, P. K., Hewitt, S. M., Wakefield, L. M. Tags: Articles Source Type: research

New Aspects of Progesterone Interactions with the Actin Cytoskeleton and Neurosteroidogenesis in the Cerebellum and the Neuronal Growth Cone
The impact of progesterone on neuronal tissues in the central (CNS) and peripheral (PNS) nervous system is of significant scientific and therapeutic interest. Glial and neuronal cells of vertebrates express steroidogenic enzymes, and are able to synthesize progesterone de novo from cholesterol. Progesterone is described to have neuroprotective, neuroreparative, anti-degenerative, and anti-apoptotic effects in the CNS and the PNS. Thus, the first clinical studies promise new therapeutic options using progesterone in the treatment of patients with traumatic brain injury. Additionally, experimental data from different animal ...
Source: Journal of Histochemistry and Cytochemistry - November 25, 2014 Category: Biochemistry Authors: Wessel, L., Olbrich, L., Brand-Saberi, B., Theiss, C. Tags: Review Source Type: research

Determination of Phosphate-activated Glutaminase Activity and Its Kinetics in Mouse Tissues using Metabolic Mapping (Quantitative Enzyme Histochemistry)
In conclusion, PAG is mainly active in mouse kidney, brain and liver, and shows different kinetics depending on which type of PAG is expressed. (Source: Journal of Histochemistry and Cytochemistry)
Source: Journal of Histochemistry and Cytochemistry - October 25, 2014 Category: Biochemistry Authors: Botman, D., Tigchelaar, W., Van Noorden, C. J. F. Tags: Articles Source Type: research

Determination of Glutamate Dehydrogenase Activity and Its Kinetics in Mouse Tissues using Metabolic Mapping (Quantitative Enzyme Histochemistry)
In conclusion, GDH activity in mice is highest in the liver with NAD+ as a coenzyme and highest GDH activity was determined at a glutamate concentration of 10 mM. (Source: Journal of Histochemistry and Cytochemistry)
Source: Journal of Histochemistry and Cytochemistry - October 25, 2014 Category: Biochemistry Authors: Botman, D., Tigchelaar, W., Van Noorden, C. J. F. Tags: Articles Source Type: research

Cystic Fibrosis Transmembrane Conductance Regulator Protein (CFTR) Expression in the Developing Human Brain: Comparative Immunohistochemical Study between Patients with Normal and Mutated CFTR
Cystic Fibrosis Transmembrane conductance Regulator (CFTR) protein has recently been shown to be expressed in the human adult central nervous system (CNS). As CFTR expression has also been documented during embryonic development in several organs, such as the respiratory tract, the intestine and the male reproductive system, suggesting a possible role during development we decided to investigate the expression of CFTR in the human developing CNS. In addition, as some, although rare, neurological symptoms have been reported in patients with CF, we compared the expression of normal and mutated CFTR at several fetal stages. I...
Source: Journal of Histochemistry and Cytochemistry - October 25, 2014 Category: Biochemistry Authors: Marcorelles, P., Friocourt, G., Uguen, A., Lede, F., Ferec, C., Laquerriere, A. Tags: Articles Source Type: research

Endothelial Matrix Assembly during Capillary Morphogenesis: Insights from Chimeric TagRFP-Fibronectin Matrix
Biologically relevant, three-dimensional extracellular matrix is an essential component of in vitro vasculogenesis models. WI-38 fibroblasts assemble a 3D matrix that induces endothelial tubulogenesis, but this model is challenged by fibroblast senescence and the inability to distinguish endothelial cell-derived matrix from matrix made by WI-38 fibroblasts. Matrices produced by hTERT-immortalized WI-38 recapitulated those produced by wild type fibroblasts. ECM fibrils were heavily populated by tenascin-C, fibronectin, and type VI collagen. Nearly half of the total type I collagen, but only a small fraction of the type IV c...
Source: Journal of Histochemistry and Cytochemistry - October 25, 2014 Category: Biochemistry Authors: Chang, F., Lemmon, C. A., Nilaratanakul, V., Rotter, V., Romer, L. Tags: Articles Source Type: research

Comparative Analysis of H&E and Prussian Blue Staining in a Mouse Model of Cerebral Microbleeds
Cerebral microbleeds are microscopic hemorrhages with deposits of blood products in the brain, which can be visualized with MRI and are implicated in cerebrovascular diseases. Hematoxylin and eosin (H&E) and Perl’s Prussian blue are popular staining methods used to localize cerebral microbleeds in pathology. This paper compared these two staining techniques in a mouse model of cerebral microbleeds. We used lipopolysaccharide (LPS) to induce cerebral microhemorrhages. C57B6 mice were treated with LPS (5 mg/kg, i.p.) or vehicle at baseline and at 24 hr. The brains were extracted 48 hr after the first injection and ...
Source: Journal of Histochemistry and Cytochemistry - October 25, 2014 Category: Biochemistry Authors: Liu, S., Grigoryan, M. M., Vasilevko, V., Sumbria, R. K., Paganini-Hill, A., Cribbs, D. H., Fisher, M. J. Tags: Articles Source Type: research

Corrigendum
In the article, Dentin Sialophosphoprotein (DSPP) Plays an Essential Role in the Postnatal Development and Maintenance of Mouse Mandibular Condylar Cartilage. Q. Liu, M. P. Gibson, Hongchen Sun, and C. Qi. J Histochem Cytochem. 61(10):749-758, there was an error in the author affiliation listing for Dr. Q. Liu. (DOI: 10.1369/0022155413502056) The correct author affiliation listing for the article is as follows: Q. Liu, M. P. Gibson, Hongchen Sun, and C. Qi Department of Oral Pathology and Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Jilin University, Changchun, China (QL, HS), and Department of Biome...
Source: Journal of Histochemistry and Cytochemistry - September 25, 2014 Category: Biochemistry Tags: Corrigendum Source Type: research

Picrosirius Red Staining: A Useful Tool to Appraise Collagen Networks in Normal and Pathological Tissues
Specific staining of the extracellular matrix components is especially helpful in studying tissue remodeling, particularly in the case of connective tissue pathologies. As developed by Junqueira and colleagues in 1979, specific staining by Picrosirius red is one of the most important stains to study collagen networks in different tissues. Under polarized light, collagen bundles appear green, red or yellow, and are easily differentiated from the black background, thus allowing for quantitative morphometric analysis. As Junqueira and colleagues point out, many studies use color staining to differentiate collagen bundles and ...
Source: Journal of Histochemistry and Cytochemistry - September 25, 2014 Category: Biochemistry Authors: Lattouf, R., Younes, R., Lutomski, D., Naaman, N., Godeau, G., Senni, K., Changotade, S. Tags: Articles Source Type: research

Ultrastructure of Cytoplasmic and Nuclear Inosine-5'-Monophosphate Dehydrogenase 2 "Rods and Rings" Inclusions
Inosine-5'-monophosphate dehydrogenase catalyzes the critical step in the de novo synthesis of guanosine nucleotides: the oxidation of inosine monophosphate to xanthosine monophosphate. This reaction can be inhibited by specific inhibitors, such as ribavirin or mycophenolic acid, which are widely used in clinical treatment when required to inhibit the proliferation of viruses or cells. However, it was recently found that such an inhibition affects the cells, leading to a redistribution of IMPDH2 and the appearance of IMPDH2 inclusions in the cytoplasm. According to their shape, these inclusions have been termed "Rods and R...
Source: Journal of Histochemistry and Cytochemistry - September 25, 2014 Category: Biochemistry Authors: Juda, P., Šmigova, J., Kovačik, L., Bartova, E., Raška, I. Tags: Articles Source Type: research

Mast Cell Function: A New Vision of an Old Cell
Since first described by Paul Ehrlich in 1878, mast cells have been mostly viewed as effectors of allergy. It has been only in the past two decades that mast cells have gained recognition for their involvement in other physiological and pathological processes. Mast cells have a widespread distribution and are found predominantly at the interface between the host and the external environment. Mast cell maturation, phenotype and function are a direct consequence of the local microenvironment and have a marked influence on their ability to specifically recognize and respond to various stimuli through the release of an array o...
Source: Journal of Histochemistry and Cytochemistry - September 25, 2014 Category: Biochemistry Authors: da Silva, E. Z. M., Jamur, M. C., Oliver, C. Tags: Review Source Type: research

Controls for Immunohistochemistry: The Histochemical Society's Standards of Practice for Validation of Immunohistochemical Assays
Immunohistochemistry is widely used in biomedical research to localize specific epitopes of molecules in cells and tissues. The validity of interpretations based on immunohistochemistry requires appropriate positive and negative controls that are often not reported in publications. This omission may lead to incorrect interpretations and irreproducible results in the literature and contribute to wasted time, effort, and resources as well as erosion of confidence in scientific investigation by the general public, legislative bodies and funding agencies. The present article summarizes essential controls required for validatio...
Source: Journal of Histochemistry and Cytochemistry - September 25, 2014 Category: Biochemistry Authors: Hewitt, S. M., Baskin, D. G., Frevert, C. W., Stahl, W. L., Rosa-Molinar, E. Tags: Perspective Source Type: research