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PubMedDate not supplied

Unusual fallopian tube neoplasm in a 10-year-old patient with Proteus syndrome: a case report and review of the literature.

We report a case of fallopian tube and peritoneal lesions resected from a 10-year-old girl with Proteus syndrome. The papillary fallopian tube tumor expressed CA-125 and demonstrated negative staining for CEA, CD15, calretinin, and CK 5/6. Nondilated portions of fallopian tube exhibited evidence of an in situ component. This patient was also found to have peritoneal mesothelial hyperplasia as well as cystic peritoneal lesions of mesothelial origin, with positive calretinin staining. This case provides direct evidence of both Müllerian and mesothelial lesions arising concurrently in a patient with Proteus syndrome and represents the 1st documented fallopian tube neoplasm. Previously reported lesions involving the female and male genital tracts in patients with Proteus syndrome are reviewed.

Biomarkers, Tumor↗
PubMed2006

Neonatal loss of gamma-aminobutyric acid pathway expression after human perinatal brain injury.

OBJECT: Perinatal brain injury leads to chronic neurological deficits in children. Damage to the premature brain produces white matter lesions (WMLs), but the impact on cortical development is less well defined. Gamma-aminobutyric acid(GABA)ergic neurons destined for the cerebral cortex migrate through the developing white matter and form the subplate during late gestation. The authors hypothesized that GABAergic neurons are vulnerable to perinatal systemic insults in premature infants, and that damage to these neurons contributes to impaired cortical development. METHODS: An immunohistochemical analysis involving markers for oligodendrocytes, GABAergic neurons, axons, and apoptosis was performed on a consecutive series of 15 human neonatal telencephalon samples obtained postmortem from infants born at 25 to 32 weeks of gestation. The tissue samples were divided into two groups based on the presence or absence of WMLs by performing routine histological analyses. The expression of GABAergic neurons was compared between the two groups by using age-matched samples. Two-tailed t-tests were used for statistical analyses. Ten infants had WMLs and five did not. Significant losses of oligodendrocytes and axons and markedly increased apoptosis were appreciated in tissue samples from the infants with WMLs. Samples from infants with WMLs also showed significant losses of glutamic acid decarboxylase-67-positive cells and calretinin-positive cells, shorter neuropeptide Y-positive neurite lengths, and losses of cells expressing GABA(A)alpha1, GABA(B)R1, and N-acetylaspartate diethylamide NR1 receptors when these factors were compared with those in samples from infants without WMLs (all p < 0.02). CONCLUSIONS: In addition to oligodendrocyte loss, axonal disruption, and excess apoptosis, a significant loss of telencephalon GABAergic neuron expression was found in neonatal brains with WMLs, compared with neonates' brains without WMLs. The loss of GABAergic subplate neurons in infants with WMLs may contribute to the pathogenesis of neurological deficits in children.

Calbindin 2↗
PubMed2004

E-cadherin and calretinin as immunocytochemical markers to differentiate malignant from benign serous effusions.

AIM: To investigate the expressions of E-cadherin and calretinin in exfoliated cells of serous effusions and evaluate their values in distinguishing malignant effusions from benign ones. METHODS: Fresh serous effusion specimens were centrifuged and exfoliated cells were collected. Cells were then processed with a standardized procedure, including paraformaldehyde fixation, BSA-PBS solution washing and smears preparation. E-cadherin and calretinin were detected by immunocytochemistry (ICC). RESULTS: In the exfoliated cells of serous effusions, most of carcinoma cells only expressed E-cadherin, and most of mesothelial cells only expressed calretinin, and benign cells (lymphocytes and granulocytes) did not express either of them. For E-cadherin, 85.7% (30/35) of malignant effusions and 8.1% (3/37) of benign fluids were ICC-positive (P<0.001). The sensitivity of E-cadherin ICC in the diagnosis of malignant effusions was 85.7%, specificity 91.9%, and diagnostic rate 88.9%. For calretinin, 94.6% (35/37) of benign effusions and 11.4% (4/35) of malignant effusions were ICC-positive (P<0.001). The sensitivity of calretinin ICC in the diagnosis of benign effusions was 94.6%, specificity 88.6%, and diagnostic rate 91.7%. For diagnosis of benign and malignant effusions by combining E-cadherin ICC and calretinin ICC, the specificities were up to 100% and 97.1%, respectively. CONCLUSION: E-cadherin ICC and calretinin ICC are sensitive and specific in differential diagnosis of benign and malignant serous effusion specimens and specificities are evidently improved when both markers are combined.

Adolescent↗
PubMed2001

Unusual clear cell variant of epithelioid mesothelioma.

Clear cell mesothelioma is an extremely rare neoplasm of the pleura, which can easily be mistaken for a metastasis of clear cell carcinoma to the pleura. We report here the histochemical, immunohistochemical, and ultrastructural aspects of a new case of clear cell pleural mesothelioma in a 52-year-old man with no known asbestos exposure. He was admitted to the hospital for recurrent pleural effusion, which was negative for neoplastic cells at the cytologic examination. A partial decortication of the right pleura was performed. The morphologic, immunohistochemical, and ultrastructural features reported for this case are consistent with the diagnosis of clear cell mesothelioma. The differential diagnosis and immunohistochemical features in comparison with other clear cell neoplasms are discussed.

Biomarkers, Tumor↗
PubMed1998

[Immunohistochemistry in the differential diagnosis of mesothelioma and adenocarcinoma. Evaluation of 5 new antibodies and 6 traditional antibodies].

The aim of this study was to evaluate three recently marketed putative mesothelioma-binding antibodies, calretinin, HBME-1 and thrombomodulin, and two putative adenocarcinoma-binding antibodies, AUA1 and MOC31, on paraffin sections from 28 mesotheliomas and 30 adenocarcinomas. Moreover, the expression of ACE, BerEP4, CA125, CA19.9, LeuM1 and vimentin was assessed. Calretinin, HBME-1 and thrombomodulin, which showed a 100%, 89% and 43% sensitivity, and a 50%, 70% and 87% specificity for mesothelioma respectively, were less efficient than vimentin (100% specificity and 67% sensitivity) for the positive identification of mesothelioma. AUA1, BerEP4 and MOC31 were 100% sensitive to adenocarcinoma, with BerEP4 and MOC31 having the highest specificity (86% each). The immunophenotype "vimentin-positive, ACE-negative, CA19.9-negative" yielded 100% sensitivity and 97% specificity for diagnosis of mesothelioma. We advocate the use of the four-marker panel of ACE, CA19.9, MOC31 (or BerEP4) and vimentin for differentiating mesothelioma from adenocarcinoma.

Adenocarcinoma↗
PubMed1999

Comparative studies of calretinin expression by WiDr cell line in vivo in xenografts in nude mice and in vitro.

WiDr cells from a human colon adenocarcinoma cultivated in vitro express the calcium binding protein calretinin. The immunoreactivity is present in some interphasic cells and decreases after seven days in culture together with the augmentation of the cell number. Calretinin expression is maintained in the undifferentiated cells of the tumoral mass developed in nude mice and in recultivated isolated tumour cells from the xenograft. From the experiments here described, the protein expression is quantitatively influenced in vitro by the addition of drugs, such as colchicine and taxol, which intervene in cytoskeleton organisation. The percentage of the calretinin immunoreactive cells increases after the addition of colchicine to the medium while the immunoblot analysis shows a higher calretinin content in the cells treated with taxol.

Adenocarcinoma↗
PubMed1999

Antibody to calretinin stains AII amacrine cells in the rabbit retina: double-label and confocal analyses.

The AII or rod amacrine cell is a critical interneuron in the rod pathway of mammalian retinae. In this report, it is shown that commercially available antibodies to the calcium binding protein calretinin may be used to label the population of AII amacrine cells selectively. Calretinin-positive amacrine cells had the morphological attributes of AII amacrine cells. Double-labeling procedures showed that calretinin-positive somata were surrounded by dopaminergic varicosities and that calretinin-positive dendrites enclosed rod bipolar terminals, both as previously described for AII amacrine cells. By analyzing the surrounding kernel for each labeled pixel in the rod bipolar image, it is shown here that AII processes are adjacent to rod bipolar terminals at a level that far exceeds the random overlap present in images in which one label was rotated out of phase. Such a spatial relationship is indicative of synaptic connections, as well described for rod bipolar input to AII amacrine cells. AII amacrine cells also were double-labeled for calretinin and parvalbumin; however, a scattergram analysis of red versus green intensity showed that the parvalbumin antibody stained additional unidentified amacrine cells. In conclusion, at the appropriate dilution, calretinin antibodies are a useful marker for AII amacrine cells in the rabbit retina.

3,3'-Diaminobenzidine↗
PubMedDate not supplied

Calretinin--sensor or buffer--function still unclear.

Calretinin (CR) is a 31.5 kDa EF-hand calcium binding protein. CR has a general neuronal localization and is a predominantly cytosolic protein. The biochemical properties of this protein are well characterized but its function is still unclear. It is often postulated that the presence of CR correlates with an increased survival ability of cells under pathological conditions connected with increased intracellular calcium levels. However, not all studies confirm such a relationship and they indicate that the presence of CR does not protect cells against calcium overload and that CR does not act as an intracellular calcium buffer. Recent results concerning CR function are discussed and the conclusion is proposed that CR acts as a calcium modulator rather than a calcium buffer.

Animals↗
PubMed1999

Prosomeric map of the lamprey forebrain based on calretinin immunocytochemistry, Nissl stain, and ancillary markers.

The structural organization of the lamprey extratelencephalic forebrain is re-examined from the perspective of the prosomeric segmental paradigm. The question asked was whether the prosomeric forebrain model used for gnathostomes is of material advantage for interpreting subdivisions in the lamprey forebrain. To this aim, the main longitudinal and transverse landmarks recognized by the prosomeric model in other vertebrates were identified in Nissl-stained lamprey material. Lines of cytoarchitectural discontinuity and contours of migrated neuronal groups were mapped in a two-dimensional sagittal representation and were also classified according to their radial position. Immunocytochemical mapping of calretinin expression in adjacent sections served to define particular structural units better, in particular, the dorsal thalamus. These data were complemented by numerous other chemoarchitectonic observations obtained with ancillary markers, which identified additional specific formations, subdivisions, or boundaries. Emphasis was placed on studying whether such chemically defined neuronal groups showed boundaries aligned with the postulated inter- or intraprosomeric boundaries. The course of diverse axonal tracts was studied also with regard to their prosomeric topography. This analysis showed that the full prosomeric model applies straightforwardly to the lamprey forebrain. This finding implies that a common segmental and longitudinal organization of the neural tube may be primitive for all vertebrates. Interesting novel aspects appear in the interpretation of the lamprey pretectum, the dorsal and ventral thalami, and the hypothalamus. The topologic continuity of the prosomeric forebrain regions with evaginated or non-evaginated portions of the telencephalon was also examined.

Acetylcholinesterase↗
PubMed2000

Serotonin-driven long-range inhibitory connections in the cerebellar cortex.

Disturbances of the serotoninergic neuromodulation in the cerebellar cortex have been involved in several types of ataxia, but the physiological action of serotonin in this structure remains poorly understood. We report that in slices of the rat cerebellar vermis, serotonin triggers the firing of an inhibitory interneuron presynaptic to Golgi cells. The Lugaro cell, a neglected interneuronal type, satisfies the expected criteria for this input, whereas basket cells, stellate cells, or Golgi cells do not. Lugaro cells are selectively excited by serotonin, and their firing behavior (sustained steady frequency in the 5-15 Hz range) resembles the pattern of occurrence of serotonin-evoked IPSCs in Golgi cells. Immunohistochemical stainings and single cell reconstructions show that Lugaro cell axons form a parasagittal plexus but also extend long transverse branches that run parallel to the parallel fibers and are partly myelinated. Electrophysiological data suggest that these transverse axons participate in synaptic contacts of the Lugaro cells with Golgi cells, and we calculated that in the intact cerebellum a given Lugaro cell contacts >100 Golgi cells. Serotonin modulation of Lugaro cells may constitute an intracortical switch involved in information patterning at the level of Golgi cells and granule cells populations, and particularly in synchronizations recorded along the transverse axis in vivo.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed2000

Embryonic and early fetal development of the human neocortex.

Early corticogenesis was studied in human embryos and early fetuses from Carnegie stages 16 to 22 (5-8 gestational weeks) by using immunohistochemistry for Reelin (Reln), calretinin (CR), and glutamic acid decarboxylase (GAD). A first population of Reln-positive cells appears in the neocortical anlage at stage 16 and increases in number at stages 17-18. At stages 19-20, a monolayer of horizontal CR- and GAD-positive, Reln-negative neurons forms in the preplate, whereas Reln-positive cells shift into a subpial position. Another cell class, the pioneer projection neuron, is CR-positive but GAD- and Reln-negative; pioneer cells contribute early corticofugal axons. Pioneer cells first appear below the monolayer at stage 20 and form a pioneer plate at stage 21. The cortical plate (CP) proper emerges at stage 21 and inserts itself within the pioneer plate, which is thus split into a minor superficial component and a larger deep component that presumably corresponds to the subplate. Initial CP neurons are radially organized and mostly CR-negative. Reln-positive cells remain consistently segregated from the pioneer cells and are thus not directly involved in preplate partition. Our data indicate that the neuronal composition of the human neocortical preplate is more complex than generally described and that various neurons participate in a sequence of events that precede the emergence of the CP.

Calbindin 2↗
PubMed2000

Developmental changes in the subcellular localization of calretinin.

Brainstem auditory neurons in the chick nucleus magnocellularis (NM) express high levels of the neuron-specific calcium-binding protein calretinin (CR). CR has heretofore been considered a diffusible calcium buffer that is dispersed uniformly throughout the cytosol. Using high-resolution confocal microscopy and complementary biochemical analyses, we have found that during the development of NM neurons, CR changes from being expressed diffusely at low concentrations to being highly concentrated beneath the plasma membrane. This shift in CR localization occurs at the same time as the onset of spontaneous activity, synaptic transmission, and synapse refinement in NM. In the chick brainstem auditory pathway, this subcellular localization appears to occur only in NM neurons and only with respect to CR, because calmodulin remains diffusely expressed in NM. Biochemical analyses show the association of calretinin with the membrane is detergent-soluble and calcium-independent. Because these are highly active neurons with a large number of Ca2+-permeable synaptic AMPA receptors, we hypothesize that localization of CR beneath the plasma membrane is an adaptation to spatially restrict the calcium influxes.

Animals↗
PubMed2000

Diagnostic utility of calretinin immunohistochemistry in cytologic cell block preparations.

BACKGROUND: Calretinin (CR) is a valuable marker in the immunohistochemical distinction between malignant mesothelioma (MM) and adenocarcinoma (ACA) in tissue sections. However, there is limited and conflicting data regarding the utility of CR in this differential diagnosis on cytologic material, especially cell block preparations. Also, the possible role of CR in the distinction of papillary serous borderline tumor (SBT) cells from reactive mesothelial cells in peritoneal washings has not been examined. METHODS: Formalin-fixed paraffin-embedded cell block specimens of cytologic fluids, washings, and aspirates with a suspicious or positive cytologic diagnosis and a confirmed diagnosis of MM (29 cases), ACA (39 cases), and SBT (10 cases) were used for CR immunohistochemistry (IHC). RESULTS: Moderate to strong staining in > 50% of tumor cells was noted in 26 of 29 (90%) MMs but in only 3 of 39 (8%) ACAs and 1 of 10 (10%) SBTs. Admixed reactive mesothelial cells (when present) were strongly positive in all fluid specimens, but the staining pattern of benign non-reactive mesothelial cells in washing specimens was less reliable. CONCLUSIONS: CR is both a sensitive and specific marker of reactive and neoplastic mesothelial cells in cytologic cell block preparations and is thus useful in the differential diagnosis of MM and metastatic ACA in malignant effusions. However, CR IHC does not appear to allow definitive identification of SBT in peritoneal washings.

Adenocarcinoma↗
PubMed2001

Calretinin-, neurocalcin-, and parvalbumin-immunoreactive elements in the olfactory bulb of the hedgehog (Erinaceus europaeus).

The distribution pattern and morphology of calretinin-, neurocalcin-, and parvalbumin-immunoreactive neurons were studied in the main and accessory olfactory bulbs of the hedgehog. The detection of these markers was carried out by using monoclonal or polyclonal antibodies and the avidin-biotin-immunoperoxidase method. Specific neuronal populations were positive for these calcium-binding proteins in the hedgehog olfactory bulb, revealing both similarities to and differences from the data reported in the olfactory bulb of rodent species. The distribution pattern of each calcium-binding protein studied in the accessory olfactory bulb was highly similar to that described in other macrosmatic species. However, in the main olfactory bulb, the markers analyzed were expressed in similar interneuronal populations as they are in the rodent olfactory bulb, whereas cell groups categorized as projecting neurons demonstrated striking differences in the expression of these calcium-binding proteins. These results suggest that the expression of calcium-binding proteins in a given brain region is not a constant feature among species despite a similar organization but that different factors could influence their expression. Thus, the accessory olfactory system involved in the processing of specific and similar olfactory cues among species demonstrates a more constant organization among species. By contrast, the functionally important role of the main olfactory system in the hedgehog is accompanied by a more complex organization, which is reflected in an increased diversity of calcium-buffering systems.

Animals↗
PubMedDate not supplied

Expression of calretinin in human mesothelioma cell lines and cell cycle analysis by flow cytometry.

Human mesothelioma cell lines were studied concerning the expression of the calcium-binding protein calretinin (CR), and the relation of the DNA index to their cell cycle. The results obtained for cell lines with different morphological characteristics, were compared to those from human mesothelial cells transfected with SV40 to escape senescence. Immunocytochemical expression of calretinin (confirmed by immunoblot) was observed in all mesothelioma cell lines but not in the control cells. It is suggested that calretinin is active in the first steps of carcinogenesis in all human mesotheliomas and during several stages in the evolution towards malignancy.

Blotting, Western↗
PubMed2001

Morphology of calretinin and tyrosine hydroxylase-immunoreactive neurons in the pig retina.

The morphology of calretinin- and tyrosine hydroxylase-immunoreactive (IR) neurons in adult pig retina was studied. These neurons were identified using antibody immunocytochemistry. Calretinin immunoreactivity was found in numerous cell bodies in the ganglion cell layer. Large ganglion cells, however, were not labeled. In the inner nuclear layer, the regular distribution of calretinin-IR neurons, the inner marginal location of their cell bodies in the inner nuclear layer, and the distinctive bilaminar morphologies of their dendritic arbors in the inner plexiform layer suggested that these calretinin-IR cells were AII amacrine cells. Calretinin immunoreactivity was observed in both A-and B-type horizontal cells. Neurons in the photoreceptor cell layer were not labeled by this antibody. The great majority of tyrosine hydroxylase-IR neurons were located at the innermost border of the inner nuclear layer (conventional amacrines). The processes were monostratified and ran laterally within layer 1 of the inner plexiform layer. Some of the tyrosine hydroxylase-IR neurons were located in the ganglion cell layer (displaced amacrines). The processes of displaced tyrosine hydroxylase-IR amacrine cells were also located within layer 1 of the inner plexiform layer. Some processes of a few neurons were located in the outer plexiform layer. A very low density of neurons had additional bands of tyrosine hydroxylase-IR processes in the middle and deep layers of the inner plexiform layer. The processes of tyrosine hydroxylase-IR neurons extended radially over a wide area and formed large, moderately branched dendritic fields. These processes occasionally had varicosities and formed "dendritic rings". These results indicate that calretinin- and tyrosine hydroxylase-IR neurons represent specific neuronal cell types in the pig retina.

Animals↗