The morphological and functional characteristics of the human aortic endothelium. I. 2 variants of the organization of the endothelial monolayer in atherosclerosis]

The en face organization of human aortic endothelium in zones of low (LP) and high (HP) probability of sudanophilia was examined in preparation impregnated with silver nitrate. It is found that the heterogeneity of endothelial cells (EC) by area is "random" or "clusterized". In the latter case the major part of small and medium-sized EC (less than 800 microns 2) is associated in distinct groups ("clusters") and form foci with high monolayer density. The luminal surface outside the clusters was formed by preferentially large and giant EC. Clusterized endothelium was found with statistically significant higher frequency in HP zones of both "normal" and "atherosclerotic" vessels. The maximum clusterization of EC was revealed on the shoulder region and on the periphery of atherosclerotic plaques which was speculated to be a growth zone of the lesion. It is suggested that the appearance of clusterized endothelium is associated with the active development of atherosclerosis.     

3H-thymidine incorporation into human aorta cells in primary culture. An autoradiographic study

Primary cell culture has been obtained from intima and media of unaffected zones of human aorta and from atherosclerotic lesions (fatty infiltration, fatty streaks, atherosclerotic plaques). Cellular polymorphism was found in these cultures. Four morphological types of aortic cells are described: elongated, asymmetric, polygonal, and stellate cells. These cell types also differed in their proliferative activity. On the 7th day of culturing, polygonal cells do not incorporate 3H-thymidine; the thymidine index of other three cell types was similar. The thymidine index of medial cells was higher than that of intimal ones.     

Inhibition of 3H-thymidine incorporation by hydroxyurea: atpical response of mycoplasma-infected cells in culture.

Contamination with Mycoplasma hyorhinis was demonstrated in long-term cultures of HeLa, BICR/M1R_K rat mammary tumor, and NV1C rat neurinoma cells, by microbiological, equilibrium sedimentation, and autoradiographic techniques. In non-infected DNA-synthesizing cells, hydroxyurea (HU) in concentrations ? 10^?4 M typically inhibits ³H-thymidine (³H-TdR) incorporation into acid-insoluble material. This effect was lacking in the contaminated cell lines, although HU did block nuclear DNA replication, as shown by pulse-cytophotometric analyses. The response to HU could be restored to normal by supplementing the culture medium either with the anti-mycoplasma agent Tylosin or with fresh rat serum. The total ³H-activity in non-infected (or anti-mycoplasma treated) versus infected cells, in the absence of HU, was up to four times higher in the former. The data indicate that (i) incorporation of ³H-TdR into the nuclear DNA of contaminated cells was strongly reduced, probably due to a ‘scavenger effect’ (i.e. utilisation and rapid cleavage) by the mycoplasma; (ii) mycoplasmal ³H-TdR incorporation, contrary to nuclear DNA replication, was insensitive to HU in concentrations ? 10^?2 M. If equally valid for other species of mycoplasma, the observed phenomenon provides a criterion (together with the possibility of a rapid test) for the presence of mycoplasmal contamination in cell cultures.     

3H-thymidine incorporation at the end of the S phase in cultured human lymphocytes

Summary Cultured human lymphocytes of female origin were exposed to 3 Ci/ml ³H-thymidine for 2, 21/4, 21/2, 23/4, and 3 hrs, respectively; a proportion of 0, 1.95, 6.4 and 16.7% of labeled metaphases were found among a total of over 3000 metaphases examined. 25 metaphases with little overall radioactivity were analyzed in detail by grain count and grain distribution. Earlier studies were confirmed that the grain distribution at the extreme end of the S phase is non-random in several regions of the chromosomal complement, but that a distinct interval (Z interval) probably does not exist.

---

Zusammenfassung In der vorliegenden Arbeit wird das Ende der DNA-Synthese mittels autoradiographischer Methoden an normalen menschlichen Lymphocyten untersucht. Das Reduplikationsverhalten der Chromosomen am Ende der DNA-Synthese wird näher beschrieben und diskutiert.Das Reduplikationsmuster der letzten 30 min der S-Phase als auch das Verhalten des späten X-Chromosoms sowie der Vergleich zum Markierungsverhalten der Autosomen sprechen gegen die Existenz eines sogenannten Z-Intervalls am Ende der Synthese-Phase.

---

---

This investigation was supported in part by grants from the Deutsche Forschungsgemeinschaft and is part of a thesis by D. B. in fulfillment of requirements for the M. D., at the University of Hamburg.     

Dynamics of 3H-thymidine and 3H-deoxycytidine incorporation into the nuclei of a rabbit kidney cell culture during the S period

Using a combined cytophotometric-autoradiographic method a study was made of 3H-thymidine and 3H-deoxycytidine incorporation rates into the interphase nuclei of rabbit kidney cell culture during the S-period. The rate of 3H-deoxycytidine (10(-4) M--10(-6) M) incorporation into nuclei increases throughout the first part of the S-period and decreases from its middle to the end. The patterns of variations of 3H-thymidine and 3H-deoxycytidine incorporation rates into the nuclei of cultured rabbit kidney cells during the S-period were identical.     

3H-thymidine incorporation into the macrophages in the process of eye regeneration in adult tritons]

Changes in the number of labelled macrophages in the regenerating eye cavity of the adult common newts were studied within 2 to 12 days following the injection of 3H-thymidine and removal of retina and lens from the eyes. A small number of labelled macrophages was found in the eye cavity (0,2--5.9%) at all regeneration stages under study upon the pulse 3H-thymidine incorporation. Their number rapidly increased and attained by the end of observation (12 days) 73%. These results suggest that mainly mature non-dividing forms of macrophages which completed the mitotic cycle S-phase in the places of their active reproduction migrate in the eye cavity. The sharp increase of the number of labelled macrophages in the eye cavity is determined by the migration of those macrophages the precursors of which were labelled as a result of both the pulse 3H-thymidine incorporation and reutilization of the labelled precursors of DNA synthesis. New portions of labelled macrophages migrate in the eye cavity within 2 to 4 days.     

The effect of estrogen on the incorporation of 3H-thymidine by PHA-stimulated human peripheral blood lymphocytes

The effect of estrogen on the incorporation of tritiated thymidine by phytohemagglutinin (PHA)-stimulated human peripheral blood lymphocytes was evaluated in 15 adult males. Varying concentrations of diethylstilbestrol diphosphate (DEP-S) were added to peripheral blood lymphocytes with and without PHA to study the effects of estrogen on blastogenesis. Maximum suppression of blastogenesis occurred after the addition of 500 mcg/ml culture of DEP-S. The absence and presence of DEP-S 500 mcg/ml culture resulted in a 52% reduction in lymphocytic reactivity (p.002). It was concluded that this reduction or suppression of lymphocytic blastogenesis in the presence of estrogen suggests that the palliative effects of estrogenic therapy in treating patients with hormone-dependent tumors may be countered by its adverse effect on the host's immunologic responsiveness to malignancy.     

A comparison between quinacrine fluorescence banding and 3H-thymidine incorporation patterns in human chromosomes

Summary Late ³H-thymidine incorporation patterns and quinacrine fluorescence banding patterns were analyzed in metaphase chromosomes prepared from human blood cultures. In general, late-labeling regions correspond to the strongly fluorescent bands in the chromosomes. However, the dully fluorescent secondary constrictions of the chromosomes Nos. 1, 9 and 16 may show late replication in some instances. In contrast, the brilliantly fluorescent distal part of the Y chromosome is not labeled during the latest phase of the DNA replication. In the male X and in the isopycnotic X of the female the labeling pattern also agrees with the quinacrine fluorescence banding. The heteropycnotic X of the female is still more strikingly late-labeling. However, the pattern of its late replication agrees also with the quinacrine fluorescence bands.

---

Zusammenfassung An aus menschlichen Blutkulturen gewonnenen Chromosomenpräparaten wurden vergleichende Untersuchungen über die späten Replikationsmuster (mittels ³H-Thymidin-Autoradiographie) und die Quinacrin-Fluorescenz-Bänderungsmuster durchgeführt. Im allgemeinen stimmen die spät replizierenden Abschnitte mit den intensiv fluorescierenden Regionen der Chromosomen überein. Allerdings können die nur schwach fluorescierenden sekundären Constrictionen der Chromosomen Nr. 1, 9 und 16 manchmal auch eine späte Replikation zeigen. Auf der anderen Seite wird der stark fluorescierende Abschnitt des Y-Chromosoms in den letzten Phasen der DNS-Replikation nicht mehr markiert. Beim X-Chromosom des Mannes und beim isopyknotischen X der Frau wird ebenfalls eine Übereinstimmung des Spät-Replikationsmusters und der Quinacrin-Bänderung gefunden. Das heteropyknotische X der Frau zeigt eine noch deutlichere Spät-Replikation; das Muster der Silberkörner stimmt aber ebenfalls mit den Quinacrine-Bändern überein.

---

---

This study was supported by the österreichischen Fonds zur Förderung der Wissenschaftlichen Forschung.     

Reduced uptake and incorporation of 3H-thymidine in fanconi anemia fibroblasts

Summary Uptake of ³H-thymidine and its incorporation into DNA was studied in fibroblastic cell lines derived from normal individuals, patients with Fanconi anemia, and those heterozygous for this genetic trait. Uptake and incorporation for the normal cells were about five and seven times higher, respectively, than for Fanconi anemia fibroblasts; mean values for heterozygotes were intermediate. This effect was dependent on the duration of cell exposure to ³H-thymidine and was not observed with other labeled compounds. Thus, a genetically-determined metabolic defect may exist in Fanconi anemia patients which can be readily studied at the cellular level. This finding may be relevant to the observed clinical, cytogenetic, biochemical, and biologic properties related to expression of the Fanconi anemia gene.     

The 3H-thymidine incorporation into G0 human lymphocytes induced by low doses of UVC radiation

The 3H-thymidine incorporation in human lymphocytes of healthy donors induced by UVC radiation under doses 0.0008-60 J/m2 was investigated. It was shown that the incorporation of 3H-thymidine increases under doses in interval 0.1-20 and is constant under doses higher than 20 J/m2. Under doses in interval 0.006-0.03 J/m2 near a half of all samples had the level of incorporation increased in comparison with control samples. We connect the presence, absence or variability of this effect with individual peculiarities of cells and with different activity of cell subpopulations that are different on morphological and physiological characteristics. The hypothesis about the role of this factor in the influence of low doses of pathogenic agents (UVC and X-radiation, chemical compounds) on human lymphocytes is discussed.     

Infra-red laser irradiation enhances interleukin-1 receptor antagonist,increases 3H-thymidine incorporation and the release of [3H]arachidonic acid in human monocytes

The effect of infra-red laser irradiation has been experimented on various biological systems and particularly in human tissues, in vitro as well as in vivo. In order to examine the influence of laser irradiation on cells of the monocytic lineage we have irradiated human peripheral blood mononuclear cells with an infra-red laser at a wavelength of 904 nm, at 2000 Hz frequency and 15 mW for 2 min. Here, we report that laser irradiation for 2 min. at different preincubation times (T = 0 and T = 30 min) enhances LPS (10 g/ml or PHA (10 g/ml, suboptimal concentration) -stimulated monocytes by modifying cell proliferation, as judged by [3H] thymidine incorporation. IL-1 receptor antagonist (IL-1ra) along with an increased release of [3H] Arachidonic acid production, is also influenced by laser irradiated monocytes when treated for 2 min after 1 h incubation. IL-1RA production increased 4-5-fold after laser irradiation, while 3H-arachidonic acid incorporated from PMA-stimulated cellsased and the effect was significant at T = 0 and T = 30 min; while at T = 1 h the effect was negligible. These results may provide new information regarding the effect of laser irradiation on the immune system.     

The effect of photosensitizing dyes on the 3H-thymidine incorporation of cells grown in vitro

The photodynamic inactivation of ³H-thymidine incorporation in mouse embryo (ME) and mouse L cells by acridine orange (AO), methylene blue (MB) or neutral red (NR) has been studied by estimating the number of nuclei capable of incorporating ³H-thymidine during a 24 h period following light exposure. In the dark NR and AO reduced the number of ME-nuclei incorporating ³H-thymidine but MB caused an increase in non-scheduled DNA synthesis. The dark effect on L cells was less but the photoinactivation of thymidine uptake was proportionally greater in these cells. Polyoma virus was shown to be capable of growing in cells whose thymidine uptake was reduced or completely stopped by photoinactivation with NR. However, if the NR damage was very great, or when AO was used to photosensitize cells, the synthesis of viral DNA was interfered with.     

Rat intermediate lobe cell groups in suspension culture: morphological and functional characteristics

Intermediate lobe cells mechanically isolated from adult rat hypophyses were maintained in histiotypic suspension culture. The cells survived as long as six weeks. The proliferation of the connective tissue cells was limited, they did not overgrow the glandular cells. The fine structure of the parenchymal cells in the epithelial spherules of the early cultures was practically identical to that in the intact tissue. The morphological sign of exocytosed secretory material could also be found. In older cultures, cells with light and dark cytoplasm were present. The former gradually degenerated and disappeared, while the latter, forming trabecules, retained their intact fine structure. Immunoactive and bioactive ACTH was detected in the culture medium.     

Intramural ganglia of the human gallbladder: morphological and functional observations]

The purpose of this study was the reinvestigation of the intrinsic innervation of human gall bladder with an immunohistochemical technique named peroxidase anti-peroxidase (PAP). The antigen demonstrated was the S100 protein normally present in the surface of glial cells, Schwann cells and satellite cells in ganglia. The tissues used were taken from 20 human gall bladders, fixed after surgery. This technique is not specific to demonstrate adrenergic or cholinergic innervation but it reveals just myelinated fibers. The current study was undertaken in order to study the organization and the function of plexus of nerves and ganglia present in the wall of the gall bladder. The neck of the gall bladder was the region in which the higher number of nerve cells and nervous fibers was present. The technique used has demonstrated ganglionated plexus and nerves in submucosal layer, fibromuscular and adventitial layer according to the enteric nervous system. All ganglia are postganglionic stations related with preganglionic cholinergic fibers. These results confirm that the intramural ganglia of the gall bladder are analogous to those of the enteric nervous system according to their common origin.