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Biomedical subjects

A Calligaro

Publications and source records attributed to A Calligaro.

At least 37 records · Page 2Linked to original sources

Ultrastructural Localization of elastin-like immunoreactivity in the extracellular matrix around human small lymphatic vessels.

The distribution of elastin-like immunoreactivity around small lymphatic vessels was investigated in three different human tissues (skin, heart and dental pulp) using high resolution immunocytochemistry. Quantitative assessment of the immunogold reaction was performed with an image analysis system. Intense and moderate elastin-immuno-reactivity was detected in the extracellular matrix around small lymphatic vessels of the skin and heart, respectively. By contrast, absence of immunostaining was observed around lymphatic vessels in the dental pulp. Although the staining was mostly detectable on the non-fibrillar amorphous component of the extracellular matrix, some microfibrils were also immunostained in close proximity to the lymphatic vessel wall. These findings support the concept that small lymphatic vessels may be heterogeneous with respect to the composition of the extracellular matrix around their wall. The observation that it is possible to observe small lymphatic vessels displaying low or no elastin-immunoreactivity in the adjoining matrix militates against the hypothesis that elastic fibers play a pivotal role in the mechanisms that regulate the function of small lymphatic vessels.

Elastin↗

Expression of c-erbB-2 proto-oncogene-encoded protein (p185erbB-2) in functional rat ameloblasts.

The c-erbB-2 proto-oncogene encodes a transmembrane glycoprotein with tyrosine kinase activity, p185erbB-2, that has been previously localized in some developing and mature epithelia. The possible occurrence of p185 in the inner enamel epithelium of rat tooth germ was here investigated immunocytochemically. Postmitotic functional ameloblasts displayed intense p185-immunoreactivity, thus suggesting that c-erbB-2 proto-oncogene is active during odontogenesis. The expression of this gene in differentiated and functional cells of the enamel organ suggests that its role is not restricted to mitotic events but may also be important in signalling pathways related to other cell activities.

Ameloblasts↗

Stimulation of DNA synthesis by endothelin-1 in primary cultures of human dental pulp.

Endothelin (ET), besides being a powerful vasoactive agent, acts as a mitogen in some cell types. ET-like immunoreactivity has been recently detected by immunocytochemistry in the vascular endothelium of human tooth germ and dental pulp, thus providing evidence for local ET production in these tissues. The effects of ET-1 on DNA synthesis in primary cultures of human dental pulp were now investigated. DNA synthesis was evaluated by flow cytometric assay and by 5-bromo,2'deoxyuridine incorporation as detected by immunocytochemistry. Cultured cells were morphologically similar to dental pulp cells and displayed vimentin immunoreactivity. Incubation of cultures with ET-1 resulted in a dose-dependent increase in the number of S-phase-traversing cells over control (unstimulated) cultures. Control skin fibroblasts were also responsive to ET. This finding raises the possibility that the multifunctional peptide ET-1 might subserve growth-promoting activity in the human tooth. It is tentatively suggested that such as an activity might be important during tooth development and in pulp inflammation and healing.

Adolescent↗

A catastrophic antiphospholipid syndrome: the importance of high levels of warfarin anticoagulation.

We report the clinical observation of a 23-year-old woman affected by the so-called 'catastrophic antiphospholipid syndrome'. Within a 3-month period she suffered a number of thrombotic events and haemolytic anaemia with thrombocytopenia and had high levels of immunoglobulins G and M and anticardiolipin antibodies associated with lupus anticoagulant activity. The severity of the clinical and laboratory changes is described and diagnostic and therapeutic difficulties are discussed. The apparent control of thrombotic events only with high levels of warfarin anticoagulation is stressed.

Adult↗

Antiphospholipid antibodies (aPL) in systemic lupus erythematosus. Are they specific tools for the diagnosis of aPL syndrome?

OBJECTIVE: Antiphospholipid antibody (aPL) specificity for aPL-related events was evaluated in systemic lupus erythematosus (SLE). METHODS: A study was carried out on 105 patients affected with SLE comparing the prevalence of lupus anticoagulant (LA) and IgG and IgM anticardiolipin antibodies (aCL) between patients with and without features of antiphospholipid syndrome (APS). Antiphospholipid antibody profile was subsequently evaluated in the aPL positive patients with and without aPL-related events, thus excluding the patients with complications of APS possibly due to factors other than aPL. RESULTS: LA showed a strong association with thrombosis and livedo reticularis, and IgG aCL with thrombosis and neurological disorders, while no clinical features were associated with IgM aCL. A considerable number of aPL positive patients with no aPL-related manifestations was also observed, suggesting the low specificity of aPL assays (54.4%). When studying the 60 aPL positive patients, LA was specific (91.3%) for the diagnosis of aPL-related thrombosis, whereas aCL were not specific, although IgG aCL mean levels were higher in patients with arterial thrombosis than in those without APS features. CONCLUSIONS: LA but not aCL positivity is a specific tool for the diagnosis of thrombotic complications due to aPL in SLE.

Adolescent↗

Tissue fixation for immunohistochemical detection of proliferating cell nuclear antigen with PC10 monoclonal antibody.

PC10 is a monoclonal antibody to proliferating cell nuclear antigen, a nuclear protein associated with the cell cycle. We have evaluated the effects of tissue fixation on PC10 immunoreactivity in sections of paraffin embedded rat tissues. Immunoreactivity was well preserved in tissues after fixation with alcohol-based solutions for 3-24 hr. Fewer PC10-positive cells were detectable in samples fixed with formaldehyde-containing solutions compared with samples fixed with alcohol for the same time. Loss of PC10 immunoreactivity in formaldehyde fixed tissues was progressive, and quantifiable as early as after 3 hr fixation. Consequently, alcohol-based fixatives are strongly recommended for any immunocytochemical prospective study using PC10 antibody. In contrast, loss of PC10-immunoreactivity is always predictable, but difficult to quantitate, using formaldehyde fixed specimens. This aspect should be considered when using PC10 antibody in retrospective studies with routinely-processed archival material.

Adult↗

PC10 monoclonal antibody to proliferating cell nuclear antigen as probe for cycling cell detection in developing tissues. A combined immunocytochemical and flow cytometric study.

Proliferating cell nuclear antigen (PCNA), also referred to as cyclin, is an auxiliary protein to DNA-polymerase delta and a proposed marker of replicating cells. We have investigated the applicability and limitations of PC10 monoclonal antibody to PCNA in a cell kinetics study of developing human and rat tissues by immunocytochemical and flow cytometric techniques. Our data demonstrate that the epitope recognized by PC10 antibody is resistant to wax embedding, but sensitive to aldehyde fixation; conversely, alcoholic fixative solutions preserve the immunoreactivity to PC10. Tissue distribution, DNA content and bromodeoxyuridine uptake confirm that PC10-immunoreactive cells in alcohol-fixed tissues are cycling (G1-, S- and G2-phases traversing) cells. It is concluded that the PC10 antibody can be regarded as a powerful tool to study cell kinetics and differentiation in developing tissues, provided that the tissue processing is adequate.

Acetates↗

Occurrence, distribution and possible role of the regulatory peptide endothelin in the nasal mucosa.

Nasal blood flow is finely regulated by local release of neurotransmitters, neuropeptides and other bioactive molecules acting via paracrine mechanisms. We have investigated the occurrence and distribution in human nasal mucosa of endothelin, a potent vasoconstrictor peptide, by immunocytochemistry and the effect of systemic administration of endothelin-1 on vascular perfusion of rabbit nasal mucosa by laser Doppler flowmetry. Endothelin-like immunoreactivity was demonstrated within vascular endothelial cells in both developing and mature human mucosa. Nasal epithelial cells and some connective tissue cells, presumed to be macrophages, also displayed specific immunostaining. In rabbits injected with endothelin-1, a potent and prolonged nasal vasoconstriction was observed. It is suggested that endothelin released locally may participate in the regulation of nasal blood flow via paracrine mechanisms. Since endothelin has growth-promoting actions on several cell types, it is also tentatively proposed that this regulatory peptide may play a role during development of the nose.

Adult↗

Morphometric computerized analysis as a method to assess skin atrophy caused by corticosteroids.

We have employed morphometric computerized analysis to quantify changes in atrophic skin. Atrophy was induced on volunteers by means of the Duhring chamber assay, using two commercial corticosteroids with a steroid-free base as control. After 20 days, atrophy was assessed by subjective examination and rated on a five-point scale. After 10 and 20 days, positive replicas of the areas under test were obtained. Photographic reproductions of these replicas were stored in the image memory of an interactive automatic computerized image analysis system, Kontron Zeiss Ibas 2, and processed with a suitable program. After selection of the measurement area by means of a circular measurement window, the 'continuous tone' image was converted to a 'binary' one by means of a dynamic discrimination function, in order to perform an automatic evaluation of the percentage area corresponding to the shadows cast by the surface markings. Morphometric computerized analysis proved to be a sensitive method for quantifying skin atrophy. A high degree of correspondence with subjective examination was shown. The method allows an objective differentiation between a mid-potency and a high-potency topical corticosteroid.

Administration, Topical↗

Phospholipid hydroperoxide glutathione peroxidase of rat testis. Gonadotropin dependence and immunocytochemical identification.

A high glutathione peroxidase activity toward phospholipid hydroperoxides is present in rat testis. The attribution of this activity to the selenoenzyme phospholipid hydroperoxide glutathione peroxidase (PHGPX) was supported by cross-reactivity with antibodies raised against pig heart PHGPX which had been purified and characterized. Rat testis PHGPX is partially cytosolic and partially linked to nuclei and mitochondria. The soluble and organelle-bound enzymes appear identical by Western blot analysis. PHGPX, but neither selenium-dependent nor non-selenium-dependent glutathione peroxidase activity, is expressed in testes only after puberty, disappears after hypophysectomy, and is partially restored by gonadotropin treatment. Specific immunostaining of testes by antiserum against PHGPX appears as a fine granular brown pattern localized throughout the cytoplasm in more immature cells but is confined to the peripheral part of the cytoplasm, the nuclear membrane, and mitochondria in maturating spermatogenic cells. As expected, immunostaining of spermatogenic cells in hypophysectomized animals was negative, but gonadotropin treatment only marginally increased the immunoreactivity. The expression of PHGPX in testes is consistent with the previously described specific requirement for selenium for synthesis of a 15-20-kDa selenoprotein which is related to the production of functional spermatozoa.

Animals↗

Lymphatic vessels of the human dental pulp in different conditions.

The characteristics of the lymphatic vessel endothelial wall have been investigated in human normal and inflamed dental pulps. In normal pulps the endothelial wall is characterized by the presence of micropinocytotic vesicles and intraparietal channels. In the inflamed pulpal tissue, where an increase in interstitial fluid pressure occurs, the distended endothelial wall presents open junctions between endothelial cells and the openings of the intraparietal channels. Moreover the micropinocytotic vesicles disappear. The cytoplasm of the endothelial cells is characterized by the presence of numerous Weibel-Palade bodies, which increase in number in the dilated vessels. In the fibrillar apparatus surrounding the lymphatic vessel wall collagen fibrils are the prevalent component, while elastic fibers are not present. The different morphological properties of the lymphatic vessels are compared and discussed with regard to the variation of the functional conditions of the tissue.

Adult↗

HLA haplotype segregation and ultrastructural study in familial immotile-cilia syndrome.

The immotile-cilia syndrome (ICS) is a congenital disorder characterized by dysmotility or even complete immotility of the cilia in the ciliated epithelia. The most frequent consequences include recurrent airway infections from early childhood. Neonatal asphyxia often occurs. Males are usually sterile, whereas females may be fertile or infertile. The disease is inherited as an autosomal recessive trait, but previous attempts to localize the ICS susceptibility gene have so far been unsuccessful. Here, we present the case of two sib pairs affected by ICS from two unrelated families. The electron microscopic investigation of nasal biopsies showed structural anomalies of the cilia, characterized by single microtubules or doublets, arranged randomly in the axoneme. Histocompatibility antigen (HLA)-genotyping of all family members revealed: 1) a significant association of ICS with the HLA-DR7; DQW2 haplotype, which is shared by all the affected sibs (P = 0.0099; RR = 25.94); 2) a possible linkage of the ICS susceptibility gene with HLA, both the affected sibs being HLA-identical, the healthy brother in family B being HLA-different (sib-pair analysis: P less than 0.001).

Child, Preschool↗

Cyclosporin A in a pregnant patient affected with systemic lupus erythematosus.

Few data regarding the use of cyclosporin A (CyA) in pregnancy are available and those available refer mainly to transplant recipients and not to patients with connective tissue diseases. We report the case of a patient with systemic lupus erythematosus (SLE), taking CyA before, during and after her pregnancy at a dose of 4 mg/kg per day. CyA was effective in controlling SLE activity and no side effects were observed in mother or baby. The lack of teratogenicity in this case was in keeping with previous reports in experimental systems, animals and human transplant recipients. If our observation is confirmed by further studies, CyA might become useful in the treatment of pregnant patients with SLE.

Adult↗

Proliferative and functional stages of rat ameloblast differentiation as revealed by combined immunocytochemistry against enamel matrix proteins and bromodeoxyuridine.

A double-staining immunocytochemical technique was used for the simultaneous detection, at the light- and electron-microscopical level, of proliferating bromodeoxyuridine (BrdU)-labelled cells and enamel protein (EP)-producing cells in the inner enamel epithelium (IEE) of rat tooth germ. BrdU-positive cells were found in the region of the IEE close to the cervical loop and never displayed EP-like immunoreactivity. BrdU-immunoreactivity was confined to the nucleus of replicating cells. In contrast, epithelial cells displaying EP-like immunoreactivity were found in the region of the forming dental cusp and were consistently BrdU-negative. EP-like immunoreactivity was detectable in the cytoplasmic compartments involved in the exocrine secretion pathway and in the extra-cellular matrix close to EP-immunoreactive cells. These data support the view that withdrawal from the cell cycle in the IEE is a temporal prerequisite for acquiring the functional competence of secreting EP. Moreover, cycling cells and secretory cells in the IEE constitute two separate compartments that are spatially defined, and that exhibit clear-cut staining patterns with respect to BrdU- and EP-immunoreactivity, respectively. We thus propose that BrdU-incorporation and EP-production may be used as specific markers of the differentiation of the IIE cells in studies of the possible role of growth factors, their receptors and oncoproteins in this tissue.

Ameloblasts↗

Antiphospholipid antibodies in mixed connective tissue disease.

We studied the prevalence and clinical significance of antiphospholipid antibodies (ab) in 28 patients affected with well-defined mixed connective tissue disease (MCTD). Forty-two patients affected with systemic lupus erythematosus (SLE) and 60 healthy subjects were also evaluated, as controls. In MCTD the prevalence of anticardiolipin (aCL) ab was: IgG high level 17.8% (p less than 0.01 versus healthy controls), IgG low level 7.1% and IgM high level 7.1%. No patients had low level of aCL IgM, lupus anticoagulant or false positive VDRL. The aCL profile was similar to that found in SLE patients, but in SLE all prevalences were higher than in MCTD. Furthermore, in MCTD patients the aCL ab were correlated with thrombocytopenia but not with recurrent thrombosis and/or abortions.

Adult↗

Detection and quantitation in rat tissues of the superparamagnetic magnetic resonance contrast agent dextran magnetite as demonstrated by electron spin resonance spectroscopy.

RATIONALE AND OBJECTIVES: The compound studies in this article is a superparamagnetic macromolecular complex of magnetite cores coated with hydrophilic dextran, which is under active investigation as a contrast agent for magnetic resonance imaging (MRI) in liver and spleen. The biodistribution of paramagnetic compounds is problematic and is usually studied by histochemical reactions or by radiolabeling the compound under study. The purpose of this article is to show how electron spin resonance (ESR) spectroscopy detects dextran magnetite (DM) particles in tissues. METHODS: DM injected intravenously in the experimental animal was detected in some reticulo-endothelial organs by ESR. The spectroscopic study was validated using electron microscopy and electron-probe microanalysis. RESULTS: DM exhibits an ESR spectrum; ESR delineated the distribution of DM distribution in liver, spleen, bone marrow, and blood as a function of time. The blood clearance was biphasic, dependent on the size of particles. CONCLUSIONS: ESR spectroscopy is a highly sensitive and reproducible method of studying DM distribution.

Animals↗

Immunohistochemical evidence for the occurrence of endothelin in the vascular endothelium of normal and inflamed human dental pulp.

The occurrence and distribution of the regulatory peptide endothelin in normal and inflamed human dental pulp was studied by immunocytochemistry. Endothelial cells in both normal and inflamed pulp displayed endothelin-like immunoreactivity. Neither pulp cells nor dental nerves were immunoreactive. No significant change in the distribution of endothelin immunoreactivity could be detected in the samples of inflamed pulp tissue, the immunoreactive material being detectable only within the endothelium. However, the intensity of the immunostaining was less intense in the samples of inflamed pulp. It is reasonable to presume that endothelin, produced and released locally by endothelial cells, may participate via a paracrine mechanism in the regulation of blood pressure and flow in normal and pathological human dental tissues.

Adolescent↗

Dermal lymphatics in myotonic dystrophy.

Myotonic dystrophy is an hereditary disorder of several organ systems. Skeletal muscle is a principal target organ, but abnormalities also occur in the peripheral microcirculation. Because morphological and functional changes in the dermal blood microcirculation may affect interstitial fluid drainage of the skin, we examined dermal lymphatic morphology in adult patients with myotonic dystrophy. Skin biopsies were taken from the big toe from patients with myotonic dystrophy (age 18-50 years) and subjected to light and electron microscopy; five healthy subjects served as controls. The salient findings in myotonic dystrophy were ultrastructural changes of the lymphatic endothelial cells and the fibrillar elements that surround the lymphatic wall. These abnormal lymphatic findings are interpreted in light of changes in the blood microvasculature and loose connective tissue in this disorder.

Adolescent↗