Sensory evaluation and chemical analysis of exhaled and dermally emitted bioeffluents.

Conditions in which exhaled and dermally emitted bioeffluents could be sampled separately or together (whole-body emission) were created. Five lightly dressed males exhaled the air through a mask to another, identical chamber or without a mask to the chamber in which they were sitting; the outdoor air supply rate was the same in both chambers. The carbon dioxide concentration in the chamber with exhaled air was 2000 ppm. Chamber temperatures were 23°C or 28°C, and ozone was present or absent in the supply airflow. When dermally emitted bioeffluents were present, the perceived air quality (PAQ) was less acceptable, and the odor intensity was higher than when only exhaled bioeffluents were present. The presence or absence of exhaled bioeffluents in the unoccupied chamber made no significant difference to sensory assessments. At 28°C and with ozone present, the odor intensity increased and the PAQ was less acceptable in the chambers with whole-body bioeffluents. The concentrations of nonanal, decanal, geranylacetone, and 6-MHO were higher when dermally emitted bioeffluents were present; they increased further when ozone was present. The concentration of squalene then decreased and increased again at 28°C. Dermally emitted bioeffluents seem to play a major role in the sensory nuisance experienced when occupied volumes are inadequately ventilated.

Identification of qRBS1, a QTL involved in resistance to bacterial seedling rot in rice.

Bacterial seedling rot (BSR), a destructive disease of rice (Oryza sativa L.), is caused by the bacterial pathogen Burkholderia glumae. To identify QTLs for resistance to BSR, we conducted a QTL analysis using chromosome segment substitution lines (CSSLs) derived from a cross between Nona Bokra (resistant) and Koshihikari (susceptible). Comparison of the levels of BSR in the CSSLs and their recurrent parent, Koshihikari, revealed that a region on chromosome 10 was associated with resistance. Further genetic analyses using an F5 population derived from a cross between a resistant CSSL and Koshihikari confirmed that a QTL for BSR resistance was located on the short arm of chromosome 10. The Nona Bokra allele was associated with resistance to BSR. Substitution mapping in the Koshihikari genetic background demonstrated that the QTL, here designated as qRBS1 (quantitative trait locus for RESISTANCE TO BACTERIAL SEEDLING ROT 1), was located in a 393-kb interval (based on the Nipponbare reference genome sequence) defined by simple sequence repeat markers RM24930 and RM24944.

Enhanced biofilm formation and 3-chlorobenzoate degrading activity by the bacterial consortium of Burkholderia sp. NK8 and Pseudomonas aeruginosa PAO1.

AIMS: To characterize biofilm formation of a chlorobenzoates (CBs) degrading bacterium, Burkholderia sp. NK8, with another bacterial species, and the biodegradation activity against CBs in the mixed-species biofilm. METHODS AND RESULTS: Burkholderia sp. NK8 was solely or co-cultured with each of five other representative bacteria in microtitre dishes. Biofilm formation involving the strain NK8 was synergistically promoted by co-culturing with only Pseudomonas aeruginosa PAO1. Epifluorescent microscopy revealed that cells of the bacterial strain NK8 were viable and distributed randomly in the mixed-species biofilms. Enumeration of the attached cells on the surface of wells revealed that cells of the strain NK8 increased approx. 10-fold by the co-culture with the strain PAO1 compared to those by monoculture of the strain NK8, and the degradation activity of 3-chlorobenzoate by the dual-species biofilms was more promoted than that by the strain NK8-monocultured biofilms. CONCLUSIONS: Enhanced biofilm formation of Burkholderia sp. NK8 by the bacterial consortium occurred, but is determined by the partner bacterial species. The mixed-species biofilms have the advantage to degrade CBs on a solid surface. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides a significance of bacterial consortia on the biofilm formation and the degradation activity of Burkholderia sp. NK8, which contribute for complete degradation of chlorinated aromatics.

Bone Subtraction Iodine Imaging Using Area Detector CT for Evaluation of Skull Base Invasion by Nasopharyngeal Carcinoma.

BACKGROUND AND PURPOSE: Conventional CT has generally lower detectability of bone marrow invasion than MR imaging due to lower tissue contrast. The purpose of this study was to compare the diagnostic performance of conventional CT alone or in combination with bone subtraction iodine imaging using area detector CT for the evaluation of skull base invasion in patients with nasopharyngeal carcinoma. MATERIALS AND METHODS: Forty-four consecutive patients who underwent contrast-enhanced CT using 320-row area detector CT and contrast-enhanced MR imaging for nasopharyngeal carcinoma staging between April 2012 and November 2017 were enrolled in this retrospective study. Bone subtraction iodine images were generated by subtracting pre- and postcontrast volume scans using a high-resolution deformable registration algorithm. Two blinded observers evaluated skull base invasion at multiple sites (sphenoid body, clivus, bilateral base of the pterygoid process, and petrous bone) using conventional CT images alone or in combination with bone subtraction iodine images. Examination of MR and CT images by an experienced neuroradiologist was the reference standard for evaluating sensitivity, specificity, and area under the receiver operating characteristic curve. RESULTS: Twenty-six patients (59%) showed skull base invasion at 84 sites on the reference standard. Conventional CT plus bone subtraction iodine images showed higher sensitivity (92.9% versus 78.6%, P = .02) and specificity (95.6% versus 86.1%, P = .01) than conventional CT images alone for evaluating skull base invasion. The area under the receiver operating characteristic curve for conventional CT plus bone subtraction iodine (0.98) was significantly larger (P < .001) than the area under the receiver operating characteristic curve for conventional CT alone (0.90). CONCLUSIONS: Conventional CT plus bone subtraction iodine performs more closely to the accuracy of combining CT and MR imaging compared with conventional CT alone.

Biological and phylogenetic characterization of Fusarium oxysporum complex, which causes yellows on Brassica spp., and proposal of F. oxysporum f. sp. rapae, a novel forma specialis pathogenic on B. rapa in Japan.

Although the causal agent of yellows of Brassica rapa (turnip, pak choi, and narinosa) in Japan was reported in 1996 to be Fusarium oxysporum f. sp. conglutinans, this classification has remained inconclusive because of a lack of detailed genetic and pathogenic studies. Therefore, we analyzed the taxonomic position of this organism using Japanese isolates of F. oxysporum complex obtained from diseased individuals of various B. rapa subspecies. Phylogenetic analyses using partial sequences of the rDNA intergenic spacer region and the mating-type gene (MAT1-1-1alpha-box) showed that B. rapa and cabbage isolates belong to different monophyletic clades that separated at early evolutionary stages. Additionally, correlations were observed between the molecular phylogeny and the vegetative compatibility groups. Isolates from turnip, komatsuna, and narinosa (B. rapa group) did not show pathogenicity against cabbage or broccoli (B. oleracea group), although they caused severe symptoms on their original host species. In contrast, cabbage isolates had significantly higher (P = 0.05) virulence on B. oleracea than on B. rapa crops. Our results indicate that F. oxysporum complex isolates from B. rapa and B. oleracea are not only phylogenetically distinct but also differ in host specificity. Therefore, we propose a novel forma specialis, F. oxysporum f. sp. rapae, which causes yellows on B. rapa, including turnip, komatsuna, pak choi, and narinosa.

Induction of differentiation of cultured human and mouse myeloid leukemia cells by alkyl-lysophospholipids.

Alkyl-lysophospholipids are synthetic analogs of naturally occurring lysophospholipids. The effects of these compounds on cell proliferation and differentiation of cultured human (HL-60) and mouse (M1) myeloid leukemia cells were studied. Both cell lines were induced to differentiate into morphologically and functionally mature granulocytes and macrophages by incubation with a wide variety of these compounds. Some alkyl-lysophospholipids induced differentiation (judged morphologically and by the appearance of abilities to reduce nitro blue tetrazolium, to phagocytize latex particles, and to induce lysozyme activity) of both the cells lines at concentrations of 1 microgram/ml. However, these compounds did not affect colony formation of normal mouse bone marrow cells even at a higher concentration, 20 microgram/ml. These results suggest that alkyl-lysophospholipids induce cell differentiation of myeloid leukemia cells without affecting proliferation and differentiation of normal bone marrow cells. Thus, these compounds could be useful in therapy of myeloid leukemia.

Spectroscopic evidence for selenium(IV) dimerization in aqueous solution.

The aqueous speciation of selenium(iv) was elucidated by a combined approach applying quantum chemical calculations, infrared (IR), Raman, and (77)Se NMR spectroscopy. The dimerization of hydrogen selenite (HSeO3(-)) was confirmed at concentrations above 10 mmol L(-1) by both IR and NMR spectroscopy. Quantum chemical calculations provided the assignment of vibrational bands observed to specific molecular modes of the (HSeO3)2(2-) ion. The results presented will provide a better understanding of the chemistry of aqueous Se(iv) which is of particular interest for processes occurring at mineral/water interfaces.

Platelet activating factor antagonists: synthesis and structure-activity studies of novel PAF analogues modified in the phosphorylcholine moiety.

New analogues of platelet activating factor (PAF), in which the phosphate and trimethylammonium moieties were replaced with an acylcarbamoyl moiety and a quaternary cyclic ammonium group, were synthesized. Their biological activities as PAF antagonists were evaluated by the inhibition of PAF-induced rabbit platelet aggregation in vitro and protective effects on PAF-induced hypotension in rats and PAF-induced death in mice. Investigation of structure-activity relationships revealed that PAF antagonist activity is strongly influenced by the acyl substituent of the nitrogen atom on the carbamoyl group and the nature of the polar head group at the 3-position of the glycerin backbone. Among the compounds tested, 2-[[N-acetyl-N-[[2-methoxy-3-[ (octadecylcarbamoyl)oxy]propoxy]-carbonyl]amino] methyl]-1-ethylpyridinium chloride (21, CV-6209) was one of the most potent compounds in the in vitro assay (IC50 = 7.5 X 10(-8) M) and the most potent and long-lasting in the in vivo assays. (R)-(-)-21 and (S)-(+)-21 were also synthesized, and no significant differences were observed in PAF antagonist activity in vitro and an inhibitory effect on PAF induced hypotension in vivo between (RS)-21 and its enantiomers.

Activation of guinea pig peritoneal macrophages by platelet activating factor (PAF) and its agonists.

The platelet activating factor (PAF: 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine) and its analogs were examined to determine their effects on guinea pig peritoneal macrophages. PAF activated macrophages, but its effect on macrophages was much weaker than that observed on platelets: the concentration required for 50% maximum activation was 8.5 X 10(-6) M for macrophages and 2.9 X 10(-10) M for platelets. Three PAF agonists, 1-O-octadecyl-2-O-(N,N-dimethylcarbamoyl)-glycero-3-phosphocholine (Compound I), 1-O-octadecyl-2-acetamido-2-deoxy-glycero-3-phosphocholine (Compound II), and 1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine (Compound III), showed higher activity in stimulating macrophage function than PAF. The abilities of these non-metabolizable PAF agonists to activate macrophage paralleled their relative potency to induce platelet activation. The sn-3 enantiomers of PAF and Compound III exhibited activity, while the sn-1 did not. By comparing the activities of derivatives of Compound III, it was shown that the long-chain alkyl-ether group in the glycerol-1 position, a relatively small size of the substituent on the hydroxy group at the sn-2 position, and the choline moiety in the glycerol-3 position must play critical roles in the process of macrophage activation. A specific PAF antagonist, CV3988, which inhibits PAF-induced platelet activation and hypotension, inhibited the activation of macrophages caused by PAF and its agonists.(ABSTRACT TRUNCATED AT 250 WORDS)

Antileukemic effect of alkyl phospholipids. I. Inhibition of proliferation and induction of differentiation of cultured myeloid leukemia cells by alkyl ethyleneglycophospholipids.

Various alkyl ethyleneglycophospholipids, i.e., alkyl phospholipids, with ethyleneglycol or its congener in place of glycerol as a molecular backcone, were synthesized and their effects on cell proliferation and differentiation of cultured human (HL-60) and mosue (Ml) myeloid leukemia cells were studied. On incubation with alkyl ethyleneglycophospholipids, proliferation of both cell lines was inhibited and the cells were induced to differentiate into morphologically and functionally mature granulocytes and macrophages. Among the compounds tested, dodecyl ethyleneglycophospholipid with a pyridinioethyl group was the most effective in induction of differentiation of both cell lines.

Antileukemic effect of alkyl phospholipids. II. Prolongation of survival times of leukemic mice by alkyl ethyleneglycophospholipids.

Alkyl ethyleneglycophospholipids induced differentiation in vitro of mouse myeloid leukemia M1 cells into mature granulocytes and macrophages. The compounds also prolonged the survival of syngeneic SL mice inoculated with M1 cells. Although in mice with florid leukemia these compounds alone scarcely affected survival, administration of dodecyl ethyleneglycophospholipid with pyridinioethyl as a polar group plus actinomycin D significantly prolonged survival.

CV-3988 - a specific antagonist of platelet activating factor (PAF).

CV-3988, rac-3-(N-n-octadecylcarbamoyloxy)-2-methoxypropyl 2-thiazolioethyl phosphate was shown to be a specific inhibitor of platelet activating factor (PAF). This compound in concentrations of 3 x 10(-6) to 3 x 10(-5)M inhibited aggregation of rabbit platelets induced by PAF (3 x 10(-8)M), while it had no effect on the aggregation induced by arachidonic acid, ADP, collagen or A-23187. CV-3988 alone even at a concentration of 10(-3)M had no effect on platelet aggregation. The inhibitory action of CV-3988 on the PAF-induced aggregation was independent of the formation of micelles. The PAF (0.1 to 1.0 micrograms/kg, i.v.)-induced hypotension in anesthetized rats was also inhibited dose-dependently by the i.v. administration of CV-3988 (1 and 10 mg/kg), while the hypotensive actions induced by the i.v. administration of acetylcholine (1 micrograms/kg), arachidonic acid (1 mg/kg), bradykinin (10 micrograms/kg), isoproterenol (1 microgram/kg) and histamine (100 micrograms/kg) were not altered by CV-3988 (10 mg/kg, i.v.). All these findings indicate that CV-3988 specifically inhibits the action of PAF in vitro and in vivo. This is the first report of a PAF antagonist which can specifically inhibit the PAF-induced hypotension as well as the PAF-induced platelet aggregation.

Reappraisal of single-sample and gamma camera methods for determination of the glomerular filtration rate with 99mTc-DTPA.

The aim of this study was to assess the clinical validity of single-sample methods and gamma camera uptake methods with 99mTc-DTPA for the estimation of the glomerular filtration rate (GFR). The study was performed in 50 patients with various degrees of renal function (29 males and 21 females; age 27-90 yrs; serum creatinine level 0.34-6.49 mg/dl). As a reference the "true" GFR (GFRt) was determined from plasma clearance by means of the two compartment model curve fitting 10 plasma samples. The GFRt of more than 30 ml/min in 46 patients was compared to the GFR which was estimated with 7 single-sample methods, two gamma camera uptake methods and 24-hour endogenous creatinine clearance (24hCcr). Close correlation was observed in all single-sample methods. The highest linear correlation was observed in the Christensen and Groth's method rewritten by Watson for a 180-min plasma sample (r = 0.991, see = 5.84 ml/min). The smallest random error was observed in the Groth and Aasted's method for a 180-min plasma (r = 0.989, see = 4.31 ml/min/1.73 m2). Our method was lowest in % absolute difference analysis (mean = 4.10%). The gamma camera uptake methods correlated significantly with the GFRt (r = 0.746-0.774), but were less reliable than any of the single-sample methods (see = 15.41 ml/min-19.14 ml/min). The lowest correlation was observed in the 24hCcr (r = 0.698, see = 50.76 ml/min/1.73 m2). The single-sample method was more accurate than the gamma camera method, and the gamma camera method was more accurate than 24hCcr. The single-sample method should be recommended for the accurate determination of the GFR with 99mTc-DTPA in a patient with mild to moderate renal dysfunction.

[Participation of regional inhabitants in cancer screening].

As part of the strategic planning for increasing participation by community residents in cancer screening, a survey about life style and attitudes toward cancer screening was carried out in Samukawa town, Kanagawa Prefecture. Subjects were males 40 years old or over, and females 30 years old or over selected by random sampling from residents. Number of subjects was 1,916 (822 males, 1,094 females). Self administered anonymous questionnaires were sent and returned by mail. The response rate was 73%. The results were as follows: (1) The reported participation rates for cancer screening under "the Health and Medical Services Law for the Aged" did not reflect the actual participation rate of residents. (2) The major reason for not participation in screening was lack of interest in cancer prevention. (3) Socio-economic status of regular participants in screening was different from persons who did not participate. (4) Persons who did not participate in screening were more passive with regard to obtaining knowledge of cancer than regular participants. (5) Results of this study show that cancer prevention education at various levels such as by clinical offices, TV and radio programs, and newspapers may be effective. In order to increase participation in screening programs, health education that considers the life style characteristics of persons who do not participate in the screening programs is vitally important.

[Single-blood-sample method for the determination of glomerular filtration rate using 99mTc-DTPA in Japanese].

The study was aimed to evaluate glomerular filtration rate with 99mTc-DTPA by means of a single-plasma-sample method in Japanese. Three were 50 patients (29 males and 21 females, age range being 25 to 91 years) with diabetes mellitus of various degree of the renal function. 99mTc-DTPA of 300 MBq/2 ml that was prepared in our hospital was injected. Dynamic renal scintigraphy was carried out and 10 blood samples were taken after the injection. Blood clearance (true GFR) of 99mTc-DTPA was determined from plasma concentration which was fitted to the biexponential curve by a non-linear least squares method. Plasma concentration (ml/min/1.73 m2) after 75 min-post injection showed very higher linear regression and linear correlation with true GFR than 0.900. The best linear regression and linear correlation was observed with 180 min-sample (r = 0.989). As general, the following equation was obtained; Y = A + Bln(X), A = 436.1217 - 3.45817t + 0.01205t2 - 0.000015t3, B = -212.601 + 1.42518t - 0.04834t2 + 0.0000062t3, X = plasma concentration at sampling time t (%ID/L/1.73 m2), t = sampling time from 75 to 300 min (r > 0.900). This is the new equation of single-plasma-sample method for the determination of GFR using 99mTc-DTPA for Japanese. The clinical feasibility should be accessed in a field of a nuclear medicine practice.

[Remyelination by Schwann cell transplantation for CNS demyelinated axons: functional comparison with developmental myelination].

Transplantation of Schwann cells(SCs) induced remyelination of demyelinated rat dorsal column(DC) axons and improved conduction. To investigate the difference between developmental oligodendrocytic myelination and SC myelination in conductive functions of axons, we compared normally developmental DCs(10 days old, 22-23 days old, and adult rat), demyelinated DCs, and remyelinated DCs by SC transplantation. DCs of adult rats were demyelinated at T 11 by X-ray irradiation and ethidium bromide, and transplanted with SCs(3 x 10(4)) of adult rats. Three weeks later, the spinal cord was removed and pinned in a recording chamber and compound action potentials (CAPs) were recorded, to investigate conduction properties(conduction velocity, amplitude of CAP and response after high frequency stimulation). Normal DCs were recorded in same manner. Following transplantation of SCs, histological examination revealed SC-like patterns of remyelination of demyelinated axons. The conduction velocities of 10 days old DCs(1.7 +/- 3.4 m/s, n = 4) increased early to 8.1 +/- 3.3 m/s(22-23 days old, n = 7) and 12.2 +/- 1.5 m/s (adult, n = 5). After SC transplantation the velocity significantly improved 7.7 +/- 1.5 m/s(n = 5) compared to demyelinated axons(1.2 +/- 0.4 m/s, n = 7), but less than adult. A 600 Hz 0.5 sec stimulus train led to an amplitude decrement of 7.1 +/- 7.5%(n = 7) in demyelinated axons. Following transplantation, amplitude decreased in 66.2 +/- 11.9%(SC, n = 5), same as 22-23 days(71.7 +/- 11.7%, n = 7) old or adult(76.6 +/- 15.6%, n = 6). The recovery properties after high frequency stimulation developed earlier than conduction velocity. Following SC transplantation, the recovery properties improved to that of normal adult, but not conduction velocity. Lower conduction velocity may be due to shorter internode distance of remyelination after transplantation, same as 22-23 days old pups, than normal adult rats. Though anatomical difference and/or time after transplantation influenced the conduction, these result suggested SC remyelination might result in insufficient for conduction velocity.

[Production of human monoclonal autoantibodies that react with islet-cell-surface antigens by EBV-transformed lymphocytes: new method of preselection of ICSA producing lymphocyte by protein A magnetic microspheres].

Because of the low titer of ICSA, as well as the polyclonality of serum, it has been difficult to determine whether there is a variety of autoantibodies, each reacting with a different organ, or whether the autoimmune response is more restricted, with only a limited number of autoantibodies recognizing common antigenic determinants in the different organs. To study this and other questions, we sought to prepare human monoclonal autoimmune ICSA. Peripheral-blood lymphocytes from patients with freshly onset IDDM or other autoimmune abnormalities were transformed by EB virus. And rapid screening on In 111 cells by an ELISA were used to identify monoclonal ICSA. But, ICSA titer was very low even in freshly onset IDDM patients, and reduced very quickly. ICSA positive wells at cloning were extremely rare. So, the methods that enhance the efficiency of cloning were required. Firstly, ultrasensitive Enzyme Immunoassay for ICSA were developed. Secondly, new preselection method of ICSA producing lymphocyte by Protein A Magnetic Microspheres were employed. Thirdly, in vitro immunization was partly employed to enhance the frequency of ICSA producing lymphocytes. Fourthly, EBV-hybridoma technique was employed to stabilize the ICSA producing clone. The efficiency of cloning was increased from 0.07% to 24% by the new preselection method. The sensitivity of the fluorometric enzyme immunoassay for ICSA was 140pg/ml. 21 times sensitivity to usual ELISA was obtained. We describe sure methods for production of Human Monoclonal Autoantibodies that react with Islet-Cell-Surface Antigens.