Multidisciplinary Approach to Complicated Crown-root Fracture Treatment: A Case Report.

Treatment of complicated crown-root fractures is one of the most challenging within the various types of dental trauma and requires a multidisciplinary approach. This paper reports the complicated crown-root fracture of a maxillary right central incisor, in which there was esthetic, functional, and biologic (endodontic and biologic width invasion) involvement. A 15-year-old male patient presented to the dental clinic one month after suffering trauma with a complicated crown-root fracture on tooth 8. The patient had previously undergone endodontic treatment and was sent to have periodontal surgery to reestablish the biological width on the palatal surface. Following the surgery, a fiberglass post was cemented, and the fragment was reattached. This approach allows the exposure of the cervical margin, adequate isolation, and subsequent fragment reattachment in the same clinical appointment. Fragment reattachment is a viable approach as it is a simple and conservative procedure that restores the natural esthetic of the tooth and has superior resistance compared to a composite restoration. The patient's cooperation in understanding the limitations of the treatment and maintaining adequate oral hygiene are very important to achieving a good prognosis of the case. After a 2-year clinical and radiographic follow-up period, the clinical protocol was found to be successful, and the tooth remained functional, esthetically favorable and asymptomatic.

Inhibition of intracellular hepatitis C virus replication by nelfinavir and synergistic effect with interferon-alpha.

Liver diseases associated with hepatitis C virus (HCV) infection have become the major cause of mortality in patients with human immunodeficiency virus (HIV) infection since the introduction of highly active anti-retroviral therapy. HCV-related liver disease is more severe in HIV-infected patients than in non-HIV-infected patients, but the standard therapies used to treat chronic hepatitis C in HCV/HIV coinfected patients are the same as those for patients infected with HCV alone. HIV protease inhibitors might have potential to down-regulate HCV load of HCV/HIV coinfected patients. In this study, we evaluated the effects of nelfinavir on intracellular HCV replication using the HCV replicon system. We constructed an HCV replicon expressing a neomycin-selectable chimeric firefly luciferase reporter protein. Cytotoxicity and apoptosis induced by nelfinavir were assessed and synergism between nelfinavir and interferon (IFN) was calculated using CalcuSyn analysis. Nelfinavir dose-dependently repressed HCV replication at low concentrations (IC(50), 9.88 micromol/L). Nelfinavir failed to induce cytotoxicity or apoptosis at concentrations that inhibited HCV replication. Clinical concentrations of nelfinavir (5 micromol/L) combined with IFN showed synergistic inhibition of HCV replication in our replicon model. Our results suggest that the direct effects of nelfinavir on the HCV subgenome and its synergism with IFN could improve clinical responses to IFN therapy in HCV/HIV coinfected patients.

HRCT shows variations in appearance in disseminated tuberculosis in adults.

OBJECTIVE: To examine patterns of high resolution computed tomography (HRCT) of lungs of adults with disseminated tuberculosis (TB). DESIGN: Disseminated TB was defined as radiological involvement of all lung lobes. RESULTS: The case series illustrated wide variation in HRCT of disseminated TB. Patterns identified on HRCT included (1) miliary TB (haematogenous dissemination), (2) miliary TB with exudative reaction, (3) bronchogenic spread, (4) miliary TB mixed with bronchogenic spread, and (5) bronchogenic spread with multiple cavity formation. CONCLUSION: The HRCT patterns described allow classification of disseminated TB according to the mechanism of spread (haematogenous and/or bronchogenic) and the degree of local lung involvement (reaction or cavitation).

Activated stellate (Ito) cells possess voltage-activated calcium current.

We previously reported stellate (Ito) cells possess voltage-activated Ca2+ current. The activation of stellate cells has been indicated to contribute to liver fibrosis and the regulation of hepatic hemodynamics. The aim of this study was to investigate the relationship between voltage-activated Ca2+ current and activation of stellate cells. Voltage-activated Ca2+ current in stellate cells isolated from rats were studied using whole-cell patch clamp technique. L-type voltage-activated Ca2+ current was hardly detected in stellate cells cultured for less than 9 days. Ca2+ current was detected 12.5 and 69% of cells at the 10th and 14th day of culture, respectively. BrdU incorporation indicated cell proliferation was recognized over 50% of cells at the 3rd and 5th day of culture, respectively, then decreased significantly in a time-dependent manner. On the other hand, the expression of alpha-smooth muscle actin indicated cell activation increased from 7th day of culture and collagen type I mRNA appeared remarkably in cells cultured for more than 10 days. In this study, we concluded L-type voltage-activated Ca2+ current was recognized in activated stellate (myofibroblast-like) cells.

Stimulation of protein kinase C inhibits bursting in disease-linked mutant human cardiac sodium channels.

BACKGROUND: Mutations in SCN5A, the gene coding for the human cardiac Na+ channel alpha-subunit, are associated with variant 3 of the long-QT syndrome (LQT-3). Several LQT-3 mutations promote a mode of Na+ channel gating in which a fraction of channels fail to inactivate, contributing sustained Na+ channel current (Isus), which can delay repolarization and prolong the QT interval. Here, we investigate the possibility that stimulation of protein kinase C (PKC) may modulate Isus, which is prominent in disease-related Na+ channel mutations. METHODS AND RESULTS: We measured the effects of PKC stimulation on Na+ currents in human embryonic kidney (HEK 293) cells expressing 3 previously reported disease-associated Na+ channel mutations (Y1795C, Y1795H, and DeltaKPQ). We find that the PKC activator 1-oleoyl-2-acetyl-sn-glycerol (OAG) significantly reduced Isus in the mutant but not wild-type channels. The effect of OAG on Isus was reduced by the PKC inhibitor staurosporine (2.5 micromol/L), ablated by the mutation S1503A, and mimicked by the mutation S1503D. Isus recorded in myocytes isolated from mice expressing DeltaKPQ channels was similarly inhibited by OAG exposure or stimulation of alpha1-adrenergic receptors by phenylephrine. The actions of phenylephrine on Isus were blocked by the PKC inhibitor chelerythrine. CONCLUSIONS: We conclude that stimulation of PKC inhibits channel bursting in disease-linked mutations via phosphorylation-induced alteration of the charge at residue 1503 of the Na+ channel alpha-subunit. Sympathetic nerve activity may contribute directly to suppression of mutant channel bursting via alpha-adrenergic receptor-mediated stimulation of PKC.

Mutation in the caveolin-3 gene causes a peculiar form of distal myopathy.

The authors describe a patient with sporadic distal myopathy associated with reduced caveolin-3 in muscle fibers in which the muscle atrophy was restricted to the small muscles of the hands and feet. Gene analysis disclosed a heterozygous 80 G-->A substitution in the caveolin-3 gene that was identical to that of reported cases of elevated serum creatine kinase. This patient further demonstrated possible clinical heterogeneity of myopathies with mutations in the caveolin-3 gene.

Effect of 4-phenyl-1,2,3,4-tetrahydroisoquinoline on ambulation induced by injection of methamphetamine into the nucleus accumbens in rats.

4-Phenyl-1,2,3,4-tetrahydroisoquinoline (4-PTIQ) has previously been shown to have antagonistic properties to methamphetamine in the spinal cord. Administration of 4-PTIQ (5 mg/kg, s.c.) reduced the ambulation induced by methamphetamine (0.5 mg/kg, s.c.) in rats. Methamphetamine (3 micrograms), injected unilaterally into the nucleus accumbens, increased ambulation. Alone, 4-PTIQ (10 micrograms) failed to elicit ambulation; however, it inhibited the methamphetamine-induced increase in ambulation. The alpha 1-antagonist prazosin (0.5 micrograms) or the beta-antagonist propranolol (3 micrograms) showed no effect on ambulation induced by methamphetamine. Haloperidol (5 ng), which possesses strong dopamine-blocking activity, abolished the ambulation induced by methamphetamine. The drug 4-PTIQ had weak affinity for dopamine D1 and D2 receptors. These results support the possibility that the inhibitory effects of 4-PTIQ on the ambulation-stimulating effects of methamphetamine, are due to blocking of the dopamine-releasing effect of methamphetamine but not due to dopamine blocking effects.

Inhibitory effect of 4-phenyltetrahydroisoquinoline on locomotion and dopamine release induced by micro-injection of methamphetamine into the nucleus accumbens of the rat.

The inhibitory effects of 4-phenyl-1,2,3,4-tetrahydroisoquinoline (4-PTIQ) on methamphetamine-induced increases in dopamine and locomotion were investigated. Methamphetamine hydrochloride (10 micrograms) microinjected into the nucleus accumbens increased both locomotor activity and extracellular dopamine levels, measured by the brain microdialysis method. 4-PTIQ hydrochloride (20 micrograms) co-injected with methamphetamine inhibited both the increase in dopamine and the locomotor activity. The uptake blocker cocaine hydrochloride (20 micrograms) co-injected with methamphetamine failed to inhibit the effect of methamphetamine. Thus 4-PTIQ but not cocaine inhibited the dopamine-releasing effect of methamphetamine, and 4-PTIQ is suggested to block methamphetamine-induced locomotion by inhibition of dopamine release.

Fluctuations in plasma levels of thrombomodulin in patients with DIC.

Plasma thrombomodulin (TM) has attracted considerable attention as a marker of endothelial cell membrane injury. We examined fluctuations in plasma TM levels in patients receiving therapy for the disseminated intravascular coagulation syndrome (DIC) using an enzyme immunoassay. Sixty healthy controls and 18 patients with DIC were studied. The mean +/- SD of the TM values initially measured immediately after the onset of DIC was 42.00 +/- 20.85 ng/ml, which was markedly increased as compared with the control value of 15.36 +/- 4.85 ng/ml (p < 0.001). Fluctuations in the TM levels over time were studied after dividing the patients according to the presence or absence of improvement in the underlying disease and improvement or lack thereof in the coagulation findings. Group I showed improvement in both categories, Group II showed improvement only in the latter, and Group III showed no improvement in either category. In Group I, the mean +/- SD of initial measured TM levels was 37.02 +/- 10.12 ng/ml and the mean of final values decreased to 58.9% of the initial value. This decrease was significant by paired Student's t-test (p < 0.01). The initial value in Group II was 45.86 +/- 18.86 ng/ml and the final values increased to 117.0% of the initial values, this difference was not significant. The initial value in Group III was 44.48 +/- 21.53 ng/ml and the final values increased to 143.4% of the former. This increase was significant by paired Student's t-test (p < 0.05). The difference in % fluctuations between Group I and Group III was significant by Wilcoxon's test (p < 0.01). These results suggest that the measurement of plasma TM can be useful in the management of DIC.

4-Phenyltetrahydroisoquinoline, but not nomifensine or cocaine, inhibits methamphetamine-induced dopamine release.

The inhibitory effect of 4-phenyltetrahydroisoquinoline (4-PTIQ) on methamphetamine-induced dopamine release in the rat nucleus accumbens was investigated using a brain microdialysis method. Methamphetamine (10(-6) M) infusion through a microdialysis probe induced the release of dopamine. Although the uptake inhibitors, cocaine (3 x 10(-6) M) and nomifensine (10(-6) M), failed to block dopamine release, 4-PTIQ (10(-6 M) inhibited the dopamine-releasing effect of methamphetamine. 4-PTIQ did not affect the elevation of the extracellular dopamine level induced by high concentrations of nomifensine (10(-5) M) and cocaine (3 x 10(-5) M). 4-PTIQ was the weakest inhibitor of [3H]dopamine uptake by rat striatal synaptosomes. These results suggest that 4-PTIQ is a selective antagonist against the dopamine-releasing effect of methamphetamine in the nucleus accumbens.

Methamphetamine antagonistic property of (+)- and (-)-4-phenyltetrahydroisoquinoline in rat anococcygeus muscle.

The antagonistic effects of (+)- and (-)-4-phenyl-1,2,3,4-tetrahydroisoquinoline (4PTIQ) on methamphetamine in the rat anococcygeus muscle were compared with those of cocaine and nomifensine. Methamphetamine contracted the anococcygeus muscle through the release of norepinephrine from noradrenergic nerve terminals. (+)-4PTIQ inhibited the methamphetamine-induced contractions more strongly than cocaine and nomifensine. (+)-4PTIQ had no potentiating effects on exogenous norepinephrine-induced contraction, which was considered to be an index of amine neuronal uptake blockade. On the other hand, (-)-4PTIQ, cocaine and nomifensine produced a significant leftward shift of the norepinephrine concentration-response curve, i.e. they showed a strong blocking effect on amine neuronal uptake. These results suggest that the inhibitory effects of (+)-4PTIQ on the action of methamphetamine are mediated by a mechanism other than inhibition of amine neuronal uptake.

Monitoring of haloperidol serum levels and it's clinical significance.

Haloperidol serum levels were determined by the recently developed radioimmunoassay technique to elucidate the relationship between serum levels and clinical effects and that between serum levels and adverse effects. The serum levels of haloperidol in divided daily doses schedule reached to the steady state more rapidly (within 7 days) than in single dose schedule. The single daily dose schedule showed significantly wider diurnal variation than the divided daily doses schedule. No remarkable differences in serum concentration was noted between different types of oral haloperidol preparation (liquid, granule, tablet). No definite relationship was established between the serum concentration of haloperidol and incidence of tardive dyskinesia. No definite positive relationship between clinical effects and serum concentration of haloperidol could obtained.

Expression of tumor necrosis factor-alpha in muscles of polymyositis.

We immunohistochemically examined biopsied muscles from nine untreated patients with polymyositis (PM) and five patients with other neuromuscular diseases (ONMD), using monoclonal antibodies to tumor necrosis factor-alpha (TNF-alpha) and lymphoid surface markers. In muscles of three patients with PM, we observed many TNF-alpha positive macrophages and lymphocytes in endomysium and around vessels in the muscles. By contrast, there were few, weakly TNF-alpha stained cells in muscles of three patients with ONMD. The ratio of TNF-alpha-positive cells to the muscle fibers and the ratio of TNF-alpha-positive cells to the mononuclear cells were significantly higher in PM compared with ONMD. In addition, we observed atrophic muscle fibers more frequently in TNF-alpha-positive muscles than TNF-alpha-negative ones. We conclude that, at least, in a part of PM patients, TNF-alpha produced locally may contribute to the pathogenesis of PM.

Cortical laminar necrosis and central pontine myelinolysis in a patient with Sheehan syndrome and severe hyponatremia.

Hypoxic encephalopathy and osmotic demyelination are independent clinical entities. We describe a rare case with these two complications as demonstrated by magnetic resonance imaging (MRI). A 58-year-old woman had adrenal crises twice a decade due to Sheehan syndrome. At the second crisis, hyponatremia was remarkable with consciousness disturbance which was rapidly corrected by intravenous administration of glucocorticoid and hypertonic saline. The maneuver improved consciousness disturbance, but resulted in hypokalemic ventricular fibrillation with circulatory failure. After the normalization of the circulation, however, her consciousness level deteriorated again. Repeated brain MRI revealed acute and chronic phases of cortical laminar necrosis and central pontine myelinolysis.

Concurrent infection with Legionella pneumophila and Pneumocystis carinii in a patient with adult T cell leukemia.

A 48-year-old woman was admitted to our hospital with high fever, chills, cough, and exertional dyspnea. On admission, the chest roentgenogram and computed tomography scan showed bilateral alveolar infiltration in the middle and lower lung fields. Microscopic examination of the bronchial lavage fluid showed flower cells typical for adult T-cell leukemia (ATL) and cysts of Pneumocystis carinii, and Legionella pneumophila serogroup 1 grew on buffered charcoal yeast extract (BCYE)-alpha agar. The patient was successfully treated with antibiotics including trimethoprim/sulfamethoxazole, erythromycin, and sparfloxacin. Remission of ATL was achieved after three courses of antileukemic chemotherapy. Mixed infection of opportunistic pathogens should be considered in patients with ATL.

Determination of deoxymononucleotides and deoxynucleosides as a tool for toxicological examination--a trial for conversion of absorbance unit into molar amount in deoxydinucleotide.

Conversion of absorbance unit into molar amount was tried as to deoxydinucleotide. In order to achieve this trial, a reversed-phase high-performance liquid chromatographic method with a linear gradient mode was established for separation and quantification of deoxymononucleotides and deoxynucleosides. Selecting 2'-deoxycytidylyl-(3'-5')-thymidylic (5') acid as an example of deoxydinucleotides, this compound dissolved in water was estimated in absorbance unit at 260 nm, and it was treated by alkaline phosphatase and furthermore by snake venom phosphodiesterase. Based on quantification of the cleaved nucleic acid constituents including 2'-deoxycytidine 5'-monophosphoric acid and thymidine by the above method, the molar amount per 1.000 absorbance unit at 260 nm in the deoxydinucleotide was determined. Thus, the conversion of absorbance unit into molar amount is considered to be applicable to other deoxyoligonucleotides.

Urinary albumin determination by gel-filtration high-performance liquid chromatography.

To determine urinary albumin in a minute amount, a gel-filtration high-performance liquid chromatographic procedure was newly established. When urine of a normal rat was eluted isocratically at 0.6 ml/min by 100 mM Na sulfate in 20 mM Na, K-phosphate (pH 7.4), approximately 6 hrs for complete elution of urinary peak-forming substances was needed. Retention time of albumin was found to be 22.9 min. To shorten the analytic time, 100 mM Na sulfate in 20 mM Na, K-phosphate (pH 7.4) was first used during a 30 min period for separation of albumin. A mixture of acetonitrile/the above solvent = 3/7 (v/v) was then flushed to wash away the peak-forming substances. By this elution mode, the analytic time could be reduced to 3 hrs. When the validity of this procedure was tested, the detection limit of albumin was 0.04 microgram/injection, and a linearity was observed between 0.2 and 50 micrograms/injection. Rats then received single subcutaneous injections of puromycin aminonucleoside, which is a nephrotoxic agent. The plasma albumin concentrations fell at 5, 10 and 15 days after the administration, and the urinary excretions of albumin rose from the 1st day up to the 15th day. The results denoted that our procedure could be a good evaluative tool for nephrotoxicity studies where albuminuria was manifested.

Misleading serological identification of Legionella anisa as Legionella bozemanii.

Identification of six Legionella species, which we previously identified by serological test as Legionella bozemanii (L. bozemanii), was performed by DNA-DNA hybridization using a commercial DNA-DNA hybridization kit (Kobayashi Pharm. Co., Japan) introduced by Ezaki et al. All strains were identified as Legionella anisa (L. anisa), this being the first identification of L. anisa in Japan. Conventional laboratory tests were performed following the DNA-DNA hybridization. In this study the results of biochemical examination obtained, corresponded closely with those described in previous reports, but the oxidase reaction was very weak and varied according to the age of the culture, indicating the unreliability of this test in our case. All strains examined under long wave ultraviolet (UV) light (366 nm) revealed a blue-white fluorescence, the intensity of which ranged from strong to weak. Serological identifications were performed by both the slide agglutination test (SAT) and indirect immunofluorescent assay (IFA). SAT using commercially available antiserum (Denka Seiken., Japan) supposedly specific for L. bozemanii showed cross-reaction between L. bozemanii and L. anisa. Hyperimmune rabbit antisera prepared in this study for both L. bozemanii and L. anisa, from which cross-reactive antibodies were removed by the absorption of each antigen, reacted only with homologous antigens. IFA using a commercially available antiserum and hyperimmune rabbit antiserum previously described, gave positive reactions with each strain.

[Mechanism of acquired resistance against Legionella--blastogenic responses of murine spleen lymphoid cells following the stimulation with Legionella antigens].

We have tried to characterize the blastogenic responses of murine spleen lymphoid cells from BALB/c mice immunized with Legionella pneumophila serogroup (SG) 1 (Philadelphia 1 strain) and non-treated mice. Lymphoid cells from immunized mice showed stronger blastogenic responses following stimulation with concanavalin A or formalin-treated L. pneumophila SG1 whole cell antigen than those showed by lymphoid cells from non-treated mice. These cells from immunized mice also responded strongly when stimulated in vitro with other SGs of L. pneumophila, while these responded weakly when stimulated with other species of Legionella. Serum antibody titers of immunized mice against each SG of L. pneumophila were examined and the cross reactions were also recognized. However, the relatedness of serum antibody titers and the blastogenic responses against each serogroup of L. pneumophila was small. The epitopes recognized by the cellular immunity might be different in part from those recognized by serum antibodies, and investigations should be made on what the cellular immunity recognizes and how it works.

[A case of tubercular liver and splenic abscesses].

A 34-years male was admitted to our hospital with a hypochondria pain and low grade fever. Abdominal CT revealed an encapsulated 8 x 3 cm low density lesion on the surface of the liver (S5, S8) and multiple low density lesions of the spleen. The patient had already been treated with anti-tuberculous drugs for the past 7 months after being diagnosed as tuberculous pleuritis. Although echo-guided percutaneous needle biopsy was tried for the hepatic lesion, no special finding was obtained. Therefore a diagnostic laparotomy was performed and the hepatic lesion was resected. Abscess formation of the resected lesion was noted. Histopathology of the lesion revealed epithelioid granuloma, but microscopy, culture and PCR for tuberculosis revealed negative results. Abdominal CT, 3 weeks after surgery, revealed enlargement of the splenic lesion. Splenectomy was carried out to avoid splenic rupture. Multiple abscess of the resected spleen was noted. Pathological finding, Ziehl-Neelsen stain and PCR for tuberculosis confirmed the diagnosis of tubercular splenic and liver abscess. Although tubercular liver and splenic abscess are very rare recently, it should be included in the differential diagnosis of unknown hepatic and splenic lesions.