Cyclical tensile force on periodontal ligament cells inhibits osteoclastogenesis through OPG induction.

The periodontal ligament (PDL) maintains homeostasis of periodontal tissue under mechanical tensile-loading caused by mastication. Occlusal load inhibits atrophic alveolar bone resorption. Previously, we discovered that continuous compressive force on PDL cells induced osteoclastogenesis-supporting activity, with up-regulation of RANKL. We hypothesized that, unlike compression, cyclical tensile force up-regulates OPG expression in PDL cells via TGF-beta up-regulation, and does not induce osteoclastogenesis-supporting activity. PDL cells were mechanically stimulated by cyclical tensile force in vitro. The conditioned media of PDL cells that had been subjected to cyclical tensile force inhibited osteoclastogenesis. Cyclical tensile force up-regulated not only RANKL mRNA expression, but also OPG mRNA expression in PDL cells. Tensile force up-regulated TGF-beta expression in PDL cells as well. Administration of neutralizing antibodies to TGF-beta inhibited OPG up-regulation under cyclical tensile-force stimulation in a dose-dependent manner. Additionally, the osteoclastogenesis-inhibitory effect of the conditioned media of PDL cells under cyclical tensile force was partially rescued by the administration of TGF-beta neutralizing antibodies. In conclusion, tensile force inhibited the osteoclastogenesis-supporting activity of PDL cells by inducing the up-regulation of OPG via TGF-beta stimulation.

An improved estimation method for amino alkyl ligand immobilized on matrix beads.

A direct spectrophotometric suspension method for the estimation of immobilized ligands on a gel matrix was developed. The method uses 0.05% agarose solution to prepare a practically stable suspension of various agarose beads. The quantity of agarose beads in the suspension was estimated turbidometrically from the attenuance at 650 nm. The determination of immobilized ligands on agarose beads was performed colorimetrically at the wavelength of the absorption maximum against a control suspension of a similar amount of underivatized agarose beads. This method is more sensitive and simpler than previously known methods. The method is applicable to the estimation of absorption spectra and the measurement of the time course of reactions on agarose beads.

Co-existence of PACAP and nitric oxide synthase in the rat hypothalamus.

The possible co-existence of pituitary adenylate cyclase-activating polypeptide (PACAP)-38 and nitric oxide synthase (NOS) in the rat hypothalamus was examined by a combination of PACAP-immunocytochemistry and NADPH-diaphorase histochemistry. Virtually all PACAP-38-immunoreactive neurones in the paraventricular and supraoptic nuclei exhibited NADPH-diaphorase activity. Since NADPH-diaphorase activity was identical to NOS-immunoreactivity in the magnocellular neurosecretory neurones, this finding indicates that the PACAP neurones synthesize NO.

Coexistence of oxytocin and NADPH-diaphorase in magnocellular neurons of the paraventricular and the supraoptic nuclei of the rat hypothalamus.

Nitric oxide (NO), which was firstly identified as an endothelium-derived relaxing factor, has recently been demonstrated to be a neurotransmitter in the central and peripheral nervous systems. In the hypothalamus, abundant nitric oxide synthase (NOS) immunoreactivity and its histochemical marker, NADPH-diaphorase activity, have been demonstrated in the hypothalamo-neurohypophyseal system. In the present study, we examined whether NOS is coexpressed with posterior pituitary hormones in the rat hypothalamus by combination of oxytocin and vasopressin immunofluorescence and NADPH-diaphorase histochemistry. Most oxytocin-immunoreactive neurons in the paraventricular and supraoptic nuclei expressed NADPH-diaphorase activity, but virtually no vasopressin-immunoreactive neurons contained NADPH-diaphorase activity. This suggests that oxytocin neurons are the main source of NO production in the hypothalamic-pituitary system.

Acute lupus peritonitis successfully treated with steroid pulse therapy.

A 21-year-old man with systemic lupus erythematosus (SLE) who developed acute lupus peritonitis is described. Acute lupus peritonitis appeared during generalized lupus flare, with nausea, vomiting, frequent diarrhea, and abdominal tenderness with rebound and guarding. The patient was afebrile and had decreased bowel sounds. Abdominal ultrasonography and computed tomography revealed marked thickening of the gastric, duodenal, and jejunal walls, massive intraluminal fluid collection, and increasing ascites. Gastrointestinal endoscopy showed edematous mucosa with multiple erosions of the stomach and duodenum. The ascitic fluid was remarkable for low complement levels and elevated anti-DNA antibody. These manifestations of acute lupus peritonitis resolved after steroid pulse therapy with methylprednisolone. We should consider acute lupus peritonitis in a patient with SLE when abdominal symptoms are severe. Experience with our patient indicates that steroid pulse therapy is effective for this rare but severe manifestation of SLE.

Selective accumulation and tumoricidal effect of cisplatin suspended in viscous ethyl oleate on hepatic cancers in animals after intraarterial infusion.

The selective accumulation and tumoricidal effects of cisplatin after intra-arterial infusion suspended in viscous ethyl oleate (VEO) on hepatic cancers of AH 272 tumor-bearing rats and VX-2 tumor-bearing rabbits were compared with those of cisplatin suspensions in ethyl oleate (EO) and Lipiodol Ultra Fluide (LP). The viscosities of VEO, EO and LP were 120, 4, and 21 centipoise (cp) respectively. Complete in vitro release of cisplatin from EO and LP occurred within 24 h, whereas only about 25% of cisplatin was released from VEO over the same period. When EO or VEO containing 3H-oleic acid were infused into the hepatic artery of rat liver inoculated with AH 272 tumor cells, radioactivity in the tumor site was higher than that in normal liver. In the case of cisplatin, concentration ratios after the infusion of EO and VEO were almost the same as those of oily carriers. Similar results were obtained in rabbit liver inoculated with VX-2 tumor cells. Cisplatin concentration in the tumor site seven days after intra-arterial infusion of VEO suspension was 5- and 1.7-fold higher, respectively, than that after EO and LP suspensions. The tumoricidal effect of cisplatin in VEO suspension on AH 272 tumor-bearing rats was higher than that after cisplatin solution and EO and LP suspensions, while VX-2 tumor growth was inhibited by the infusion of all cisplatin-containing oily carriers. VEO suspension thus appears very promising in intra-arterial infusion therapy.

Histopathologic findings of cornea verticillata in a woman heterozygous for Fabry's disease.

PURPOSE: To report the histopathologic findings of the cornea verticillata observed in a woman who was heterozygous for Fabry's disease. METHOD: A 67-year-old woman was found to have a whorl-like corneal opacity on her visit to the Department of Ophthalmology. Aichi Saiseikai Hospital. Her visit was because of a sudden loss of vision in her right eye owing to a central retinal artery occlusion in association with an ophthalmic artery occlusion. The patient died suddenly of an acute heart failure; with family consent, an autopsy was performed and the right eye was removed for histopathologic examination by light and electron microscopy. RESULTS: Low levels of alpha-galactosidase in the leukocytes together with the corneal finding led to the diagnosis of heterozygous Fabry's disease. Light microscopy revealed a 0.3- to 0.5-microm thick layer between the epithelial and Bowman's layers. Oil red O positive deposits were accumulated in the subepithelial layer, and the density varied in different regions. Electron microscopy showed that subepithelial layer differed in thickness, and the basal lamina reduplicated regionally. We were not able to determine the structure that correlated with the "ridge" in the central part of the cornea. CONCLUSION: The oil red O positive deposits and their variation in density in the subepithelial area of the cornea may have caused the characteristic whorl-like corneal opacity in this woman who was heterozygous for Fabry's disease.

Surface ultrastructure of collagen fibrils and their association with proteoglycans in human cornea and sclera by atomic force microscopy and energy-filtering transmission electron microscopy.

PURPOSE: We aimed to investigate the possible association of proteoglycans with D-periodic collagen fibrils in the human cornea and sclera, using energy-filtering transmission electron microscopy (EF-TEM) and atomic force microscopy (AFM). METHODS: Human cornea and sclera were digested with keratanase to eliminate keratan sulfate proteoglycans (KSPGs). For EF-TEM observation, surface proteoglycans were detected by cupromeronic blue (CB) staining. For AFM observation, cornea and sclera were treated with sodium hydroxide before and after keratanase digestion, and the surface topology of collagen fibrils was analyzed. RESULTS: With CB staining, numerous CB-positive short filaments of surface proteoglycans (proteoglycan filaments) were observed in the interfibrillar spaces of cornea and sclera associated with collagen fibrils. AFM imaging showed that the depth and periodicity of D-periodic collagen fibrils in keratanase-treated corneal collagens were deeper and more regular than in untreated ones. Moreover, the depth and periodicity of keratanase-untreated corneal collagens were shallow and irregular in comparison with keratanase-untreated scleral collagens. On the other hand, there was no difference in depth or regularity between keratanase-treated and -untreated scleral collagen fibrils. Using AFM imaging, additional thin grooves sub-bands were detected on the surface of keratanase-treated corneal collagen fibrils. The grooves were not detected in keratanase-untreated collagen fibrils nor in scleral collagen fibrils with or without keratanase digestion. Comparing densitometry waves, the grooves of D-periodic corneal collagen sub-bands corresponded to a and c bands. CONCLUSION: Using AFM and EF-TEM to study corneal and scleral collagen fibrils and their association with proteoglycans, we conclude that KSPG is found in ample amounts in the human cornea in comparison with sclera. Moreover, we topologically detected KSPG attached to a and c bands of collagen fibrils.

Rapidly progressive pneumonia due to Aeromonas hydrophila shortly after near-drowning.

An 87-year-old woman died of rapidly progressive pneumonia due to Aeromonas hydrophila shortly after a near-drowning event. Autopsy showed necrotizing pneumonia and postmortem cultures of both blood and lung revealed the organism. Fulminant pneumonia should be considered in patients of a near-drowning event.

Thalassemia incidentally found by marked erythrocytosis due to an ochre mutation at codon 35 in a Japanese man.

We present a patient with beta-thalassemia minor diagnosed on the basis of the incidental findings of marked hypochromic and microcytic erythrocytosis. Hemoglobin analysis revealed increased hemoglobin (Hb) A2 levels and decreased beta/alpha synthesis ratio in both the propositus and his mother. Further molecular studies identified a single base substitution of TCA to TAA within codon 35 in heterozygous state, which creates a premature terminator resulting in a defect of effective beta globin synthesis. This is the first report of beta-thalassemia due to a nonsense mutation at codon 35 of beta-thalassemia gene in the Japanese population.

Surface topology of collagen fibrils associated with proteoglycans in mouse cornea and sclera.

PURPOSE: To clarify the histological basis for the optical difference in the cornea and sclera, we investigated the surface ultrastructure of D-periodic collagen fibrils. METHODS: The fibril arrangement and topology of D-periodic collagen fibrils from corneas and scleras of mice were examined by transmission and scanning electron microscopy, as well as by atomic force microscopy (AFM). The banding patterns were estimated by densitometry and compared with the profile made by computer simulation from the protein sequence of amino acids. RESULTS: Considerable association of proteoglycans/glycosaminoglycanson XI bands of corneal D-periodic collagen fibrils was seen with ruthenium red staining. Atomic force microscopy imaging showed that the depth of the groove in the D-periodicity of corneal collagen fibrils was shallow compared with that of the sclera. CONCLUSIONS: Corneal collagen fibrils are associated with many extracellular matrix components on both elevated and depressed surfaces of D-periodic bands, which may serve to maintain interfibrillar spaces, resulting in corneal transparency.

The effect of calcium channel antagonist administered by iontophoresis on the pain threshold.

Three calcium (Ca) channel antagonists were administered to healthy adult volunteers by iontophoresis alone or in combination with lidocaine to study their effects on the pain threshold. Nicardipine, verapamil or diltiazem administered alone elevated the pain threshold to the same degree as lidocaine. When Ca channel antagonists were administered in combination with lidocaine, the pain threshold elevation was not changed, but the duration of the analgesic action was prolonged when compared with lidocaine alone. These results suggest a role of the calcium channel in the pain control mechanism and the possible clinical application of Ca channel antagonists delivered by iontophoresis in pain management.

[A case of acute myelogenous leukemia accompanied with myelofibrosis and megakaryocyte-like giant bizarre blasts].

A 45-year-old man was admitted with high fever and leukocytosis in August 1993. The diagnosis of acute myelogenous leukemia (AML; M2) was made on the basis of morphological, cytochemical and immunological characteristics of the blasts in the bone marrow. The induction therapy with BHAC, daunorubicin, 6-MP was unsuccessful in achieving remission; the bone marrow biopsy specimen revealed the proliferation of the remaining leukemic cells and massive fibrosis accompanied with unusual megakaryocyte-like giant bizarre cells. These megakaryocyte-like giant cells were positive for myeloperoxidase and CD34, but not GPIIIa and factor VIII, indicating that those were derived from myelogenous stem cells. Following the low-dose Ara-C therapy, improvement of fibrosis and disappearance of these giant cells were observed in the bone marrow. After the reinduction therapy with high-dose Ara-C and MIT against markedly increased blasts, the patient died of systemic fungal infection. The presence of myelofibrosis and giant atypical blasts might allow resistance to therapy and poor prognosis.

[Cerebral aspergillosis as a cerebral vascular accident].

Cerebral aspergillosis is one of the most common mycotic infections in the central nervous system causing different clinical features such as brain abscess, granuloma, meningitis, and encephalitis. Cerebral aspergillosis, however, may lead to a cerebral vascular accident such as intracranial hemorrhage or cerebral infarction. In this report, we present two patients with cerebral aspergillosis accompanied by intracranial hemorrhage. A total of 124 reported cases of cerebral aspergillosis are reviewed to ascertain the pathogenesis of the associated vascular lesion. The first patient was a 9-year-old girl, who developed drowsiness with a headache during the medical treatment for acute myelocytic leukemia. CT disclosed subarachnoid and intraventricular hemorrhage. The autopsy revealed that the aspergillus arteritis was the cause of repeated hemorrhage. The second patient was a 15-year-old boy with allergic purpura and renal failure, who suddenly developed a stupor with convulsive seizure. CT disclosed an intracerebral hemorrhage in the right parieto-occipital area. The patient gradually deteriorated and died in spite of the surgical removal of the hematoma. The autopsy revealed that the hemorrhage was caused by the aspergillus arteritis. Cerebral aspergillosis has two routes of infection to the central nervous system: hematogenous dissemination from the distant site (usually the lung) and direct extension from the contiguous site (usually the paranasal sinuses or orbit). The primary mechanism of neuropathology is different between these two types. Primary cerebral arteritis is most often seen in patients with the former type, whereas primary basal meningitis occurs in the latter. The incidence of clinico-pathological features is different between hematogenous dissemination type and direct extension type.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effective pentostatin-based treatment of adult T cell leukemia in a patient with severe arthritis].

We report a 48-year-old woman with adult T-cell leukemia who had refractory arthralgia, intense headaches, and fever. Leukemic cell infiltration of the cerebrospinal fluid was detected but no other acute signs were observed. Abnormal lymphocytes with lobulated nuclei were found in the synovial fluid, and a histologic examination revealed proliferation into the synovium. Because combination chemotherapy did not elicit a favorable response, the patient was treated with a pentostatin bolus injection. The articular symptoms disappeared and complete remission was obtained. Six months later, she experienced arthralgia again together with a gradual increase of abnormal lymphocytes in peripheral blood. Sixteen months later, the patient was given pentostatin and achieved a complete remission again. She is still free from relapse without further therapy after 36 months, and her articular symptoms have not returned either. There were no adverse effects due to pentostatin. The patient's serum IL-6 level was elevated, suggesting that IL-6 may play a role in arthropathy.

[Reversal by the calcium ion of the pain threshold elevated by calcium channel blockers].

We investigated calcium ion effects on pain thresholds by iontophoresis in healthy adult volunteers. The pain thresholds were measured by the pain thermometer to evaluate the effect of iontophoresis. At first, one of calcium channel blockers (nicardipine, verapamil or diltiazem) was administered by iontophoresis with 1.0 mA for 10 minutes. Then, next to calcium channel blockers, calcium chloride or saline was locally administered after the elevation of pain threshold had been recognized and the pain threshold had been measured again. Calcium ion decreased the elevation of pain threshold significantly, but saline had no effect. These results suggest that not only sodium channel but also calcium channel influences pain mechanism during iontophoresis.

[Anesthetic consideration of a patient with congenital insensitivity to pain with anhidrosis].

Congenital insensitivity to pain with anhidrosis is a rare disorder but several clinical anesthetic experiences have been reported in Japan. We anesthetized a patient with this disorder for orthopedic operation with a low concentration of sevoflurane under monitoring of body temperature and we measured the concentrations of plasma ADH, cortisol and catecholamines levels before, during and after the operation. There were no problems during anesthesia and the patient awoke rapidly. ADH, cortisol and epinephrine concentrations showed slight elevations during and after the operation. The management of intraoperative body temperature and the necessary level of anesthesia were also discussed.

[The analgesic effect of iontophoresis with magnesium sulfate].

The analgesic effect of iontophoresis with magnesium sulfate was studied in fourteen healthy adult volunteers. Magnesium sulfate 0.5 mg.5 ml-1 was locally administered by the iontophoresis with 1.0 mA electric current for 10 min. The pain recognition time was measured by the pain thermometer before and after 5, 15, 30, 60 and 120 min of iontophoresis. The pain recognition time was significantly extended and lasted for 60 min. These results support the view that the iontophoresis with magnesium sulfate has analgesic effect and produces a good pain relief clinically.

[The effect of iontophoresis with several Ca channel blockers for PHN patients].

We performed iontophoresis with Ca channel blockers for healthy adult volunteers. In this clinical study, we used iontophoresis with Ca channel blockers. Ten out patients with PHN treated at our pain clinic were treated with iontophoresis. They were randomly assigned to one of the following four treatments: (1) 5 ml of 4% lidocaine HCl, (2) 2 mg of nicardipine HCl + 5 ml of distilled water, (3) 2 mg of verapamil HCl + 5 ml of distilled water, and (4) 2 mg of diltiazem HCl + 5 ml distilled water. Iontophoresis was performed using the above four drugs on the positive pole. Using a VAS, each patient was evaluated concerning the analgesic effect. The pain before treatment (10 points) was used as the base line. Compared with the scores before treatment, VAS scores decreased significantly after iontophoresis in all four groups. In the lidocaine group, a significant decrease in VAS scores occurred immediately after iontophoresis and lasted up to 24 hours, reaching the nadir at 2 hours. In the nicardipine group, the decrease occurred immediately after iontophoresis and lasted up to one day, reaching the nadir at four hours. In the verapamil group, the decrease started 1 hour after iontophoresis and lasted up to 48 hours, reaching the nadir at 8 hours. In diltiazem group, the decrease started 1 hour after iontophoresis and lasted up to 48 hours, reaching the nadir at 4 hours. Iontophoresis with Ca channel blockers produced a prolonged analgesic effect in PHN patients. Previously we had observed the same effect in adult volunteers. Therefore, we believe that this therapy will be clinically useful.

[Influence of calcium channel blockers and beta-blockers on pain relief with iontophoresis].

We studied usefulness of iontophoresis on pain relief using several Ca channel blockers, propranolol and guanethidine. Subjects were 18 healthy adult volunteers. We used 4% lidocaine with/without several drugs (2 mg of nicardipine, verapamil, diltiazem, propranolol and 10mg of guanethidine), and evaluated the pain relief effect with Nakahama's algesimeter. In all groups except for propranolol group, the pain recognition time was elongated significantly in comparison with control. In making comparison between each Ca channel blocker, we did not observe any significant differences, but, in propranolol group, elongation of pain threshold time was observed in some subjects. During these processes, systemic blood pressure and heart rate showed no remarkable changes. Our results suggest that it is possible to achieve more prolonged analgesic effect by the Ca channel blockers.