Immunohistochemical analysis of human neuronectin expression in normal, reactive, and neoplastic tissues

Neuronectin (NEC1) is a human extracellular matrix protein of central nervous system (CNS) parenchyma found throughout the white matter of rostral brain segments (telencephalon, diencephalon, some areas of mesencephalon), but not in rostral CNS gray matter, most areas of mesencephalon, pons, cerebellum, medulla, spinal cord, or peripheral nerves. The present immunohistochemical study, using two monoclonal antibodies to distinct epitopes on the NEC1 molecule, examined NEC1 expression in normal non-neural tissues, malignant tumors of diverse histological types, non-malignant skin lesions, and dermal incision wounds. We show that (a) NEC1 is expressed in normal fetal precartilage blastemas and fetal and adult vascular and visceral smooth muscle, but not in most loose connective tissues and skeletal or cardiac muscle; (b) NEC1 is found along epithelial-mesenchymal junctions, with marked differences in prevalence and histological patterns in different organs; and (c) mesenchymal activation associated with wound healing, actinic keratosis, psoriasis, and neoplasia leads to strong induction of NEC1 expression. Parallel studies with cultured human cells suggest that region-specific NEC1 expression in normal developing tissues and localized induction in wound healing and disparate diseases is under the control of extrinsic signals provided by regulatory polypeptides.     

Erythrocyte insulin binding in normal infants, children and adults

To establish normal insulin binding criteria, we studied the binding of insulin to erythrocytes from normal subjects of different ages. Insulin binding to cord erythrocytes and to erythrocytes from infants aged 2-7 days was significantly higher at tracer and physiological insulin concentrations than was binding to cells from children aged 1-15 years and adults. In infants aged 1-12 months the maximum insulin binding to erythrocytes was significantly higher than that to erythrocytes from children, and in addition, it correlated negatively with age. An increase in receptor concentration was found in cord erythrocytes whereas an increased receptor affinity for insulin was found in erythrocytes from infants. Insulin binding characteristics in erythrocytes from prepubertal and pubertal children were basically similar to those in women. Erythrocytes from men bound significantly higher amounts of insulin than did those from women. This difference was associated with changes in receptor affinity for insulin. There was no correlation between the insulin binding characteristics and the circulating concentration of insulin or C-peptide. The increased erythrocyte insulin binding at birth persisted over the neonatal period. There was an overall negative correlation between the maximum insulin binding and age in the subjects studied, but the major decrease in erythrocytes insulin binding occurred during the first year of life past the neonatal period. These observations stress the importance of using age-matched controls in studies on erythrocyte insulin binding in disease states.     

LHRH in the human hypothalamus. Immunohistochemical mapping in normal newborn infants or in sudden death infants

The topographical distribution of neurons containing LHRH has been investigated in newborn hypothalamus using the peroxidase anti-peroxidase technique. In control subjects, LHRH immunoreactive (LHRH-IR) perikarya have been mainly observed essentially in the infundibular nucleus. The preoptic region displayed a moderate density of LHRH-IR cell bodies. High LHRH innervation was observed in the anterior hypothalamus in the lamina terminalis and in the mediobasal hypothalamus in the median eminence, and in the peri- and paraventricular regions. In sudden death infant syndrome, a comparable mapping was observed, except a low density in the mediobasal peri- and paraventricular areas.     

Biochemical analysis of normal and osteoarthritic human cartilage

Non-collagenous proteins from the articular cartilage of normal subjects and patients with degenerative joint disease were extracted sequentially. Proteoglycans and the other glycoproteins were more extractable from the osteoarthritic cartilage at lower ionic strength than those from the normal cartilage. A 50-kD protein which seems specific to osteoarthritic cartilage was identified. Three different populations of proteoglycans were purified from normal and only two from osteoarthritic cartilage. Moreover, greater amounts of albumin and fibronectin were found in the pathological cartilage. No differences were observed between link proteins from normal and osteoarthritic cartilage, nor in their molecular weight or the amounts extracted.     

Variations in karyotypes of normal premature babies,new-born babies and infants

Summary 4754 lymphocyte metaphases of 36 male and female premature babies, new-born babies and infants were analyzed for the occurrence of numeric and structural chromosome aberrations, of associations and visible satellites of the acrocentric chromosomes. The cell cultures were done after a micro-method according to Hirschhorn and were interrupted after 48 and 72 hrs of culturing. Aneuploidy. 1.87% of all the cells analyzed showed an abnormal number of chromosomes. In the single individual aneuploidic nuclei occurred between 0 and 5% without there being a difference in group of age, sex or time in culture. Structural Abnormalities. In 4.75% of the total number of mitoses analyzed, and between 1to 8% in the single individual, structural abnormalities were recorded. Gaps were found most frequently in chromosomes of group D, breaks in those of group A. The difference between age-groups,sexes, culturing time and culture series has no significance for the occurrence of structural abnormalities. Associations of Acrocentric Chromosomes. Their occurrence depends on the individual, the age-group and the time in culture. Within one age-group with a certain culturing time, an individual is characterized by their occurrence. Satellites. The frequency of D-satellites as well as of G-satellites is characteristic of an individual. Their occurrence with one subject remained the same for both culture periods.

---

Zusammenfassung 4754 Lymphocytenmetaphasen von 36 männlichen und weiblichen Frühgeborenen, Neugeborenen und Kleinkindern wurden nach dem Vorkommen von numerischen und strukturellen Aberrationen, von Assoziationen und sichtbaren Satelliten der akrozentrischen Chromosomen untersucht, wobei wir die Zellkulturen, die wir nach einer Mikromethode nach Hirschhorn durchführten, nach 48 und 72 Std Kulturzeit abbrachen. Aneuploidie. Wir beobachteten in 1,87% sämtlicher untersuchter Zellen eine abnorme Chromosomenzahl. Bei den einzelnen Individuen kamen aneuploide Kerne zwischen 0 und 5% vor, ohne daß ein Unterschied zwischen den Altersgruppen, den Geschlechtern oder den Kulturzeiten bestand. Strukturelle Abnormalitäten. Sie traten in 4,75% aller untersuchten Mitosen, bei den einzelnen Probanden zwischen 1 und 8% auf, wobei Gaps am häufigsten auf den Chromosomen der gruppe D, Brüche auf denen der Gruppe A gefunden wurden. Die Altersgruppen, die Geschlechter, die Präparate der verschiedenen Kulturzeiten und Serien unterschieden sich im Vorkommen von Strukturabnomalitäten nicht voneinander. Assoziationen der akrozentrischen Chromosomen. Ihr Vorkommen ist abhängig von: Individuum, Altersgruppe, Kulturzeit. In einer Altersgruppe für eine bestimmte Kulturzeit charakterisiert ihre Häufigkeit ein Individuum. Satelliten. Sowohl die Häufigkeit der D-Satelliten wie auch die der G-Satelliten ist für ein Individuum charakteristisch. Ihr Vorkommen bei einem Probanden war bei beiden Kulturzeiten gleich.

---

---

Presented in part at the Second Colloquium of the Section Cytogenetic of the Gesellschaft für Anthropologie und Humangenetik, June 11th, 1970, in Kiel, and at the symposium on Chemical Mutagenesis as a Problem for Medicine, December 11th, 1970, in Freiburg i. Br.     

Choline status in newborns, infants, children, breast-feeding women, breast-fed infants and human breast milk

This study assessed the choline status in newborns, infants, children, breast-feeding women, breast milk, infant formula, breast-fed and formula-fed infants. The serum free choline level was 35.1+/-1.1 micromol/L at birth and decreased to 24.2+/-1.6, 18.1+/-0.8, 16.3+/-0.9, 14.3+/-0.8, 12.9+/-0.6 or 10.9+/-0.6 micromol/L at 22-28, 151-180, 331-365, 571-730, 731-1095 or 4016-4380 days after birth, respectively. The serum phospholipid-bound choline level was 1997+/-75 micromol/L at birth and increased gradually to 2315+/-190 or 2572 +/-100 micromol/L at 571-730 or 4016-4380 days after birth, respectively. In breast-feeding women, serum free and phospholipid-bound choline levels were doubled at 12-28 days after birth, they decreased toward the control values with time. Free choline, phosphocholine and glycerophosphocholine were major choline compounds in breast milk. Their concentrations in mature milk were much greater than in colostrum and serum. Choline contents of breast milk varied greatly between mothers, and milk free choline levels were correlated with serum free choline (r=.541; P<.001), phospholipid-bound choline (r=.527; P<.001) and glycerophosphocholine (r=.299; P<.01) concentrations and lactating days (r=.520; P<.001). In breast-fed infants, serum free choline concentrations were correlated with free choline (r=.47; P<.001), phosphocholine (r=.345; P<.002), glycerophosphocholine (r=.311; P<.01) and total choline (r=.306; P<.01) contents of breast milk. Serum free choline concentration in formula-fed infants was lower than breast-fed infants. These data show that (a) circulating choline status is elevated during infancy and lactation, (b) choline contents of breast milk vary between mothers and milk free choline contents are influenced by maternal circulating choline status, and (c) the choline contents of breast milk can influence infants' circulating choline status.     

Cellular heterogeneity in normal human urothelium: an analysis of optical properties and lectin binding

Flow cytometry was used to study the optical properties of normal urothelial cells in suspension. Narrow-angle light scatter, which is a function of cell size, defined one major and one minor cell population, and 90 degrees light scatter, a function of intracellular structure, showed three distinct cell populations. These properties were displayed as a 2-dimensional dot plot or fingerprint which proved to be characteristic and reproducible from one specimen of urothelium to the next. Cell sorting on the basis of these two parameters demonstrated that the small cells of the basal layer occupy the low narrow angle, low 90 degrees light-scatter region; the giant cells of the superficial layer lie in the high narrow angle, high 90 degrees scatter region; and the pyramidal cells of the intermediate layer lie in an intermediate zone. Studies of tissue sections using the galactose-specific, FITC-conjugated Maclura Pomifera lectin (MPA) demonstrated preferential binding to the superficial layers of intact urothelium. In order to quantify the apparent differences in lectin binding between the superficial and basal layers, urothelial cell suspensions were labeled with FITC-conjugated MPA and studied by flow cytometry. The resolution obtained on the basis of light scatter made it possible to quantify the difference in lectin binding to the three morphologically recognized cell types present in normal urothelium.     

Social smiling in normal and down syndrome infants: A comparative study

The aim of this work has been to compare social smiling in Down syndrome (mongolism) and normal infants, attending especially to the brow movements that appear before it. Facial responses of eight Down syndrome and eight normal infants from three to five months were analized by means of an anatomically based measurement technique during face-to-face interactions with their mothers. Despite their mental retardation, Down syndrome infants showed identical muscle movements as normal infants before and during smiling. However, some differences were found in smile frequency and leght, as well as in the brow movements frequency before smiling. Results are discussed in terms of the psychophisiological dysfunction of Down syndrome infants that are originated by a chromosome imbalance.     

Antigenic analysis of human cytomegalovirus isolates from Japanese women and infants.

The antigenic differences among human cytomegalovirus (CMV), two laboratory strains (Davis, AD 169), isolates from pregnant women's cervical secretions, mother's milk, infants' throat swabs and urine were analyzed by means of the plaque reduction assay using human sera which were assumed to contain monotypic antibodies by primary infection and boosted antibodies by reinfection or reactivation of the latent virus. In the cross-neutralization tests, there were no remarkable differences among the CMV strains examined. Moreover, in the neutralization kinetics, normalized kappa values among the strains constantly exceeded 80. Therefore, it is suggested that the human CMV strains examined in the study were serologically identical or very closely related.