Morphological detection of X- and Y-chromosomes in smears and paraffin-embedded tissues using a non-isotopic in situ hybridization technique (NISH)

Pharyngeal smears and paraffin-embedded tissue specimens (skeletal muscle, kidney) obtained from 10 male and 10 female individuals were evaluated using non-isotopic in situ hybridization (NISH) with commercial X- and Y-specific biotinylated probes which recognize the pericentromeric regions DXZ1 and DYZ1/DYZ3 of the X- and Y-chromosome, respectively. The results provide evidence that the morphological sex determination of a single cell can be performed by critical application of this staining method leading to one nuclear signal in ldquomalerdquo cells using the Y-specific probe whereas ldquofemalerdquo cells are negative. In situ hybridization of ldquofemalerdquo tissues with an X-specific probe results regularly in 2 signals whereas ldquomalerdquo cells show only one spot in the nucleus

The immunohistochemical analysis of fibronectin, collagen type III, laminin, and cytokeratin 5 in putrified skin

Fibronectin, collagen type III, laminin, and cytokeratin 5 were visualized in normal skin and in skin showing early or advanced signs of autolytic decomposition to prove whether the immunohistochemical analysis of these antigens can provide useful information for an age-estimation of skin wounds obtained from putrified corpses. In cases with early signs of decomposition (visible course of veins, greenish discoloration) and without microscopic alterations like relaxation of the epidermal cell layers or destruction of the blood vessel structures, the staining pattern was identical to that found in normal, non-putrefied skin. In skin already showing microscopic alteration of the tissue structure, fibronectin and collagen type III could not be localized unambiguously. The distribution of laminin and cytokeratin 5, however, was well preserved. In advanced putrefied skin no reliable staining results could be obtained for fibronectin, collagen type III, and laminin. Even though cytokeratin 5 was still detectable in remnants of decomposition-resistant skin appendages, no information useful for an age-estimation of skin wounds can be obtained due to the autolytic detachment of the epidermal layers

Diffuse somatostatin-immunoreactive D-cell hyperplasia in the stomach and duodenum

This paper presents the first case of extensive, diffuse, somatostatin- immunoreactive D-cell hyperplasia in the human stomach and duodenum. It occurred in a 37-yr-old woman, who showed clinical signs of dwarfism, obesity, dryness of the mouth, and goiter. The density of the distribution of D cells was increased 39-fold in the stomach fundus, 23- fold in the proximal antrum, 25-fold in the distal antrum, and 31-fold in the upper duodenum in comparison with normal values. At the same time, the gastrin-immunoreactive cells were increased 2.3-fold in the antrum. Although the range in size of the D cells was within normal limits in all regions examined, the G cells showed pronounced hypertrophy of up to 127%. A possible relationship between the immuno- histochemical findings and the clinical picture is discussed

Expression of the insulin-like growth factor-II/mannose-6-phosphate receptor in multiple human tissues during fetal life and early infancy

The insulin like growth factor-II/mannose-6-phosphate (IGF-II/M6P) receptor has been detected in many cells and tissues. In the rat, there is a dramatic developmental regulation of IGF-II/M6P receptor expression, the receptor being high in fetal and neonatal tissues and declining thereafter. We have systematically studied the expression of the human IGF-II/M6P receptor protein in tissues from 10 human fetuses and infants (age 23 weeks gestation to 24 months postnatal). We have asked 1) whether there is differential expression among different organs, and 2) whether or not the human IGF-II/M6P receptor is developmentally regulated from 23 weeks gestation to 24 months postnatal. Protein was extracted from human tissues using a buffer containing 2% sodium dodecyl sulfate and 2% Triton X-100. Aliquots of the protein extracts were analyzed by sodium dodecyl sulfate- polyacrylamide gel electrophoresis and immunoblotting using an anti-IGF- II/M6P receptor antiserum (no. 66416) and 125I-protein A or an immunoperoxidase stain. IGF-II/M6P receptor immunoreactivity was detected in all tissues studied with the highest amount of receptor being expressed in heart, thymus, and kidney and the lowest receptor content being measured in brain and muscle. The receptor content in ovary, testis, lung, and spleen was intermediate. The apparent molecular weight of the IGF-II/M6P receptor (220,000 kilos without reduction of disulfide bonds) varied among the different tissues: in brain the receptor was of lower molecular weight than in other organs. Immunoquantitation experiments employing 125I-protein A and protein extracts from human kidney at different ages revealed a small, albeit not significant, difference of the receptor content between fetal and postnatal tissues: as in other species, larger amounts of receptor seemed to be present in fetal than in postnatal organs. In addition, no significant difference of the receptor content between human fetal liver and early postnatal liver was measured employing 125I-protein A- immunoquantitation in three fetal and five postnatal liver tissue samples. The distribution of IGF-binding protein (IGEBP) species, another abundant and major class of IGF binding principles, was also measured in human fetal and early postnatal lung, liver, kidney, muscle, and brain using Western ligand blotting with 125I-IGF-II: as with IGF-II/M6P receptor immunoreactivity there was differential expression of the different classes of IGFBPs in the various organs