Effect of splenectomy on establishment of hematopoietic chimerism in nonirradiated mice.

When spleen cells (1.5 x 10(8)) from beige C57BL/6 (bgJ/bgJ, Chediak-Higashi syndrome) mice were injected into nonirradiated (C57BL/6 x CBA)F1 hybrid mice, the hematopoietic stem cells of the hybrid host were eradicated by graft-versus-host (GVH) reaction, and peripheral neutrophils of the host changed from normal type to beige type with giant sudanophilic granules in 20 days after the cell injection. Effect of splenectomy on the establishment of such hematopoietic chimerism was investigated. The splenectomy carried out either before or after the cell injection reduced the proportion of mice in which more than 90% of peripheral neutrophils became of beige type. Since the mortality of mice which received splenectomy at various days prior to or after the cell injection paralleled the proportion of establishment of chimerism, the splenectomy does not seem to affect directly the establishment of chimerism but indirectly through the modification of the intensity of GVH reaction.

Survival of host mast cells after establishment of hematopoietic chimerism by graft-versus-host reaction in nonirradiated F1 hybrid mice.

Since the tissue mast cell has been shown to be progeny of the multipotential hematopoietic stem cell (CFU-S), and the CFU-S is a sensitive target of graft-versus-host (GVH) reaction, we examined whether or not the mast cell is also the target of GVH reaction. Giant granules of C57BL/6-bgJ/bgJ mice were used as a marker of donor cells. When 10(8) spleen cells of C57BL/6-bgJ/bgJ mice were injected into nonirradiated (C57BL/6 X CBA)F1 hybrid mice, erythrocytes and neutrophils became of donor type in about one-half of the recipient mice. In the bone marrow and spleen of the chimeric mice, the CFU-S was of donor type as well. In contrast, mast cells of host type remained in many tissues of the chimeras. Moreover, mast cell precursors with capabilities of proliferation and differentiation were preserved in the skin of chimeras. The present results suggest that the effect of systemic GVH reaction on mature mast cells and the mast cell precursor fixed in the skin is significantly less severe than that on the CFU-S itself.

Scanning electron and light microscopic observations on the healing process after sintered bone implantation in rats.

The healing process after implantation of sintered bone in the rat parietal bone was compared with that of synthetic hydroxyapatite using both scanning electron and light microscopy. The results showed that the differences between the sintered natural bone and the synthetic hydroxyapatite implantations were in the states of bone union and the bioresorbability of the implanted materials, even though both materials consist of the same hydroxyapatite. In the sintered bone implantation, the newly formed bone invaded into the material at 1 to 2 weeks after implantation. The sintered bone surface on the dura mater side was completely covered by the new bone at 5 weeks. It is noteworthy that bone resorbing areas characterized by Howship's lacunae were observed on the sintered bone surface at 2 weeks and the material was replaced by new bone. Light microscopy, which revealed the invasion and the development of the new bone into the sintered bone, supported the scanning electron microscopic observations. In the synthetic hydroxyapatite, the new bone adhered closely to the material just like the sintered bone implantation. The new bone did not invade into the synthetic hydroxyapatite. There was no evidence of the resorption of the hydroxyapatite. This shows that the natural and the biological structures of the sintered bone offer an advantageous environment to fluid circulation and ingrowth after implantation.

Morphological changes of autoclaved autogenic bone implantation and autoclaved autogenic bone supplemented with allogenic demineralized bone matrix in rat parietal bone.

The healing process of resected, autoclaved (121 degrees C, 20 minutes) and re-implanted bone in the rat parietal bone was compared with that of autoclaved bone that was supplemented with allogenic bone matrix (AAA-bone), using a scanning electron microscope and a light microscope. In the implant without AAA-bone, bone union and replacement of the autoclaved bone was seen at 2 weeks after implantation. There was no evidence of any inflammatory reaction around the autoclaved bone. The implant was gradually replaced by the new bone. In the implant with AAA-bone, the new bone formation around the implanted bone was more abundant than that of the implant without AAA-bone. An inflammatory reaction was also observed after 1 week. The replacement of the implant with AAA-bone was inferior to the nonsupplemented group. The reason for the poor replacement was the disturbance of the blood supply in the implant by abundant new bone formation. In these results, the autoclaved bone re-implantation was an excellent bone substitute with osteoconductive ability and biocompatibility. The implantation with AAA-bone was good for the new bone formation, but the position and the technique of supplement with AAA-bone have to be more deeply investigated.

Retinopathy of prematurity (1) birth weight, gestational age and maximum PaCO2.

The eyes of neonates who were premature or had been exposed to increased ambient oxygen from May 1979 to February 1985 were examined. Twenty out of 256 infants were found to have retinopathy of prematurity (ROP) of stage 3 or worse in at least one eye. Using uivariate statistics (chi-square analysis or the Wilcoxon test), the variables representing the maximum arterial carbon dioxide tension (PaCO2), birth weight and gestational age were found to be significantly associated with ROP. It is recommended to maintain PaCO2 values under 50 mmHg, especially in less mature newborns (less than 1,300g in birth weight or 210 days of gestational age).

Evaluation of the isolated perfused heart of mice, with special reference to vasoconstriction caused by intracoronary acetylcholine.

A perfusion model of isolated hearts (left in situ) was developed in mice. The heart was perfused retrogradely, according to the Langendorff technique, under a fixed flow rate of about 4 ml/min with physiological saline solution. Left ventricular (LV) cavity pressure, LV dP/dt max, heart rate, and arterial perfusion pressure were measured. Stable levels of mechanical parameters were reached within 30 min following the start of perfusion and were maintained for more than three hours. Single doses of acetylcholine (ACh, 0.003-1 microgram) administered into the coronary perfusion system elicited a dose-dependent increase (vasoconstriction) followed by a decrease (vasodilation) in perfusion pressure. Coronary vasoconstriction in response to ACh was especially prominent. LV systolic pressure, LVdP/dt max, and heart rate resulted in a decrease followed by an increase. A single injection of atropine (10 micrograms) antagonized both cardiac and vascular (vasoconstrictor and dilator) effects of ACh. The present studies substantiate the validity of this experimental model for the assessment of the action of drugs on the heart and coronary vasculature.

Origin of the marrow cells in bones induced by implantation of osteosarcoma-derived bone-inducing factor in mice.

Bone marrow cells (10(7)) of beige mice were injected into 800-rad-irradiated normal (C57BL/6 - +/+ or -bgJ/+) mice. After establishment of chimerism, bone formation including bone marrow was induced by implantation of the bone-inducing substances extracted from the murine osteosarcoma. The origin of marrow cells was investigated by observing giant granules of beige (C57BL/6-bgJ/bgJ, Chediak-Higashi syndrome) mice, as a marker for the origin of the cells. The marrow cells in the induced bones were of the beige type. These results indicate that the induced bone marrow cells are not the progeny of undifferentiated mesenchymal cells in situ, but that of hemopoietic stem cells circulating in the peripheral blood.

Different effects of dimethylbenz(alpha)anthracene and tetradecanoylphorbol acetate on differentiation of mast cells in the skin of mice.

The effects of the application of 7,12-dimethylbenz(alpha)anthracene (DMBA) and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on the differentiation of tissue mast cells were investigated. Giant granules of beige (C57BL/6-bgJ/bgJ, Chédiak-Higashi syndrome) mice were used to distinguish different populations of mast cells. In the normal C57BL/6 mice that had been irradiated and injected with bone marrow cells of beige C57BL/6 mice (bgJ/bgJ leads to +/+ chimeras), mast cells in the skin remained normal in type and mast cell precursors in the bone marrow and peripheral blood were of the beige type. When DMBA was painted onto the skin of such bgJ/bgJ leads to +/+ chimeras, the number of beige mast cells increased, but the number of normal mast cells did not change. On the other hand, the number of both normal and beige mast cells increased when TPA was applied to the bgJ/bgJ leads to +/+ chimeras. Autoradiographic study showed that beige mast cells proliferated more than normal mast cells after DMBA applications, whereas the proliferations was comparable in both types of mast cells after TPA applications. The present results indicate that the effects of DMBA applications on differentiation of mast cells are different from those of TPA applications.

Comparison of the isometric cervical extension strength and a cross-sectional area of neck extensor muscles in college wrestlers and judo athletes.

Increasing neck muscle strength can play an important role in preventing neck injuries in contact sports. The purpose of this study was to examine the actual conditions of the isometric cervical extension strength (ICES) and the cross-sectional area (CSA) of neck extensor muscles in male athletes participating in college wrestling and judo. The subjects comprised 18 wrestlers and 37 judo athletes from Nippon Sports Science University in Japan. The ICES was measured at eight angles (126 degrees, 108 degrees, 90 degrees, 72 degrees, 54 degrees, 36 degrees, 18 degrees, 0 degrees ). Transverse slices of 10 mm thickness were obtained at the position of each intervertebral disc between C2 and C3, C3 and C4, C4 and C5 and C5 and C6 using magnetic resonance imaging. The ICES of the wrestlers were significantly higher than those of the judo athletes. The ICES curve against the angle in wrestlers tended to differ from that of judo athletes. The CSA of neck extensor muscles in the wrestlers was significantly larger at all intervertebral levels examined than those of the judo athletes. A significant difference was observed in the CSA of the deepest area of neck extensor muscles between the groups although the difference was not significant in the superficial area. In this study, the ICES and the CSA in wrestlers were shown to be significantly higher and larger respectively than in the judo athletes, indicating a significant difference between these two sports.

Joint laxity, vitreoretinal degeneration, facial abnormalities, and generalized skeletal alterations: a new syndrome?

A Japanese girl with a hitherto unknown combination of malformations is reported. The cardinal features included hyperextensibility of the joints, vitreoretinal degeneration with cataracts, and facial abnormalities, comprising hypertelorism, prominent eyes, downslanting of the palpebral fissures, mid-face recession with a short nose, deformed auricles, and microretrognathia with a high arched palate. Skeletal survey revealed multiple wormian bones, hypoplastic facial bones and mandible, narrow thorax with wavy ribs, narrow ilia, and coxa valga with slight broadening of the proximal femora, findings of which were individually minor, but the assemblage of which assisted in the syndromic identification. Although skin biopsy did not contribute to the causal clarification, it was tempting to speculate that the syndromic constellation of the present disorder resulted from an underlying defect of connective tissues.

Plasma Aluminum levels of patients on long term sucralfate therapy.

Plasma aluminum levels of 17 patients with gastric and/or duodenal ulcers who had received four grams of sucralfate daily for eight to ten weeks were compared with those of six control subjects. Aluminum levels were determined by flameless atomic absorption spectrometry. Plasma of patients on long term sucralfate therapy contained 8.41 +/- 6.00 (SD) micrograms aluminum per liter and was not significantly different than that of the control subjects which contained 6.50 +/- 7.87 micrograms per liter. These values agreed well with normal values reported previously. In conclusion, long term ingestion of sucralfate in therapeutic doses does not result in an increase in plasma aluminum concentration.

Presence of pluripotent haemopoietic precursors in vitro (CFU-mix) in haemopoietic tissues from mice of W/Wv genotype.

The presence or absence of haemopoietic precursors, which produce mixed colonies in vitro (CFU-mix) was examined in the bone marrow and spleen of (WB X C57BL/6) F1-W/Wv mice. Despite the failure of macroscopically evident colony-formation in the spleens of irradiated mice, haemopoietic cells of W/Wv mice did produce macroscopically-evident mixed colonies containing erythroid cells, macrophages, and often megakaryocytes, in culture medium. The size and constitution of mixed colonies derived from W/Wv mice were comparable to those of mixed colonies from congenic +/+ mice. The present results appear consistent with in vivo haemopoiesis in the W/Wv mice, which is obviously deficient, but sufficient for survival.

Increase in histidine decarboxylase activity in skin of genetically mast-cell-deficient W/Wv mice after application of phorbol 12-myristate 13-acetate: evidence for the presence of histamine-producing cells without basophilic granules.

Histidine decarboxylase (HisDCase, EC 4.1.1.22) activity in mouse skin increased by a factor of more than 10 after a single application of phorbol 12-myristate 13-acetate. The cell type that was responsible for the increase in HisDCase activity was examined by using (WB X C57BL/6)F1-W/Wv mice, which are genetically deficient in tissue mast cells. In contrast to a report that increase of ornithine decarboxylase (EC 4.1.1.17) activity occurs in the epidermis [O'Brien, T. G., Simisiman, R. C. & Boutwell, R. K. (1975) Cancer Res. 35, 2426-2433], the HisDCase activity was found to increase in the dermis. Although most of the histamine in the dermis was present in mast cells, the increase in HisDCase activity in the skin in W/Wv mice was comparable with that in congeneic +/+ mice. This increase of HisDCase activity in the skin of W/Wv mice was abolished by prior x-ray irradiation (800 rads; 1 rad = 0.01 gray) but was restored by subsequent bone marrow transplantation. Because mice, in general, are known to lack basophilic leukocytes, the present results suggest the existence of histamine-producing cells without basophilic granules that are derived from the bone marrow.

Serum C1q levels as a prognostic guide to articular erosions in patients with rheumatoid arthritis.

C1q was measured serially by single radial immunodiffusion in 54 rheumatoid arthritis (RA) patients over a period of more than 5 years, and values were correlated with laboratory, radiographic, and clinical findings. The number of joints with erosion (NJE) was determined retrospectively from radiographs of patients who had RA of greater than 7 years duration. In patients with clinically "burned out" RA, C1q levels were not statistically different from those of healthy adults. During the period of active disease, each patient's C1q level remained very constant, irrespective of erythrocyte sedimentation rate, C-reactive protein (CRP) level, or whether the RA was active or in remission. No sustained correlation was found between the C1q level and the other 2 acute phase reactants, but patients with C1q levels of at least 250 micrograms/ml showed a positive CRP over a period of years, in contrast to those with C1q levels below 250 micrograms/ml. Patients with an initial C1q above 250 micrograms/ml had more erosive RA when compared with those having C1q levels below 250 micrograms/ml. These data suggest that active RA can be classified into two subsets by C1q levels, one with persistent inflammation and a high NJE and another without persistent inflammation and with a low NJE.

Fluorimetric and high-performance liquid chromatographic determination of D-lactate in biological samples.

D-Lactate in biological samples was converted into a strongly fluorescent substance in a one-vial reaction. It was first converted into the pyruvate hydrazone in the presence of D-lactate dehydrogenase, an NADH-reoxidation system using diaphorase, D,L-6,8-thioctamide and hydrazine. This hydrazone was then converted into 2-hydroxy-6,7-dimethoxy-3-methylquinoxaline by 1,2-diamino-4,5-dimethoxybenzene in 1 M hydrochloric acid, and the quinoxaline was extracted and measured fluorimetrically at 432 nm (excitation at 365 nm). The calibration curve for D-lactate was linear up to at least 100 nmol/ml of the assay mixture, with a determination limit of 2 nmol/ml. The quinoxaline was also analysed by high-performance liquid chromatography with fluorimetric detection. The calibration curve for D-lactate was linear from 500 fmol to 75 nmol in the reaction mixture. This method was 4000 times more sensitive than the fluorimetric method, and could determine D-lactate in blood plasma volumes of less than 1 microliter.