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Biomedical subjects

A Miki

Publications and source records attributed to A Miki.

At least 73 records · Page 4Linked to original sources

A differential staining method for adenohypophyseal cells.

A modification of azan staining (Heidenhain, 1915) for a better differential demonstration than previously of adenohypophyseal cells is reported. Human hypophyses were fixed in Bouin's solution, washed in 70% alcohol, dehydrated and then embedded in paraffin. Sections of about 5 microns thickness were cut and mounted on albumin-coated glass slides. These sections were gently oxidized with a mixture of potassium permanganate and sulphuric acid, and then stained according to the original azan technique; thereafter a procedure of staining with aniline blue was added. With this improvement, human adenohypophyseal cells were clearly classified into six groups on the basis of the color and intensity of staining of the cytoplasm. The alpha-acidophilic cells were stained clear red with azocarmine and the epsilon-acidophilic cells, orange-red with orange G, respectively. With aniline blue, beta-basophilic cells stained a deep blue, while delta-basophilic cells were more weakly stained. The gamma-chromophobic cells were faintly stained red, and the ACTH-cells very faintly blue. Collagenous fibers developed a blue color, while erythrocytes were orange-red to red. This modified method produces much better results in differential staining of the adenohypophyseal cells than either the original azan or Masson-Goldner staining methods.

Adult↗

[Functional magnetic resonance imaging of the human primary visual cortex during visual stimulation].

Signal changes in the human primary visual cortex during visual stimulation were evaluated using non-invasive functional magnetic resonance imaging (fMRI). The experiments were performed on 10 normal human volunteers and 2 patients with homonymous hemianopsia, including one who was recovering from the exacerbation of multiple sclerosis. The visual stimuli were provided by a pattern generator using the checkerboard pattern for determining the visual evoked potential of full-field and hemifield stimulation. In normal volunteers, a signal increase was observed on the bilateral primary visual cortex during the full-field stimulation and on the contra-lateral cortex during hemifield stimulation. In the patient with homonymous hemianopsia after cerebral infarction, the signal change was clearly decreased on the affected side. In the other patient, the one recovering from multiple sclerosis with an almost normal visual field, the fMRI was within normal limits. These results suggest that it is possible to visualize the activation of the visual cortex during visual stimulation, and that there is a possibility of using this test as an objective method of visual field examination.

Adult↗

Functional magnetic resonance imaging of the primary visual cortex: evaluation of human afferent visual system.

The authors evaluated signal changes in the human primary visual cortex during visual stimulation using functional magnetic resonance imaging (MRI) scanner at 1.5 Tesla. Experiments were performed on 10 normal volunteers and 2 patients with homonymous hemianopsia. In the normal volunteers, a signal increase was observed on the bilateral primary visual cortex during hemifield stimulation. In one patient with homonymous hemianopsia after cerebral infarction, the signal change was clearly decreased on the affected side. In the other patient, who was recovering from multiple sclerosis to an almost normal visual field, the fMRI results were within normal limits. These results suggest that it is possible to map noninvasively the activation of the visual stimulation with a clinical MRI system, and that this test might be useful as an objective method of visual field examination.

Adult↗

Plasmodium chabaudi: association of reversal of chloroquine resistance with increased accumulation of chloroquine in resistant parasites.

The effects of tricyclic antidepressants, desipramine and imipramine, and phenothiazines, chlorpromazine and trifluoperazine, on chloroquine (CQ)-resistant and CQ-sensitive lines of P. chabaudi were examined in vivo. In mice that received daily injections of these drugs the growth of CQ-resistant and CQ-sensitive parasites was unaffected or affected very slightly, if at all. A combination of CQ and each drug suppressed the growth of CQ-resistant parasites in a dose-dependent manner. In addition, in CQ-sensitive parasites each drug also increased the susceptibility to CQ. Measurements of CQ levels by high-performance liquid chromatography showed that CQ accumulated in sensitive parasites to more than twice the level in resistant parasites at 2 to 4 hr after an injection of CQ. Verapamil and desipramine substantially increased CQ levels in both CQ-resistant and CQ-sensitive parasites. These results suggest that not only Ca2+ antagonists but tricyclic antidepressants reverse CQ resistance in CQ-resistant parasites and enhance the inhibitory effect in sensitive parasites by increasing CQ levels in those parasites. The effects of Ca2+ antagonists, tricyclic antidepressants, and phenothiazines on a pyrimethamine-resistant line of P. chabaudi were also studied. None of the Ca2+ antagonists (verapamil, nicardipine, and diltiazem) affected the growth of the parasite in combination with 20 mg/kg pyrimethamine. Tricyclic antidepressants and phenothiazines suppressed pyrimethamine-resistant parasites to some extent. However, the extent of this suppression was less pronounced as compared with that of suppression of CQ resistance by the same drugs.

Animals↗

Traumatic degeneration of transected myelinated fibers of the mouse sciatic nerve.

Traumatic degeneration of myelinated fibers was studied by electron microscopy over 5 days following transection of mouse sciatic nerve. Special attention was paid to the mechanism which separates the degenerating part, while preserving the viable part of the axon. Immediately after transection, the opened end of the proximal stump revealed extensive subcellular changes including the disorganization of neurofilaments, and disruption of mitochondria and axonal endoplasmic reticulum (SER). Subsequently, vesicles of round and tubular profiles filled up the whole area of the stump end, and proximal to it appeared a neurofilament-predominant area characterized by randomly oriented neurofilaments and normally appearing mitochondria and SER. Characteristic membranous demarcations occurred in early periods at the border between the vesicle accumulation and the neurofilament-predominant areas, and later also within these areas. The demarcation membranes formed both by invagination of the surface plasma membrane and, probably, by fusion of the large vesicles. These became prominent with time, dividing the axoplasm into compartments of varying sizes, which gradually underwent degeneration and were liberated from the parent axon. Occurrence of autophagic vacuoles was characteristic of the degenerating portions of the parent axon. Thus, by the function of demarcation membranes, the parent axon to be preserved could remain membrane-bound, while the degenerating parts were shed off.

Animals↗

Ultrastructural changes associated with reversal of chloroquine resistance by verapamil in Plasmodium chabaudi.

Reversal of chloroquine (CQ) resistance by verapamil, a Ca2+ antagonist, has been shown in CQ-resistant human and rodent malaria parasites. Here, we report ultrastructural changes associated with this phenomenon in CQ-resistant Plasmodium chabaudi (AS strain) after infected mice were administered CQ and verapamil. At parasitaemias of 5-7%, CQ at 6 mg/kg caused little morphological effect on CQ-resistant parasites. In contrast, co-administration of CQ and verapamil at 50 mg/kg induced swelling of food vacuoles with clumped pigment at 2.5 h. Morphological changes other than food vacuole enlargement occurred at 21 and 45 h: disappearance of endoplasmic reticulum, formation of myelin structures, focal cytoplasmic vacuolization and coarse clumping of electron-dense material in nuclei. These structural changes appeared to be very similar to those observed in CQ-sensitive P. chabaudi in mice injected with CQ alone or CQ plus verapamil. On the other hand, verapamil at 50 mg/kg alone did not induce any effect on both CQ-sensitive and CQ-resistant P. chabaudi. These results suggest that swelling of the food vacuoles is an initial event associated with reversal of CQ-resistance by verapamil.

Animals↗

Morphological analysis of selective destruction of pancreatic beta-cells by cytotoxic T lymphocytes in NOD mice.

Interactions of pancreatic islets and islet-associated mononuclear cells (IAMCs) from the nonobese diabetic (NOD) mouse were morphologically investigated. To obtain IAMCs, pancreatic islets isolated from adult NOD mice were cultured for 7 days with interleukin 2. Noted by light microscopy, interactions between IAMCs and freshly isolated islets from young NOD mice began 30 min after the initiation of the coculture, and 6 h later, normal cellular array of the islets was lost. By electron microscopy, most IAMCs had low nucleus-cytoplasm ratio, the nucleus was notched and exhibited condensed chromatin along the nuclear membrane, and well-developed Golgi complexes and several mitochondria were distributed in the cytoplasm. These IAMCs adhered to beta-cells, but not to alpha- or delta-cells, with their pseudopods and caused cytolysis of beta-cells. Immunohistochemical study with antibodies specific for pancreatic hormones demonstrated that only cells reacting with anti-insulin antibody were selectively lost as the incubation time proceeded. Electron immunohistochemistry by immunogold technique showed that effector cells in IAMCs reacted with anti-CD8 (Lyt-2) antibody, but not anti-CD4 (L3T4) or anti-asialogangliosideM1 antibody. In addition, the concentration of pancreatic hormones in the culture medium, used as a marker of cytolysis, also demonstrated that insulin was significantly increased after 6 h of culture, whereas glucagon and somatostatin were not. These results suggest that CD8+ cytotoxic T lymphocytes are involved in the selective destruction of pancreatic beta-cells in the NOD mouse.

Animals↗

Membrane potential of Plasmodium falciparum gametocytes monitored with rhodamine 123.

The membrane potential of Plasmodium falciparum gametocytes was monitored with the cationic permeant fluorescent dye rhodamine 123 (R123) as a probe. Epifluorescence microscopy revealed that R123 at 1 microgram/ml rather selectively partitioned into structure resembling large mitochondria. Treatment of R123-loaded gametocytes with various inhibitors including those of respiration resulted in disappearance of fluorescence from what appeared to be the mitochondria, but not from the cytosol. These results indicate that P. falciparum gametocytes have the mitochondrion maintaining an inside negative membrane potential.

Animals↗

[Experimental study on a mechanical root canal enlargement].

It is difficult for us to enlarge a root canal by using hand-instruments in a molar tooth, so we experimented by using mechanical equipment. We examined the human extracted premolar which we had preserved in physiological saline solution by using a speculative root canal enlarging machine. Then we observed the time and condition of enlarging root canal by X-ray. After slicing off, we examined with our eyes and with an electronic microscope. Afterward, we compared hand-instruments with the enlarging machine. As the results of this experiment; 1. Time by hand filing instruments took three or four times to make a root canal enlargement by a speculative enlarging machine. 2. After enlarging a root canal, we made a root canal filling. Observing them by X-ray and ground specimen, we recognized a good result in the root canal enlargement and in the condition of the root canal filling. 3. Observing the figure by electronic microscope, in the case of using a speculative machine, we found that the slicing aspects of dentin which had been enlarged by root canal were thoroughly smoothed in the center part of the root canal, but it was not smoothed in the root apex region. 4. It had a tendency to remain part of the cutting dentin near the apical area both by using hand cutting instruments and by using a speculative machine.

Bicuspid↗

Immunological characterization of papain-induced fragments of Clostridium botulinum type A neurotoxin and interaction of the fragments with brain synaptosomes.

After treatment of Clostridium botulinum type A neurotoxin with papain, three fragments (Mrs, 101,000, 45,000, and 43,000) were purified by hydrophobic and ion-exchange chromatography with a high-performance liquid chromatographic system. Immunoblotting analyses with monoclonal antibodies showed that the 101,000-dalton fragment consisted of the light chain and a part of the heavy chain (H-1 fragment) linked together by a disulfide bond, and the other two fragments were correlated to the remaining portion of the heavy chain (H-2 fragment). The 45,000- and 43,000-dalton fragments effectively competed for binding of the 125I-labeled neurotoxin to synaptosomes, while no inhibition was observed with the 101,000-dalton fragment. The results indicate that the H-2 fragment interacts with the binding site on the neural membrane. The binding of the neurotoxin was impaired by treatment of synaptosomes with neuraminidase. Incorporation of gangliosides into neuraminidase-treated synaptosomes resulted in the restoration of binding. The results suggest that gangliosides are one of the components of the toxin-binding site.

Animals↗

Inflammatory cell changes in haversian canals. A possible cause of osteoporosis in rheumatoid arthritis.

We studied the morphology of the haversian canals in the osteopenic cortical bone of the medial femoral neck from patients with rheumatoid arthritis and compared the findings with those in patients with osteoarthritis and with uncomplicated coxa valga. In the rheumatoid bone, the diameters of the canals were larger and many more contained osteoclasts. Fewer haversian canals showed only lining cells than in the osteoarthritic or coxa valga patients. In bone from rheumatoid patients, especially in canals with osteoclasts, small blood vessels were frequently lined by tall endothelial cells with an infiltration of mononuclear cells. These morphological differences are discussed with reference to the possible mechanisms of loss of cortical bone in rheumatoid arthritis and other conditions.

Adolescent↗

Histochemical study of the differentiation of microglial cells in the developing human cerebral hemispheres.

Applying nucleoside diphosphatase (NDPase) histochemistry, the appearance and differentiation of microglial cells in the developing human cerebral hemispheres were investigated by light and electron microscopy. In the pallium of the 38 days old human embryo, a few round NDPase-positive cells (round cells) were observed in the expanding zone. Although distinct blood vessels had not yet formed within the wall of the pallium, some cellular elements resembling haemopoietic cells were noticed in the expanding zone. In the 51 days old fetus, blood vessels displaying NDPase activity were seen in the mantle and marginal layers, and some invaded the matrix. Several round NDPase-positive cells were distributed, mainly around the vascular sprouts (primitive blood vessels) in the matrix. In the marginal layer, NDPase-positive cells exhibiting short cytoplasmic processes were encountered (poorly ramifying cells). In the 58, 66 and 82 days old fetuses, the round NDPase-positive cells were seen mainly in the matrix or subcortical layer where vascular sprouts were conspicuous and the poorly ramifying cells were in the subcortical and marginal layers. In the two latter fetuses, NDPase-positive cells showing long highly ramifying cytoplasmic processes (highly ramifying cells) were noted mainly in the marginal layer and sometimes in the subcortical layer. In the 5 months old fetuses, numerous NDPase-positive cells were distributed in the mantle, subcortical and marginal layers, and most of them appeared to belong to the populations of the poorly or highly ramifying cells. On the basis of the ultrastructural features, the round cells and highly ramifying cells were regarded as amoeboid cells and microglial cells, respectively. These findings suggest that at least some amoeboid cells are transformed into microglial cells via the stages of poorly ramifying microglial cells, and also that, in the human cerebral hemispheres, appearance of the microglial elements is closely related with vascularisation, especially in the early developmental stages.

Brain↗

Histochemical studies of enzymes of the energy metabolism in postimplantation rat embryos.

Using histochemical procedures for the detection of lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), and cytochrome c oxidase (cytox), we investigated the levels of these enzymes of the energy metabolism in postimplantation rat embryos (9.5-12.5 days of gestation). On day 10.5 of gestation, the neural tube, somites, myocardium, and mesenchyme displayed moderate levels of LDH activity; this activity gradually increased in strength, so that, on day 12.5 of gestation, intense LDH activity was uniformly distributed in these intraembryonic tissues. In contrast to LDH, distinct regional differences in the distribution of SDH and cytox were detected. On day 10.5 of gestation, the myocardium exhibited weak to moderate SDH and cytox activity, and on day 11.5, the myocardial activity of these enzymes had become moderate to intense. However, in all other embryonic tissues, e.g., the neural tube and somites, only weak SDH and cytox activity was present. On day 12.5 of gestation, the myocardium displayed very intense SDH and cytox activity, whereas the mantle layer of the neural tube, the spinal ganglia, and the myotomes exhibited only moderate levels of SDH and cytox activity. In the matrix of the neural tube and mesenchyme, these enzyme activities remained at low levels. At electron microscopy, cytox activity was detectable in the spaces between the inner and outer membranes as well as in the intracristal spaces of mitochondria. In general, cytox activity increased in parallel with the differentiation of mitochondria (i.e., increased mitochondrial numbers and size, and the development of mitochondrial cristae), but when the distribution of the cytox activity was considered in detail, it was found to differ among mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of oxygen concentration on embryonic development in rats: a light and electron microscopic study using whole-embryo culture techniques.

By using a whole-embryo culture technique (New 1978), the effects of oxygen concentration (5%, 20% and 95% oxygen) on embryonic development in the rat were investigated by light and electron microscopy. The best embryonic development occurred when the 9.5-day-old embryos were cultured for 24 h with 5% oxygen, and the 10.5-day-old embryos with 20% oxygen (optimum oxygen concentration). When the 9.5- and 10.5-day-old embryos were cultured for 24 h with too little or too much oxygen, retardation of the embryonic growth and abnormal development was observed. Using light microscopy, numerous degenerating cells, exhibiting granular deposits in the cytoplasm, were seen, but the distribution of the degenerating cells was quite different between the two groups. With electron microscopy, the most striking feature of the degenerating cells in the embryos cultured with too little oxygen, was the extreme swelling of the mitochondria without any morphological alterations of the nucleus or the other cell organelles. On the other hand, the characteristic feature of the degenerating cells in the embryos exposed to too much oxygen, was the formation of phagolysosomes in the cytoplasm. Morphological alterations of the nucleus or mitochondria were not evident. In the present study, the possible teratogenic mechanism of too much or too little oxygen in the whole-embryo culture of the rat embryo is discussed.

Animals↗