Bioaccumulation and toxicity of gold nanoparticles after repeated administration in mice.

Gold nanoparticles (GNPs) offer a great promise in biomedicine. Currently, there is no data available regarding the accumulation of nanoparticles in vivo after repeated administration. The purpose of the present study was to evaluate the bioaccumulation and toxic effects of different doses (40, 200, and 400 microg/kg/day) of 12.5 nm GNPs upon intraperitoneal administration in mice every day for 8 days. The gold levels in blood did not increase with the dose administered, whereas in all the organs examined there was a proportional increase on gold, indicating efficient tissue uptake. Although brain was the organ containing the lowest quantity of injected GNPs, our data suggest that GNPs are able to cross the blood-brain barrier and accumulate in the neural tissue. Importantly, no evidence of toxicity was observed in any of the diverse studies performed, including survival, behavior, animal weight, organ morphology, blood biochemistry and tissue histology. The results indicate that tissue accumulation pattern of GNPs depend on the doses administered and the accumulation of the particles does not produce sub-acute physiological damage.

Transport of prion protein across the blood-brain barrier.

The cellular form of the prion protein (PrP(c)) is necessary for the development of prion diseases and is a highly conserved protein that may play a role in neuroprotection. PrP(c) is found in both blood and cerebrospinal fluid and is likely produced by both peripheral tissues and the central nervous system (CNS). Exchange of PrP(c) between the brain and peripheral tissues could have important pathophysiologic and therapeutic implications, but it is unknown whether PrP(c) can cross the blood-brain barrier (BBB). Here, we found that radioactively labeled PrP(c) crossed the BBB in both the brain-to-blood and blood-to-brain directions. PrP(c) was enzymatically stable in blood and in brain, was cleared by liver and kidney, and was sequestered by spleen and the cervical lymph nodes. Circulating PrP(c) entered all regions of the CNS, but uptake by the lumbar and cervical spinal cord, hypothalamus, thalamus, and striatum was particularly high. These results show that PrP(c) has bidirectional, saturable transport across the BBB and selectively targets some CNS regions. Such transport may play a role in PrP(c) function and prion replication.

Brain receptor binding characteristics and pharmacokinetic-pharmacodynamic analysis of thyrotropin-releasing hormone analogues.

Thyrotropin-releasing hormone (TRH) receptor binding in the rat brain after intravenous (i.v.) injections of novel TRH analogues, taltirelin and montirelin, was examined and the data were analyzed in relation to their plasma concentrations which were simultaneously determined. Taltirelin and montirelin inhibited specific [3H]-Me-TRH binding in the rat brain and their Ki values were 311 and 35.2 nM, respectively. The i.v. injection of taltirelin and montirelin (0.1-3 mg/kg) produced a significant reduction in [3H]-Me-TRH binding sites (Bmax values) in the rat brain. The reduction by both agents tended to reach a maximum after 60 min and lasted up to at least 120 min. On the other hand, the i.v. injection of both agents had little significant effect on the apparent dissociation constant (Kd) for [3H]-Me-TRH in the rat brain. Plasma concentrations of taltirelin and montirelin in rats peaked immediately after i.v. injection, and thereafter they decreased with t 1/2 of 23.0 and 14.1 min, respectively. Counter-clockwise hysteresis between the plasma concentration and receptor occupancy of these agents was observed after the i.v. injection of taltirelin and montirelin, and the temporal delay between plasma concentration and brain receptor occupancy was successfully minimized using the "effect compartment" model in combination with the "linear-effect" model. We concluded that taltirelin and montirelin exert a fairly potent effect following sustained occupation of brain TRH receptors under in vivo condition. Thus, both agents could be clinically useful for the treatment of CNS disorders.

Ameliorating effects of amlodipine on plasma and myocardial catecholamines in BIO 53.58 Syrian hamsters, a model of dilated cardiomyopathy.

Our previous study has shown that the concentrations of norepinephrine, epinephrine and dopamine in the plasma of BIO 53.58 hamsters (a model of dilated cardiomyopathy: DCM) at 18 weeks of age (severe cardiomyopathic stage) were twice those of age-matched F1B control and conversely the myocardial norepinephrine level was decreased. The present study was undertaken to examine the effect of amlodipine on catecholamine concentration, myocardial receptors and histopathological changes in BIO 53.58 hamsters. Oral administration of amlodipine (10 mg/kg/day) for 7 weeks in 11 week-old-BIO 53.58 hamsters brought about marked decreases in the concentrations of norepinephrine, epinephrine and dopamine in the plasma, compared with those in vehicle-treated BIO 53.58 hamsters. This was accompanied by a concomitant increase in the concentration of myocardial catecholamine concentration. In other words, the concentrations of catecholamines in plasma and myocardium of amlodipine administered BIO 53.58 hamsters approximated to the control level in age-matched F1B. In addition, amlodipine administration caused a significant reduction of calcium deposition with a tendency toward a decrease in the myocardial necrosis, and it had little effect on the affinity and number of specific binding for (+)-[3H]PN 200-110, (-)-[125I]iodocyanopindolol (CYP) and [3H]prazosin in the myocardium. In conclusion, the present study shows that administration of amlodipine in BIO 53.58 hamsters may exhibit ameliorating effect on plasma and myocardial catecholamines with a significant reduction of calcium deposition. These data may offer further support for the use of amlodipine in patients with DCM.

Improvement in the histopathology of hearts from cardiomyopathic BIO TO-2 hamsters following long-term administration of amlodipine and cilnidipine.

The effect of long-term administration of amlodipine and cilnidipine was examined on the histopathology and 1,4-dihydropyridine (DHP) calcium channel antagonist receptors in the left ventricle of BIO TO-2 hamsters, a model of dilated cardiomyopathy (DCM). Oral administration of amlodipine (3 and 10 mg/kg/d, 19 weeks) in 7 week-old BIO TO-2 hamsters produced a significant reduction in calcium deposition and necrosis with little change in the cavity area and fibrosis. A reduction of calcium deposition and necrosis in the myocardium of BIO TO-2 hamsters was also seen following similar administration of cilnidipine (10 mg/kg/d). The long-term administration of amlodipine (3 and 10 mg/kg/d) caused a significant increase (36.6 and 21.7%, respectively) in the Bmax for specific (+)-[3H]PN 200-110 binding in the myocardium from BIO TO-2 hamsters, compared with that in control hamsters. In conclusion, the present study has shown that long-term administration of amlodipine and cilnidipine improves calcium deposition and necrosis in the myocardium from BIO TO-2 hamsters. Thus, these data suggest that both agents may be effective pharmacological treatments of DCM.

Increase in receptor binding affinity for nimodipine in the rat brain with permanent occlusion of bilateral carotid arteries.

The permanent occlusion of bilateral common carotid arteries (2VO) in rats has been shown to cause progressive and long-lasting cognitive deficits which may be due to impairment of memory retention and/or memory recall process. To clarify the function of voltage dependent calcium channels and the receptor binding of nimodipine by chronic cerebral ischemia, we examined specific (+)-[3H]PN 200-110 binding and the effect of oral administration of nimodipine in brain regions and hearts of rats, at 2 weeks to 4 months after permanent 2VO. There was no significant difference in either dissociation constant (Kd) or maximal number of binding sites (Bmax) for (+)-[3H]PN 200-110 in the cerebral cortex, hippocampus, corpus striatum and thalamus between 2VO and sham rats. In addition, in vitro inhibitory effect of nimodipine on cerebral cortical (+)-[3H]PN 200-110 binding in 2VO rats was similar to that in sham rats. Compared to control rats, oral administration of nimodipine to both 2VO and sham rats at 2 months after permanent 2VO brought about a significant increase in Kd values of specific (+)-[3H]PN 200-110 binding in the cerebral cortex, hippocampus, thalamus and myocardium, and the increase in Kd values was much larger in brain regions of 2VO rats than sham rats. However, the increase in Kd values in the myocardium did not differ between 2VO and sham rats. This observation suggests an increased in vivo binding affinity for nimodipine in chronic ischemic brain. In conclusion, the present study has shown that oral administration of nimodipine may cause a greater occupation in vivo of 1,4-dihydropyridine (DHP) calcium channel antagonist receptors in brains of permanent 2VO rats than in sham rats. Thus, nimodipine may be pharmacologically effective in preventing brain dysfunction due to cerebral ischemia in vivo.

Comparison of conventional vs self-etching adhesive bonds to caries-affected dentin.

The mechanism of resin adhesion to caries-affected dentin is still unclear. This study evaluated the interfacial morphology of two bonding systems to caries-affected dentin, coupled with the measurement of microtensile bond strengths (mu TBS). Carious human molars were prepared as previously described in Nakajima and others (1995) and were bonded with Single Bond (SB) or FluoroBond (FB) according to the manufacturer's instructions, followed by creation of AP-X composite buildups. After one day of storage in 37 degrees C water, the teeth were serially sectioned vertically into 0.8 mm slabs, trimmed to yield a 1 mm2 test area, and tested to failure in a Bencor device used in an Instron machine operated at 1 mm/min. Resin-dentin interfaces were observed with SEM before or after acid/base challenge. Bonding to normal dentin with the two bonding systems (SB and FB) showed tensile bond strengths significantly higher than those to caries-affected dentin. The moist bonding technique significantly increased bond strength of SB to normal and caries-affected dentin. SEM examination revealed that typical hybrid layer and resin tags could not be formed to caries-affected dentin. The results suggested that resin penetration may be prevented by occlusion of dentinal tubules by mineral deposits that may also impart acid-resistance to the intertubular matrix of caries-affected dentin.

Brain receptor binding characteristics and pharmacokinetics of JTP-2942, a novel thyrotropin-releasing hormone (TRH) analogue.

JTP-2942 competed with [3H]-Me-TRH for the binding sites in rat brain in vitro, and its inhibitory effect was approximately 17 times less potent than TRH, as shown by Ki values of 673 and 39.7 nM, respectively. Both JTP-2942 and TRH significantly increased apparent dissociation constant (Kd values) for brain [3H]-Me-TRH binding. Intravenous injection of JTP-2942 (0.3-3 mg/kg) and TRH (3 and 10 mg/kg) produced a significant reduction of [3H]-Me-TRH binding sites (Bmax values) in rat brain. Although the decrease by TRH was maximal 10 min after the injection and declined rapidly with time, the decrease by JTP-2942 (1 and 3 mg/kg) tended to be maximal at 30 min later and it lasted until 120 min. The intravenous injection of JTP-2942 was at least 3 times more potent than that of TRH in decreasing Bmax values for brain [3H]-Me-TRH binding. Plasma concentration of JTP-2942 (0.3-3 mg/kg) after intravenous injection in rats rose with the increase of dose, and it peaked immediately after the injection, thereafter decreasing with t1/2 of 19.3-29.9 min. It is concluded that JTP-2942, compared to TRH, may exert fairly potent and sustained occupation of brain TRH receptors under in vivo condition. Thus, JTP-2942 could be clinically useful for the treatment of CNS disorders.

In vivo receptor occupancy and plasma concentration of pranidipine, a potent and long-acting dihydropyridine calcium antagonist.

The occupancy of 1,4-dihydropyridine (DHP) receptors by pranidipine was characterized in tissues of spontaneously hypertensive rats (SHR). Oral administration of pranidipine (1 and 3 mg/kg) in SHR produced significant (26-67%) decreases in the number of specific (+)-[(3)H]PN 200-110 binding sites (B(max)) with 2- to 4-fold increases in the apparent dissociation constant (K(d)) in myocardial tissues at 1, 3 and 6 h later. In these rats, there was a reduction (16-37%) of B(max) in cerebral cortical (+)-[(3)H]PN 200-110 binding. Occupancy of myocardial DHP receptors after oral administration of pranidipine correlated well with its plasma concentration. Oral administration of nifedipine (10 mg/kg) in SHR caused significant increase in K(d) values for (+)-[(3)H]PN 200-110 binding in myocardium and cerebral cortex at 1 h later. In vivo specific (+)-[(3)H]PN 200-110 binding in particulate fractions of SHR aorta was markedly (59-78%) reduced at 1, 3 and 12 h after oral administration of pranidipine (3 mg/kg), while myocardial (+)-[(3)H]PN 200-110 binding was decreased by 46-48% at 1 and 3 h later. In these rats, there was a significant decrease (34%) in cerebral cortical (+)-[(3)H]PN 200-110 binding at 3 h later. In contrast, nifedipine administration produced a similar degree of reduction (71-84%) of in vivo (+)-[(3)H]PN 200-110 binding in the myocardium, aorta and cerebral cortex. It is concluded that pranidipine may exert more selective and sustained occupation in vivo of DHP receptors in vascular tissues of SHR than in myocardial and brain tissues.

Adhesion of a new commercial self-etching/self-priming bonding resin to human caries-infected dentin.

We have examined the adhesive properties of a new commercial self-etching/self-priming bonding resin (Unifil Bond, UB) to normal and caries-infected dentin of human extracted molars using scanning electron microscopy (SEM) and a micto-tensile bonding strength (MTBS) test. In this study, 7 human extracted molars with moderate occlusal caries were used, and flat surfaces including occlusal dentin caries were prepared from the teeth. After the application of UB to the surfaces, a composite resin was built up, and subjected to the measurement of MTBS and SEM observation of the interfacial morphology between UB and dentin. The MTBS of UB to normal dentin was 33.4 MPa, but that to caries-infected dentin was 11.0 MPa. There was a significant difference between the MTBS to normal and carious dentin. SEM observation revealed that the typical hybrid layer was not formed in caries-infected dentin. These results suggested that resin infiltration into caries-infected dentin was not sufficient to allow perfect sealing of the restoration.

[A nation-wide survey on the occurrence of Vogt-Koyanagi-Harada disease in Japan].

A nation-wide survey was performed to estimate the frequency and distribution of Vogt-Koyanagi-Harada's disease (Harada's disease hereinafter). A questionnaire was mailed to the departments of ophthalmology in hospitals with at least 200 beds. The response rate was 489 out of 1,077 institutions, or 45.4%. The annual prevalence was estimated to be 15.5 per million and the incidence was 6.5 per million, based on the number of reported cases and the response rate per prefecture. It was estimated that 1,800 patients were treated for this disease annually and that there were about 800 new cases. Geographical distribution showed no special characteristics such as these seen with Behçet's disease. The results of the analysis of 1,833 cases showed that females were slightly more affected than males, with an age distribution on first visit which showed a peak in the fourth decade and with the median age being 42.3 years.

A nationwide survey on the occurrence of Vogt-Koyanagi-Harada disease in Japan.

A nationwide survey was carried out to estimate the frequency and distribution of Vogt-Koyanagi-Harada disease in Japan. A questionnaire was mailed to the Departments of Ophthalmology in hospitals with 200 or more beds. Responses were received from 489 out of 1,077 institutions, or 45.5%. The annual prevalence was estimated to be 15.5 per million and the incidence was 6.5 per million, based on the number of reported cases and the response rate per prefecture. It was estimated that 1,800 patients were treated for this disease annually and that there were about 800 new cases each year. Geographical distribution showed no special characteristics such as those seen with Behçet's disease. The results of the analysis of 1,833 reported cases showed that females were slightly more often affected than males. For both male and female patients, the age distribution on the first visit showed a peak in the fifth decade and a median age of 42.3 years.

Double-masked trial of cyclosporin versus colchicine and long-term open study of cyclosporin in Behçet's disease.

The efficacy and safety of oral cyclosporin 10 mg/kg per day in Behçet's disease were compared in a randomised double-masked study with those of colchicine, 1 mg orally per day, and were also investigated in a long-term open study. The double-masked study showed that cyclosporin was effective in treating not only the ocular manifestations of Behçet's disease but also oral aphthous ulcer, dermal lesions, and genital ulceration. Efficacy did not weaken during long-term treatment.