Web-mediated interspecific competition among spider mites.

Some spider mites, such as Tetranychus spp. and Amphitetranychus spp., create complicated webs (CWs), whereas others, such as Panonychus spp., produce little webs (LWs). We verified whether interspecific competition occurred between CW and LW mites via habitat arrangement under laboratory conditions. The complicated webs produced by CW mites clearly inhibited juvenile development in LW mites, whereas there was no effect of LW mites on CW mites. In oviposition site choice tests, both CW and LW females preferred the lower surface of leaves to the upper surface. The preference of LW mites for the lower leaf surface, even in the presence of CW mite webs, suggests that the costs of amensalism are outweighed by the possible benefits, such as avoiding rain. These findings show that the shift in mite species composition from LW to CW mites can occur as a consequence of the interspecific association between spider mites via their webs, without pesticide applications or the presence of natural enemies.

Bone-anabolic effect of salmon calcitonin on glucocorticoid-induced osteopenia in rats.

The anabolic effect of salmon calcitonin (SCT) on skeletal tissue was examined on glucocorticoid-induced osteopenia in female rats (12 weeks old). By the administration of methylprednisolone acetate (MPA: 0.1 mg/kg, s.c., 3 times/week) for 8 weeks, histomorphometrically detectable osteopenia with the characteristics of decreased bone formation and increased bone resorption developed in proximal tibia metaphysis. Serum osteocalcin level was also decreased by MPA treatment. Subsequent treatment with SCT (10 U/kg, s.c., 5 times/week) was found to reverse once developed osteopenia with the return of the osteocalcin level, though rats were still on MPA. Histomorphometry revealed that SCT decelerated bone resorption but augmented bone formation in this osteopenic model. After a single injection of SCT (2.5 U/kg--40 U/kg, s.c.), the serum level of parathyroid hormone (PTH) which had a potent anabolic on bone formation increased in a dose-dependent manner. These results indicate that SCT has a stimulating effect on osteoblastic bone formation and this anabolic effect may at least in part be due to its indirect effect to increase endogenous PTH secretion.

Bone anabolic effects of PTH(1-34) and salmon calcitonin in ovariectomy- and ovariectomy-steroid-induced osteopenic rats: a histomorphometric and biomechanical study.

Using an experimental model of type 1 osteoporosis under the chronic therapy with an anti-inflammatory steroid, the bone anabolic effect of PTH(1-34) was evaluated by histomorphometrical and biomechanical analysis. Wistar female rats (12-week-old) were ovariectomized and allowed to develop an osteopenic model in the presence or absence of methylprednisolone acetate (MPA: 0.1 mg/kg, s.c., 3-days-a-week basis from the 5th week after ovariectomy (OVX)). The osteopenia that developed for the first 12 weeks after OVX was almost completely normalized by subsequent PTH pulsing (20 microg/kg, s.c., 5-days-a-week) for 8 weeks starting at the 13th week; the following characteristics were observed: 1) proximal tibial metaphysis: recovered bone volume, rather increased trabecular thickness and osteoid volume, and normalized eroded surface; 2) 5th lumbar vertebra (L-5): partially recovered trabecular connectivity; 3) femur and 4th lumbar vertebra (L-4): recovered mechanical strength in maximum elastic load and maximum elastic energy. The anabolic effect of PTH(1-34) was not substantially modified by MPA. Salmon calcitonin (SCT: 10 U/kg per day, s.c., 5-days-a-week, for 8 weeks) was anabolic in limited parameters: decreased number of osteoclasts, recovered maximum elastic load in femur, and partially recovered maximum elastic load in L-4. The results suggest that PTH(1-34) pulsing is able to recover OVX-induced osteopenia in the structure and mechanical strength not only of the cancellous bone but also of the cortical bone, and the anabolic effect can be clinically expected even under steroid medication.

Identification of tranilast-binding protein as 36-kDa microfibril-associated glycoprotein by drug affinity chromatography, and its localization in human skin.

To elucidate the molecular mechanism involved in the suppression of keloids and hypertrophic scars by tranilast, we investigated the target protein of tranilast in bovine skin and aorta. A specific tranilast-binding protein was isolated from both tissues by drug affinity chromatography and was identified as 36-kDa microfibril-associated glycoprotein (36-kDa MAGP). Binding of 36-kDa MAGP to tranilast seemed to be specific since 36-kDa MAGP could be eluted from the drug affinity column by tranilast itself and also binding of 36-kDa MAGP to other anti-allergy drugs (amlexanox and cromolyn) is significantly weaker than that to tranilast. Light and electron microscopic immunohistochemistry detected the protein at the periphery of elastic fibers in normal human skin. In hypertrophic scar tissue, however, 36-kDa MAGP was located on small bundles of microfibrils. These findings provide support for the concept that elastogenesis occurs in scar tissue and 36-kDa MAGP might be one of the targets for tranilast.

Ultrastructural distribution of 36-kD microfibril-associated glycoprotein (MAGP-36) in human and bovine tissues.

We observed the ultrastructural distribution of MAGP-36 by immunoelectron microscopy in human and bovine tissues. MAGP-36 was present in microfibrils associated with tropoelastin in skin, aorta, and spleen. It was not detected in microfibrils from the ocular zonule and kidney mesangium that were not associated with tropoelastin. In skin, MAGP-36 was present in both early immature elastic fibers and mature elastic fibers. In mature elastic fibers, MAGP-36 was localized around amorphous elastic cores at the elastin-microfibril interface and in electron-dense bundles. Localization of MAGP-36 in elastic fibers coincided with the distribution of lysyl oxidase, an enzyme that plays a pivotal role in the deposition of tropoelastin. These findings suggest that MAGP-36 may be involved in elastogenesis.

Three distinct anti-allergic drugs, amlexanox, cromolyn and tranilast, bind to S100A12 and S100A13 of the S100 protein family.

To investigate the roles of calcium-binding proteins in degranulation, we used three anti-allergic drugs, amlexanox, cromolyn and tranilast, which inhibit IgE-mediated degranulation of mast cells, as molecular probes in affinity chromatography. All of these drugs, which have different structures but similar function, scarcely bound to calmodulin in bovine lung extract, but bound to the same kinds of calcium-binding proteins, such as the 10-kDa proteins isolated in this study, calcyphosine and annexins I-V. The 10-kDa proteins obtained on three drug-coupled resins and on phenyl-Sepharose were analysed by reversed-phase HPLC. It was found that two characteristic 10-kDa proteins, one polar and one less polar, were bound with all three drugs, although S100A2 (S100L), of the S100 family, was bound with phenyl-Sepharose. The cDNA and deduced amino acid sequence proved our major polar protein to be identical with the calcium-binding protein in bovine amniotic fluid (CAAF1, S100A12). The cDNA and deduced amino acid sequence of the less-polar protein shared 95% homology with human and mouse S100A13. In addition, it was demonstrated that the native S100A12 and recombinant S100A12 and S100A13 bind to immobilized amlexanox. On the basis of these findings, we speculate that the three anti-allergic drugs might inhibit degranulation by binding with S100A12 and S100A13.

[Effect of liver hydrolysate on ethanol- and acetaldehyde-induced deficiencies].

Since it has been reported that amino acids have alleviating effects on ethanol- and acetaldehyde-induced toxicity, we investigated the effect of liver hydrolysate derived from bovine liver on ethanol- or acetaldehyde-induced toxicity and deficiency models of mice and rats in the present study. Liver hydrolysate improved the deficiencies of beam walking and food intake of mice in a dose-dependent fashion when challenged with ethanol at the dose of 5 ml/kg, p.o. According to the analysis using selective inhibitors for alcohol dehydrogenase and acetaldehyde dehydrogenase, it has been suggested that this improvement effect of liver hydrolysate is mainly due to the reduction of acetaldehyde toxicity. No effect of liver hydrolysate was found in coma and death produced by orally treated ethanol at 10 ml/kg. In contrast, liver hydrolysate dose-dependently decreased the coma and death of mice administered acetaldehyde at 1.8 ml/kg, p.o. Furthermore, an increase in serum GPT activity, which was caused by twice oral administration of acetaldehyde at 1.2 ml/kg at interval of 1 hr, was inhibited by liver hydrolysate. These results suggest that liver hydrolysate has a protective effect against ethanol- and acetaldehyde-induced toxicity.

Two cases of Merkel cell carcinoma cured by intratumor injection of natural human tumor necrosis factor.

Two patients were treated with intratumor injection of natural human tumor necrosis factor for recurrent or primary Merkel cell carcinoma. In both patients, local chemotherapy achieved complete tumor regression without causing ulceration or scarring. These results suggest that intratumor injection of natural human tumor necrosis factor may be very effective for the treatment of Merkel cell carcinoma.

Treatment of nevus Ota: combined skin abrasion and carbon dioxide snow method.

Thirty-seven patients with nevus Ota were treated by skin abrasion-carbon dioxide snow therapy. Data obtained from 24 patients (including 5 infants) who completed treatment were analyzed to determine the number of treatment courses and to assess the outcome by color and histologic type. The 5 infants completing treatment received a mean of 10 courses of carbon dioxide snow therapy. Excluding these infants, the mean number of treatment courses was 3 for skin abrasion and 16 for carbon dioxide snow therapy. The therapeutic outcome was satisfactory, being graded as "excellent" in 6 patients, "excellent to good" in 6 patients, "good" in 10 Patients, and "fair" in 2 patients. None of the patients had a "poor" outcome. Darker lesions were generally less responsive to treatment. Lesions with melanocytes in the superficial dermis showed a better response with fewer courses of treatment. Those with melanocytes throughout the whole dermis generally had a poorer outcome. The results confirm that the benefits of this procedure are limited by the associated pain and the need for great skill and a long treatment duration.

The effect of Beraprost sodium on the survival of subcutaneous transferred jejunum after vascular pedicle interruption in a rat model.

Beraprost sodium, a stable PGI2 analog, having antiplatelet aggregation and vasodilating actions, was tested in a rat subcutaneous heterotopic jejunal model for its ability to improve survival after vascular pedicle interruption. Forth Sprague-Dawley rats were divided into four groups: Group 1 (control, ligation of pedicle on postoperative day 5); Group 2 (Beraprost sodium, ligation on day 5); Group 3 (control, ligation on day 7); and Group 4 (Beraprost sodium, ligation on day 7). The resulting viability rates were: Group 1 = 0 percent, Group 2 = 40 percent, Group 3 = 30 percent, Group 4 = 90 percent. These results indicate that the administration of Beraprost sodium facilitates the neovascularization of the transferred intestine and shortens the time required for viability of the transferred tissue, after interruption of the vascular pedicle.

Pectoralis major myocutaneous flaps in rats.

A new neurovascular, island, myocutaneous flap, including the pectoralis major, was created in a rat model. This model is useful for the observation of muscle degeneration and skin changes due to ischemia or denervation. Although this model requires a delay procedure, it allows the flap, that can be used as a free myocutaneous flap, to be raised reliably.

[Effect of efonidipine hydrochloride, a calcium channel blocker, on the experimental cerebral ischemia/anoxia].

The anti-ischemic and anti-anoxic effects of efonidipine, a dihydropyridine calcium antagonist, were studied in several models for cerebral ischemia and anoxia in mice and rats, and the effects were compared with those of nicardipine and flunarizine. Both efonidipine and flunarizine showed protective effects in the models of KCN-induced anoxia and complete ischemia induced by decapitation in mice 6 hr after the treatment, while nicardipine did not show such a long-lasting effect. Efonidipine (1 mg/kg, i.p.), but not nicardipine and flunarizine, prolonged the tolerance times in the asphyxic anoxia model. In mice, efonidipine (4 mg/kg, i.p.) significantly reduced the cumulative mortality rate after bilateral carotid artery ligation. The survival rates at 20 hr after bilateral carotid artery ligation were 33% in the group treated with efonidipine, significantly higher than that in the control group, 0%. On the other hand, the treatment with nicardipine or flunarizine did not increase the rates at 20 hr after the ligation. Moreover, efonidipine attenuated the disturbance of cerebral energy metabolism induced by decapitation in rats. These effects of efonidipine observed in this study were on the whole superior to those of the reference drugs, strongly suggesting the improving effect of efonidipine on cerebral ischemia and anoxia.

Effects of citalopram, a synthetic serotonin uptake inhibitor, on indoleamine and catecholamine concentrations in the cerebrospinal fluid of freely moving rats.

We studied changes in the concentrations of 5-hydroxytryptamine (5-HT), other indoleamines, and catecholamines in the cerebrospinal fluid (CSF) of freely-moving rats that had been administered citalopram, +/-1-[3- (Dimethylamino)propyl)-1-(4-fluorophenyl)-1, 3-dihydro-5-isobenzo-furancarbonitrile hydrobromide), a selective inhibitor of 5-HT uptake. In a microdialysis experiment, the intracerebral extracellular free 5-HT increased significantly, peaking 60 to 90 min after citalopram (30 mg/kg p.o.) was administered. The 5-HT concentrations in CSF from the cisterna magna increased significantly, reaching a maximum 6 hours after a single dose of citalopram (30 mg/kg p.o.) was given. Six hours after this dose, the CSF 5-HT concentration in the cisterna magna was significantly increased, and the 5-hydroxyindoleacetic acid (5-HIAA) concentration was significantly decreased. There were non-significant changes in the other indoleamines (tryptophan, 5-hydroxytryptophan, and kynurenine) and in the catecholamines (dopamine, homovanillic acid, normetanephrine, and 3-methoxy-4-hydroxyphenethyleneglycol). The 5-HT/tryptophan ratio was correlated significantly with the kynurenine/tryptophan ratio before treatment with citalopram (r = 0.81, p = 0.051), indicative that there is coordination of the serotonin and kynurenine pathways in normal rats. In the animals posttreatment there was no such correlation, suggesting that the changes in 5-HT are independent of the kynurenine system at least within the 6 hours postreatment. These CSF results appear to reflect selective inhibition of 5-HT uptake in brain tissues by citalopram that is not associated with changes in catecholamines.

Comparative study of auricular dimensions for the normal auricles of microtia patients, their parents, and normal individuals.

We measured the dimensions of the normal auricle in microtia patients and those of both auricles in their parents, and compared the findings with data obtained in normal individuals. The physiognomical auricular length and width dimensions of the normal auricles in microtia patients were significantly smaller than those in normal individuals. The auricular length of the parents of microtia patients was also significantly smaller than in normal individuals. The auricles of the parents on the same side as those affected by microtia in the patients tended to be smaller than those on the contralateral side.

[Effects of NC-1100, a calcium channel blocker, on experimental cerebral ischemia/anoxia in rodents].

The anti-ischemic and anoxic effects of NC-1100, a piperazine type calcium channel blocker, were investigated in various cerebral ischemia and anoxia models in mice, gerbils and guinea pigs. Minimal effective doses of NC-1100 were 8 mg/kg, i.p. and 30 mg/kg, p.o. for KCN-induced anoxia; 16 mg/kg, i.p. for decapitation-induced gasping; 30 mg/kg, i.p. for cerebral ischemia induced by occlusion of bilateral carotid arteries in gerbils; and 10 microM for the in vitro ischemic model in hippocampal slices. Moreover, NC-1100 attenuated the disturbance of cerebral energy metabolism induced by decapitation in mice. These results suggest that NC-1100 has a cerebral protective effect, and that attributable to its ability to improve the cerebral energy metabolism disturbance.

Experimental study of subcutaneously transferred intestine: effect of the presence of serous membrane on acceptance rate.

The serous membrane is considered suspect as one of the obstacles responsible for delaying the acceptance of free jejunal grafts. A model was created of subcutaneous transfer of jejunum from which the serous membrane had been experimentally removed and the authors compared acceptance rates for intestine with and without a serous membrane. Results showed acceptance rates of 0 percent, 37.5 percent, 87.5 percent, and 100 percent respectively, for intestine devoid of serous membrane in which the vascular pedicle had been ligated 4, 5, 7 and 14 days after transfer. When these findings were compared with the results of an experiment on intestine with an intact serous membrane, statistically significant differences were detected between the acceptance rates of the groups ligated 5 and 7 days after transfer. The acceptance time required for intestine devoid of serous membrane was shown to be shorter.

An experimental study of ischemia following subcutaneous heterotopic intestinal graft: influence of arterial or venous occlusion.

A subcutaneously transferred intestinal model was prepared in the rat and experimental observations were conducted of serial histologic changes in the intestine and of acceptance rates, when the artery or vein in the pedicle was ligated at various times after transfer. Results showed 50 percent take in the artery-ligated group and 12.5 percent in the vein-ligated group, when performed 5 days after transfer; 87.5 percent take in the artery-ligated group and 50 percent in the vein-ligated group, when performed 7 days after transfer; and 100 percent take for both groups, when ligation was performed 14 days after transfer. Consequently, it was concluded that venous occlusion constitutes a greater danger to successful transfer of intestine than arterial occlusion.

Interleukin 1-induced sequential myelorestoration: dynamic relation between granulopoiesis and progenitor cell recovery in myelosuppressed mice.

The myelorestorative effect of recombinant human interleukin 1 alpha (IL-1 alpha) was studied in mice treated with anticancer drugs. The treatment of mice with 5-fluorouracil (5-FU; 250 mg/kg body weight) or cyclophosphamide (CPA; 100 mg/kg) considerably decreased bone marrow or splenic colony-forming units in culture (CFU-C) or neutrophils in blood. The daily administration of IL-1 alpha (100 ng/mouse/day) after 5-FU treatment markedly accelerated the recovery of bone marrow CFU-C. This increase was followed by the recovery of splenic CFU-C and neutrophils in blood. In the CPA-treated mice the recovery of bone marrow CFU-C over the normal level was observed regardless of the administration of IL-1 alpha 3 days after CPA treatment. The daily administration of IL-1 alpha after CPA treatment markedly increased splenic CFU-C or neutrophils over the normal level following the increase of bone marrow CFU-C. Thus, in both 5-FU- and CPA-treated mice, the increase of bone marrow CFU-C after the administration of IL-1 alpha was observed several days earlier than the increase of splenic CFU-C and neutrophils in blood. The increase of splenic CFU-C and neutrophils in blood was observed concomitantly. The experiments, in which effective timing of IL-1 alpha injection was examined, indicated that the administration of IL-1 alpha within a few days after treatment with anticancer drugs was necessary for the accelerated recovery of bone marrow progenitor cells. On the other hand, the effective recovery of splenic progenitor cells and peripheral neutrophils required the administration of IL-1 alpha after the increase of bone marrow progenitor cells. Thus, the administration of IL-1 alpha daily or every other day was the most effective for recovery from myelosuppression induced by anticancer drugs.

Accelerated recovery of antigen-presenting cell activity by the administration of interleukin 1 alpha in 5-fluorouracil-treated mice.

In this study, we investigated the effect of human recombinant interleukin-1 alpha (IL-1 alpha) on antigen-presenting cell (APC) activity of spleen cells in mice treated with 5-fluorouracil (5-FU). APC activity was determined by the antigen-specific proliferation of T cell clone D10.G4.1 cells. When mice were injected with 5-FU, APC activity of spleen cells was suppressed. The administration of IL-1 alpha accelerated the recovery from this suppression. The most accelerated recovery was observed when these mice were administered with IL-1 alpha both before and after the 5-FU treatment. The recovery was also accelerated when the mice were injected with IL-1 alpha after the 5-FU treatment, but not when injected before the 5-FU treatment. The injection of 5-FU also decreased the cell numbers of whole spleen cells, B cells, and non-T non-B cells (Ig- and Thy-1- cells). The administration of IL-1 alpha accelerated the recovery of the decreased cell numbers. Both B cells and non-T non-B cells possessed APC activity, but most APC activity of unseparated spleen cells was carried by non-T non-B cells. B cells possessed only 1/20 of the APC activity of non-T non-B cells. The injection of 5-FU decreased APC activity of both B cells and non-T non-B cells, but the administration of IL-1 alpha accelerated its recovery. Thus, the accelerated recovery of APC activity by IL-1 alpha was suggested to be due to the recovery in the numbers of APC activity-bearing cell subpopulations and also due to the recovery of the APC activity of each subpopulation. Possible mechanisms for the recovery were discussed.

A specific 70K protein found in epileptic rat cortex: induction of bursting activity and negative resistance by its intracellular application in Euhadra neurons.

The effect of 70-kD protein (P70, a specific protein found in cobalt-induced epileptogenic focus of rat cerebral cortex) on membrane properties was examined in identified neurons of the snail, Euhadra peliomphala, using the pressure injection method combined with the voltage-clamp technique. In neurons that normally exhibited spontaneous regular firing, intracellular injection of P70 elicited bursting activity and a negative slope resistance (NSR) region in their current-voltage (I-V) curve in a manner corresponding to the duration of its injection. These responses were suppressed by prior injection of an antibody to P70 into the neurons, and were markedly inhibited by a reduction of extracellular Na+ ions and the anticonvulsant agent phenytoin, but not by Co(2+)-substituted Ca(2+)-free saline. In addition, intracellularly applied P70 potentiated both bursting activity and the NSR induced by a Na channel activator, veratridine. However, prior application of a saturating dose of this activator occluded the effect of P70. These results suggest that P70 elicits a Na(+)-dependent negative resistance, which may contribute to the generation of bursting activity.