Factors of importance for decreased early embryo survival in female mice treated neonatally with diethylstilboestrol.

Neonatal female mice of the NMRI strain were treated s.c. with 5 micrograms of the synthetic oestrogen diethylstilboestrol (DES) or vehicle only (olive oil; controls) for the first 5 days after birth. Embryos at the two-cell stage from control females were transferred to the oviducts of control females or DES-treated females (6-8-weeks-old). Immediately after transfer, the oviducts were removed and incubated in vitro for 6 h. After incubation, slightly fewer embryos were recovered from oviducts exposed to DES compared with control oviducts (81% versus 92%; 0.05 > P > 0.01). When the recovered embryos were cultured in vitro, 64% of the embryos from control oviducts reached the blastocyst stage in contrast to only 24% of those from oviducts of DES-treated mice; slightly fewer of the latter showed trophoblastic outgrowth (76% versus 93%; 0.05 > P > 0.01). Oviductal transport and uterine attachment was studied by introduction of blue-stained dextran microspheres into the oviduct. The microspheres appeared earlier in the uteri of DES-treated females than in the uteri of controls. Moreover, the spheres were not captured in the uterus in most DES-treated females, but they were trapped when a piece of Spongostan was placed in the uterine lumen. Scanning electron microscope studies showed differences between controls and DES-treated females in uterine mucosal lining of possible importance for embryo attachment to the surface epithelium. These studies showed that neonatal DES treatment caused changes in the adult reproductive tract as revealed by a detrimental effect upon embryo development, more rapid oviductal transport and loss of embryos from the uterus.

Ultrastructural cerebrovascular changes in a model of subarachnoid hemorrhage in baboon based on triple cisternal blood injection.

In a subarachnoid hemorrhage model in the baboon, achieved through three cisternal blood injections with 1-day intervals, the cerebral arteries were dissected out 7 days after the first blood injection for electron microscopy All the animals showed ultrastructural changes in the cerebral arteries: two with severe, one with moderate, and three with mild alterations in the vessel walls. The most constant findings were seen in the muscle cells of the media layer. Fragmentation of the nuclei was frequently observed together with cytoplasmic vacuoles. Scattered groups or single degenerated muscle cells were also noted. In the intima the changes included rounding of the nuclei along with the appearance of cytoplasmic vacuoles. Desquamation or flattening of the endothelium and loss of tight junctions were encountered in some vessel areas. Degenerating mitochondria were a common finding.

Localization of eosinophil cationic protein, major basic protein, and eosinophil peroxidase in human eosinophils by immunoelectron microscopic technique.

An immunoelectron microscopic technique using protein A-gold as a specific marker was used for precise intracellular localization of eosinophil granule proteins. Eosinophils from healthy individuals were isolated in metrizamide gradients. Eosinophil cationic protein (ECP) and eosinophil peroxidase (EPO) were clearly located in the matrix of the large crystalloid-containing granules. In addition, ECP was probably present in the small granules of eosinophils. Major basic protein (MBP) was present in the crystalloid structure of specific granules. This method can be applied in studies of eosinophil degranulation to trace the release of biological effector molecules.

Scanning electron microscopy and transmission electron microscopy of the ciliated cells of the trachea of the rabbit treated with misonidazole alone and in combination with ionizing radiation.

The trachea is often located in the treatment volume when irradiating malignant tumours in the thorax. In order to evaluate possible synergism between misonidazole and irradiation on this tissue, the following studies were made. Fifty rabbits were treated with daily injections of 100 mg misonidazole given i.p. on consecutive days from 1 to 10 days. Morphological investigations of the trachea were made with scanning electron microscopy (SEM), transmission electron microscopy (TEM) and light microscopy (LM). Physiological examinations were performed with recording of the ciliary beat frequency. The results were compared with those from a group of 100 rabbits given misonidazole in a similar manner and exposed to irradiation (2 Gy) 15-30 min after each injection. Ten rabbits were used as controls. The results are compared to the effect of fractionated irradiation alone with 2 Gy/day. Fractionated irradiation of the ciliary epithelium in the trachea of the rabbit has shown dose-dependent physiological and morphological effects. Misonidazole potentiates these effects of radiation with a more pronounced change of the ciliary beat frequency and an increased metabolic activity as could be visualized on TEM. The combination of drug and irradiation also induced a hyperplasia of the ciliary epithelium. Misonidazole itself had no effect on the ciliary beat frequency, but caused a hypoplasia of the ciliary epithelium.

Life cycle of the endoparasitic nematophagous fungus Meria coniospora: a light and electron microscopic study.

The obligate endoparasitic fungus Meria coniospora lives its entire vegetative life within infected nematodes. Conidia of M. coniospora infect the nematode Panagrellus redivivus mainly in the mouth region. The infection, starting with adhesion of conidia to the nematode surface, growth of trophic hyphae, production of conidiophores and conidia, was followed using light, scanning and transmission electron microscopy.

The effects of 10 Gy single-dose irradiation on the ciliated epithelium measured during and one-to-ten days following irradiation. A comparative physiological and morphological study.

Experiments recently done in this laboratory have shown that ionizing irradiation can result in an increase in the ciliary beat frequency of the trachea in vitro even during ongoing irradiation, implying an immediate stimulation of the physiological activity of the cells. This phenomenon is not fully understood and is by itself contradictory to general radio-biological concepts. Ultrastructural investigations on specimens taken immediately after irradiation (10 Gy) have, so far, not shown any changes. It has therefore seemed useful to extend the investigations by examining the effects of in vivo- irradiation (10 Gy) on rabbit trachea during the first 10 days after treatment. The following results have been obtained: Phase I: On days 1-3 after irradiation the beat frequency showed a slight increase (10%) within the irradiated part in comparison with a non-irradiated area of the trachea. No pronounced ultrastructural changes were found these days. Phase II: On days 4-7 after irradiation the mucociliary activity was reduced in those places where it could be measured. The surface of the cilia assumed a generally destructed appearance with elongated cilia and disorganisation. TEM-pictures showed an increased amount of goblet cells and signs of membrane damage such as cytoplasmic extrusions at the apical end of the cells. Phase III: On days 8-10 after irradiation a normalization of the tissue gradually took place with a beat frequency returning to normal. The ciliary surface showed a recovery process which also could be observed in the TEM-pictures.

Scanning electron microscopy and evans blue staining for assessing endothelial trauma and reconstitution after venous thrombectomy.

Thrombectomy and a temporary arteriovenous fistula as treatment for deep venous thrombosis has been shown to cause a severe endothelial trauma in an experimental dog study. In vivo staining with Evans blue has been compared to scanning electron microscopy findings to assess the extent of the trauma and the time necessary for endothelial reconstitution. A good correlation between these techniques was found. After the initial almost complete denudation of the veins, new endothelium grew out from small branch openings and valve pockets within four weeks except in the proximity of the arteriovenous fistula where endothelialization was still incomplete after four months.

Scanning electron microscopy and recording of the physiological activity of tracheal ciliated cells treated by fractionated irradiation.

The ciliated epithelium of the rabbit's trachea was irradiated with daily fractions of 2 Gy to an accumulated dose of 20 Gy. The beat frequency of the cilia was recorded and specimens were taken for SEM-, TEM- and LM-investigations. Examination was made 1-10 days after each fractionation schedule. An increased ciliary beat frequency was recorded at 2 Gy and 4 Gy. With increasing dose, there was an inverted relationship to the frequency. Light-microscopy showed edema and an increased amount of goblet cells in relation to the increasing dose. With SEM an increased number of ciliary blebs could be seen. These could be classified according to size and number, and showed a positive correlation to the dose. TEM-investigations showed signs of increased intracellular activity at higher doses in the form of multilobulated nuclei and an increasing number of nuclear pores. At lower doses, an increased amount of mitochondria appeared in the apical part of the cell. It is at present difficult to evaluate any correlation between the physiological activity and the morphology. More biological data are needed to explain the early irradiation effects.

The effects of ionizing irradiation on the ciliated cells of the central nervous system (CNS) in man--a scanning electron microscopy study.

In order to investigate the effects of radiotherapy on the ciliated ependymal lining of the CNS, samples were taken at autopsy from the lateral ventricles down to the IVth ventricles in patients irradiated for Glioblastoma multiforme in one hemisphere. Doses higher than 20 Gy affected the cilia and the lesions were more pronounced the heavier the dose. No regeneration of cilia were found in those regions which had received more than 20 Gy. In the areas irradiated with 20 - 15 Gy the cilia had a more normal form and cilia irradiated with less than 15 Gy were considered to be normal.

Chlamydia trachomatis infection of the Fallopian tubes. Histological findings in two patients.

In two patients with acute salpingitis, C. trachomatis was isolated from the cervix. In one of the patients, the organism was also recovered from the Fallopian tubes, and in the other, chlamydial inclusions were found in Giemsa-stained tubal epithelial cells. A significant change in micro-immunofluorescence antibodies to C. trachomatis occurred in both patients during the course of the disease. The Fallopian tubes of both patients were removed and studied by conventional histological techniques and, in the case of one of them, by transmission electron microscopy.

Effect of human leukocyte enzymes on tracheal mucosa and its mucociliary activity.

Human neutrophil granulocytes contain proteolytic enzymes. In purulent bronchial and paranasal secretions these enzymes have been found extra cellulary in complex with enzyme inhibitors as well as in free form indicating saturation of the inhibiting capacity. Isolated human leukocyte enzymes, elastase and neutral protease, were found to arrest the mucociliary activity and subsequently cause superficial tissue destruction. Elastase was found to be the most potent of the enzymes. Experimental studies with elastase together with specific inhibitor indicated the importance of the enzyme inhibitors for the integrity of the mucous membrane.

The mucociliary activity of the upper respiratory tract. III. A functional and morphological study on human and animal material with special reference to maxillary sinus diseases.

Local mucociliary activity (mucociliary wave frequency) was recorded in vivo and in vitro in maxillary sinus diseases. Activity was fairly normal even in cases with chronic purulent sinusitis. Scanning electron microscopy (SEM) sometimes showed a quantitative reduction of cilia. The mucociliary wave frequency was mostly about 1000 waves/min, and there was no in vivo/in vitro discrepancy. Antral mucosa was compared to adenoids, nasal polyps and healthy rabbit trachea. The mucociliary wave frequency was about the same in all materials.

Scanning electron microscopy of cells in the lymph of the human thoracic duct in advanced malignancies.

At scanning electron microscopy of cells from the lymph of the thoracic duct, collected by cannulation of patients with malignant tumours of different types, lymphocytes and red blood cells were found, but few malignant cells. The latter came from patients with pulmonary carcinoma and mammary carcinoma with lung metastases. Various abnormal shapes of the red blood cells were observed, possibly due to extrinsic factors such as administered drugs of abnormal metabolites from the malignant disease. The lymphocytes had no consistent pathologic shape, but appeared to act as immunologic, active cells against the malignant cells in the thoracic lymph.

Studies on ciliated epithelia of the human genital tract. I. Swelling of the cilia of Fallopian tube epithelium in organ cultures infected with Mycoplasma hominis.

Organ cultures of human Fallopian tubes were infected with Mycoplasma hominis. Scanning and transmission electron microscopy revealed swelling of the cilia of the tubal epithelial cells in infected cultures. In some, the entire cilia were swollen; in others, only the tips. Uninfected cultures kept for up to 7 days showed no structural changes in the cilia or other surface structures. M. hominis multiplied in organ cultures, but not in culture medium without tissue. A practical organ culture technique for the preparation of specimens for electron microscopy is described.

The ultrastructure of spontaneous radial segmentation of the nuclei in bone marrow cells from 3 patients with acute myeloid leukaemia.

Bone marrow cells with spontaneous radial segmentation of the nuclei from three patients with acute myeloid leukaemia were analysed by means of electron microscopy. The centriole was found in the centre of the cluster of nuclear segments. Cytoplasmic microtubules were found in the intersegmental clefts and in the pericentriolar area. The present observations support the hypothesis that spontaneous radial segmentation of the nuclei in tumour cells is due to a microtubule-associated contractile process, by which the nucleus is folded into segments.