Crystalline egg albumin denatured by exposure to ozone was analyzed by ultraviolet spectrophotometry. Rabbits were injected with denatured egg albumin to induce specific antibody. Native egg albumin (NEa) and ozone denatured egg albumin (O3Ea) cross-reacted with antinative egg albumin antibody (anti-N) and anti-ozone denatured egg albimin antibody (anti-O3), respectively. Heat denatured egg albumin (HEa) and O3Ea cross-reacted with anti-heat denatured antibody (anti-H). The precipitin lines in immuno-double diffusion tests between NEa and anti-N and between O3Ea and anti-N were fused with spur formation. The precipitin lines of O3Ea with anti-O3 and HEa with anti-O3 were fused with slight spur formation. Two precipitin lines of O3Ea with anti-O3 were observed by immunoelectrophoresis. The faster line had similar majority to the line of NEa and anti-N. The other line had a slightly slower mobility than the former line, indicating that this slower line was due to ozone specific antigen.
The present experiment was designed to examine the effect of vasogenic brain edema produced by exposure to air on the electrical activity of the thalamocortical pathway and local cerebral blood flow (1CBF) measured by the hydrogen clearance method. A large area of unilateral cerebral hemisphere of anaesthetized cats was exposed to air for 12 hours, and vasogenic brain edema was produced in the cortical and subcortical structure. Water content of the primary sensorimotor cortex, white matter, and thalamus, which was measured by the specific gravity method, significantly increased by 1.9, 4.1, and 0.7% g water/g tissue, respectively, after 12 hours of exposure to air. After six hours of exposure, the blood flow in the cortex, white matter, and thalamus significantly decreased from control levels of 56.8±12.7, 21.7±5, 2, 44, 1±13.4 ml/100g/min to 46.5±13.2, 16.3±5.0, and 31.3±11.9 ml/100g/min, respectively. These decreases were accompanied by diminution in the amplitude of the direct cortical responses (DCRs) and prolongation of N1 latencies of the somatosensory evoked responses (SERs) recorded at the primary sensorimotor cortex. Both electrical activities were progressively suppressed in proportion to the reduction of 1CBF after six hours of exposure. These results suggest that thalamocortical ischemia is probably responsible for the neural electrical suppression observed in this experimental edema. There was a significant correlation between the amount of edema and the degree of ischemia in the white matter. Accumulation of the edema fluid may induce ischemia affecting sensory conduction through the white matter as suggested by prolongation of the N1 latency of SER.
The present study was carried out to evaluate the effects of methylprednisolone (MP, 30 mg/kg) on brain edema and subcortical neural activity in anaesthetized cats with the cerebral cortex exposed to the air. MP was administered intravenously immediately after cortical exposure. After treatment with MP, significant reduction of edema was demonstrated in the primary sensorimotor cortex, white matter, and thalamus compared to the control. MP significantly increased local blood flow in the cortex and thalamus, but not in the white matter, and also protected the cortex against the ischemic suppression of the direct cortical response. However, MP did not affect the prolongation of the N1 latency of the somatosensory evoked response. The effect of steroids on cortical neural activity, including their beneficial effect on cortical edema and ischemia, may explain the rapid clinical improvement following administration of steroids.
Vectorcardiograms were recorded with the Frank lead system during laparoscopy. Pneumoperitoneum produced the following significant vectorcardiographic changes: (1) The maximum spatial QRS vector increased in magnitude and shifted horizontally and slightly anteriorly. (2) The L/W ratio of the frontal QRS loop decreased. (3) The scalar data of each X, Y, Z lead of the QRS loop increased. In particular, the increases in R of all three leads and in S of lead Z were significant. These results suggest that the changes in the cardiac anatomic position due to lift of the diaphragma and a decrease of left ventricular end-diastolic volume might produce these vectorcardiographic changes during laparoscopy.
The effects of antioxidants on cell proliferation and lipid peroxide formation of cultured aortic smooth muscle cells obtained from normal, hypercholesterolemic and aged rabbits were studied in vitro. α-Tocopherol and coenzyme Q10 were used as antioxidants. The lipid peroxide content of cells was generated in a time dependent manner with cell proliferation, it remained within the cells and did not accumulate in the media. α-Tocopherol promoted cell proliferation and suppressed the intracellular lipid peroxide formation expressed in terms of DNA content. These effects were significant only in HL-SMC, and were not significant in NR-SMC or Aged-SMC. Coenzyme Q10 did not effect cell proliferation or lipid peroxide formation.
The effects of antioxidants on the fatty acid composition of cultured aortic smooth muscle cells obtained from normal, hypercholesterolemic and aged rabbits were studied in vitro. α-Tocopherol and coenzyme Q10 were used as antioxidants. Cell lipid extracts were separated into cholesterol ester (CE), triglyceride (TG) and phospholipid (PL) fractions by thin layer chromatography. The fractions were analyzed by gas-liquid chromatography for fatty acid composition. Upon addition of α-tocopherol, the proportion of oleate (18:1) increased and that of linoleate (18:2) decreased in the CE and TG fractions, but that of stearate (18:0) increased and those of linoleate and arachidonate (20:4) decreased in the PL fraction. These effects of α-tocopherol were most significant in HL-SMC, and almost no change in Aged-SMC was observed. Addition of coenzyme Q10 resulted small changes in the fatty acid composition of the cultured cells and culture media.
The copper, zinc and metallothionein contents were measurded in various tissues of mutant (hemizygous and homozygous) macular, heterozygous macular and normal mice. The copper contents in the kidney and small intestine of 7-day-old mutants and heterozygotes were much higher than in normal mice, whereas the copper contents in the other tissues (liver, brain spleen and serum) were low. Renal copper contents of 1-day-old mutants were not different from contents of normal mice, whereas those of 7-, and 14-day-old mutants were significantly elevated. Hepatic and brain copper contents were extremely low at all stages compared to normals. Copper therapy was applide to 7-day-old mutant mice. One day after injection, hepatic copper contents in mutants increased slightly compared to the normal controls, whereas renal copper contents increasded greatly. Seven days after injection, hepatic copper contents in mutants decreased greatly compared to the normal control, whereas the increase in renal copper contents remained. One and 7 days after injectin, brain copper contents in mutants and normal mice increasde slightly compared to untreated mice. The renal MT-Cu contents in 7- and 8-day-old mutants and heterozygotes were significantly greater than that of normal mice. The marked elevation in renal MT-Cu contents was also observed in 8- and 9-week-old mutants and heterozygotes. In contrast to the kidney, hepatic MT-Cu contents in 7- and 8-day-old mutants and heterozygotes were much lower than in normal mice. In contrast to copper, no significant differences in zinc levels in any tissue were evident.
Timm's sulfide silver method was employed in studies of zinc localization in the hippocampal formation of the rat. Both the technique reported by Haug and that of Ibata and Otsuka have been used to perform Timm's method. Various problems with Timm's method, including fixation, duration of immersion in Timm's developing solution, temperature and illumination were evaluated in order to obtain the best possible results. The technique described in this communication afforded the following results. The silver particles were deposited preferentially in the synaptic vesicles of the mossy fiber terminals. None were seen in the mitochondria or in the dendritic spine. No silver granules were noted either in the pyramidal cells of the hippocampus or in the granular cells of the dendate gyrus. The preferntial localization of zinc in the mossy fiber ending is discussed.
In mycoplasmal pneumonia, both bacteriological and serological diagnoses are retrospective and, therefore, offer little help to clinicians. The isolation and identification of M. pneumoniae (Mp) require more than 1-2 weeks, and the serum antibody titer takes 1-2 weeks to rise after onset of the disease. I have developed a rapid test which applies the indirect immunofluorescent technique, for patient throat smears. Anti-Mp rabbit IgG was prepared by immunizing a rabbit with a concentrated cell suspension of Mp. Purified anti-Mp rabbit igG (primary antibody) and FITC-labeled anti-rabbit IgG goat IgG were applied to the throat smears which were pretreated with normal goat IgG to eliminate background non-specific fluorescence. Specific immunofluorescence was seen in 9 out of 10 throat smears from patients with Mp infection. Both granular and diffuse types of fluorescence were seen in the smears; the former was found in mucus and the latter on the entire surface of the epithelium. No specific immunofluorescence was seen in smear taken from patients with other respiratory infections or from normal volunteers. I consider that the method is useful for the early and rapid diagnosis of mycoplasmal infection.
Cathepsin B and L activities in cancer tissue and non-cancerous mucosal tissue were determined with small amounts of tissue homogenate from 29 operated patients with gastric cancer. Both enzyme activities were significantly higher in cancer tissues than in noncancerous mucosal tissues. The level of cathepsin B activity was higher in specimens of poorly differentiated adenocarcinomas, deeply invaded ps (+) and extensively metastasized regional lymph nodes (n2 or n3) than in specimens of tubular adenocarcinomas, invaded ps (-) and no metastasized regional lymph nodes. No significant correlation was observed between the elevated levels of cathepsin L activity in cancer tissues and any of the histological findings. These results suggest a close relationship of this enzyme to local invasion and metastasis to regional lymph nodes.
I investigated the possibility of production of antibody to bacterial membrane-cardiolipin using cardiolipin micelles, Staphylococcus aureus (S. aureus) and L-form of S. aureus. I also investigated and the reactivity of anti-cardiolipin (CL) antibody with bacteria and DNA. I confirmed that the bacterial membrane-cardiolipin induced the production of anti-CL antibody, which was present mainly in the IgM fraction, but also in the IgG fraction. The anti-CL antibody reacted with L-form of bacteria and bacterial membrane fractions, but it did not react with the vegetative form of bacteria other than S. aureus nor with DNA. On the basis of these findings, I considered that the L-form of bacteria or fragments of bacterial membranes can be detected by immunofluorescence antibody technique or nitrocellulose membrane-enzyme immunoassay using the anti-CL antibody.
I have developed a method for measuring the anti-osmotic stability of liposomes prepared with lipids extracted from bacteria, using the release of carboxyfluorescein trapped in liposomes as an indicator. I also examined the sub-second physical changes of liposomes submerged in a solution of low osmotic pressure with a stopped flow spectrophotometer. The trapped carboxyfluorescein was released when the liposomes burst upon the inflow of excess water. Liposomes prepared with the lipids of a stable S. aureus L-form strain or Mycoplasma orale were more resistant to osmotic pressure than those prepared from the wild strain of S. aureus. It was found that cardiolipin enhanced the membrane-stability in S. aureus and cholesterol (about 30% of the total membrane lipids) in Mycoplasma orale. By the present method, the resistibility of membranes to low osmotic pressure could be determined precisely with extracted and purified bacterial membrane lipids.
The distribution of zinc and postnatal development of the rat amygdala were investigated by light and electron microscopy with Timm's sulphide silver method. By Timm's reaction and toluidine blue staining, the rat amygdala could be divided into cortical (CO), lateral (L), basolateral (ABL), basomedial (ABM), central (C), medial (M) and intercated (I) nuclear regions. CO was the most strongly positive to Timm's reaction, L, ABM and C were strongly positive, ABL was also positive, and I and M were weakly positive. In newborns, no positive Timm's reaction could be observed, but a very weakly positive reaction was observed from the 5th postnatal day. The reaction became stronger with development, and reached the strength of that of the adult on the 30th postnatal day. Electron microscopically, the deposits of silver grains were only located in the nerve fiber endings containing many small clear vesicles. This finding is quite similar to that described in the mossy fiber endings in the hippocampal formation. Deposits of silver grains could not be found in newborns, and only very few could be observed after the 5th postnatal day. The number of deposits increased with age, and reached that of the adult from the 20th to the 30th postnatal days. It was elucidated that a significant increase in the number of silver grains per square 10-8 centimeter of Timm's positive nerve fiber endings occurred between the 10th and 20th postnatal days. These findings suggest that zinc exists in the nerve fiber endings in the rat amygdala and the amount of zinc increases with age. Zinc may play an important role in synaptic transmission.
The distribution of monochlorobenzene among the organs of mice was investigated by gas chromatographic equilibration after inhalation of 500ppm of monochlorobenzene for 1 hour, 300ppm for 1 hour, 100ppm for 1 hour and 100ppm for 3 hours. Monochlorobenzene was detected just after exposure at concentrations in the descending order of that in the adipose tissue, liver, kidneys, blood, heart and brain. The biological half-lives in the second phase of monochlorobenzene accumulation in the organs were in the descending order of length in the adipose tissue, brain, liver, spleen, kidneys and blood. There was a correlation between the amounts of lipid in the organs and the biological half-lives in the second phase. Biological half-lives in the second phase of monochlorobenzene accumulation in the blood, brain, liver and kidneys were in the descending order of exposure: 500ppm for 1 hour, 300ppm for 1 hour and 100ppm for 1 hour. Concentrations of monochlorobenzene in the blood, brain, liver and kidneys just after exposure to 300ppm of monochlorobenzene for 1 hour were higher than that after exposure to 100ppm for 3 hours. However, the biological half-life in the second phase after exposure to 100ppm for 3 hours was longer than that after exposure to 300ppm for 1 hour. The biological half-life in the second phase after exposure to 100ppm for 3 hours was longer than that after exposure to 100ppm for 1 hour.
Pregnant mice were exposed to 500ppm of monochlorobenzene, 500ppm of trichloroethylene and 1000ppm of 1, 1, 1-trichloroethane for 1 hour. The transfer of each solvent to the fetus through the placenta was investigated by gas chromatographic equilibration. The distribution of solvents among organs of the mother was also investigated. Monochlorobenzene just after exposure was in the desending order of concentrations in the adipose tissue, liver, kidneys and brain of mothers. This order was the same as that of non-pregnant mice. It was shown that these three organic solvents transferred from the mother to the fetus. Concentrations of monochlorobenzene and trichloroethylene were higher in the placenta than in the blood. The concentration of 1, 1, 1-trichloroethane was slightly lower in the placenta than in the blood. The ratio of transfer of these three organic solvents from the placenta to fetuses was about 1/3. The transferability to the placenta and fetuses, and to the liver, kidneys and brain, of monochlorobenzene was higher than that of trichloroethylene.
Cerebral vasospasm is the most severe complication of subarachnoid hemorrhage. Although the etiology of vasospasm is still obscure, it has been suggested that some Ca antagonists can relieve cerebral vasospasm. The effects of Ca antagonists (Nifedipine, Nicardipine, Verapamil, Cinnarizine, Diltiazem) on the diameter of the basilar artery and rCBF in the brain stem were investigated in cats with experimental cerebral vasospasm. Three days after an intracisternal injection of blood (3ml), the basilar artery was exposed by the transclival approach. Experimental cerebral vasospasm was induced by the topical application of a blood CSF mixture. The rCBF was measured by the heat clearance method, and the diameter of the basilar artery was examined in serial photographs. Whenever Ca antagonists were given, the mean arterial blood pressure (MABP) decreased dose dependently. Ca antagonists can dilate (from 7.1% to 45% 10 minutes after the administration of Ca antagonists) the normal basilar artery, but can hardly change the basilar artery of cats with experimental cerebral vasospasm. The response of the rCBF in the brain stem after the administration of Ca antagonists was of 3 types: continuous increase, transient increase and no response. The greater the MABP decreased, the higher the possibility of continuous increase and transient increase among all cases was. The possibility of continuous increase and that of transient increase were higher in the control group than that in the vasospasm group. However, even in the vasospasm group the possibility of continuous rCBF increase and that of transient rCBF increase in the brain stem were not above 50%. Therefore, the clinical application of Ca antagonists to the treatment of vasospasm after subarachnoid hemorrhage may be difficult. But, on the other hand, it is possible that the administration of Ca antagonists before the appearance of vasospasm would be the prophlaxis for vasospasm after subarachnoid hemorrhage.
An image processing and 3-D display method was developed for MRI of pituitary gland hematomas. MRI was performed with a TOSHIBA MRT-15A resistive magnetic system. The visualized anatomical region was recorded on film in constant slice intervals. Image data were taken with a TV camera and digitized into 256 gray levels on a 512×512 array of points. The boundaries of the pituitary gland, ventricles and brain were extracted by region segmentation. The boundaries of the pituitary gland in 7 sagittal sections and 2 axial sections were displayed three-dimensionally. The boundaries of the brain tumor, ventricles and brain were extracted well by the image processing techniques. This method may provide valuable diagnostic and physiologic information.
The three dimensional architecture of the rat femur bone marrow stroma was observed by scanning electron microscopy. The bone marrow was fixed and vibrated by ultrasonication to dislodge the hematopoietic cells embedded in the meshes of the marrow reticulum. The stromal reticulum was exposed, while its three dimensional architecture was preserved. The marrow stromal reticulum consisted of intersinusoidal reticular cells and perivascular reticular cells (adventitial cells), and a small amount of reticular fibers. Two types of meshwork were noted in the bone marrow reticulum. Each mesh of one type was the size of one hematopoietic cell. This meshwork consisted of short and broad or stout cytoplasmic processes. A hematopoietic cell was enwrapped by attenuated cytoplasmic sheaths of the reticular cells and sequestered from the surrounding hematopietic cells. The other type of meshwork was composed of straight running, long and slender filopodia of the reticular cells. Several hematopoietic cells were tightly packed in each mesh of the meshwork of this type, thus presenting a polyhedral appearance. Attenuated adventitial cells enveloped the marrow blood vessels, and were connected with each other by their sheath-like cytoplasmic processes but round, variously sized intercellular spaces remained. The intersinusoidal reticular cells extended their processes to the adventitial cells and the adventitial cells also stretched their processes to the intersiusoidal reticulum.
The organization of the elastic tissues of the human and rabbit aorta was studied by scanning and transmission electron microscopy of tissues treated with 88% formic acid at 45°C. When the tissues were treated with formic acid until they became semitransparent, fixed tissues showed the same organization of the elastic tissues as unfixed tissues. When tissues were treated with formic acid for a short time, the intimal elastic lamina was observed as a plate with numerous small pores or fenestrae, whereas long treatment revealed that the lamina consisted of a network of elastic fibers. The tunica media consisted of concentrically arranged elastic lamellae. There were 50-60 elastic lamellae in humans and about 30 in the rabbit. The elastic fibers, that consisted of elastic microfibrils, ran longitudinally, repeatedly branching and anastomosing, and formed the network of the elastic lamella. Between the adjacent elastic lamellae stretched interlamellar elastic fibers. Thus, the elastic tissues in the aorta formed an integral network, which seems to be highly associated with the aortic function of distributing the pressure applied to the vascular wall. Disruptions of the integrity of the elastic tissue architecture may be responsible for the increased incidence of hypertension with age.
Snapping thumb of the infant is usually caused by the disturbance of movement of the flexor tendon in its sheath. Few reports have dealt in detail with the histopathology of the snapping thumb. In the present study, light and electron microscopic observations were carried out on parts of the tendon and tendon sheath excised from snapping thumbs. The tendons and tendon sheaths were obtained from 21 fingers of 19 infants operated on over the last 2 years, under the diagnosis of snapping thumb. There were 10 males and 9 females, ranging in age from 1 to 6 years old. Sectioned samples were studied histochemically, using periodic-acid Schiff, toluidine-blue and high-iron-diamine alcian blue stains. The ultrastructure of the tendon nodule was also studied by scanning and transmission electron microscopy. Increase of the amorphous material and collagen fibrils was observed at the site of tendon nodule formation. The amorphous material was basophilic with hematoxylin-eosin staining, metachromatic with toluidine blue staining, and became dark violet with high-iron-diamine alcian blue, which indicates that it contains acidic mucopolysaccharide with a sulfate radical. The tendon sheath did not contain much amorphous material, but connective tissue cells proliferated. Synovial lining cells were not stratified and no inflammatory cells were found. By scanning electron microscopy, bundles of collagen fibrils of 0.1-0.15μ in diameter, probably newly synthesized, were observed in the middle portion of the tendon nodule. Transmission electron microscopy confirmed that ruthenium-red positive materials exist inside and outside of the fibroblasts. These fibroblasts seem to very actively produce acid mucopolysaccharide and collagen fibrils. Accumulation of the mucopolysaccharide and collagen fibrils by metaplasia of the fibroblasts in the endotenon and epitenon might make the nodule in the tendon. This tendon nodule formation is the primary cause of the snapping thumb of infants.