1,721,013 research outputs found
IMMUNOLOCALIZATION OF PROTEIN KINASE C ALPHA, DELTA, EPSILON, ZETA DURING PRE-OSTEOCLASTS DIFFFERENTIATION INDUCED BY RANKL AND M-CSF
Selective distribution of multiple protein kinase C isoforms in mouse cerebellar cortex.
An immunohistochemical study concerning the distribution of protein kinase C isoforms, a lipid-regulated serine/threonine kinase essential for signal transduction, was performed in mice cerebellar cortex, with particular emphasis on the localization of -iota and -lambda isozymes. By the means of immunoblotting analyses we detected the presence of 11 PKC subspecies in whole cerebellar extracts. Immunoreactivity on cryostat sections revealed, using polyclonal and monoclonal antibodies, that a few isoforms were widely but discretely distributed in all three cortical layers (molecular, granular and Purkinje cells) whereas other isozymes were present in a limited neuronal compartment. Overall, the distribution of several isoforms was in agreement with data obtained by other authors using rat cerebellum. As far as -iota and -lambda isozymes were concerned, we found them abundantly expressed in endothelial cells. Moreover, protein kinase C-lambda was also present in the body of Purkinje cell, conceivably associated with a 200-kDa neurofilament component. In all, these results hint at the possibility that in the cerebellar cortex at least some protein kinase C isoforms are involved in functions other than signal transduction at the synaptic level
Atypical isoenzymes of PKC-iota, -lambda, -mu: relative distribution in mouse foetal and neonatal organs.
On the presence of a secondary cartilage in the mental symphyseal region of human embryos and fetuses.
The presence of a secondary cartilage in the mental symphyseal region was examined in this study. A double-staining method with alcian blue and alizarin red S was performed on both whole human embryos and fetuses (developmental age between 8 and 17 weeks, crown-rump length, CRL, between 37 and 124 mm) and their disjointed mandibles. Histological and histochemical techniques were applied to transverse serial sections of whole disjointed fetal heads. The ossification process observed in the mental symphysis is quite different from that of the mandibular body, whose membranous ossification is induced by the contiguous Meckel's cartilage. No evidence of any fusion of Meckel's cartilage with the symphyseal cartilage, that lies within the symphyseal space, was detected. On the basis of these findings, we suggested that the mental secondary cartilage is able to change into bone according to an endochondral ossification process. Moreover, the role of mechanical causes in the development of the mental symphysis was hypothesized
An immunohistochemical study of protein kinase C distribution in fetal mouse vertebral column.
Using polyclonal antibodies we have studied the distribution of protein kinase C in fetal mouse low thoracic vertebrae. By means of a pan protein kinase C antiserum recognizing the catalytic domain of the enzyme, we show that protein kinase C is markedly expressed in chondrocytes before birth. The enzyme seems to be very abundant in the more mature cells that are close to ossification centres as well as the periphery of the intervertebral disc, although it can also be detected in chondrocytes. In order to establish which protein kinase C isoenzyme(s) the chondrocytes produce, we employed polyclonal isoenzyme-specific antisera developed against three calcium-dependent isoforms (alpha, beta, gamma) and three calcium-independent isoforms (delta, epsilon, zeta). Secondary antibody conjugated to alkaline phosphatase revealed that chondrocytes markedly express the beta-isoform. Cells were also weakly stained by the anti-epsilon serum. The immunostaining was completely abolished by pre-incubating primary antibodies with the peptide antigens to which they were raised. These results suggest that protein kinase C (and particularly the beta isoform) could play an important role in mouse fetal chondrogenesis of the vertebral column
Histochemical study on catecholamines in human coronary endothelium.
Specific catecholaminergic granules had been previously described in the endothelial cells of blood and lymphatic vessels. 2 histochemical techniques were used in this work for detecting catecholamines in human coronary vessels: both the postfixation with OsO4-KI mixture and the formaldehyde induced fluorescence (FIF) reaction. Ultrastructural examinations of bioptic specimens processed with the OsO4-KI staining showed a marked positivity in the coronary endothelial cells, as well as in the smooth muscle fiber cells of the coronary arteriolae and the adventitial nerve endings. These findings were confirmed by a high level of fluorescence in the same structures, obtained using the FIF reaction. Myocardial fiber cells never reacted. Therefore, an important role of the endothelium of human coronary vessels in the turn-over of catecholamines could have been supposed
, "REGULATION OF BREAST CANCER MIGRATION TO BONE: ROLE OF RANK/RANKL/OPG". ATTI DEL CONVEGNO "32 ND NATIONAL CONGRESS OF THE ITALIAN SOCIETY OF HISTOCHEMISTRY",
A quantitative study on the spatial and temporal ossification patterns of vertebral centra and neural arches and their relationship to the fetal age.
A double-staining technique on 37 human embryos and fetuses (crown-rump length, CRL, between 38 and 116 mm) has been performed to study the ossification patterns of the vertebral column. Different growth sequences for centra and neural arches were observed. The survey of ossified centers suggested it was possible to relate significantly their appearance with the CRL. On the basis of already known data defining the developmental age in relationship to the latter parameter, we suggest their numerical evaluation as a further parameter for the assessment of the fetal age. Therefore, we have worked out a table that may be used either to determine the normal fetal growth, or when other parameters cannot be relied upon (i.e. in morphological diseases) for this aim
Subtraction of autofluorescent dead cells from the lymphocyte flow cytometric binding assay.
Flow cytometry allows the quantitative analysis of lymphocyte-target cell
conjugates and the identification of the lymphocyte subset involved in the
binding phenomenon. We recently described a methodology to identify the effector
cells bound to K562 targets based on target cell autofluorescence coupled with
lymphocyte staining by means of fluorescent monoclonal antibodies. Here we
describe an implementation of the methodology that allows the subtraction of
spontaneously dead targets to which lymphocytes may or may not adhere, thereby
preventing the overestimation of the binding phenomenon and limiting its
evaluation to living effector-target conjugates, thus preserving the specificity
of the phenomenon
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