Negative motor networks: Electric cortical stimulation and diffusion tensor imaging.

INTRODUCTION: This study investigated the networks of Negative motor areas (NMAs) using electric cortical stimulation and diffusion tensor imaging (DTI). METHODS: Twelve patients with intractable focal epilepsy, in which NMAs were identified by electrical cortical stimulation, were enrolled in this study. Electric stimulation at 50Hz was applied to the electrodes during motor tasks to identify the NMAs. DTI was used to identify the subcortical fibers originating from the NMAs found by electrical stimulation. RESULTS: NMAs were found in lateral frontal areas (premotor area (PM) and precentral gyrus) in all 12 patients, in pre-supplementary motor areas (pre-SMAs) in four patients, and in posterior parietal cortices (PPCs) in four. DTI detected fibers connecting to the ipsilateral PMs, PPCs and temporal regions via U-fibers, superior longitudinal fasciculus (SLF), and arcuate fasciculus (AF) from the lateral frontal NMAs. Pre-SMA-NMAs had connections with ipsilateral PMs and contralateral pre-SMAs via the frontal aslant tract and transcallosal commissural fibers, and PPC-NMAs with ipsilateral PMs via SLF and AF. CONCLUSION: This study found the characteristic cortical network of each NMA, and especially revealed new insight of pre-SMA-NMA and PPC NMA. These NMAs might be associated with different mechanism of negative motor response.

Missense variants of the alanine: glyoxylate aminotransferase 2 gene correlated with carotid atherosclerosis in the Japanese population.

Alanine:glyoxylate aminotransferase 2 (AGXT2; EC 2.6.1.44) degrades asymmetric dimethylarginine (ADMA), a competitive inhibitor of nitric oxide (NO) synthase. Increased ADMA, reduced NO, and hypertension are shown in Agxt2 knockout mice. There are four single nucleotide polymorphisms (rs37370, rs37369, rs180749, and rs16899974) with which AGXT2 activity changes in humans and may be related to vulnerability of vascular sclerosis. To examine the relationship between them, we studied the functional haplotypes of the AGXT2 gene and decided their relationship with arteriosclerotic changes via carotid intima-media thickness (carotid IMT) in Japanese subjects. Genotyping of those polymorphisms and the carotid IMT in 1,426 Japanese subjects were then evaluated. Subjects with C-A-A-A haplotype (rs37370, rs37369, rs180749, rs16899974) showed low AGXT2 activity (P<0.0001; Pearson’s correlation coefficients: 0.497). The C-A-A-A haplotype was significantly associated with mean carotid IMT (P=0.049) and max carotid IMT (P=0.004). Subjects with two C-A-A-A haplotypes exhibited thicker mean carotid IMT (P=0.022) and maximum carotid IMT (P=0.001). In multiple regression analysis, subjects with two C-A-A-A haplotypes were independently and positively associated with mean carotid IMT (P=0.02) and maximum IMT (P=0.005) after correction. There was a significant correlation between the functional variants in the AGXT2 gene and carotid IMT in Japanese. The AGXT2 genotype may be an important factor underlying atherosclerosis.

Absorption, distribution and excretion of radioactivity by male rats after a single intravenous or subcutaneous dose of 14C-azacitidine.

The absorption, distribution and excretion of radioactivity were investigated in male Sprague-Dawley rats after a single intravenous or subcutaneous dose of 14C-azacitidine. After subcutaneous administration, 14C-azacitidine was rapidly absorbed and the radioactivity was distributed to the tissues. The absorption of radioactivity after subcutaneous administration was 76.6% of that observed after intravenous administration. There were no marked differences in the tissue distribution of the radioactivity between administration routes. The concentrations of radioactivity in most tissues, including the spleen and bone marrow, which are sites of action of azacitidine, were higher than those in the plasma. Particularly high concentrations of radioactivity were detected in the spleen, kidney and liver. The accumulation of radioactivity in blood cells increased from 0.5 to 48 h. The binding of azacitidine to serum protein was low at <9%, and the cumulative urinary and fecal excretion of radioactivity for 168 h after intravenous or subcutaneous administration was >95% of the administered dose, indicating that radioactivity did not accumulate in the tissues. The radioactivity was mainly excreted in the urine.

Extraction of nonylphenol and nonylphenol ethoxylates from sewage sludge using the pressurized fluid extraction method and the supercritical fluid extraction method.

To establish endocrine disruptor (ED) monitoring and control methods for wastewater treatment plants, it is necessary to clarify the fate of EDs in each process of a sludge treatment system. However, the ED analysis method for sewage sludge containing large quantities of organic matters has not been established. So, a highly reliable analytical method that accurately measures EDs in sewage sludge must be developed. This paper reports on the results of applying the Pressurized Fluid Extraction (PFE) method and the Supercritical Fluid Extraction (SFE) method to the extraction of nonylphenol (NP) and nonylphenol ethoxylate (NPnEO) existence of which has often been confirmed in past surveys. To clarify the quantity of EDs in sludge specimens and the fate of EDs in the sludge treatment process, appropriate analytical methods should be developed. In this study, the PFE method and the SFE method were tested to extract NP and NPnEO from sewage sludge specimens. The findings indicate that the extraction from dried sludge by the PFE method can extract a larger quantity of NP and NPnEO in a shorter time than the widely used heating reflux method. Regarding the extraction by the SFE method, conditions for extraction of NP and NPnEO at a higher rate than the heating reflux method were not found.

Effect of tyrosine kinase inhibition by imatinib mesylate on mast cell tumors in dogs.

BACKGROUND: Imatinib mesylate is a small molecule targeted at dysregulated protein-tyrosine kinase. Mutation of c-kit exon 11, which induces constitutive phosphorylation of KIT, is one of the mechanisms for the development or progression of mast cell tumor (MCT) in dogs. The purpose of this study was to examine the therapeutic potential of imatinib mesylate in canine MCT. HYPOTHESIS: Imatinib mesylate has activity against MCT in dogs, and response to treatment can be correlated to presence of mutation within exon 11 of c-kit. ANIMALS: Twenty-one dogs with MCT with gross tumor burden and median tumor size of 7.2 cm (range, 1.0-25.3 cm) before treatment. METHODS: Tumors were analyzed for mutation of c-kit exon 11. Imatinib mesylate was administered PO to the dogs at a dose of 10 mg/kg daily for 1-9 weeks. RESULTS: Ten of 21 dogs (48%) had some beneficial response to imatinib mesylate treatment within 14 days of treatment initiation. All 5 dogs with a demonstrable c-kit mutation in exon 11 responded to the drug (1 complete remission, 4 partial remission). CONCLUSIONS AND CLINICAL IMPORTANCE: Imatinib mesylate has clinical activity against MCT in dogs. Response could not be predicted based on presence of absence of a mutation in exon 11 of c-kit.

Fate of nonylphenol and 17beta-estradiol contained in composted sewage sludge after land application.

Many environmental problems caused by endocrine disrupters (EDs) have been reported. Because little is known about the fate of EDs accumulated in sewage sludge, we carried out a study to clarify the fate of EDs in composted sludge after its application to soil. Nonylphenol (NP) and 17beta-estradiol (E2) were measured for leachate and soil. High concentrations of NP and E2 were detected in the leachate at the early stage, but they decreased rapidly. Also, the high contents of NP and E2 in soil decreased significantly within 300 days. Because the decrease of NP and E2 in the soil was much larger than that of NP and E2 in the leachate, there must have been a physicochemical or biological decomposition mechanism in the soil layer. We also tried to clarify the transfer of NPs to plants from compost. In the experimental conditions of this study, the transfer of NPs to plants from compost was not observed.

Fate of nonylphenol polyethoxylates and nonylphenoxy acetic acids in an anaerobic digestion process for sewage sludge treatment.

Many environmental problems caused by endocrine disruptors (EDs) have been reported. It is reported that EDs flow into sewage treatment plants, and it has been pointed out that these may be shifted from the wastewater treatment process to the sludge treatment process. Little is known about the fate of EDs accumulated in sewage sludge, so we carried out a study to clarify the fate of EDs in sewage sludge treatment processes, especially in an anaerobic digestion process. In this study, nonylphenol (NP) was selected as a target ED. Nonylphenol ethoxylates (NPnEO) or nonylphenoxy acetic acids (NPnEC), which were the precursor of NP, were added to an anaerobic digestion process, and mass balance was investigated. The following results were obtained from the anaerobic digestion experiments. (1) NP1EO was injected to an anaerobic digestion testing apparatus that was operated at a retention time of approximately 28 d and a temperature of 35 degrees C with thickened sludge sampled from an actual wastewater treatment plant. Approximately 40% of the injected NP1EO was converted to NP. (2) NP1EC was injected to an anaerobic digestion testing apparatus with thickened sludge. As a result, almost all injected NP1EC was converted to NP. When NP2EC was injected, NP2EC was not converted to NP until the 20th day.

Favorable outcomes with tacrolimus in two patients with refractory interstitial lung disease associated with polymyositis/dermatomyositis.

Two cases of progressive interstitial lung disease associated with polymyositis/dermatomyositis are presented. Both patients were refractory to conventional therapy with high-dose corticosteroids, cyclosporine, and intermittent pulse cyclophosphamide, and thus a therapeutic trial of tacrolimus was instituted. Tacrolimus was markedly effective in achieving subjective, laboratory and radiographic improvement in both patients.

Clostridium perfringens epsilon-toxin increases permeability of single perfused microvessels of rat mesentery.

Epsilon-toxin, the primary virulence factor of Clostridium perfringens type D, causes mortality in livestock, particularly sheep and goats, in which it induces an often-fatal enterotoxemia. It is believed to compromise the intestinal barrier and then enter the gut vasculature, from which it is carried systemically, causing widespread vascular endothelial damage and edema. Here we used single perfused venular microvessels in rat mesentery, which enabled direct observation of permeability properties of the in situ vascular wall during exposure to toxin. We determined the hydraulic conductivity (L(p)) of microvessels as a measure of the response to epsilon-toxin. We found that microvessels were highly sensitive to toxin. At 10 microg ml(-1) the L(p) increased irreversibly to more than 15 times the control value by 10 min. At 0.3 microg ml(-1) no increase in L(p) was observed for up to 90 min. The toxin-induced increase in L(p) was consistent with changes in ultrastructure of microvessels exposed to the toxin. Those microvessels exhibited gaps either between or through endothelial cells where perfusate had direct access to the basement membrane. Many endothelial cells appeared necrotic, highly attenuated, and with dense cytoplasm. We showed that epsilon-toxin, in a time- and dose-dependent manner, rapidly and irreversibly compromised the barrier function of venular microvessel endothelium. The results conformed to the hypothesis that epsilon-toxin interacts with vascular endothelial cells and increases the vessel wall permeability by direct damage of the endothelium.

Role of women in medicine: a look at the history, the present condition and the future status of women in the surgical field, especially neurosurgery.

We have analyzed the historical background of women's progress in medicine in Japan and the role of female neurosurgeons as models for the next generation. Female neurosurgeons were asked to complete a questionnaire regarding their professional life in detail and the problems they are facing while managing their personal life after getting married and having a child. Some feel that there remain some constraints at work for being a female, due to their male colleagues who are not so understanding in nature. The younger generation is not so keen on joining the neurosurgical branch as their life career due to hard work and complete dedication demanded by neurosurgery. It is not easy for all to manage a neurosurgical career along together with a married life and children. Hence it is now time for those successful female neurosurgeons to become role models. Government can play an important role in these social reforms by coming up with programs to give social security to females and initiate programs for child care for married females pursuing such a demanding profession. Certain measures to encourage females to take up surgery are providing more time by arranging care for babies and families, flexibility in working hours, in addition to having a considerate husband and a considerate chief of department and senior staff. Departmental policies need to be completely impartial and should promote everyone based on their skills and knowledge. Women neurosurgeons need to get together and discuss all these issues so that the younger generation will not hesitate to take up this profession and become stalwarts of neurosurgery like their male counterparts.

Characterization of bacteriocin N15 produced by Enterococcus faecium N15 and cloning of the related genes.

Enterococcus faecium N15 was isolated from nuka (Japanese rice-bran paste), which is utilized as starter in the fermenting of vegetables, and was found to produce a bacteriocin that exhibited a broad spectrum of activity, including activity against Listeria monocytogenes and Bacillus circulans JCM2504. The bacteriocin was sensitive to proteases (alpha-chymotrypsin, proteinase K, trypsin, and pepsin) and alpha-amylase, but it was resistant to lipase. The bacteriocin was resistant to heat treatment at 100 degrees C for 2 h, but its activity was completely lost after autoclaving at 121 degrees C for 15 min. It was active over a wide pH range from 2.0 to 10.0. The bacteriocin showed bactericidal activity against Lactobacillus sake JCM1157 at a concentration of 40 AU/ml. Its molecular weight was estimated by SDS-PAGE to be about 3-5 kDa. PCR primers were designed based on the conserved amino acid sequences of class IIa bacteriocins. A 3-kb DNA fragment was amplified and three open reading frames (ORFs) were found. The first encodes a probable immunity protein of 103 amino acid residues and shows complete homology with the putative immunity protein of E. faecium DPC1146. The second and third ORFs respectively encode a probable transposase gene and an inducing factor. The upstream region of the immunity gene, in which the bacteriocin structural gene is located, was amplified. A homology search revealed that the bacteriocin produced by E. faecium N15 exhibits complete identity to enterocin A, a bacteriocin produced by E. faecium DPC1146. PCR using the primers designed in this study is a rapid and sufficient method of screening for bacteriocin-producing strains.

A new implant designed to maximize contact with trabecular bone: survival to 18 months.

PROBLEM: A roughened, commercially pure titanium (CP-Ti) implant design has been developed that features a different length and pitch for each screw thread to direct functional stresses away from cortical bone and to the more resilient trabecular bone. Abutment-implant connection is made using a conical taper to provide a seal against invasion by microorganisms. PURPOSE: To assess short-term (18 months) clinical performance of this innovative implant design. METHODS: A total of 1,419 implants were placed in 313 patients to support 419 prostheses in a multidisciplinary, multicentered, prospective clinical study conducted by the Ankylos Implant Clinical Research Group (AICRG). More than 100 dentists at 32 centers in the United States, 1 in Korea, and 1 in Taiwan are involved in the study. Failure was defined as implant removal for any reason. The influence of mobility at placement, implant length and diameter, incision type, augmentation, crestal bone reduction, bone density, and the use of the operating room or dental clinic on survival were evaluated over 18 months. Crestal bone loss between placement and uncovering was also determined. RESULTS: Crestal bone loss ranged from 0.2 to 0.5 mm. The overall success rate from placement to 18 months was 96.6%. Implants mobile at placement failed more frequently (16.9%) compared with stable implants (3.1%). Wide-diameter implants and longer implants exhibited higher survival rates. Incision design and surgery location did not influence survival. Bone density was important to clinical survival.

A 48-month multicentric clinical investigation: implant design and survival.

This report is based on a total of 2,955 implants of 6 different designs, randomized and placed in 829 patients and followed for 48 months. Implant failure was defined as nonintegration at uncovering or removal due to mobility, persistent pain, infection, and evidence of radiographic bone loss. Failures were reported for 3 phases of treatment: implant placement to uncovering (phase 1), uncovering to loading (phase 2), and postloading (phase 3). Differences in survival were compared with Kaplan-Meier survival curves. The maxillary single tooth application resulted in 95.2% survival for the hydroxyapatite-coated grooved implants. In the maxillary completely edentulous application, survival of hydroxyapatite grooved and screw implants were considerably better compared with the titanium screw implants. The hydroxyapatite-coated cylinder had better survival than the titanium basket and screw designs in the mandibular completely edentulous application. The hydroxyapatite-coated cylinder and grooved implants in the maxillary posterior partially edentulous application had similar survival rates. The survival of the hydroxyapatite-coated cylinder exceeded that of the titanium basket in mandibular posterior partially edentulous applications. Analyses by phase of treatment indicated a pattern of early failure for nonhydroxyapatite-coated implants compared with hydroxyapatite-coated implants. The implant with the highest survival at all phases of treatment was the hydroxyapatite-coated press-fit cylinder. Two hydroxyapatite-coated implant designs performed well in the challenging posterior maxillary region.

Up-to-date modification of the anaerobic sludge digestion process introducing a separate sludge digestion mode.

Sewage treatment plants in Japan are subjected to advanced treatment to remove nutrients and hence control eutrophication problems in lakes and bays. This paper discusses the advantages and disadvantages of the separate digestion treatment mode for sludge generated from advanced wastewater treatment. In the separate digestion only primary sludge is digested and the excess activated sludge is directly dewatered. Separate digestion can reduce the return load of nutrients to approximately one third, and has major potential for the beneficial use of sludge.

Role of tryptophan-1 in hemolytic and phospholipase C activities of Clostridium perfringens alpha-toxin.

Replacement of the Trp-1 in Clostridium perfringens alpha-toxin with tyrosine caused no effect on hemolytic and phospholipase C (PLC) activities or on binding to the zinc ion, but that of the residue with alanine, glycine and histidine led to drastic decreases in these activities and a significant reduction in binding to the zinc ion. The hemolytic and PLC activities of W1H and W1A were significantly increased by the preincubation of these variant toxins with zinc ions, but the preincubation of W1G with the metal ion caused little effect on these activities. Gly-Ile-alpha-toxin, which contained an additional Gly-Ile linked to the N-terminal amino acid of alpha-toxin, did not show hemolytic activity, but showed about 6% PLC activity of the wild-type toxin. A mutant toxin, which contained an additional Gly-Ile linked to the N-terminus of a protein lacking 4 N-terminal residues of alpha-toxin, showed about 1 and 6% hemolytic and PLC activities of the wild-type toxin, respectively. Incubation of the mutant toxin with zinc ions caused a significant increase in PLC activity. These observations suggested that Trp-1 is not essential for toxin activity, but plays a role in binding to zinc ions.

Characterization of the enzymatic component of Clostridium perfringens iota-toxin.

The iota(a) component (i(a)) of Clostridium perfringens ADP ribosylates nonmuscle beta/gamma actin and skeletal muscle alpha-actin. Replacement of Arg-295 in i(a) with alanine led to a complete loss of NAD(+)-glycohydrolase (NADase) and ADP-ribosyltransferase (ARTase); that of the residue with lysine caused a drastic reduction in NADase and ARTase activities (<0.1% of the wild-type activities) but did not completely diminish them. Substitution of alanine for Glu-378 and Glu-380 caused a complete loss of NADase and ARTase. However, exchange of Glu-378 to aspartic acid or glutamine resulted in little effect on NADase activity but a drastic reduction in ARTase activity (<0.1% of the wild-type activity). Exchange of Glu-380 to aspartic acid caused a drastic reduction in NADase and ARTase activities (<0.1% of the wild-type activities) but did not completely diminish them; that of the residue to glutamine caused a complete loss of ARTase activity. Replacement of Ser-338 with alanine resulted in 0.7 to 2.3% wild-type activities, and that of Ser-340 and Thr-339 caused a reduction in these activities of 5 to 30% wild-type activities. The kinetic analysis showed that Arg-295 and Ser-338 also play an important role in the binding of NAD(+) to i(a), that Arg-295, Glu-380, and Ser-338 play a crucial role in the catalytic rate of NADase activity, and that these three amino acid residues and Glu-378 are essential for ARTase activity. The effect of amino acid replacement in i(a) on ARTase activity was similar to that on lethal and cytotoxic activities, suggesting that lethal and cytotoxic activities in i(a) are dependent on ARTase activity.

The efficacy of a counter-rotational powered toothbrush in the maintenance of endosseous dental implants.

BACKGROUND: Although most patients with implants have lost their natural teeth because of poor oral hygiene, limited data exist to guide practitioners in their recommendations of home-care regimens for their patients' endosseous dental implants and maintenance of peri-implant soft-tissue health. The authors conducted a study to compare the home-care effectiveness of a counter-rotational powered tooth-brush with that of conventional home-care regimens. METHODS: Before starting the six-year study, the authors trained 85 clinical investigators at 32 dental research centers across the United States in gathering periodontal data. Data for 2,966 implants were entered into a centralized database. Outcomes were derived from 24-month observations of a subset of the implants studied. RESULTS: Repeated-measures analysis of the toothbrushing methods used on 2,966 implants showed that the counter-rotational powered toothbrush removed plaque significantly better than manual methods (P < .0001 Wald statistic) from all implant surfaces and at all recall intervals up to 24 months. Similar results were demonstrated for the gingival index. CONCLUSIONS: The counter-rotational powered brush appears to be well-suited for home-care regimens aimed at maintaining optimal peri-implant soft-tissue health in patients with dental implants. CLINICAL IMPLICATIONS: The importance of maintaining the health of the peri-implant tissues is well-recognized by the dental profession. The counter-rotational powered toothbrush is an effective tool in meeting the oral hygiene challenges associated with implant prosthesis maintenance.

The ankylos endosseous dental implant: assessment of stability up to 18 months with the Periotest.

Osseointegration is an ongoing histometric process that may vary during clinical function. The implant must be stable at uncovering, which reflects the status of bone-implant interface. The physiology of bone healing associated with endosseous implants suggests that this process occurs between 8 and 12 months, and Periotest values (PTVs) tend to reflect changes in the stability of the bone-implant interface. Stability generally increases gradually from the time of uncovering to an optimal PTV that occurs at a point close to 12 months. This stable interface must remain intact for long-term clinical survival. Rapid development of this optimal PTV is highly desirable in order to prevent premature overloading of the bone-implant interface. The Ankylos implant is a new screw-type implant design in which the thread pitch and length vary to maximize trabecular bone contact. The purpose of this report is to evaluate to 18 months the stability (PTVs) of this implant design. More than 457 implants were placed and followed for a period of 18 months by the multicentered, multidisciplinary Ankylos Implant Clinical Research Group (AICRG). Implant stability (PTVs) was assessed using the Periotest at abutment connection and at 3, 6, 9, 12, and 18 months after uncovering. The Periotest values for all implants rapidly reached an optimal status between uncovering (-3.1 PTVs) and 3 months (-3.4 PTVs). This rapid increase in stability has not previously been reported for other implant designs. The mandibular arch was more negative (-3.8 PTVs) at uncovering as compared with the PTVs for the maxillary arch (-1.7 PTVs). Negative PTVs were recorded (1) as length and diameter increased, (2) as bone density increased, (3) in certain jaw regions, (4) as the number of implants/case increased, and (5) for implants stabile at placement. The Ankylos screw implant design produced rapid stabilization 3 months after uncovering.

Survival of various implant-supported prosthesis designs following 36 months of clinical function.

BACKGROUND: The use of endosseous dental implants to replace natural teeth lost to trauma, dental caries, or periodontal disease has become a predictable form of prosthetic treatment since gaining popularity in the early 1980s. While numerous clinical studies have focused on the survival of implants, few address the survival of different prosthesis designs. METHODS: Beginning in 1991, 882 prostheses supported by more than 2,900 implants (687 patients) were placed by the Department of Veterans Affairs Dental Implant Clinical Research Group (DICRG). These prostheses were divided into five research strata based on arch location. The recommended design for each stratum was: bar-supported overdenture (maxillary completely edentulous); screw-retained hybrid denture (mandibular completely edentulous); screw-retained fixed partial denture (mandibular and maxillary posterior partially edentulous); and cemented single crown (maxillary anterior single tooth). Alternative overdenture designs were utilized in the edentulous arches when the recommended prosthesis could not be fabricated. Prosthesis success rates for the research strata were calculated for an observation time of up to 36 months following prosthesis placement. RESULTS: Success rates for the maxillary edentulous stratum ranged from 94.6% for the bar-retained overdenture supported by five to six fixtures to 81.8% for the cap-retained overdenture. The mandibular edentulous strata produced success rates of 98.1% for the fixed hybrid prosthesis to 91.7% for the cap-retained prosthesis. Success rates for maxillary and mandibular posterior fixed partial dentures were 94.3% and 92.6%, respectively, while the maxillary anterior single-tooth prosthesis yielded a success rate of 98.1% for the 36-month observation period. CONCLUSIONS: The recommended prosthesis designs investigated in this study proved to be reliable, with encouraging success rates for an observation period of 36 months following placement.