Oral environment and taste function of Japanese HIV-infected patients treated with antiretroviral therapy.

The aim of the study was to evaluate the oral environment and the taste function of Japanese HIV-infected patients treated with antiretroviral therapy. Their median age of 73 patients taking anti-HIV drugs was 46 years. The median period of taking anti-HIV drugs was 30 months. The oral condition was evaluated by measurement of oral moisture, amount of saliva secretion, the number of oral bacteria, presence of oral candida, a taste test, and the number of missing teeth. The levels of oral moisture and secreted saliva were significantly lower in the HIV-infected group than in the healthy volunteer (control) group. The HIV-infected group showed a more robust decrease in taste sensation than the control group. The number of missing teeth was significantly higher in the HIV-infected group than in the control group. Furthermore, all of the evaluated oral conditions were worse in the HIV-infected patients whose CD4+ T lymphocyte counts were less than 500/mm3 than in the control group. It became clear that the patients taking anti-HIV drugs, especially the CD4+ count < 500/mm3 group, had a deteriorated oral environment and dysgeusia, suggesting that the management of oral hygiene is necessary to maintain oral health, which leads to systemic health.

Rapid multiresidue determination of pesticides in livestock muscle and liver tissue via modified QuEChERS sample preparation and LC-MS/MS.

Many multiresidue methods for the determination of pesticides in vegetables and fruits have been reported to date. However, few such methods have been employed to investigate pesticide residues in animal tissue. In this study, an LC-MS/MS multiresidue method coupled with modified QuEChERS extraction was developed and validated for the investigation of eight pesticide residues: prallethrin (PR), resmethrin (RMT), imidacloprid (IMC), diflubenzuron (DFB), cyromazine (CYR), etofenprox (EFP), dinotefuran (DNT) and phthalthrin (PTLT). This method involves initial extraction in a water/acetone system, the addition of salts and a subsequent extraction/partitioning step and, finally, a clean-up step utilising dispersive solid-phase extraction (SPE). The mean recoveries of seven of the pesticides (the exception being CYR) ranged between 74.7% and 113.5%, and the CVs of the livestock tissue - bovine, swine, and chicken muscle and liver tissue spiked at 10 ng g-1 (50 ng g-1 for RMT and DNT) and 100 ng g-1 - were < 13.8%. The recoveries of CYR in all muscle and liver spiked samples ranged from 56.9% to 78.3%, while those of RMT in swine liver were > 120%. Therefore, this method was considered as being unsuitable for the investigation of these samples. The limits of quantitation (LOQs) of seven of the investigated pesticides (the exception being swine liver) in the tissue samples ranged from 0.9 to 15.2 ng g-1. We therefore concluded that this LC-MS/MS multiresidue method is a valid and suitable for the investigation of seven pesticides in animal tissue, but it is unsuitable for the analysis of CYR in all animal tissues and RMT in swine liver tissue.

Decreased cortical thickness, as estimated by a newly developed ultrasound device, as a risk for vertebral fracture in type 2 diabetes mellitus patients with eGFR of less than 60 mL/min/1.73 m2.

UNLABELLED: Cortical porosity is increasingly recognized as an important risk for fracture in DM patients. The present study demonstrated that decreased cortical thickness, assessed using a newly developed quantitative ultrasonic bone densitometry, is a significant risk factor for vertebral fractures in type 2 diabetes mellitus patients with stage 3 or higher chronic kidney disease, but not in those without. INTRODUCTION: Cortical porosity is increasingly recognized as an important risk factor for fracture in type 2 diabetes mellitus (T2DM) patients as well as in stage 3 chronic kidney disease (CKD) patients in whom serum parathyroid hormone (PTH) starts to increase. The present study aimed to clarify whether the coexistence of CKD might affect the relationship of decreased cortical thickness (CoTh) in the development of vertebral fractures (VF) in T2DM patients. METHODS: In this cross-sectional study, trabecular bone mineral density (TrBMD), elastic modulus of trabecular bone (EMTb), and CoTh were estimated with a new quantitative ultrasound bone densitometry in 173 T2DM patients. VFs were identified radiographically. RESULTS: Thirty-nine patients (22.5%) had VF. Those with estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m(2) (low eGFR) showed a significantly higher VF rate (32.4%) than those with eGFR ≥60 mL/min/1.73 m(2) (high eGFR, 16.2%). Serum PTH was significantly higher with low eGFR than with high eGFR. In those with high eGFR, EMTb was significantly lower in VF(+) than VF(-). In those with low eGFR, TrBMD, EMTb, and CoTh were significantly lower in VF(+) than in VF(-). In a multivariate logistic regression analysis, EMTb was independently and significantly associated with VF in T2DM patients with a high eGFR, in contrast to those with only CoTh with VF in T2DM with low eGFR. CONCLUSION: This study demonstrated CoTh as a factor independently associated with VF in T2DM patients with low eGFR and increasing serum PTH levels.

All-optical flip-flop operation based on asymmetric active-multimode interferometer bi-stable laser diodes.

We demonstrate fast and low energy all optical flip-flop devices based on asymmetric active-multimode interferometer using high-mesa waveguide structure. The implemented devices showed high speed all-optical flip-flop operation with 25 ps long pulses. The rising and falling times of the output signal were 121 ps and 25 ps, respectively. The required set and reset pulse energies were only 7.1 fJ and 3.4 fJ, respectively.

Investigation on virtual palpation system using ultrasonic elasticity imaging.

In the field of medical ultrasound, ultrasonic elasticity imaging has been developed and used in practical diagnosis. However, unfortunately, elasticity information is now displayed on ordinary 2D screen (for example, CRT). Since elasticity information is haptic information, not visible information, the information is expected to be displayed in "the sense of haptic". In this research, it is attempted that elasticity information is displayed on haptic display for virtual palpation system. In this system, a surface of diseased part is expressed by one rigid plane which has elasticity information. The elasticity information is a function of 2D coordinates of a haptic device on the plane. 3D elasticity information of the diseased part which is measured by ultrasonic elasticity imaging equipment is rendered to the plane by a way which is similar to volume rendering technique, to reduce computational cost and to realize human haptic sense easily in virtual palpation in real time. It is also important to reproduce the haptic sense in the case where a physician slides fingers on the surface in real palpation. The proposed system enables to reproduce the haptic sense in not only the perpendicular direction to the surface but the direction of horizontal movement of fingers on the surface. This system can realize virtual palpation system.

Ultrasonic diffraction tomography by pulse-plane wave: experimental result by frequency synthesis method.

Diffraction tomography is a technique for imaging with acoustic fields in which parameter, such as reflective index, sound velocity, etc., can be mapped from scatter wave resulting from insonifying the object with a plane wave at a single temporal frequency. By solving the direct scattering problem, the scattered field can be presented in term of scattering parameters. Different inversion techniques can be applied to take advantage of the linearization process of the non-linear wave equation describing wave propagation in heterogeneous media under a limited class of scattering. Specifically, when the scattering effect is weak, one can invoke the Born or Rytov approximation and thus derive the generalized Fourier slice theorem to reconstruct the cross-section of the insonified object. Although diffraction tomography is a promising technology for medical application as it provides a quantitative ultrasonic image, its realization toward medical use is still far-to-go, and this may be due to the complexity of the hardware involved. In this research we investigate a potential use of diffraction tomography for medical application by using a delicate-designed ultrasonic computerized tomographic system. The contribution of the paper is that we have purposed to frequency synthesis method (FSM) that uses 3 frequency components of the Fourier transform of scattered filed, i.e. the center frequency and the -3 dB from center frequency of both side, to fill Fourier space for the generalized Fourier slice theory. The reconstructed ultrasonic image from such theory provides a very promising result.

International harmonisation of anthelmintic efficacy guidelines (Part 2).

The "International Co-operation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products (VICH)" is an international programme of co-operation between regulatory authorities and the animal health industries of the European Union, Japan and the United States of America which aims to harmonise the technical requirements for the registration of veterinary medicinal products. Australia and New Zealand participate as active observers. The objective of this second paper is to present additional guidelines established by the Working Group on anthelmintic guidelines: (1) efficacy of anthelmintics: specific recommendations for equine (VICH GL15), (2) efficacy of anthelmintics: specific recommendations for porcine (VICH GL16), (3) efficacy of anthelmintics: specific recommendations for canine (VICH GL19), (4) efficacy of anthelmintics: specific recommendations for feline (VICH GL20) and (5) efficacy of anthelmintics: specific recommendations for poultry (VICH GL21). These guidelines do not consist of rigid stipulations, but make clear recommendations on the minimal standards needed. To the veterinary profession, livestock producers and animal owners, harmonisation should mean quicker access to safer and more effective veterinary anthelmintics. In general, products should be relatively more affordable because of the reduction in registration costs and efficient use of resources by the regulatory authorities.

Kinetic analyses of alcohol-induced potentiation of the response of GABA(A) receptors composed of alpha(1) and beta(1) subunits.

To investigate the kinetics of both the potentiation and desensitization of the response of ionotropic GABA receptors (GABA(A) receptors) in the presence of various compounds, we expressed receptors composed of alpha(1) and beta(1) subunits by injecting cells with the cRNAs synthesized from cloned bovine GABA(A) receptor cDNAs and measured the electrical responses of the cells electrophysiologically with or without the compounds. The potentiation of the GABA(A) receptor-mediated response was quantitatively analyzed using a simple model with the assumption that the receptors have two identical binding sites for GABA molecules with a dissociation constant of K(1), and one potentiation site for the compound with a dissociation constant of K(p), and that the binding of the compound to the potentiation site only increases the affinity of the GABA binding sites, changing K(1) to K(1p). The estimated K(p) and K(1p) were dependent on the functional groups and the chain length of the compounds. These results could be satisfactorily analyzed using this simple model. The potentiation of the GABA(A) receptor-mediated response by the components of essential oils used for aromatherapy was also examined. These compounds accelerated the decay of the response, possibly due to desensitization of the receptors, which was also analyzed on the basis of the model.

The effect of soybean oil refuse powder used as vehicle on the absorption of oxolinic acid in chickens under fasting and nonfasting conditions and the correlation with in vitro dissolution.

The effect of soybean oil refuse powder (SOR) when used as a vehicle on the absorption of oxolinic acid (OXA) powder in chicken, the dissolution profile of OXA and the correlation between in vivo and in vitro study were examined. To examine in vivo bioavailability, chickens fed or fasted were studied using a 2 x 2 crossover design. The OXA was administered OXA or OXA-SOR (1 : 9) mixture 20 mg OXA/kg. In vitro dissolution rates for OXA and OXA-SOR were measured using the paddle (PD) and the rotatory dialysis cell dissolution (PTSW) methods. Maximum plasma concentration (Cmax) and area under the curve (AUC) were significantly increased by the addition of SOR to OXA. Differences between OXA and OXA-SOR were more remarkable under fasted as compared with fed condition. In vitro dissolution rates of OXA-SOR pH 1.2, 6.5 and 7.2 as determined by the PD and the PTSW methods were increased in the presence of SOR vehicle. Differences between OXA and OXA-SOR in vitro dissolution rates were greater than in vivo bioavailability. Correlation between in vitro release (%) and in vivo absorption (%) showed good linearity (gamma=0.8805-0.9999).

Novel compound heterozygous mutations for lipoprotein lipase deficiency. A G-to-T transversion at the first position of exon 5 causing G154V missense mutation and a 5' splice site mutation of intron 8.

We systematically investigated the molecular defects causing a primary LPL deficiency in a Japanese male infant (patient DI) with fasting hyperchylomicronemia (type I hyperlipoproteinemia) and in his parents. Patient DI had neither LPL activity nor immunoreactive LPL mass in the pre- and post-heparin plasma. The patient was a compound heterozygote for novel mutations consisting of a G-to-T transversion at the first nucleotide of exon 5 [+1 position of 3' acceptor splice site (3'-ass) of intron 4] and a T-to-C transition in the invariant GT at position +2 of the 5' donor splice site (5'-dss) of intron 8 (Int8/5'-dss/t(+2)c). The G-to-T transversion, although affecting the 11 nucleotide of the 3'-consensus acceptor splice site, resulted in a substitution of Gly(154) to Val (G154V; GG(716)C(-->)GTC). The mutant G154V LPL expressed in COS-1 cells was catalytically inactive and hardly released from the cells by heparin. The Int8/5'-dss/t(+2)c mutation inactivated the authentic 5' splice site of intron 8 and led to the utilization of a cryptic 5'-dss in exon 8 as an alternative splice site 133 basepairs upstream from the authentic splice site, thereby causing joining of a part of exon 8 to exon 9 with skipping of a 134-bp fragment of exon 8 and intron 8. These additional mutations in the consensus sequences of the 3' and 5' splice sites might be useful for better understanding the factors that are involved in splice site selection in vivo.

In vitro activities of moxifloxacin and other fluoroquinolones against Mycoplasma pneumoniae.

A total of 105 isolates of Mycoplasma pneumoniae were evaluated for susceptibility to moxifloxacin, sparfloxacin, levofloxacin, and ciprofloxacin. Moxifloxacin, a newly synthesized compound, showed the greatest activity. The MICs and MBCs at which 50 and 90% of isolates were affected were 0.15 (MIC(50) and MBC(50)) and 0.3 microg/ml (MIC(90) and MBC(90)) respectively. The results indicate that moxifloxacin might be promising an antimycoplasmal agent.

International harmonisation of anthelmintic efficacy guidelines.

The "International Co-operation on Harmonisation of Technical Requirements for Registration of Veterinary Medicinal Products (VICH)" is an international programme of co-operation between regulatory authorities and the animal health industries of the European Union, Japan, and the United States of America which aims to harmonise the technical requirements for the registration of veterinary medicinal products. Australia and New Zealand participate as active observers. The objective of the present paper is to present the guidelines established by the working group on Anthelmintic Efficacy Guidelines: (1) efficacy of anthelmintics: general requirements (VICH GL7); (2) efficacy of anthelmintics: specific recommendations for bovines (VICH GL12); (3) efficacy of anthelmintics: specific recommendations for ovines (VICH GL13); (4) efficacy of anthelmintics: specific recommendations for caprines (VICH GL14). These guidelines do not consist of rigid stipulations, but make clear recommendations on the minimal standards needed. To the veterinary profession, livestock producers and animal owners, harmonisation should mean quicker access to safer and more effective veterinary anthelmintics. In general, products should be relatively more affordable because of the reduction in registration costs and efficient use of resources by the regulatory authorities.

Bioavailability of amprolium in fasting and nonfasting chickens after intravenous and oral administration.

The bioavailability of amprolium (APL) was measured after intravenous (i.v.) and oral (p.o.) administration to chickens. Twelve healthy chickens weighing 1.28-1.41 kg received a dose of 13 mg APL/kg intravenously, and 13 or 26 mg APL/kg orally in both a fasted and a nonfasted condition in a Latin square design. Plasma samples were taken from the subwing vein for determination of APL concentration by HPLC method. The data following intravenous and oral administration were best fitted by 2-compartment and 1-compartment models, respectively, using weighted nonlinear least squares regression. The half-life beta t(1/2)beta, volume of distribution (Vd) and total body clearance (Cl) after intravenous administration were 0.21 h, 0.12 L/kg and 1.32 L/h.kg, respectively. The elimination half-life (t(1/2) Kel) after oral administration was 0.292-0.654 h which is 1.5-3.2 times longer than after intravenous administration, suggesting the presence of a 'flip-flop' phenomenon in chickens. The maximum plasma concentration (Cmax) of 13 mg/kg APL administered orally to chickens during fasting was significantly (about four times) higher than that during nonfasting (P < 0.05). Bioavailability during nonfasting was from 2.3 to 2.6%, and 6.4% during fasting.

Potentiation of GABAA receptors expressed in Xenopus oocytes by perfume and phytoncid.

To study the effects of perfume and phytoncid on GABAA receptors, ionotropic GABAA receptors were expressed in Xenopus oocytes by injecting mRNAs that had been prepared from rat whole brain. Essential oil, perfume and such phytoncid as leaf alcohol, hinokitiol, pinene, eugenol, citronellol and citronellal potentiated the response in the presence of GABA at low concentrations (10 and 30 microM), possibly because they bound to the potentiation-site in GABAA receptors and increased the affinity of GABA to the receptors. Since it is known that the potentiation of GABAA receptors by benzodiazepine, barbiturate, steroids and anesthetics induces the anxiolytic, anticonvulsant and sedative activity or anesthetic effect, these results suggest the possibility that the intake of perfume or phytoncid through the lungs, the skin or the intestines modulates the neural transmission in the brain through ionotropic GABAA receptors and changes the frame of the human mind, as alcohol or tobacco does.

Expression of functional tissue factor on small vesicles of lipopolysaccharide-stimulated human vascular endothelial cells.

We examined tissue factor expression on lipopolysaccharide-stimulated endothelial cells and their small vesicles by using specific antibodies and flow cytometry. Tissue factor functional activity was also assessed by activation of factor X. Endothelial cells were stimulated with 10 microg/ml of lipopolysaccharide in M-199/bovine serum albumin. Flow cytometry showed that expression of tissue factor on endothelial cells reached a maximum at 6 hours after stimulation, whereas that on small vesicles reached a maximum after 12 hours. Factor X activation mediated by factor VIIa and tissue factor was observed over a similar time course and was inhibited by the addition of antitissue factor antibody. Immunoelectron microscopy suggested that small vesicles with expression of some tissue factor were produced from the surface of endothelial cells. Our findings thus showed that tissue factor on endothelial cells produced by lipopolysaccharide stimulation was partly released to small vesicles. This may cause disseminated intravascular coagulation and related coagulation disorders.

Fluorescence in situ hybridization analysis of 12;21 translocation in Japanese childhood acute lymphoblastic leukemia.

Fluorescence in situ hybridization (FISH) analysis was applied to detect t(12;21) using two yeast artificial chromosome probes and cosmid probes covering the TEL(ETV6) and the AML1 gene to clarify the incidence of abnormality of t(12;21) in Japanese childhood acute lymphoblastic leukemia (ALL). We detected seven TEL/AML1 fusion positive patients (9.5%), all of whom were diagnosed as B-lineage ALL, among 74 childhood ALL. On the other hand, no TEL/AML1 fusion positive patients were found among 37 adult ALL. The incidence among Japanese seemed to be lower than that among other nations. Of the seven patients with the TEL/AML1 fusion, five exhibited normal karyotype, one was t(8;12)(q11;p13), i(21q) and the remaining one exhibited a near-triploid karyotype in conventional G-banding. The FISH method clearly demonstrated that all patients with the TEL/AML1 fusion had subpopulations of leukemic cells with deletion of the normal TEL allele, which is significant for understanding the progression of leukemia with t(12;21).

Prognostic value of dipyridamole-thallium myocardial scintigraphy in patients with Kawasaki disease.

BACKGROUND: Although coronary artery lesions are critical complications of Kawasaki disease, their long-term outcome is still unclear. It is sometimes difficult to monitor progressive changes from aneurysms to stenotic lesions because coronary angiography (CAG) cannot be repeated very often, especially in infants. Our prospective study was designed to evaluate the prognostic value of dipyridamole-thallium single-photon-emission CT (SPECT) in the long-term follow-up of patients with Kawasaki disease. METHODS AND RESULTS: Of 459 consecutive patients with Kawasaki disease, coronary aneurysms were detected in 90 cases by echocardiography during the acute stage. After paired studies of selective CAG and SPECT were conducted, all patients were followed up and monitored for the occurrence of any cardiac events for > or =8 years. During the follow-up interval, there were 15 cardiac events (1 death, 5 infarctions, 2 coronary artery bypass graft operations, and 7 occurrences of unstable angina). Of patients who had some event, thallium redistribution was found on SPECT in 14 (93%, P<0.001). Of the various clinical and scintigraphic image variables, the presence of thallium redistribution was the best multivariate independent predictor of a late cardiac event (chi2=57.8, P<0.0001). The number of aneurysms detected on CAG added minimal statistical improvement to the model (chi2=1.9, P=0.0009). CONCLUSIONS: Dipyridamole-thallium SPECT is safely performed and is useful and important for risk stratification in the long-term follow-up of patients with Kawasaki disease.

[Splenic lymphoma with villous lymphocytes and complex chromosomal abnormality].

We report a 70-year-old Japanese man who had splenic lymphoma with villous lymphocytes and a complex chromosomal abnormality. No monoclonal gammopathy was present. The peripheral blood film showed lymphocytes with thin and short villi arising from one or two poles of the cells. These cells were negative for tartrate-resistant acid phosphatase stain. Immunophenotyping of peripheral blood lymphocytes showed moderate to strong expression of surface membrane IgM, IgD, IgA, and lambda as well as CD19, CD20, CD21, CD24, and HLA-DR. In addition, there was weak CD5, CD22, and CD25 expression, but no CD10, CD11c, CD23, CD38, or B-ly-7 expression. All 20 metaphases obtained from peripheral blood cells cultured for 5 days with lipopolysaccharide showed an abnormal karyotype: 47, XY, +der(3) t(3; 13) (q26; q12) inv(3) (?), t(7; 14), (q21; q11), der(13) t(3; 13) (q26; q12). Our patient followed a relatively benign clinical course and splenectomy was not performed.

Rapid and sensitive determination of amprolium in chicken plasma by high-performance liquid chromatography with post-column reaction.

A rapid, sensitive and reproducible reversed-phase HPLC assay was developed for the determination of amprolium (APL) in chicken plasma. Protein in plasma sample was precipitated with 0.33 M perchloric acid and supernatant solution was injected into the HPLC system. Following the chromatographic separation of APL and the beclotiamine (I.S.) on a C18 column, the derivatives of APL and I.S. were formed by post-column reaction and detected by fluorescence detection (excitation at 400 nm, emission at 460 nm). The method showed excellent precision, accuracy and speed with a detection limit of 2 ng/ml. The intra- and inter-assay variance of this method were less than 11.2%. This method has been successfully applied to plasma determinations after oral administration of APL to chicken.