We found Encephalitozoon-like organisms in an in vitro culture of a human liver lesion which was due to larval Echinococcus multilocularis. The organisms developed in the same fashion as an Encephalitozoon cuniculi. The spores that developed in parasitophorous vacuoles were 2.0-2.6 x 1.1-1.5 microns; each contained a single nucleus and 4-5 polar tubule coils, closely resembling E. cuniculi in its ultrastructure. Mature spores were collected from the supernatants by the use of Percoll centrifugation resulting in the banding of the spores on continuous gradients. We prepared three sorts of spores which were different in buoyant density in 0.15 M NaCl: 1.05-1.07 g/ml spores, 1.12 g/ml spores, and spores of over 1.14 g/ml. Polyclonal antibodies to a pool of each spore preparation were produced in a rabbit and applied to the detection of microsporidian antigen in situ. The histoimmunoperoxidase (HIP) procedure was used to detect the microsporidian antigen in echinococcal liver lesions from patients with alveolar hydatid disease (AHD). Ten echinococcal liver lesions from different AHD patients were examined and four were found to be positive in the HIP test. The Percoll-separated spores were also used as an antigen to detect for antibodies in the sera from the patients with AHD by Western blotting. Antibodies were detected in 62 (52%) of the 119 AHD patients and in only 8 (5%) of the 159 normal healthy individuals.

Adolescent↗

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Primers designed for amplification of Echinococcus multilocularis DNA amplify the DNA of Encephalitozoon-like spores in the polymerase chain reaction.

Microsporidian spores were developed from cells which were grown in vitro from a human liver lesion which was due to larval Echinococcus multilocularis. The microsporidian spores developed in the same fashion as an Encephalitozoon cuniculi. The Encephalitozoon-like spores were completely separated on Percoll gradients. The separated spores contained DNA capable of amplification by two different primer sets designed for the polymerase chain reaction (PCR) of E. multilocularis DNA. However, the cell DNA from which microsporidium developed was thoroughly insensitive to the PCR using the E. multilocularis primer sets. The results strongly suggested that Encephalitozoon should be taken into consideration, when DNA isolated from larval E. multilocularis is analyzed.

Animals↗

PubMedDate not supplied

Nucleotide sequences of DNA fragments of Encephalitozoon cuniculi amplified by polymerase chain reaction with primers regarded as specific for Echinococcus.

Encephalitozoon-like spores were separated from a human echinococcal liver lesion, which was caused by Echinococcus multilocularis. They were found to fall into the species Encephalitozoon cuniculi, which was shown to have En. cuniculi-specific DNA by way of polymerase chain reaction (PCR). We also used PCR to genetically discriminate between the En. cuniculi spores and the Ec. multilocularis larvae. Two primer sets, known to be specific for Echinococcus, were examined. These primers were expected to work normally when the two quite different DNA preparations were tested as templates, i.e. only Echinococcus DNA could give a positive signal in the PCR tests. However, it was found that the two Echinococcus-specific primer sets could amplify not only Ec. multilocularis DNA, but also En. cuniculi spore DNA. We then tried to determine the order of nucleotides in the Echinococcus-specific primers-amplified En. cuniculi PCR products and compared the determined sequences with those of Ec. multilocularis. The results clearly indicated that sequencing made little difference between En. cuniculi and Ec. multilocularis.

Animals↗

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Role of the liver in alloimmune response following inoculation of donor spleen cells.

The liver is thought to be an immunologically privileged organ in the response to inoculated antigens. We previously demonstrated that it is possible to localize inoculated antigens to the liver alone or only to extrahepatic tissue using orthotopic syngeneic liver transplantation (OSLT). In this study, we analyzed more detailed mechanisms of the anti-alloimmune response in the liver. DA rat spleen cells were systemically injected into WS rats (donor spleen cell inoculation, DSI). In the sensitized liver-grafted (SLG) group, after DSI, liver grafts were retrieved from sensitized WS rats, then transplanted into naive WS rats. In the sensitized liver-removed (SLR) group, after DSI, WS rats were totally hepatectomized and given livers transplanted from naive WS rats. All the rats were challenged with heterotopic heart grafts 10 days after DSI. Mean heart graft survival in the control, DSI, SLG, and SLR groups were 11.6+/-1.6, 10.7+/-2.4, 4.4+/-1.0, and 24.6+/-6.3 days, respectively. Accelerated rejection in the SLG group as well as graft prolongation in the SLR group disappeared when OSLT was performed 2 days after DSI or later. Irradiation of DA splenocytes before inoculation did not alter graft survival in SLG. However, pretreatment with gadolinium chloride prior to DSI attenuated the antidonor response in the SLG group. In conclusion, a vigorous antidonor response occurred in the liver after systemic inoculation of spleen cells. It peaked I day after DSI and disappeared rapidly. Kupffer cells seemed to play an important role in this phenomenon.

Animals↗

PubMedDate not supplied

Paracoccidioidomycosis in Japan. Report of a case.

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Adult↗

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Effects of a human urinary kininogenase (SK-827) on cerebral microcirculation after glass bead-induced cerebral embolism in rabbits.

The pharmacological effects of a human urinary kininogenase (SK-827) on the cerebral microcirculation were studied intravital-microscopically in a rabbit with cerebral microvessel injury induced by glass bead injection into an internal carotid artery. Intravenous administration of SK-827 dilated pial arteries of the brain and increased cerebral hemoglobin content (IHb). SK-827 tended to dilate smaller arterioles more markedly than larger arterioles, and larger arterioles than small arteries. SK-827 appeared to have a vasodilating effects qualitatively different from those of nicardipine and CDP-choline. Findings obtained by concurrent hematological and biochemical examinations also suggested its beneficial effects on the function of platelets and erythrocytes.

6-Ketoprostaglandin F1 alpha↗

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Morphological changes during survival, cellular transformation, and calcification of the embryonic mouse: Meckel's cartilage transplanted into heterotopic sites.

Meckel's cartilages obtained from 18-day-old embryonic mice were transplanted into heterotopic sites to examine their site-specific effects on cellular modification, calcification, and long-term survival. The explants were isografted into the liver, subcutaneous tissue, anterior chamber of the eye, kidney, peritoneal cavity, and intrafemoral muscle of mouse for up to 2 weeks. In addition, Meckel's cartilages were wrapped in a Nucleopore filter and transplanted into the spleen. Grafted Meckel's cartilages were observed by light and electron microscopy, including application of von Kossa's reaction and osmium-potassium ferrocyanide fixation. It was demonstrated by von Kossa's reaction that when Meckel's cartilage was grafted in liver parenchyma, initial calcification appeared on the territorial matrix as spotted deposits. The chondrocytes gradually transformed into small ovoidal cells and showed a morphology closely resembling that of osteocytes. Transplants in the kidney subcapsular site contained active surviving chondrocytes, and subcutaneously grafted Meckel's cartilage revealed three types of chondrocytes: nonhypertrophic and calcification-inducing hypertrophic chondrocytes, as well as cells showing osteocytelike phenotypes. Intraocular and Nucleopore filter-wrapped explants showed degenerative changes, and peritoneal-cavity transplants showed cellular hypertrophy, but calcified precipites were not observed. The present investigation demonstrated that in blood-rich tissues Meckel's cartilage showed long-term survival and could modulate calcified precipitation. Thus, this adaptation of Meckel's cartilage to the environment of various heterotopic sites suggests that it has the ability to induce calcification.

Animals↗

PubMedDate not supplied

Effects of a human urinary kininogenase (SK-827) on cerebral and cutaneous microcirculation in rabbits.

Pharmacological effects of a human urinary kininogenase (SK-827) on the cerebral and cutaneous microcirculation were studied in anesthetized normal rabbits. Intravenous administration of SK-827 had no effect on the diameter of pial arterioles or the hemoglobin content (IHb) of the brain. On the other hand, SK-827 increased IHb of the skin, dose-dependently and dilated cutaneous arterioles transiently with increased capillary red blood cell velocity. These findings suggest that the pharmacological effects of SK-827 on the microcirculation of the brain and the skin differ from each other due to the activity of the autoregulatory mechanism.

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