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[Presence of calretinin in neurons of the human intestine].

Calcium-binding proteins are present in different neuron populations in the Central Nervous System. As concerns the Enteric Nervous System, only a few studies have been performed. In the present work we investigated immunohistochemically the localization of Calretinin in neurons of the human intestinal wall. Our results showed the presence of stained cell bodies and fibers with antibodies against Calretinin in the Auerbach plexus. Since most of the enteric neurons are characterized by a slow phase after hyperpolarization caused by Ca2+ dependent K+ channels, the role of Calretinin could be to modulate this particular electrophysiological behaviour.

Calbindin 2↗

[Significance of combining detection of E-cadherin, carcinoembryonic antigen, and calretinin in cytological differential diagnosis of serous effusion].

BACKGROUND & OBJECTIVE: The morphological characteristics of metastatic adenocarcinoma cells,malignant mesothelioma cells,and reactive mesothelial cells in serous effusions are similar. Sometimes, it is difficult to make a correct diagnosis just based on the cytological characteristics. Although now immunocytochemistry has been widely used in this field,yet only a few antibodies were reported in China,which can't help to make a correct differential diagnosis. This study was to evaluate the value of combining detection of E-cadherin,carcinoembryonic antigen (CEA),and calretinin in cytological differential diagnosis of serous effusion. METHODS: A total of 93 effusion cytology specimens (66 pleural fluid samples, 24 ascites samples, 3 pericardial fluid samples)from 55 cases of metastatic adenocarcinoma, 6 cases of malignant mesothelioma, and 32 cases of reactive mesothelium were collected. All cases were confirmed histologically or clinically. Materials from each case were prepared in HE staining smears,and cell blocks,immunocytochemical staining was performed on cell block sections. RESULTS: In diagnosing metastatic adenocarcinoma, the sensitivities of E-cadherin, and CEA were 85.5%(47/55), and 79.6% (43/55), the specificities were 100% (38/38), and 97.4% (37/38);if E-cadherin and CEA were used together,the positive rate was 96.4%(53/55). The sensitivity of calretinin for mesothelial cells was 81.6%(31/38),and the specificity was 87.2% (48/55). CONCLUSION: The combination of E-cadherin, CEA, and calretinin is a very useful antibody panel in cytological differential diagnosis for metastatic adenocarcinoma and mesothelial cells in serous effusion.

Adenocarcinoma↗

A cytokeratin- and calretinin-negative staining sarcomatoid malignant mesothelioma.

Malignant Mesothelioma, or mesothelioma, is a mesothelial-based malignancy that may occur in the pleura, pericardium and peritoneum. Mesothelioma is a very aggressive cancer with limited treatment, and a median survival of about 1 year. At times, the diagnosis of mesothelioma may be problematic. The final diagnosis of mesothelioma relies on histology and often is dependent upon immunohistochemistry. It is generally assumed that mesotheliomas must stain positive for cytokeratin and calretinin and negative staining for these markers would rule out the diagnosis. We encountered a patient with a pleural-based, cytokeratin- and calretinin-negative sarcomatoid malignancy. These negative stainings would rule out the diagnosis of mesothelioma but, after careful consideration of the patient's clinical records, and additional histological and immunohistochemical studies, we conclude that this patient suffered from mesothelioma of the sarcomatoid type.

Aged↗

Mice lacking IGF-I and LIF have motoneuron deficits in brain stem nuclei.

We analyzed the neural embryonic phenotype of single and double-mutant mice for insulin-like growth factor-I (IGF-I) and leukemia inhibitory factor (LIF). The anatomical structure of the hippocampus, the cerebellum, and the olfactory epithelium, regions showing expression of both factors and their receptors, appeared largely normal in all mutant mice. In the fimbria and the spinal cord, similar patterns of glial fibrillary acidic protein (GFAP)-expressing astrocytes were found in wild-type and mutant mice. In contrast, single Igf-I and double-mutant mice showed a significant reduction in the number of trigeminal and facial motoneurons, whereas mice lacking LIF showed a significant reduction of trigeminal motoneurons. These results suggest that IGF-I and LIF regulate cooperatively motoneuron numbers in specific brain stem nuclei.

Animals↗

Identification of calretinin and the alternatively spliced form calretinin-22k in primary pleural mesotheliomas and in their metastases.

BACKGROUND: Antibodies against calretinin represent an established, powerful and reliable immunohistochemical marker in the differential diagnosis between mesothelioma and adenocarcinomas. However, in studies published so far, the exact molecular identity of the immunoreactive protein(s) detected in mesothelioma sections has not yet been determined. MATERIALS AND METHODS: Tissue biopsies from ten mesothelioma samples, primary and metastatic, were analyzed by Western blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR), in addition to immunohistochemical staining. RESULTS: Besides the selective positive immunoreaction in all 10 samples, calretinin was identified by Western blot analysis and by identification of its mRNA by RT-PCR. In addition, we identified the alternatively spliced form calretinin-22k and the corresponding mRNA, previously detected only in neoplastic cells in colon carcinomas and derived cell lines. CONCLUSION: Our findings demonstrate that the immunoreactivity observed in the mesotheliomas and their metastases investigated in this report is due to the concomitant presence of calretinin and the alternatively spliced form calretinin-22k.

Adult↗

Epithelial splenic cysts.

BACKGROUND: Epithelial splenic cysts (ESC) are uncommon lesions of the spleen. The etiopathogenesis of these cysts is controversial, even if Burrig's theory is the most accredited. The histological distinction between epidermoid and mesothelial cysts may be difficult, particularly for monostratified epithelium. PATIENTS AND METHODS: In the period between January 1986 and February 2004, 11 patients with ESC were studied. The history, physical findings, all relevant diagnostic studies and treatment were reviewed. All histological material was reviewed in detail with immunohistochemistry for CEA, CA 19-9, cytokeratin and calretinin. RESULTS: Epidermoid cysts were positive for CEA, CA 19-9, and cytokeratin, but negative for calretinin. Mesothelial cysts were positive for cytokeratin and calretinin, but negative for CEA and CA 19-9. CONCLUSION: Immunohistochemistry allows differential diagnosis between epidermoid and mesothelial cysts. With regard to etiopathogenesis, these data could mean that epidermoid and mesothelial cysts have distinct origins, though at variance with Burrig's theory. Although the ESC in this series were treated by open splenectomy, the recent approach by conservative and laparoscopic techniques offers great promise.

Adolescent↗

Analysis of calretinin immunoreactivity in the rat piriform cortex after open field stress during postnatal maturation.

In our study we used c-Fos protein to identify whether cells containing calretinin (CR) in the rat piriform cortex are engaged in the response to stress stimulation and to find out how this expression changes during maturation (PC). The material consisted of Wistar strain rats of between 0 and 120 days of age divided into 9 groups. Each group consisted of 5 experimental and 3 control rats. Animals from the experimental groups were exposed to the open field test throughout 10 minutes. The control animals were kept in a home cage. In all age-related control rats weak c-Fos immunoreactivity was observed. Our results showed that cells containing c-Fos following an acute open field test were observed predominantly in layers II and III of the PC just after birth. Their number then increased and stabilised on P30. We had already observed immature CR-ir cells at birth. In the 4th week of life these neurons achieved maturity. Their number increased to P90 and decreased in older animals. CR-ir neurons were localised mainly in layer II and to a lesser degree in layers III and I of the PC. Double immunostaining c-Fos/CR revealed that the level of co-localisation was low. Only small differences were observed between the anterior and posterior parts of the PC. In the anterior part a higher number of CR-ir neurons was found. The difference in the level of co-localisation between the anterior and posterior parts was age-related and differentiated. Our results may suggest that during maturation CR-ir neurons of the piriform cortex are not the main population engaged in response to the open field test.

Age Factors↗

Expression of mesothelial markers in malignant mesotheliomas: an immunohistochemical evaluation of 173 cases.

OBJECTIVE: To see the distribution of Calretinin, thrombomodulin, CK5/6 and HBME-1 markers in various subtypes of mesotheliomas and extend the published data on this topic. The positivity of adenocarcinoma specific markers (CEA and BerEP4) in malignant mesotheliomas have also been evaluated. METHODS: Various markers in 173 cases of malignant mesotheliomas received over a period of 8 years were evaluated by immunohistochemistry. RESULTS: In majority of malignant mesotheliomas i.e., epithelioid and biphasic types, the positive staining patterns complement the gold standard histologic diagnosis. However, in a small minority mainly sarcomatoid variant, heavy reliance cannot be placed on these markers. CEA and BerEP4 are useful negative markers of mesotheliomas, although occasionally these are positive in clear cut mesotheliomas. CONCLUSIONS: Specificity of various markers in malignant mesotheliomas should be assessed according to histologic subtypes. The existing generation of markers is not reliable in diagnosis of sarcomatoid mesotheliomas. Fortunately this forms only a small group of mesothelial malignancy. In common epithelioid and biphasic variants calretinin, thrombomodulin, CK5/6, HBME-1 are sensitive positive markers whereas CEA and BerEP4 are negative markers of malignant mesotheliomas.

Antigens, Neoplasm↗

[Differential diagnostic value of B72.3, Ber-EP4 and calretinin in serous effusions].

OBJECTIVE: To determine the diagnostic value of B72.3, BerEP4 and calretinin in differentiating metastatic carcinoma cells from reactive mesothelial cells (RMC) in serous effusions by using immunocytochemical method (ICC), and to investigate the feasibility of ThinPrep (TP) preparation for ICC. METHODS: One hundred fifty eight serous effusion specimens were examined by ICC on cell block (CB) sections (CB-ICC) using antibodies against of B72.3, BerEP4 and calretinin. Fourty-nine of the samples, ICC on ThinPrep slides (TP-ICC) and CB-ICC were performed concurrently. RESULTS: The sensitivities of B72.3 and Ber-EP4 for detecting carcimoma cells were 76.9% and 69.2% respectively, and when combined the sensitivity was increased to 89.7%. The sensitivity and specificity of Calretinin for detecting mesothelial cells were 90.9% and 87.2% respectively. The sensitivity of B72.3 in differentiating cancer cells from reactive mesothelial cells by CB-ICC and TP-ICC was 78.9% and 68.4%. It was 78.9% and 68.4% of BerEP4 respectively. No statistical significance was observed between CB-ICC and TP-ICC in differentiating metastatic carcinoma cells from reactive mesothelial cells. CONCLUSION: The combination of antibodies of B72.3, Ber-EP4 and calretinin is quite helpful as an auxiliary in differentiating metastatic carcinoma cells from reactive mesothelial cells. ThinPrep preparation slides may effectively replace the cell block sections for ICC in differential diagnosis of serous effusions.

Antibodies, Monoclonal↗

[Clinicodiagnostic significance of gastroesophageal reflux and structural changes in the esophagus and stomach in bronchial asthma].

Four hundred patients with bronchial asthma (BA) were questioned. Complex clinical and instrumental examination of 60 patients with BA and endoscopic-positive gastroesophageal reflux disease was carried out; the control groups included 30 patients with BA alone and 30 patients with GERD alone. Clinical symptoms of GERD were revealed in 58% of BA patients; endoscopic-positive GERD was diagnosed in 48.6% of BA patients. The study proved the connection between the degree of respiratory disturbances, on the one hand, and the degree of esophagitis and the level of intraesophageal pH, on the other, in patients with BA plus GERD. The study also established the role of esophageal and gastric epitheliocytes, producing endothelin-1, melatonin, NO-synthase, and calretinin, in the occurrence of mutual exacerbation syndrome.

Adult↗

[Adenomatoid tumor].

Adenomatoid tumour is a neoplastic process of discussed origin, but the immunohistochemical phenotype leads a mesothelial derivation. The preferential site of origin is the genital apparatus of both sexes, however extragenital cases have been described. The histological pattern varies from tubular formation, to solid growth, to cystic areas. In the present report we described a case of Adenomatoid tumour of the uterus body in a 46 years old patient.

Actins↗

Expression of calcium binding proteins in the developing spine: preliminary report.

Previous ultrastructural studies on the developing spine induced the Authors to analyse the presence of Ca-binding proteins in the vertebral anlagen of the chick embryo, by immunostaining for calretinin. At 7 incubation days, intense positivity for calretinin at level of the intervertebral region is detectable. From the 7th day the positivity is localized in particular to the intervertebral menisci and it appears also in the degenerating chondrocytes of the chondrifying vertebrae and at level of the peripheral zones of vascular erosion in the ossifying vertebrae. The intervertebral positivity for calretinin can be related to the presence at this level of fibroblast-like cells, that for such immunoreactivity are distinguishable from true fibroblasts. Since other Authors showed presence of S-100 protein in fibroblast-like cells during histogenesis of white adipose tissue, the significance of the expression of some Ca-binding proteins in the differentiation of these cell types can be supposed. The meaning of the immunoreactivity in the vertebral bodies is still uncertain and needs further immunohistochemical studies.

Animals↗

Interneurons containing calretinin are specialized to control other interneurons in the rat hippocampus.

Spine-free calretinin-immunoreactive (CR-IR) interneurons form a subpopulation of GABAergic cells in the rat hippocampus. A characteristic feature of these cells--located in all areas and layers--is the frequent dendro-dendritic and axo-dendritic contacts they form with each other. In this study we examined in detail the connectivity of these neurons by reconstructing their dendritic and axonal arbor and by identifying their postsynaptic targets. Radially running dendrites of CR-IR cells, located in different layers, intermingled into long braids. An average cell was in contact with dendrites of three to seven other CR-IR cells. Reconstruction of the dendritic trees from six consecutive sections demonstrated that at least 15 cells may participate in a dendro-dendritically connected cluster. Electron microscopical examination revealed that regularly spaced zonula adherentia connect the touching dendrites. The postsynaptic targets of CR-IR neurons have been examined using postembedding immunogold staining for GABA. CR-containing GABA-immunoreactive axons of local origin formed multiple symmetrical synaptic contacts (two to five) exclusively on GABAergic dendrites (CR-negative as well as CR-positive). Two to 10 CR-IR axons may converge onto a single CR-IR neuron, often from cells belonging to the same dendro-dendritically connected cluster. Using double immunocytochemistry, CR-IR cells were shown to heavily innervate calbindin D28k-containing interneurons and VIP-containing basket cells but avoided the parvalbumin-containing basket and axo-axonic cells. The unique connectivity of CR-IR cells may enable them to play a crucial role in the generation of synchronous, rhythmic hippocampal activity by controlling other interneurons terminating on different dendritic and somatic compartments of principal cells.

Animals↗

Calretinin immunoreactivity in the monkey and cat cerebellum: cellular localisation and modular distribution.

We have studied the morphology and distribution of calretinin-containing neurons in the cat and monkey cerebellum immunocytochemically. Immunoreactivity is detectable in cortex and white matter, with cell bodies mainly in the granular layer, the commonest of which are granule cells. Labelled Lugaro cells and a type of bipolar neuron are also occasionally seen. Other larger cells in the granular layer are strongly labelled. They are characterised by a spherical cell body giving rise to a thick primary dendrite with a spray of terminal appendages, like the unipolar brush cells (UBC) described in several species. They are distributed according to a mediolateral gradient of numerical density, with most in parasagittally congruent cell clusters in the vermis and the paravermal parts of the hemispheres. The axons of UBCs originate from the soma or primary dendrite, and are a major source of labelled fibres in the granular layer. Positively stained climbing fibres are radially oriented in the white matter and cortex, and form longitudinal bands, especially in the vermis. Climbing fibre bands are congruent with UBC clusters in the vermis but less so in the hemispheres. Both mossy and climbing fibres seem likely to synapse on UBSs. We postulate that a pathway from afferent fibres to UBCs to Purkinje cells may represent a short circuit for rapid cerebellar reflexes without passing through much of the multisynaptic circuits of the cortex. Calretinin may be involved in signal processing in the cerebellum, perhaps regulating the amount and form of intracellular calcium.

Animals↗

Distribution of calretinin-immunoreactivity in the rat entorhinal cortex: coexistence with GABA.

Inhibitory neurons in the entorhinal cortex control information flow between the cortical areas and the hippocampus. We characterized the inhibitory circuits in the rat entorhinal cortex by analyzing the distribution of calretinin-immunoreactivity and its colocalization with glutamate decarboxylase (GAD) and gamma-aminobutyric acid (GABA). The location of calretinin-immunoreactive (IR) neurons and terminals varies between the different layers and subfields of the entorhinal cortex. The immunopositive neurons can be divided into two major morphological classes: bipolar and multipolar, which have two or more long, aspiny or sparsely spiny dendrites that extend through several layers. In addition, there are unclassified immunopositive neurons that have large lightly stained somata. They are located primarily in layer V. Colocalization analyses with GAD and GABA revealed that approximately 40% (657 out of 1,777) of all calretinin-IR cells within the entorhinal cortex contain GAD or GABA. In layers I-III, over 90% of the calretinin-IR neurons contain GAD or GABA. In layers V-VI, however, most of the calretinin-IR neurons do not colocalize with either GAD or GABA. The distribution patterns of calretinin-immunoreactivity in the entorhinal cortex is consistent with the partitioning of the rat entorhinal cortex into six subfields. Furthermore, calretinin is expressed in a morphologically heterogeneous population of cells in the rat entorhinal cortex which includes both GABAergic and non-GABAergic neurons.

Animals↗

Heterogeneity of expression of the calcium-binding protein calretinin in human colonic cancer cell lines.

We searched for the presence of calretinin (CR) in 12 colonic cancer cell lines using immunohistochemistry, Western blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). We were able to demonstrate that calretinin is expressed in the cell lines HT-29, WiDr, LoVo, LS180, CO112, CO115. SW480, SW620, COLO205 and SK-CO-1, while no detectable amounts were found in the cell lines SW1116 and Caco-2. In general, rapidly proliferating cell lines expressed calretinin, whereas the protein was absent from cell lines with a low multiplication rate. A possible role for calretinin in maintaining the proliferative cycle of tumor cells is discussed.

Adenocarcinoma↗