Long-term skeletal stability in the treatment of mandibular prognathism with a physiological positioning strategy.

Our aim was to evaluate the long-term skeletal stability of the mandible in 21 patients after orthognathic surgery with physiological positioning. The measurement points SNB, B point (X, Y), Pog (X, Y), and the angle of the ramus were measured on cephalometric photographs to assess skeletal stability preoperatively, immediately after operation, and one and two years postoperatively. In addition, we evaluated the clinical symptoms of disorders of the temporomandibular joint (TMJ). The analysis of the cephalometric photographs showed that SNB, B point X, and Pog X showed no significant differences among the postoperative time points. On the other hand, B point Y and Pog Y showed no significant differences throughout the study period. We compared the angle of the ramus before operation and two years postoperatively, and no significant difference was found. In addition, no cases showed any pathological symptoms of disorders of the TMJ two years postoperatively. The long-term stability after orthognathic surgery with physiological positioning was confirmed, and it seems to be a reliable orthognathic treatment in patients with mandibular prognathism.

Behavior of pharmaceuticals in waste water treatment plant in Japan.

The fate of pharmaceuticals in a wastewater treatment plant (WWTP) in Kumamoto, Japan with activated sludge treatment is reported. Selected pharmaceuticals were detected in influent. Results from the present study confirmed that Acetaminophen, Amoxicillin, Ampicillin and Famotidine were removed at a high rate (>90% efficiency). In contrast, removal efficiency of Ketoprofen, Losartan, Oseltamivir, Carbamazepine, and Diclofenac was relatively low (<50%). The selected pharmaceuticals were also detected in raw sludge. In digestive process, Indomethacin, Atenolol, Famotidine, Trimethoprim and Cyclofosamide were removed at a high (>70% efficiency). On the other hand, removal of Carbamazepine, Ketoprofen and Diclofenac was not efficient (<50%).

Identification of triclosan intermediates produced by oxidative degradation using TiO2 in pure water and their endocrine disrupting activities.

The photodegradation pathways of 2-(2,4-dichlorophenoxy)-5-chlorophenol (triclosan) in water were studied. The main purposes were to identify structures of intermediates derived by radical reaction using TiO(2) advanced oxidation processes and to evaluate the endocrine disrupting activities in treated triclosan during oxidative reactions. Intermediates such as dichlorophenols, 2,8-dibenzo-p-dioxin, tetrachlorinated diphenyl ether (tetraclosan) and hydroxylated triclosan were produced by photoreaction. The estrogen, thyroid hormone and retinoid X receptor activities of the treated triclosan were measured with the yeast two-hybrid assay. It was found that tetraclosan and 2,4-dichlorophenol have stronger thyroid hormone activities than triclosan in the presence of S9.

Genotoxicity of dental resin polymerization initiators in vitro.

The polymerization initiators for resins cured using visible light usually consist of a photosensitizer, primarily camphorquinone (CQ), and a reducing agent, which is often a tertiary amine (DMPT, DMAEMA), while the initiator used for self-curing resins consists of benzoyl peroxide (BPO) and a tertiary amine (DMPT). The genotoxicities of camphorquinone (CQ), benzoyl peroxide (BPO), dimethyl-para-toluidine (DMPT), 2-dimethylamino-ethyl-methacrylate (DMAEMA), and 1-allyl-2-thiourea (ATU) were examined using the bioluminescent bacterial genotoxicity test. 4-Nitroquinoline-N-oxide (4NQO) was prepared for comparison with these chemicals. Acetone solutions of the five polymerization initiators and 4NQO were prepared. Benzoyl peroxide (BPO), dimethyl-para-toluidine (DMPT), and 1-allyl-2-thiourea (ATU) showed significant genotoxic activity at 24 h in the bioluminescent bacterial genotoxicity test, at concentrations of approximately 5 microM, 4 mM, and 1 mM, respectively. 2-Dimethyloamino-ethyl-methacrylate (DMAEMA) did not have genotoxic activity and CQ had questionable genotoxic activity. In comparison, 4NQO had strong genotoxicity, at 4 microM, roughly the same as that of BPO. Therefore, BPO should be used carefully in clinical dentistry.

Effects of dental resin metabolites on estrogenic activity in vitro.

Three monomers (Bis-GMA, UDMA, and TEGDMA) and five polymerization initiators (CQ, BPO, DMPT, DMAEMA, and ATU) commonly used in dental composite resins were tested for estrogenic activity using a reporter gene assay (yeast two-hybrid system) in vitro, and compared with bisphenol-A (BPA). Estrogenic activity was indicated by agonist and antagonist activity, with (+S9) and without (-S9) metabolic activation using rat liver cells. No estrogenic agonist activity was seen for each monomer and polymerization initiator in either the -S9 and +S9 tests in the concentration ranges examined in this study. On the other hand, estrogen antagonist activity was found with BPO and DMPT. BPO showed antagonist activity at a concentration of approximately 1800 nM with the -S9 test, but not with the +S9 test. With DMPT, antagonist activity was not seen with the -S9 test, but it was seen at a concentration of approximately 610 nM using the +S9 test. With BPA, the +S9 test indicated antagonist activity at a concentration of approximately 780 nM. The estrogen antagonist activities of DMPT and BPA appeared to be similar. CQ, DMAEMA, ATU, and the three monomers did not show antagonist activity as demonstrated by the -S9 or +S9 tests within the concentration range tested in this study.

Organochlorine pesticides and polychlorinated biphenyl residues in foodstuffs and human tissues from china: status of contamination, historical trend, and human dietary exposure.

Concentrations of persistent organochlorine pesticides such as DDTs, hexachlorocyclohexanes (HCHs), chlordane compounds (CHLs), hexachlorobenzene (HCB), and polychlorinated biphenyls (PCBs), were determined in a wide variety of foodstuffs and human tissues collected from Shanghai and its vicinity in China in 2000-2001. Among the organochlorines analyzed, DDT and its metabolites were prominent compounds in most of the foodstuffs. In particular, mussels contained noticeable residues of DDTs (34,000 ng/g lipid weight), which are one to three orders greater than those reported levels in bivalves from other Asian countries. Concentrations of HCHs, CHLs, HCB, and PCBs in foodstuffs were generally low, suggesting small amounts of inputs into the environment. Temporal trends examined by comparing the results of previous studies of organochlorine levels in Chinese foodstuffs in 1970s and 1992 revealed a greater amounts of declines of DDTs and HCHs residues and the average daily intakes during the past 30 years. In contrast, very high concentrations of DDTs and HCHs were detected in human tissues from Shanghai, with the maximum values as high as 19,000 ng/g lipid weight (mean: 7,600 ng/g) and 17,000 ng/g (mean: 7,400 ng/g), respectively. Considering that foodstuffs are a main source of human exposure to contaminants, the greater concentrations of DDTs and HCHs in Chinese people might be due to past extensive usage of these compounds as agricultural pesticides. Continuous monitoring and epidemiological studies of organochlorine pesticides in humans are warranted in China. To our knowledge, this is the first report to present the residue levels of persistent organochlorine pesticides and PCBs in human tissues of China.

A direct competitive enzyme-linked immunosorbent assay (ELISA) for determination of praziquantel concentration in serum.

A simple and reproducible enzyme-linked immunosorbent assay (ELISA) for the determination of the concentration of praziquantel in the serum was developed. Since praziquantel has no functional group to conjugate with carrier protein, praziquantel was first converted to a compound with an amino group similar to praziquantel. This compound was then conjugated to bovine serum albumin for use as an immunogen, and to horseradish peroxidase, as enzyme-labeled praziquantel, respectively. The conjugate of praziquantel-bovine serum albumin conjugate was used to raise anti-praziquantel antiserum in mice. The direct competitive ELISA was conducted by simultaneously incubating praziquantel and horseradish peroxidase-labeled praziquantel conjugate with anti-praziquantel antiserum over a second antibody and the enzyme activity of the remaining horseradish peroxidase-labeled praziquantel conjugate was measured. The intra- and inter-assay coefficient of variation was < 10% in the range of 1.0 to 30 ng ml(-1), and the limit of the detection was 0.3 ng ml(-1). The cross reactivities of anti-praziquantel antibody with compounds related to praziquantel were negligible. Using this ELISA, serum levels of praziquantel were easily determined in male Wistar rats up to 8 h following a single intraperitoneal injection at 2 mg kg(-1) of body weight.

Determination of cytochrome P450 1A activities in mammalian liver microsomes by high-performance liquid chromatography with fluorescence detection.

A sensitive method for the determination of cytochrome P450 (P450 or CYP) 1A activities such as ethoxyresorufin O-deethylase (EROD) and methoxyresorufin O-demethylase (MROD) in liver microsomes from human, monkey, rat and mouse by high-performance liquid chromatography with fluorescence detection is reported. The newly developed method was found to be more sensitive than previous methods using a spectrofluorimeter and fluorescence plate reader. The detection limit for resorufin (signal-to-noise ratio of 3) was 0.80 pmol/assay. Intra-day and inter-day precisions (expressed as relative standard deviation) were less than 6% for both enzyme activities. With this improved sensitivity, the kinetics of EROD and MROD activities in mammalian liver microsomes could be determined more precisely. EROD activities in human and monkey liver microsomes, and MROD activities in liver microsomes from all animal species exhibited a monophasic kinetic pattern, whereas the pattern of EROD activities in rat and mouse liver microsomes was biphasic. In addition, the method could determine the non-inducible and 3-methylcholanthrene-inducible activities of EROD and MROD in rat and mouse liver microsomes under the same assay conditions. Therefore, this method is applicable to in vivo and in vitro studies on the interaction of xenobiotic chemicals with cytochrome CYP1A isoforms in mammals.

Decreased hepatobiliary secretion of inorganic mercury, its deposition and toxicity in the Eisai hyperbilirubinemic rat with no hepatic canalicular organic anion transporter.

Eisai hyperbilirubinemic (EHB) rats, a new mutant strain inbred from Sprague-Dawley (SD) rats, show no inherent expression of the canalicular multidrug resistance protein (cMrp) and lack canalicular multispecific organic anion transporter (cMOAT) activity. A sample of 203Hg (40 microCi with 40 microg Hg/kg) was injected intravenously (i.v.) into four male SD and EHB rats. Biliary excretion of reduced-glutathione (GSH) was 426 and 2 microg/bile for 15 min in the SD and EHB rats, respectively. Biliary excretion of 203Hg for 45 min in EHB rats significantly decreased to 1/4 of that of the SD rats. However, there was no difference in the hepatic uptake of 203Hg between the two strains. Other rats were injected subcutaneously (s.c.) with HgCl2 solution (at 0.2 and 1.6 mg/kg) containing 203Hg. Some 4 days after the injection of 0.2 mg/kg, about 3 and 13% of the total dose was found in the liver in SD and EHB rats, respectively. The hepatic supernatant Hg was recovered mainly in the void volume of a Sephadex column. Some 2 days after the injection of 1.6 mg/kg, these values were 3 and 23% in SD and EHB rats, respectively. The increased retention stimulated hepatic metallothionein (MT) induction and increased the proportion of Hg in the MT region on the Sephadex column. On the other hand, biliary excretion of 203Hg for 15 min in EHB rats was about 1/6-1/4 of that in SD rats. With the injection of 1.6 mg/kg, hepatic and renal functions worsened in EHB rats. In particular, severe necrosis was found in the renal tubules. Our results suggest that biliary secretion of inorganic Hg may be partly regulated by the ATP-dependent transport system, the glutathione S-conjugate export pump (GS-X pump) composed of Mrp and MOAT. Significantly decreased excretion stimulates hepatic retention of inorganic Hg. However, the hepatic lesions are less predictive. The MT induction may reduce the toxicity of metal to the liver cells.

Development and Application of an Effective Detection Method for Fish Plasma Vitellogenin Induced by Environmental Estrogens.

Vitellogenin is a protein induced by estrogens, including environmental chemicals with estrogenic activity. To measure the effects of environmental estogens, we developed an effective and rapid one-step method of detecting and purifying fish plasma vitellogenin using a high-performance anion-exchange chromatography column, POROS-HQ. Vitellogenin in a plasma of estradiol-treated male fish (mummichog and red sea bream) was eluted as a single peak with a retention time of 10 minutes from the column, which gives an almost pure preparation as assessed by SDS-PAGE. The lowest detectable amount of vitellogenin was 2 µg per assay. The method was used to analyze the plasma vitellogenin level of aquacultured red sea breams caught in August, when the spawning season is over, and usually no vitellogenin is detected in either females or males, physiologically. However, the data showed that in addition to a few females, some male fish synthesized vitellogenin, suggesting that some chemicals or unknown factors with estrogenic activity have induced fish in the ocean to produce vitellogenin.

In-vivo microdialysis study of the distribution of cisplatin into brain tumour tissue after intracarotid infusion in rats with 9L malignant glioma.

Simultaneous brain microdialysis in tumour and non-tumour tissues has been used for kinetic determination of the local distribution of an anticancer agent, cisplatin, in rats. Rat brain was implanted with 9L malignant glioma and cisplatin (3.5 mg kg-1) was administered as a selective intracarotid infusion for 30 min to rats prepared for brain microdialysis. The amount of platinum in the dialysate collected from tumour and non-tumour brain tissues was determined by atomic absorption spectrophotometry, as representative of cisplatin. Total and free platinum concentrations in plasma were also measured. Free platinum is accumulated preferentially in the tumour tissue and the brain tumour distribution coefficient (the ratio of brain tumour platinum AUC to plasma free platinum AUC, where AUC is the area under the platinum concentration-time curve) was 0.69, although there was little distribution into normal brain tissue. Drug binding to plasma proteins was 65%. It is concluded that simultaneous microdialysis is an easy and available method for assessing in-vivo local pharmacokinetics and distribution of cisplatin in tumour and non-tumour tissues of the brain.

Lack of biliary excretion of Cd linked to an inherent defect of the canalicular isoform of multidrug resistance protein (cMrp) does not abnormally stimulate accumulation of Cd in the Eisai hyperbilirubinemic (EHB) rat liver.

A new mutant, the Eisai hyperbilirubinemic (EHB) rat, shows no inherent expression of the canalicular isoform of the multidrug resistance protein (cMrp) in the liver. It has defective biliary secretion of organic anions such as bilirubin glucuronides, bromosulfophthalein (BSP), cysteinyl leukotrienes, glutathione (GSH) and bile acid sulfate and glucuronides. When rats were injected intravenously with CdCl2, biliary excretion of Cd over 30 min was 0.28% and 0.004% of the total dose in Sprague-Dawley (SD) and EHB rats, respectively. Six SD rats and five EHB rats were fed a diet containing Cd. Bile Cd was detected at the level of 2 ng/20 min in SD rats, but not in EHB rats. There was no significant difference of hepatic Cd concentration between SD and EHB rats. Furthermore, there were no significant differences of renal and intestinal Cd, and hepatic and renal metallothionein (MT) concentrations between the SD and EHB groups. Biliary excretion of reduced-GSH for 20 min was 1.3 +/- 0.3 mg and 3.6 +/- 0.9 micrograms in SD and EHB rats, respectively. Our results suggest that hepatobiliary excretion of exogenous Cd is mediated mainly via carrier transport, including a cMrp or GSH carrier, but that the lack of the transport pathway does not contribute to abnormal accumulation of Cd in the liver.

Biliary excretion of copper, manganese, and horseradish peroxidase in Eisai hyperbilirubinemic mutant rats (EHBRs) with defective biliary excretion of glutathione.

A mixture of copper (Cu) (0.38 mg/kg), manganese (Mn (0.038 mg/kg), and horseradish peroxidase (HRP) (5.0 mg/kg) was injected intravenously (i.v.) into mature Eisai hyperbilirubinemic rats (EHBRs) and Sprague-Dawley rats (SDRs). Bile was collected at 10-min intervals before and after the injection, under anesthesia. The liver, kidneys, and blood were removed 40 min after the injection. The serum conjugated bilirubin concentration was 0.85 mg/dL in the EHBRs, but was below detection limits in the SDRs. The bile-reduced glutathione (GSH) concentration was much lower in the EHBRs (0.04 mg/mL) than in the SDRs (1.30 mg/mL). However, the hepatic GSH concentration was about 1.6 times higher in EHBRs (2.26 mg/g liver) than in SDRs (1.43 mg/g liver). The low excretion of biliary GSH was not caused by the activity of GGT in the liver, since there was no significant difference in the activity between the two groups (5.8 +/- 3.4 and 4.6 +/- 2.4 mumol p-nitroaniline/g protein/30 min in SDR and EHBR groups, respectively). There was a delay of initial biliary excretion of Cu in EHBRs compared to SDRs. The biliary concentration of Mn was slightly lower in EHBRs than in SDRs. Forty min after the injection of metals, however, there was no difference between hepatic concentrations of the two metals in the two groups. Our results suggest that abnormal deposition of the two metals is not observed naturally in EHBRs. Injected HRP was excreted rapidly and notably in the EHBRs compared to SDRs. Furthermore, the biliary concentration of beta-N-acetyl-D-glucosaminidase (beta-NAG) was significantly higher in EHBRs than in SDRs, Rapid biliary excretion of Cu, but not of Mn, may be related to the hepatobiliary transport of GSH, but the transport and lysosomal function do not originally regulate the biliary excretion of Cu.

Biliary excretion of exogenous cadmium, and endogenous copper and zinc in the Eisai hyperbilirubinuric (EHB) rat with a near absence of biliary glutathione.

Mutant Eisai hyperbilirubinuric (EHB) rats derived from an inbred strain of Sprague-Dawley (SD) rats are characterized by a near absence of biliary excretion of glutathione (GSH) due to inherently impaired ATP-driven organic anion transport. Cd (0.1 mg/kg bw from CdCl2) was injected intravenously into EHB rats and control SD rats. Output of biliary excretion of Cd was followed over 15-min intervals up to 60 min. Cd was excreted rapidly and reached the maximum level (73.2 ng/15 min) in the period from 15 to 30 min in SD rats. Its excretion in EHB rats, however was one-fortieth (only 1.8 ng/15 min) of that in SD rats. Biliary concentrations of two endogenous metals, Cu and Zn were also measured. The output of Cu in EHB rat bile (50 ng/15 min before Cd injection) was about one-fifth of that in SD rat bile (270 ng/15 min). The output was not influenced by the Cd injection in the two groups. There was a slight difference of Zn output between the two groups. The biliary excretion of GSH was 500 to 700 micrograms/15 min and only 1 to 2 micrograms/15 min in SD and EHB rats, respectively. Sixty min after Cd injection, the Cd concentrations in the serum, liver and kidney were slightly higher in EHB rats than in SD rats. There was no difference in the hepatic metallothionein (MT-I and-II) concentration between SD (34 micrograms/g liver) and EHB (33 micrograms/g liver) rats. The renal Cu concentration was about four times in the higher in the EHB rat than in the SD rat. These results suggest that reduced biliary excretion of Cd is mainly, but that of Cu is only partly, based in reduced canalicular transport of GSH due to lack of an ATP-driven organic anion transport system, not MT induction in EHB rats. It seems likely that biliary excretion of Cd is regulated mainly by the canalicular anion transport in rats.

[Computed tomographical evaluation of diabetic nephropathy].

Diabetic nephropathy can be regarded mainly as a type of microangiopathy, but is a disease that may also include aspects of macroangiopathy. This is especially true of renal disease in non-insulin dependent diabetes mellitus (NIDDM), which is characterized not only by diabetic glomerulosclerosis, but also by atherosclerosis. We performed morphological studies on the kidney, using computed tomography (CT), focusing on such points as: (1) abdominal aortic calcifications at the level of kidney, (2) calcifications in the renal artery, and (3) wedge-shaped defects on the renal surface. We noted that these findings became more prominent in NIDDM patients during end-stage renal failure than during normal renal function, and were significantly more common in those two NIDDM groups than in age-matched nondiabetic patients without hypertension, hyperlipidemia or gout. NIDDM patients exhibited these features more frequently than IDDM patients.

Nitric oxide mediated metallothionein induction by lipopolysaccharide.

The roles of calcium and/or of the other cellular transduction pathways, and of nitric oxide (NO) on the induction of metallothionein (MT) mRNA by lipopolysaccharide (LPS) has been studied in rat primary cell culture, using inhibitors of protein kinase pathways (H-7, W-7 and TMB-8) and NO production inhibitors (L-NAME, PTIO). LPS exposure led to a rapid increase of MT-mRNA and a peak level revealed 2.5-fold induction as compared to control for 6h incubation at a dose of 3.0 mg/L. A dose of 5.0 and 10.0 mg/L of LPS also provided the same level of MT-mRNA induction. The inhibition of MT induction by LPS was observed with L-NAME, PTIO, but not H-7, W-7. These findings indicate that the alteration of cellular calcium concentration and distribution does not relate to the induction of MT-mRNA by LPS in hepatocytes and that protein kinase C and calmodulin dependent protein kinase pathways have not contributed to MT-mRNA induction by LPS. Finally, the present results show that NO plays an important role in MT induction by LPS.