A finite volume approach to dosimetric calculations in diffusing alpha-emitters radiation therapy.

Objective.In diffusing alpha-emitters radiation therapy ('Alpha DaRT'), the diffusion-leakage (DL) model is used to determine the spatial distributions of the emitters and the corresponding alpha dose, critical for a successful treatment. This work first introduces a finite volume (FV) approach to develop numerical schemes to simulate the DL model in one, two and three dimensions then presents how variations over realistic ranges of the DL model parameters related to desorption, diffusion and leakage processes affect the alpha dose distribution and the position of the clinically significant alpha particle10Gy isodose. This work also presents the effects of three modeling approximations: two source geometry approximations (solid cylinder instead of hollow, pixelized cross section instead of circular), and one dosimetric approximation (single-source dose superposition instead of multiple-sources direct dose calculation).Approach.The introduced FV approach was used to obtain spatial distributions of the emitters, from which the corresponding alpha dose distributions were calculated under the assumption of a local deposition of the alpha particles' energies. Variation ranges of the DL model parameters were based on previously published data. For each modeling approximation studied, the error and relative error on the alpha dose distribution were calculated and the displacement of the10Gy isodose was evaluated.Main results.Over realistic ranges, the desorption probabilities, diffusion lengths, and leakage probabilities affect the position of the alpha particle10Gy isodose by∼0.1mm,∼1.5mm and∼0.5mm, respectively. The three modeling approximations studied have a negligible effect on the alpha particle10Gy isodose position, with displacements⩽0.01mm.Significance.This work quantitatively evaluates the relative importance of different parameters and approximations in Alpha-DaRT alpha dose calculations based on their impact not only on the dose variation at a given distance from the source but also on the displacement of clinically significant isodoses.

Quantitative autistic traits ascertained in a national survey of 22 529 Japanese schoolchildren.

OBJECTIVE: Recent epidemiologic studies worldwide have documented a rise in prevalence rates for autism spectrum disorders (ASD). Broadening of diagnostic criteria for ASD may be a major contributor to the rise in prevalence, particularly if superimposed on an underlying continuous distribution of autistic traits. This study sought to determine the nature of the population distribution of autistic traits using a quantitative trait measure in a large national population sample of children. METHOD: The Japanese version of the Social Responsiveness Scale (SRS) was completed by parents on a nationally representative sample of 22 529 children, age 6-15. RESULTS: Social Responsiveness Scale scores exhibited a skewed normal distribution in the Japanese population with a single-factor structure and no significant relation to IQ within the normal intellectual range. There was no evidence of a natural 'cutoff' that would differentiate populations of categorically affected children from unaffected children. CONCLUSION: This study provides evidence of the continuous nature of autistic symptoms measured by the SRS, a validated quantitative trait measure. The findings reveal how paradigms for diagnosis that rest on arbitrarily imposed categorical cutoffs can result in substantial variation in prevalence estimation, especially when measurements used for case assignment are not standardized for a given population.

Modulation in concentrative nucleoside transporters-mediated intestinal absorption of mizoribine, an immunosuppressive agent, in lipopolysaccharide-treated rats.

The characteristics of intestinal absorption of mizoribine and cephalexin, that are mediated by concentrative nucleoside transporters (CNTs) and PEPT1, respectively, was examined in lipopolysaccharide (LPS)-treated rats. LPS treatment is known to modify the expression of some transporters and induce cholestasis. At 24 h after the LPS treatment, averaged concentrations of IL-6 and total bile acids in plasma were 15-fold and 2-fold that in untreated control rats, respectively, and bile flow rate decreased by 40% of control, indicating the induction of inflammatory and cholestatic states. The oral bioavailability, estimated by urinary excretion percentage of unchanged form, of mizoribine in LPS-treated rats was 1.5-fold higher than that in control rats, whereas the bioavailability of cephalexin remained unchanged. When mizoribine and cephalexin were administered into in-situ jejunum loops, there were no differences in the absorption rates between control and LPS-treated rats. These results indicated that the functional expression of CNT1, CNT2, and PEPT1 were not modulated by LPS treatment. When mizoribine (a CNT1/CNT2 substrate) and gemcitabin (a CNT1 substrate) were administered as a solution dissolved in bile into the intestinal loop, their absorption rates decreased significantly. In contrast, the absorption rate of ribavirin (a CNT2 substrate) remained unchanged. In conclusion, LPS treatment exerted no significant effect on the expression of CNT1 and CNT2 in the intestine. Bile was found to suppress the CNT1-mediated intestinal absorption of mizoribine and gemcitabin. The increased oral bioavailability of mizoribine in LPS-treated rats could be ascribed to the less amount of bile or bile acids in the intestine under cholestatic state of rats.

Increased intestinal absorption of mizoribine, an immunosuppressive agent, in cholestatic rats.

The intestinal absorption of mizoribine, an imidazole nucleoside, is mediated by concentrative nucleoside transporter (CNT)1 and CNT2 in rat. Previously, bile and bile salts such as sodium glycocholate were found to suppress the intestinal absorption of mizoribine. In the present study, the contribution of bile on the intestinal absorption of mizoribine was further evaluated in rats. Cholestatic states were induced by an intraperitoneal injection (2 ml/kg) of 50% carbon tetrachloride (CCl4) dissolved in olive oil, or oral administration (100 mg/2 ml/kg) of alpha-naphthylisothiocyanate (ANIT) dissolved in olive oil. The animals were subjected to absorption studies 24 h after treatment. Cholestatic states were confirmed by measuring plasma concentrations of bile acids and bile flow rates. When oral bioavailability of mizoribine was estimated by the recovery amount in the urine, rats under cholestatic states exhibited significantly higher oral bioavailabilities than untreated control rats. In contrast, the intestinal absorption percentages of mizoribine from in-situ lavaged intestinal loops were the same magnitudes among untreated control, CCl4- and ANIT-treated rats. These results indicated that the increased oral bioavailability of mizoribine in cholestatic rats was not ascribed to the modulation of nucleoside transporter's expression. In conclusion, various diseased states accompanied with cholestasis may increase the oral bioavailability of mizoribine, possibly due to its less amounts of bile in the intestinal lumen.

A surgical case of pulmonary adenocarcinoma complicated with pulmonary infarction presenting as an intrapulmonary metastasis.

Pulmonary adenocarcinoma complicated with a pulmonary infarction presenting as an intrapulmonary metastasis is relatively rare. We present a case of pulmonary infarction manifesting as intrapulmonary metastases of lung cancer. A previously healthy 59-year-old woman was admitted to our hospital on May 16, 2002 for evaluation of multiple abnormal radiographic shadows in the right lower lung field. Laboratory tests showed no abnormalities except for a slight elevation of carcinoembryonic antigens. Computed tomography of the chest revealed a hilar mass lesion with parenchymal lesions in the periphery of the right lower lobe, highly suspected to be a pulmonary adenocarcinoma with intrapulmonary metastases. A diagnosis of pulmonary adenocarcinoma was confirmed by a transbronchial brushing examination. A right middle and lower bilobectomy with mediastinal lymph node dissection was needed by hilum lymphadenopathy and a lower lobe invasion of the main tumor. Histopathological findings of the resected specimens revealed poorly differentiated adenocarcinoma of the lung with N1 (#11i) disease and multiple pulmonary infarctions with coagulation necrosis and recanalization. Pulmonary infarctions are demonstrated on chest x-rays as round or polygonal in shape, and located at the periphery of the same lobe as the primary tumor. Computed tomography is more sensitive than conventional radiography in the detection of pulmonary infarction. Our case suggests that pulmonary infarction associated with lung cancer should be considered as one important cause of peripheral pulmonary nodules.

Attention-deficit/hyperactivity disorder and physical multimorbidity: A population-based study.

BACKGROUND: There has been little research on the association of attention-deficit/hyperactivity disorder (ADHD) with co-occurring physical diseases. The aim of this study was to examine the association between possible ADHD and physical multimorbidity (i.e.≥2 physical diseases) among adults in the English general population. METHODS: Data were analyzed from 7274 individuals aged≥18 years that came from the Adult Psychiatric Morbidity Survey 2007. ADHD symptoms were assessed with the Adult Self-Report Scale (ASRS) Screener. Information was also obtained on 20 self-reported doctor/other health professional diagnosed physical health conditions present in the past 12 months. Multivariable logistic regression and mediation analyses were conducted to assess the associations. RESULTS: There was a monotonic relation between the number of physical diseases and possible ADHD (ASRS score≥14). Compared to those with no diseases, individuals with≥5 diseases had over 3 times higher odds for possible ADHD (odds ratio [OR]: 3.30, 95% confidence interval [CI]: 2.48-4.37). This association was observed in all age groups. Stressful life events (% mediated 10.3-24.3%), disordered eating (6.8%), depression (12.8%), and anxiety (24.8%) were significant mediators in the association between possible ADHD and physical multimorbidity. CONCLUSION: Adults that screen positive for ADHD are at an increased risk for multimorbidity and several factors are important in this association. As many adults with ADHD remain undiagnosed, the results of this study highlight the importance of detecting adult ADHD as it may confer an increased risk for poorer health outcomes, including physical multimorbidity.

Examining cross-cultural differences in autism spectrum disorder: A multinational comparison from Greece, Italy, Japan, Poland, and the United States.

Autism spectrum disorder (ASD) is characterized by social and communication impairments as well as restricted, repetitive behavior patterns. Despite the fact that ASD is reported worldwide, very little research exists examining ASD characteristics on a multinational scale. Cross-cultural comparisons are especially important for ASD, since cultural differences may impact the perception of symptoms. Identifying behaviors that are similarly reported as problematic across cultures as well as identifying behaviors in which there is cultural variation could aid in the development and refinement of more universally effective measures. The present study sought to examine similarities and differences in caregiver endorsement of symptom severity through scores on the Baby Infant Screen for Children with aUtIsm Traits (BISCUIT). The BISCUIT was utilized to examine ASD core symptomology in 250 toddlers diagnosed with ASD from Greece, Italy, Japan, Poland, and the United States. Significant differences in overall ASD symptom severity and endorsement were found between multinational groups. Implications of the results are discussed.

Field trial of time-spreading and wavelength-hopping OCDM transmission using FBG en/decoders.

In this paper, field trial on optical code division multiplexing through the commercial-used fiber line is presented. We fabricated fiber Bragg grating en/decoders with time-spreading and wavelength-hopping scheme, considering environmental fluctuation of transmission fiber. 200 km-long transmission of 2-channel x 10 Gb/s signals was achieved on the field photonic network test bed of JGN II. Error free transmission was demonstrated in real field deployed single-mode transmission fiber.

Outer membrane of Salmonella typhimurium. Identification of proteins exposed on cell surface.

Proteins exposed on the outer surface of the outer membrane of Salmonella typhimurium were identified by reacting intact cells with a covalent labeling reagent. Since the outer membrane permitted the free diffusion of small hydrophilic molecules, we used a macromolecular reagent, CNBr-activated dextran, as the non-penetrating labeling agent. We also used a mutant producing a lipopolysaccharide with a very short (i.e. hexasaccharide) carbohydrate chain, in order to avoid steric hindrance by the carbohydrates on membrane surface. Results showed that out of the four "major" proteins of molecular weight around 35 000, three were exposed, and that at least six other proteins were also exposed on cell surface. Only two or three outer membrane proteins consistently did not react with the reagent in intact cells.

Purification of human plasma lecithin:cholesterol acyltransferase and its specificity towards the acyl acceptor.

A simple and convenient method for the purification of human plasma lecithin-cholesterol acyltransferase was developed. The method involves the adsorption of the enzyme from diluted human plasma on DEAE-Sephadex, treatment with 1-butanol in the presence of (NH4)2SO4, DEAE-Sephadex chromatography, treatment with dextran sulfate in the presence of Ca2+, and hydroxyapatite chromatography. The enzyme purified showed a single main band by polyacrylamide gel electrophoresis in the presence and absence of sodium dodecyl sulfate. In addition, the enzyme obtained was stable for more than four weeks, when it was kept at 4 degrees C under N2 in a buffer of low ionic strength. The purified enzyme was used to study its specificity toward the acyl acceptor. This specificity was found to be broad in that not only sterols but also long chain primary alcohols exhibited considerable acceptor activity. Furthermore, in agreement with our previous observations with crude enzyme (Piran, U. and Nishida, T. (1976) J. Biochem. (Tokyo) 80, 887-889), the purified enzyme was found to be capable of hydrolyzing the ester linkage at the carbon-2 position of phosphatidylcholine. The transesterification, as well as the hydrolytic reaction, required the presence of the cofactor polypeptide, apolipoprotein A-I.

[Completion pneumonectomy 9 years after middle lobectomy for adenocarcinoma].

Completion pneumonectomy (CP) is a difficult operation in which the surgeon must use techniques such as intrapericardial ligation of the pulmonary vessels. We report herein a case of CP for a patient with recurrent lung cancer. A 63-year-old man was admitted to our hospital for evaluation of abnormal shadows in the right lung field in October 2002. Right middle lobectomy with mediastinal lymph node dissection had been performed in February 1993. Computed tomography (CT) revealed a hilar mass in the right upper lobe the day after admission. Bronchofiberscopic cytology revealed squamous cell carcinoma. Right completion pneumonectomy was performed on suspicion of metachronous multiple lung cancers 4 days later. Histopathologically, resected specimens represented adenosquamous carcinoma similar to the prior lesion from the middle lobe, and examination revealed that the tumor represented a recurrence following middle lobectomy. The patient remains well as of 19 months postoperatively.

[Aneurysmal bone cyst arising in the rib; report of a case].

We experienced with a relatively rare case of an aneurysmal bone cyst (ABC) arising in the left rib. A 34-year-old female, had experienced chest discomfort on the left anterior side and pain for 1 year. A chest X-ray suggested a left chest wall tumor involving the ribs. Computed tomography (CT), magnetic resonance imaging (MRI) and a bone scintigram revealed an expansive tumor of the anterior portion of the left 4th rib involving the 3rd and 5th rib with "blow out appearance" and "fluid-fluid level". Wide excision of the tumor and adjacent muscle tissue was performed with an antero-axillary incision. Chest wall reconstruction was performed with prolene mesh (140 x 90 mm). The resected specimen showed an encapsulated bony mass (75 x 60 x 35 mm) with multiple blood-filled spaces. Histopathological diagnosis was an ABC originating in the left 4th rib. She has been doing well with no evidence of recurrence 12 months postoperatively.

Complete nucleotide sequence and molecular characterization of the temperate staphylococcal bacteriophage phiPVL carrying Panton-Valentine leukocidin genes.

The staphylococcal Panton-Valentine leukocidin (PVL) genes, [lukS-PV-lukF-PV], exist on the genome of a temperate bacteriophage phiPVL isolated from mitomycin C-induced Staphylococcus aureus V8 (ATCC 49775) (Kaneko, J., Kimura, T., Kawakami, Y., Tomita, T., Kamio, Y., 1997b. Panton-Valentine leukocidin genes in phage-like particle isolated from mitomycin C-treated Staphylococcus aureus V8 (ATCC 49775). Biosci. Biotechnol. Biochem. 61, 1960-1962). In this study, the complete nucleotide sequence of the phiPVL genome was analyzed, and the att sites (attL, attR, and attB) required for site-specific integration of phiPVL into the host chromosome were also determined. The linear double-stranded phiPVL genome comprised 41401bp with 3' staggered cohesive ends (cos) of nine bases and contained 63 ORFs, among which the regulatory proteins involved in DNA replication, structural proteins, a holin, a lysin, an integrase, and dUTPase, were tentatively identified by the comparison of the deduced amino acid sequences and by the analysis of the proteins isolated from phiPVL particles. The [lukS-PV-lukF-PV], attP, and int (integrase gene) of phiPVL were all located very close to one another within a 4.0-kb segment on the genome in the order given, and this segment was located at the center from the left and the right cos sites. In addition, the attP region contained five direct repeat sequences that showed a high degree of homology with the recombinase-binding sites of some other S. aureus bacteriophages. The phiPVL genome was found to integrate into an ORF encoding an unknown protein comprising 725 amino acid residues with two leucine zipper-like motifs.

Phage conversion of Panton-Valentine leukocidin in Staphylococcus aureus: molecular analysis of a PVL-converting phage, phiSLT.

Staphylococcal Panton-Valentine leukocidin (PVL) is an important virulence factor, which causes leukocytolysis and tissue necrosis. Our previous report on the existence of the PVL genes (lukS-PV and lukF-PV) on the genome of prophage phiPVL in the Staphylococcus aureus strain V8 suggested the horizontal transmission of PVL genes by temperate bacteriophage among S. aureus (Kaneko, et al., 1998. Gene 215, 57-67). Here, we demonstrated the phage conversion of S. aureus leading to the production of PVL by discovery of a novel PVL-carrying phage, phiSLT (Staphylococcal Leukocytolytic Toxin) from a clinical isolate of S. aureus. phiSLT was able to lysogenize several clinical isolates of PVL-negative S. aureus strains as well as strain RN4220 at the conserved 29-bp sequence (attB) and all the lysogenized S. aureus strains had the ability to produce PVL. phiSLT had an elongated head of about 100x50 nm and a flexible tail of 400 nm long, that was quite different from phiPVL which had an isometric hexagonal head of about 60 nm diameter. The linear double-stranded phiSLT genome comprised 42,942 bp with 29-bp attachment core sequences and contained 62 open reading frames. Only 6.4 kbp region containing lysis cassette, PVL genes, attP, integrase, and orf204 of phiSLT was identical to that of phiPVL, while other regions were different from those of phiPVL. Thus, it can be concluded that PVL genes are carried by different temperate phages, which have the same attachment site.

Assembly of Staphylococcus aureus gamma-hemolysin into a pore-forming ring-shaped complex on the surface of human erythrocytes.

Staphylococcal gamma-hemolysin consists of Hlg1 (or Luk F) of 34 kDa and Hlg2 of 32 kDa, which cooperatively lyse human erythrocytes. Since gamma-hemolysin caused swelling of human erythrocytes prior to lysis, we studied pore-forming nature of the toxin by use of polyethylene glycols as osmotic protectants and determined the functional diameter of the pore. To elucidate the molecular architecture of the membrane pore formed by gamma-hemolysin, we solubilized the pore complex with 2% sodium dodecyl sulfate, separated it from erythrocyte membrane proteins by sucrose gradient ultracentrifugation, and observed the isolated complex under an electron microscope. Our data showed that Hlg1 and Hlg2 of gamma-hemolysin assemble into a ring-shaped 195 kDa complex in a molar ratio of 1 : 1, which may form a membrane pore with a functional diameter of 2.1-2.4 nm.

The C-terminal region of the S component of staphylococcal leukocidin is essential for the biological activity of the toxin.

The Staphylococcal toxin leukocidin consists of two protein components, F and S. From a culture medium of Staphylococcus aureus RIMD 310925, we isolated a truncated form of S (LS2), of which the C-terminal 17-residue segment is missing. Unlike intact S, LS2, showed neither leukocytolytic activity in the presence of F nor affinity for monosialoganglioside GM1 (GM1). When excited at 280 nm, both S and LS2 exhibited intrinsic tryptophan fluorescence with an emission maximum at 318 nm. Upon binding to GM1, the emission maximum of S underwent a blue shift to 310 nm, whereas no change in fluorescence took place on mixing GM1 with LS2. We conclude that the C-terminal region of S is essential for its biological activity as well as for its binding to GM1 and that this binding is accompanied by a conformational change of the S protein.

Identification of the minimum segment in which the threonine246 residue is a potential phosphorylated site by protein kinase A for the LukS-specific function of staphylococcal leukocidin.

Staphylococcal leukocidin and gamma-hemolysin consist of LukF and LukS for leukocidin and LukF and Hlg2 for gamma-hemolysin. In this report, we identify the minimum segment responsible for the LukS-specific function of leukocidin. After chemical analysis and homology study of the amino acid sequence of the C-terminal region between LukS and Hlg2, we found a unique 5-residue sequence I242K243R244S245T246 in LukS in which the 4-residue KRST is identical with that of the phosphorylated segment of a protein phosphorylated by protein kinase A. To elucidate whether the 5-residue segment is essential for the LukS function, we created plasmids containing a series of mutant genes corresponding to the 5-residue sequence and expressed them in Escherichia coli. The mutant proteins were purified and assayed for their leukocytolytic activity with LukF. The mutant MLS-TS, in which the T246 in the 5-residue sequence was replaced by S, showed leukocidin activity 10 times higher than that of the intact LukS. However, neither mutant MLS-TY nor MLS-TA, in which T246 was replaced by Y or A, respectively, showed leukocidin activity. The 5-residue segment was found to be deleted in Hlg2. The mutant of Hlg2, in which the 5-residue segment was inserted at the position that the segment is deleted, showed leukocidin activity. The boiled LukS, MLS-TS, and MHS-Z were strongly phosphorylated with [gamma-32P]ATP in the presence of protein kinase A in a cell-free system. Thus, we conclude that the 5-residue segment 1242K243R244S245T246 is the pivotal segment of LukS responsible for the LukS function of staphylococcal leukocidin.

Existence of a new protein component with the same function as the LukF component of leukocidin or gamma-hemolysin and its gene in Staphylococcus aureus P83.

Staphylococcal toxins, leukocidin and gamma-hemolysin, consist of two protein components, i.e. LukF and LukS for leukocodin and H gamma I and H gamma II for gamma-hemolysin. From a culture fluid of Staphylococcus aureus strain P83, a new protein component of leukocidin or gamma-hemolysin which was designated as LukM was isolated. This component showed the same biological activity as that of LukF component for leukocidin or H gamma I component of gamma-hemolysin in combination with LukS or H gamma II. However, the LukM component cross-reacted with the anti-LukS antibodies but not with the anti-LukF antibodies. On the basis of chemical analysis of the LukM component and the cloning and nucleotide sequencing of the lukM gene of S. aureus P83, we have demonstrated that LukM is an entirely new protein component of leukocidin or gamma-hemolysin. The deduced amino acid sequence of LukM from the lukM gene showed 66.7% and 67% identity to that of LukS and H gamma II, respectively. However, the amino acid sequence of LukM and LukF showed only 29% homology.

Vitronectin and its fragments purified as serum inhibitors of Staphylococcus aureus gamma-hemolysin and leukocidin, and their specific binding to the hlg2 and the LukS components of the toxins.

Staphylococcal gamma-hemolysin and leukocidin are bi-component cytolysins, consisting of LukF (or Hlg1)/Hlg2 and LukF/LukS, respectively. Here, we purified serum inhibitors of gamma-hemolysin and leukocidin from human plasma. Protein sequencing showed that the purified inhibitors of 62, 57, 50 and 38 kDa were the vitronectin fragments with truncation(s) of the C-terminal or both N- and C-terminal regions. The purified vitronectin fragments specifically bound to the Hlg2 component of gamma-hemolysin and the LukS component of leukocidin to form high-molecular-weight complexes with them, leading to inhibition of the toxin-induced lysis of human erythrocytes and human polymorphonuclear leukocytes, respectively. Intact vitronectin also showed inhibitory activity to the toxins. The ability of gamma-hemolysin and leukocidin to bind vitronectin and its fragments is a novel function of the pore-forming cytolysins.

The two Staphylococcal bi-component toxins, leukocidin and gamma-hemolysin, share one component in common.

Staphylococcal bi-component toxins, leukocidin and gamma-hemolysin, consist of two protein components, i.e. F and S for leukocidin and H gamma I and H gamma II for gamma-hemolysin. In this study we purified H gamma I and H gamma II to homogeneity from the culture medium of Staphylococcus aureus RIMD 310925 and compared their properties with those of F and S purified from the same source. The N-terminal 59- and C-terminal 2-residue amino acid sequences, apparent molecular mass, and isoelectric point of purified H gamma I were the same as those of F. In an Ouchterlony double diffusion test a fused line without spur was formed between F and H gamma I using either anti-F or anti-H gamma I antibodies. A synergistic action of F and H gamma II caused hemolysis of human red blood cells, and H gamma I acted synergistically with S to exhibit leukocidin activity. We conclude that the two toxins share one protein component (F = H gamma I) in common and leukocidin- and gamma-hemolysin-specific activities are determined by S and H gamma II, respectively. It is also reported that the N-terminal 58-residue sequence of H gamma II is 72% similar to the corresponding sequence of S.