Association of the -31C/T functional polymorphism in the interleukin-1beta gene with the intractability of Graves' disease and the proportion of T helper type 17 cells.

Interleukin (IL)-1beta is a proinflammatory cytokine and has been implicated in the pathogenesis of several autoimmune diseases. To evaluate the hypothesis that the functional -31C/T polymorphism (rs1143627) in the gene encoding IL-1beta is associated with the intractability and the severity of autoimmune thyroid diseases, we genotyped this polymorphism in 64 patients with intractable Graves' disease (GD), 28 GD patients in remission, 49 patients with Hashimoto's disease (HD) who developed hypothyroidism (severe HD), 28 untreated euthyroid HD patients (mild HD) and 59 healthy volunteers. The -31T allele, which is related to the high producibility of IL-1beta, was significantly more frequent in patients with intractable GD than in those with GD in remission (P = 0.0017; odds ratio 2.8; 95% confidence interval 1.5-5.3), although there was no difference in this frequency between two groups of HD patients. We showed additionally that the proportion of IL-17-producing T helper type 17 (Th17) cells, whose differentiation and proliferation are promoted by IL-1beta, was higher in autoimmune thyroid disease patients with the T allele than in those with CC genotypes. In conclusion, our data indicated that the T allele of -31C/T polymorphism in the IL1B gene was involved in the intractability of GD, and this involvement may arise through the differentiation and proliferation of Th17 cells.

Involvement of functional polymorphisms in the TNFA gene in the pathogenesis of autoimmune thyroid diseases and production of anti-thyrotropin receptor antibody.

The severity of Hashimoto's disease (HD) and intractability of Graves' disease (GD) varies among patients. Severity of HD is associated with the functional +874A/T polymorphism for interferon-gamma, an inflammatory cytokine. To clarify the association between functional polymorphisms in two other inflammatory cytokine genes [tumour necrosis factor (TNF)-alpha and interleukin (IL)-2] and the severity of autoimmune thyroid disease (AITD), we examined the TNF-alpha-1031T/C, TNF-alpha-857C/T and IL-2 -330T/G polymorphisms in genomic DNA samples. We genotyped 41 patients with intractable GD, 34 patients with GD in remission, 41 patients with severe HD, 36 patients with mild HD and 70 healthy controls. The frequency of carriers of TNF-alpha-1031C (CT + CC), which correlates with higher TNF-alpha production, was significantly higher in HD and GD patients than in controls, but was not associated with the severity of HD. In GD patients, the levels of anti-thyrotropin receptor antibody (TRAb) at onset of the disease was higher in patients with the TNF-alpha-857T (CT + TT) genotype, which correlates with higher TNF-alpha production, than in those with the -857CC genotype. We found no differences in the IL-2 -330T/G polymorphism among groups of AITD patients. In conclusion, the functional -1031T/C polymorphism of the TNFA gene is associated with the development of AITD and the functional -857C/T polymorphism is associated with the levels of TRAb in active GD patients.

Pressure tuning of an ionic insulator into a heavy electron metal: an infrared study of YbS.

Optical conductivity [sigma(omega)] of YbS has been measured under pressure up to 20 GPa. Below 8 GPa, sigma(omega) is low since YbS is an insulator with an energy gap between a fully occupied 4f state and an unoccupied conduction (c) band. Above 8 GPa, however, sigma(omega) increases dramatically, developing a Drude component due to heavy carriers and characteristic infrared peaks. It is shown that increasing pressure has caused an energy overlap and hybridization between the c band and 4f state, thus driving the initially ionic and insulating YbS into a correlated metal with heavy carriers.

Comparison of formation of thiolactones and active metabolites of prasugrel and clopidogrel in rats and dogs.

Prasugrel and clopidogrel are antiplatelet prodrugs that are converted to their respective active metabolites through thiolactone intermediates. Prasugrel is rapidly hydrolysed by esterases to its thiolactone intermediate, while clopidogrel is oxidized by cytochrome P450 (CYP) isoforms to its thiolactone. The conversion of both thiolactones to the active metabolites is CYP mediated. This study compared the efficiency, in vivo, of the formation of prasugrel and clopidogrel thiolactones and their active metabolites. The areas under the plasma concentration versus time curve (AUC) of the thiolactone intermediates in the portal vein plasma after an oral dose of prasugrel (1 mg kg(-1)) and clopidogrel (0.77 mg kg(-1)) were 15.8 +/- 15.9 ng h ml(-1) and 0.113 +/- 0.226 ng h ml(-1), respectively, in rats, and 454 +/- 104 ng h ml(-1) and 23.3 +/- 4.3 ng h ml(-1), respectively, in dogs, indicating efficient hydrolysis of prasugrel and little metabolism of clopidogrel to their thiolactones in the intestine. The relative bioavailability of the active metabolites of prasugrel and clopidogrel calculated by the ratio of active metabolite AUC (prodrug oral administration/active metabolite intravenous administration) were 25% and 7%, respectively, in rats, and 25% and 10%, respectively, in dogs. Single intraduodenal administration of prasugrel showed complete conversion of prasugrel, resulting in high concentrations of the thiolactone and active metabolite of prasugrel in rat portal vein plasma, which demonstrates that these products are generated in the intestine during the absorption process. In conclusion, the extent of in vivo formation of the thiolactone and the active metabolite of prasugrel was greater than for clopidogrel's thiolactone and active metabolite.

The +869T/C polymorphism in the transforming growth factor-beta1 gene is associated with the severity and intractability of autoimmune thyroid disease.

The severity of Hashimoto's disease (HD) and the intractability of Graves' disease (GD) vary among patients. To clarify whether the +869T/C polymorphism in the transforming growth factor-beta1 (TGF-beta1) gene, which is associated with TGF-beta1 expression, is involved in the intractability of GD and severity of HD, we genotyped the TGF-beta1 +869T/C polymorphism by polymerase chain reaction-restriction fragment length polymorphism method in genomic DNA samples from 33 patients with HD who developed hypothyroidism before they were 50 years old (severe HD) and 30 untreated, euthyroid patients with HD who were older than 50 years (mild HD). We also examined 48 euthyroid patients with GD who had been under treatment and were still positive for anti-thyrotropin receptor antibodies (intractable GD), 20 euthyroid patients with GD in remission and 45 healthy controls. The frequency of the T allele and the TT genotype were higher in patients with severe HD than in those with in mild HD. In contrast, the frequency of the CC genotype was higher in patients with intractable GD than in patients with GD in remission. In conclusion, the +869T/C polymorphism in the TGF-beta1 gene is associated with the severity and intractability of autoimmune thyroid disease.

Pseudogap formation in the intermetallic compounds (Fe1-xVx)3Al

Optical conductivity data of the intermetallic compounds (Fe1-xVx)3Al ( 0

A membrane-bound ATPase from Halobacterium halobium: purification and characterization.

An ATPase was newly identified on the inner face of the plasma membrane of the extremely halophilic archaebacterium Halobacterium halobium. The enzyme was released into an alkaline EDTA solution and purified by several chromatographic steps in the presence of sulfate at 1 M or over. The molecular weight of the native enzyme was around 320,000; it is most likely composed of two pairs (alpha 2 beta 2) of 86,000 (alpha) and 64,000 (beta) subunits. The enzyme hydrolyzed ATP and other nucleoside triphosphates but neither ADP nor AMP. The enzyme required divalent cations, among which Mn2+ was most effective (Mg2+ activated 35% of Mn2+). The ATPase activity was optimum at pH between 5.5 and 6, particularly in a nearly saturated Na2SO4 (or Na2SO3) solution, while it was very low in a chloride salt solution even at 4 M at any pH. The Km value for ATP was 1.4 mM and the K1 value for ADP (competitive to ATP) was 0.08 mM. Neither azide (a specific inhibitor for F0F1-and F1-ATPase) nor vanadate (for E1E2-ATPase) inhibited the enzyme. The ATPase was stable at high concentrations of sulfate. At low concentrations of salts, or at low temperatures even in high NaCl concentrations, the enzyme was inactivated. Although the ATPase isolated here from halobacterial membrane has such unusual characteristics, it is the most probable candidate for the (catalytic part of) halobacterial ATP synthase, which differs from F0F1-ATPase/synthase (Mukohata et al. (1986) J. Biochem. 99, 1-8; Mukohata and Yoshida (1987) J. Biochem. 101, 311-318).

Effect of axotomy on the cyclic GMP increase induced by preganglionic stimulation and high extracellular K+ concentration in superior cervical sympathetic ganglion of the rat.

Cyclic GMP generation, induced by preganglionic nerve stimulation or by high extracellular potassium ion concentration (70 mM) in the medium, was studied during aerobic incubation of the excised superior cervical sympathetic ganglion of the rat with and without axotomy, and the results were compared with that of preganglionic denervation. Both axotomy and denervation of the ganglion for a week caused complete loss of increase of cyclic GMP content in the ganglion in response to the preganglionic nerve stimulation. However, the increase of the ganglionic cyclic GMP content evoked by raising the extracellular potassium ion concentration was maintained at a level about two-thirds of the control after axotomy for 1-4 days, while it was abolished within a day after denervation. Ganglionic choline acetyltransferase activity was maintained for several days after axotomy, but it was decreased rapidly by denervation. Acetylcholinesterase, dopamine-beta-hydroxylase and muscarinic cholinergic receptor sites had been lost almost completely a week after axotomy. These results suggest that the increase of ganglionic cyclic GMP content caused by depolarization with high extracellular K+ is associated with the preganglionic nerve terminals rather than with postganglionic receptor.

Distribution and different activation of adenylate cyclase by NaF and of guanylate cyclase by NaN3 in neuronal and glial cells separated from rat cerebral cortex.

Distribution of adenylate cyclase and guanylate cyclase activities in neuronal perikarya and glial cells separated from rat brain, and cellular differences in activation between of adenylate cyclase by NaF and of guanylate cyclase by NaN3 have been studied. Adenylate cyclase activity was higher in the glial cells than in the neuronal fraction, while guanylate cyclase activity was equally detected in both cell fractions. Adenylate cyclase was mainly derived from the particulate fraction of both brain cell homogenates, whereas the major portion of guanylate cyclase activity was found in their soluble rather than in the particulate fractions. Although bulk-separated neurons and glial cells almost failed to change intracellular cyclic nucleotide levels in response to some putative neurotransmitters, activation of adenylate cyclase by NaF was found to be greater in neuronal than in glial cell fractions, and was observed more clearly in the soluble than in the particulate fractions. Sodium azide greatly increased guanylate cyclase in the particulate fraction, but did not affect it considerably in the soluble one. Addition of catalase to the reaction mixture together with NaN3 further stimulated guanylate cyclase both int he soluble and the particulate fractions. These results suggest that adenylate cyclase and guanylate cyclase without intimate coupling to the transmitter-receptor system, but with activation by NaF or NaN3, may be distributed ubiquitously in the cells separated from rat cerebral cortex.

Synthesis and anthelmintic activity of 13-alkoxymilbemycin derivatives.

The synthesis of milbemycin derivatives having alkyloxy groups at the C-13 position was studied and a series of these analogs of milbemycin A4 and A3 was prepared by the reaction of 13-iodomilbemycin with a variety of alcohols. 13 beta-Phenethyloxy derivatives were found to possess excellent anthelmintic activity. Especially, the activities of derivatives with N-substituted 4-aminophenethyloxy groups were comparable to or superior to ivermectin against Nippostrongylus brasiliensis in rats.

Milbemycin derivatives: modification at the C-5 position.

Protection of the hydroxyl group at the C-5 position of milbemycin A4 and D was carried out to investigate the influence of the C-5 hydroxyl group on the anthelmintic potency of these derivatives. Moreover, the hydroxyl group was converted into amide groups as bioisosters. Biological activities of these derivatives were measured against Nippostrongylus brasiliensis in vitro, and minimal concentrations which induce 100% immotility in worms were determined for each derivative. Biological testing revealed that the hydroxyl group at C-5 is a structural requirement for retaining anthelmintic activity.

[Toxicity studies of landiolol hydrochloride (ONO-1101) (1). Single intravenous toxicity study in rats and dogs].

Single dose toxicity studies of landiolol hydrochloride (ONO-1101), a novel ultra short acting beta-blocker, were conducted in Sprague-Dawley (SD) rats and beagle dogs. ONO-1101 was administered intravenously at a dose level of 37.5, 75, 150 or 300 mg/kg to rats of both sexes and 25, 50 or 100 mg/kg to male dogs. In the rat study, 5/6 males in the 150 mg/kg group and all animals in the 300 mg/kg group died during or right after administration. Survivors in the 150 mg/kg group showed temporal hypoactivity, bradypnea, dyspnea, tremor, loss of righting reflex and reddish lacrimation up to 5 min after injection. One male in the 150 mg/kg group had a tendency of suppression on body weight gain. No effects on clinical signs and body weight gain were seen in the 75 mg/kg group or lower. Necropsy findings showed only red tear in the majority of the decedents. In the dog study, all animals died within 6 min after administration in the 100 mg/kg group, showed ataxic gait, rolling and tachypnea followed by bradypnea and gasping/apnea. Incontinence of urine, defecation and vocalization were also seen in each one of two animals before death. Temporal hypoactivity was seen 1 min after administration in the 50 mg/kg group. No clinical signs were seen in the 25 mg/kg group. ONO-1101 did not affect bodyweight or food consumption. Necropsy findings of the decedents showed no abnormalities. It is indicated that the minimum lethal doses are 150 mg/kg in rats and 100 mg/kg in dogs.

[Toxicity study of sodium N-[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino] benzoyl] aminoacetate tetrahydrate (ONO-5046.Na) (4). 6-month repeated dose intravenous toxicity study in rats with 1-month recovery test].

A 6-month repeated dose toxicity study with 1-month recovery test of sodium N-[2-[4-(2,2-dimethylpropionyloxy) phenylsulfonylamino] benzoyl] aminoacetate tetrahydrate (ONO-5046.Na), a novel neutrophil elastase inhibitor, was conducted in Sprague-Dawley (SD) rats. The rats of both sexes were administered ONO-5046.Na intravenously at a daily dose of 0 (vehicle control), 18.75, 37.5 or 75 mg/kg. ONO-5046.Na did not affect clinical signs, body weight, food consumption, opthalmology, urinalysis, hematology, blood chemistry, organ weight, necropsy or histopathology at any dose. These results indicate that the NOAEL of ONO-5046.Na in rats is 75 mg/kg/day for both sexes in this study.

Molecular cloning, sequencing and expression of core and NS3 fragments of HCV from patients with HCV infection.

Fragments of core and NS3 of hepatitis C virus-Hunan (HCV-Hun) were cloned by RT-PCR and gene recombinant techniques from blood samples collected in Hunan Province, China. In comparison with sequences of our samples with those of HCV-US and HCV-J, the homologies of nucleotides and amino acids were about 90%, indicating that fragments of core and NS3 of HCV-Hun were in a relative conserved region of HCV. Two fusion proteins containing the peptides coded by HCV core (MBP-HCV core) and HCV. NS3 (MBP-HCV. NS3-Gal) were expressed by Escherichia Coli with recombinant plasmids. The specific HCV antigenicity of the two fusion proteins were identified by western blotting. Therefore, MBP-HCV. core and MBP-HCV.NS3-Gal were found useful for anti-HCV assay.

Dynamic MRI with slow injection of contrast material for the diagnosis of pituitary adenoma.

Magnetic resonance (MR) imaging of pituitary adenoma is usually carried out in dynamic studies with bolus contrast material injections, with the result that few strong images are obtained. Dynamic studies using a slowly injected contrast material were carried out in 14 cases of pituitary adenomas. The examinations were performed with a 1.5 Tesla superconducting MR imaging system using the spin-echo technique. Gd-DTPA (0.1 mmol/kg) was slowly injected (within 90 sec) by hand, providing seven to nine dynamic images during 350 sec from the start of injection. The average time to reach the maximum signal intensity was 170.6 sec in adenoma and 156.2 sec in normal gland tissue. The maximum contrast signal intensity ratio of adenoma to normal gland was 0.527 in the fifth image. The contrast of adenoma to normal gland tissue was calculated by the following formula: ASII (adenoma signal intensity index) = (adenoma signal intensity-tissue signal intensity)/tissue signal intensity. The most remarkable contrast between adenoma and normal tissue was obtained from the fourth to eighth images. In other words, we could obtain the strongest contrast at 144.8 sec to 299.6 sec from the start of contrast injection. Our results with slow injection suggest that stronger images can be obtained a longer period after contrast injection.

Assessment of breast cancer with dynamic gadolinium-enhanced MR imaging combined with magnetization transfer contrast using a newly developed breast surface coil for the supine position.

To assess the value of dynamic gadolinium-enhanced MR imaging combined with magnetization transfer contrast (MTC) of breast cancer by SPGR sequence, 15 patients with breast cancer were imaged in the supine position with the newly developed breast coil in a 1.5 Tesla imager. Dynamic gadolinium-enhanced MR imaging combined with MTC (10 cases) and conventional dynamic gadolinium-enhanced MR imaging (5 cases) were performed after the administration of Gd-DTPA (0.2 ml/kg). Sagittal images were obtained every 22 seconds during the first 264 seconds. Thus, a total of 15 images were obtained in each lesion. The signal intensity ratio (SIR) of lesion to mammary glarid was calculated for each image as follows: (SI lesion/SI mammary gland)/(SI (pre)lesion/SI (pre)mammary gland). Dynamic gadolinium-enhanced MR imaging with MTC always allowed better SIR at 88 seconds than conventional dynamic gadolinium-enhanced MR imaging after bolus injection (p < 0.01). Dynamic gadolinium-enhanced MR imaging with MTC improved detection of the boundary of the lesion and mammary gland.

Cystic radio-lucency of carpal bones in haemodialysis patients. An early indicator of the onset of carpal tunnel syndrome.

Patients receiving haemodialysis for more than 10 years were selected for this study in order to clarify an apparent sequential association of cystic lesions of carpal bones and carpal tunnel syndrome. X-rays and computed tomographs of 138 hands of 69 patients revealed cystic radiolucency of carpal bones in 35% of the hands. Radiographs were classified into three groups: Group A--cyst growing, Group B--cyst not growing, and Group C--cyst absent. The prevalence of carpal tunnel syndrome was 100% (27/27) in Group A, 5.6% (1/18) in Group B, and 6.5% (6/93) in Group C. Growth of the cyst precedes the development of carpal tunnel syndrome by about 2 or 3 years. Growth of the bone cyst indicates that inflammation had already extended to the tenosynovium and median nerve. Cystic radio-lucency of the carpal bones appears to be a useful indicator of the onset of carpal tunnel syndrome.

Acoustic tumor with hearing loss of sudden onset and recovery.

In many cases of acoustic tumor the hearing loss is gradually progressive, but some cases of acoustic tumor show a sudden onset of hearing loss. In this paper we report two interesting cases of acoustic tumors which were diagnosed as sudden deafness at the first examination. Being treated with steroid hormones, their hearing was partially recovered. But the hearing loss recurred in both cases. At the second examination, CT showed the cerebellopontine angle tumors. So it was emphasized that careful questioning as well as the caloric test and the auditory brainstem response (ABR) was important. If one of these tests shows abnormality, the patient should be examined by the CT scan. The same may be said of patients who show recovery due to steroid hormone therapy.

[A case-control study on factors relating to discontinuation of domiciliary care for the bedridden elderly in a metropolitan area].

A case-control study was conducted to examine factors relating to discontinuation of domiciliary care for the bedridden elderly in Shinagawa-ku, Tokyo. Cases were bedridden residents aged 65 years and over who had abandoned home care and applied for admission to live in a special nursing home for the aged between April and September in 1990 after being recipients of welfare allowances for disabled elderly. Controls were bedridden residents who continued to be given home care and matched to cases by sex, age and beginning month of the receiving of allowances. Among 50 cases and 94 controls interviewed, we obtained responses from 31 cases (62%) and 60 controls (64%). The main results were as follows: 1. During the home-care period, ADL (activities of daily living) of cases, especially walking ability, deteriorated more severely than in controls. Night delirium also appeared more frequently in cases. 2. The primary caregivers of cases were older than those of controls. Remarkable differences between cases and controls were observed in the family structure, the number of family members and the number of sub-caregivers. Cases tended to live alone or live with a spouse only, and with smaller number of family members and caregivers. 3. Case lived more frequently in houses with small numbers of rooms and without rooms of their own. 4. As regards utilization of domiciliary care services, cases used dispatch of home helpers more frequently and used day services less frequently.

[Coronary artery bypass grafting for a patient with bronchial asthma seceeded from cardiopulmonary bypass by additional bypass for spasm of radial artery graft: a case report].

A 78-year-old male who had a bronchial asthma underwent coronary artery bypass grafting (CABG) using the left internal thoracic artery and the radial artery. The patient could not be weaned from the cardiopulmonary bypass because the radial artery which anastomosed to the obtuse marginal artery (OM) had a spasm after CABG. An additional bypass using a long saphenous vein to OM was carried out immediately. It brought a weaning from cardiopulmonary bypass. If the cardiac function after CABG is insufficient in patients with bronchial asthma, CABG must be re-done immediately, considering that they cause the arterial spasm more than patients without bronchial asthma.