Atoxoplasma (Apicomplexa: Eimeriorina: Atoxoplasmatidae) in the greenfinch (Carduelis chloris).

Merozoites of an Atoxoplasma species were present within parasitophorous vacuoles in the cytoplasm of leucocytes in the blood vessels in the small intestine of the greenfinch (Carduelis chloris). The merozoites indented the nucleus of the leucocytes. Merozoites, macrogametes, and microgamonts were present in the epithelial cells of the intestines. No merogony was observed. Experiments provided circumstantial evidence linking disporocystid-octozoic oocysts with the blood and intestinal infections.

Crohn's disease: pathogenesis and persistent measles virus infection.

The Inflammatory Bowel Disease Study Group at the Royal Free Hospital School of Medicine has tested the hypothesis that the primary pathological abnormality in Crohn's disease is in the mesenteric blood supply. Early morphological studies involved arterial perfusion-fixation and either resin casting and scanning electron microscopy or vascular immunostaining of resected intestine affected by Crohn's disease. Granulomatous and lymphocytic damage to intramural blood vessels, even in macroscopically normal areas, was observed. We put forward possible mechanisms by which a chronic ischemic process might account for many of the idiosyncracies of Crohn's disease. It was proposed that persistent viral infection of the mesenteric microvascular endothelium might underly this vasculitic process; based on certain behavioral characteristics of measles virus, including its tropism for the submucosal endothelium of the intestine, this agent was investigated further. This report reviews the preliminary evidence from both epidemiological and basic scientific data for persistent measles virus in the intestine of patients with Crohn's disease. Possible mechanisms for virus persistence and subsequent reactivation are discussed. In conclusion, we believe that Crohn's disease may be a chronic granulomatous vasculitis in reaction to a persistent infection with measles virus within the vascular endothelium. This granulomatous inflammation, perhaps aggravated by either a hypercoagulable state or mechanical stress, results in the clinical features of Crohn's disease.

Ultrastructural observations on the third-generation merozoites of Eimeria tenella in chicks.

The fine structure of the third-generation merozoites of Eimeria tenella is described from caecal enterocytes of chicks 120 h after experimental infections. The merozoites conformed to the general eimerian pattern except that they were bounded by two unit membranes. Young gamonts also conformed to the general eimerian pattern, and were limited by an outer unit membrane and an interrupted inner membrane. Enterocytes invaded by third-generation merozoites showed a loss of microvilli and large blebs that protruded into the caecal lumen. These blebs appeared to contain only cytoplasm and not any cell organelles.

Ultrastructural observations on caecal epithelial cells invaded by first-generation merozoites of Eimeria tenella in vivo.

Transmission electron microscopy was used to study the effects of invasion of caecal epithelial cells by first-generation merozoites of Eimeria tenella. At 53 h post-infection, merozoites were found within mature meronts and in enterocytes of the crypt epithelium. Following merozoite invasion, the terminal web of microfilaments of the invaded cell was disrupted and cell extensions were present on the enterocyte surface. There was a marked morphological alteration of the host cell resulting in loss of microvilli and extensive bulging of cytoplasm into the crypt lumen. Multiple invasion of enterocytes and invasion of goblet cells were observed. Large numbers of mast cells infiltrated the mucosa, identified by the metachromatic staining of their granules, and were found within the lamina propria, epithelium and lumen. Merozoites were found within mast cells and lymphocytes within the lumen.

Evidence of persistent measles virus infection in Crohn's disease.

Transmission electron microscopy was used to examine the microvasculature of perfusion-fixed tissues from Crohn's disease and control patients. Paramyxovirus-like particles, and inclusions consisting of condensations of nucleocapsid, in giant cells and endothelium at foci of vascular injury were identified in all 9 Crohn's disease patients. Tissues from patients with Crohn's disease were also examined by either in situ hybridisation (n = 10) or immunohistochemistry (n = 15), and compared to inflammatory and noninflammatory controls (n = 22). Hybridisation for measles virus N-protein genomic RNA was positive in all cases of Crohn's disease localising to foci of granulomatous vasculitis and lymphoid follicles. Positive immunohistochemical staining for measles virus nucleocapsid protein was positive in 13 of 15 patients with Crohn's disease, localising to foci of granulomatous inflammation. Hybridisation for measles virus RNA was positive in a minority of control intestinal tissues; viral inclusions were not seen ultrastructurally. Immunostaining was negative in control cases of intestinal tuberculosis. These observations suggest that measles virus is capable of causing persistent infection of the intestine and that Crohn's disease may be caused by a granulomatous vasculitis in response to this virus.

Desaggregation by Verapamil, Sodium Nitroprusside and PGE(1), of Rabbit Platelets Aggregated by PAF-acether. A Morphological and Biochemical Study.

The action of sodium nitroprusside, an activator of guanylate cyclase, on rabbit platelets stimulated with PAF-acether, was studied singly or in combination with verapamil. Not only did sodium nitroprusside in the range 10(-6) to 10(-4) M inhibit PAF-acether induced aggregation, but it also brought about desaggregation of previously aggregated platelets, as also did verapamil. The results were compared with those obtained with PGE(1), when used alone and in combination with verapamil. Morphological studies were carried out on control and aggregated platelets, and on platelets treated with verapamil either to inhibit aggregation or to bring about desaggregation. The platelets that had been desaggregated by verapamil were discoid in shape, had very few pseudopodia and exhibited a near normal internal morphology, except that the open canalicular system was swollen and contained an electron dense material resembling the contents of the alpha granules. The morphology of platelets desaggregated by PGE(1) was similar to that of platelets desaggregated by verapamil, including a swollen canalicular system containing electron dense material.

Granulomatous vasculitis in Crohn's disease.

This study investigated a possible vascular origin for granulomas in Crohn's disease. Twenty-four consecutive resected specimens of small and large intestinal Crohn's disease were preserved by arterial perfusion-fixation with 10% formol saline, at mean arterial pressure (100 mm Hg). Fifteen specimens contained granulomas on routine examination of H&E-stained sections. These 15 specimens were examined in detail using a range of immunohistochemical staining techniques to identify vascular structures and granulomas. A total of 485 granulomas were found, 85% of which were identified as being directly involved in vascular injury. The majority (77%) of granulomas were deep to the mucosa; they were found most frequently in the submucosa (42%). The techniques used in this study enhanced the recognition of granulomatous vasculitis. The results suggest that the majority of granulomas in Crohn's disease form within walls of blood vessels. Vascular localization of granulomatous inflammation suggests that the intestinal microvasculature contains an early element in the pathogenesis of Crohn's disease.

Ultrastructural studies of the effects of the ionophore lasalocid on Eimeria tenella in chickens.

The ultrastructural development of Eimeria tenella was studied in experimentally infected chicks fed 90 ppm lasalocid, an ionophorous anticoccidial antibiotic. Drug treatment was timed to target-specific endogenous stages. At 6 h after infection, many sporozoites within the epithelium showed degradation as a result of drug action. Only a few intact sporozoites were seen. The drug caused outer-membrane blistering, large surface swellings and enlarged mitochondria in both first- and second-generation merozoites. No effect on the gamonts was discerned.

Nicotine and cigarette smoking: effects on the ultrastructure of aortic endothelium.

The aortic endothelium from rats has been examined by scanning (SEM) and transmission electron microscopy (TEM) following nicotine administration or cigarette smoking. Nicotine was administered to rats by subcutaneous injections at concentrations of 1, 5, 10 and 20 mg/kg or by continuous subcutaneous pumps delivering a solution of 0.54 g/ml of nicotine at the rate of 1 microliter/h for 7 days. Animals were exposed to fresh smoke from eight cigarettes, each yielding 19 mg tar and 1.5 mg nicotine. Twenty minutes after subcutaneous injections, no endothelial abnormalities were evident by SEM, apart from at the high dose where minimal endothelial damage was seen. Limited endothelial damage was discerned following nicotine infusion and this consisted of microvillus formation, platelet adhesion and some cell separation. TEM revealed no endothelial changes in any of the animals. Cigarette smoking, however, resulted in considerable endothelial morphological changes consisting of the formation of blebs, microvillus-like projections, increased numbers of plasmalemmal vesicles and an increase in the numbers of Weibel-Palade bodies. The results suggest that components of cigarette smoke other than nicotine are responsible for the endothelial cell changes associated with smoking.

Effect of lasalocid on the development of Eimeria tenella in chicken embryos.

Sporozoites of Eimeria tenella were treated with lasalocid in vitro and their viability tested by inoculating them into the allantoic cavity of 11-day-old chicken embryos. Concentrations of 0.1, 1.0 and 10.0 micrograms ml-1 reduced sporozoite viability, as judged by oocyst production. Injections into the embryos of 5, 50 or 500 micrograms of lasalocid 92-93 h after infection also reduced oocyst production, indicating activity against the later stages of the life cycle.

Repopulation of guinea-pig skin by melanocytes during wound healing: a morphometric study.

Epidermal keratinocytes and melanocytes have a close functional interrelationship. In order to study this relationship we used computer-assisted three-dimensional morphometry (CAM) to investigate the shape and size changes of the cutaneous melanocyte in healing guinea-pig skin. The combination of CAM with osmium iodide staining and resin embedding of tissue gave excellent results and allowed qualitative and quantitative morphometric assessment of melanocytes in vertical epidermal sections. The changes in melanocytes and keratinocytes during healing of a standard 1 cm full thickness wound in the guinea-pig were studied. After an initial decrease, more melanocytes per mm2 of epidermis were seen (from 36 days). These were smaller in volume with shorter, less branched dendrites compared to controls. An unexpected finding was a late phase of melanocyte proliferation, at the end of our study period (99 days). Clearly, the complex changes in the melanocyte-keratinocyte relationship during wound healing continue throughout and beyond the period of our study.

Pathogenesis of Crohn's disease: multifocal gastrointestinal infarction.

In a prospective study, specimens of resected small and large intestine from fifteen patients with Crohn's disease were prepared by heparin-saline vascular perfusion, followed by either resin casting of the mesenteric vascular supply and tissue maceration or glutaraldehyde perfusion-fixation, resin casting, and tissue clearance. The specimens were examined by macrophotography, histopathology, and either scanning or transmission electronmicroscopy. A pathogenetic sequence of events in Crohn's disease was seen--vascular injury, focal arteritis, fibrin deposition, arterial occlusion mainly at the level of the muscularis propria, followed by tissue infarction or neovascularisation. These features were confined to segments of intestine affected by Crohn's disease and did not occur in normal bowel. The findings suggest that Crohn's disease is mediated by multifocal gastrointestinal infarction. This pathogenetic process is compatible with many of the clinical features of Crohn's disease, and its recognition has important implications for the identification of the primary cause of the illness and advances in clinical management.