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It is known that different breeds of cattle display differential susceptibilities to Trypanosome congolense infections, and that N'Dama cattle remain more productive after infection than Boran cattle which are more susceptible to T. congolense. Macrophages from both breeds were cultured in vitro and the expressions of a number of cytokines and iNOS mRNA were analyzed using real time RT-PCR after stimulation with antibody-opsonized trypanosomes. No significant difference was seen between the responses of the two breeds. However, RNA levels of TNF-alpha in the IFN-gamma-primed macrophages were about 100-fold higher than those in the non-primed macrophages. A significant ten-fold decrease was seen for the anti-inflammatory cytokine IL-10. These results indicate that priming of the cells with IFN-gamma cause a serious shift toward an inflammatory response.

To estimate canine interleukin-8 (cIL-8) levels in blood plasma samples, a sandwich enzyme linked immunosorbent assay (ELISA) was established. For the development of the sandwich ELISA, polyclonal anti-cIL-8 (capturing), biotinylated anti-cIL-8 (developing) antibodies and glutathione-S-transferase/cIL-8 (GST/cIL-8) fusion protein as an antigen were used. cIL-8 in the fusion protein of GST/cIL-8 was detected in a dose dependent manner. The lowest limit of GST/cIL-8 detectable by this method was 2 ng/ml of GST/cIL-8 (containing; 0.470 ng/ml of cIL-8). IL-8 levels in the plasma samples from apparently healthy dogs were less than 0.470 ng/ ml. Higher levels of IL-8 were detected in the plasma samples of dogs with cystitis, dermatitis, and gastric cancer. These results suggest that the determination of cIL-8 by the sandwich ELISA is useful in diagnosis of inflammatory diseases in dogs.

In this study, the authors measured the ratio of the peak area of photoelectrons from s-orbitals to that of the p-orbitals for six elements (Al, Si, Cu, Mo, Ag and Au) resulting in eight s : p ratios. We used two angles (90° and 54.7°) between the incident X-ray beam and the photoelectron detector to investigate elastic scattering effects on the distribution of photoemission from the samples. The data obtained were compared to calculated values. When elastic scattering effects are accounted for, the ratio of s- to p-orbitals by XPS should be independent of angle. The experimental results from five of the eight elemental ratios showed this angular independence. We believe that other effects, such as electron diffraction, will be need to be considered in order to get complete agreement.

It has been reported that the concentrations of both pyrraline and pentosidine, well-characterized advanced glycation end products, are increased in the urine of diabetic patients. To determine factors that influence the urinary excretion of pyrraline or pentosidine, we compared pyrraline or pentosidine concentrations with glycemic-control indexes, urinary albumin excretion, and urinary beta2-microglobulin in patients with type 2 diabetes. The study was conducted in 39 age-matched healthy control subjects and 50 diabetic patients, including 22 patients with normoalbuminuria, 15 with microalbuminuria, and 13 with macroalbuminuria. Both urinary pyrraline and pentosidine were measured in early-morning urine specimens with the use of high-pressure liquid chromatography. The urinary pentosidine concentration was significantly higher in diabetic patients than in control subjects (P

A pig interleukin-21 (IL-21) cDNA was successfully cloned and sequenced from porcine peripheral blood lymphocytes (PBL) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin P (PHA), 50 ng/ml phorbol 12-myristate 13-acetate (PMA), and 0.5 microg/ml anti-porcine CD3 antibody for 48 hr. The open reading frame of the porcine IL-21 cDNA is 459 base pairs in length and encodes 152 amino acids. The predicted amino acid sequence of the porcine IL-21 shows 86.2%, 77.7%, and 58.4% identity to the bovine, human, and murine IL-21, respectively. The porcine IL-21 gene was mapped to porcine chromosome 8 (8q22--q23) by means of fluorescence in situ hybridization and radiation hybrid mapping, where the porcine IL-2 gene had been mapped nearby. The recombinant porcine mature IL-21 expressed by E. coli induced dose-dependent proliferation and IFN-gamma production from a human NK cell line, NK0. The porcine IL-21 identified in this study will be helpful for the enhancement of innate immune responses of pigs.

Antibody responses to the hapten dinitrophenyl (DNP) and the concentration of catecholamine in chickens received a single injection of 6-Hydroxydopamine (6-OHDA) into the chorioallantoic cavity at the embryonal stage were evaluated. A significantly enhanced anti-DNP IgA response and a markedly decreased level of noradrenaline in the peripheral blood were observed in chickens treated with 400 micrograms of 6-OHDA at 14 days of incubation. These results suggested the immunomodulatory influence of the sympathetic nervous system.

We established an enzyme-linked immunosorbent assay (ELISA) system for the quantitation of bovine macrophage colony-stimulating factor (M-CSF) and used it to measure the serum M-CSF levels in bovine fetuses and calves. The average serum M-CSF level was 2.7+/-1.5 ng/ml in 39 calves under 100 days old, and 1.8+/-0.8 ng/ml in 15 cattle between 101 and 418 days old. Fetal sera samples (n = 6) prepared from cattle between 150 and 280 days of gestational age had a higher average level of M-CSF (8.8+/-1.4 ng/ml). Alteration in serum M-CSF levels in each individual calf was also measured. The serum levels of M-CSF in calves at 0-1 day after birth ranged from 0.52 to 7.3 ng/ml. During the period 113-125 days after birth, serum levels were around 1.4+/-0.39 ng/ml. Although serum M-CSF levels generally decreased as the age of calves advanced, differences among individuals, especially among newborn calves, were observed.

Increased production of advanced glycosylation end products (AGEs) and augmented oxidative stress may contribute to vascular complications in diabetes. Little is known about the formation and accumulation of AGEs in young patients with type 1 diabetes. The aim of the present study was to investigate whether AGE production and oxidative stress are augmented in young patients with type 1 diabetes at early clinical stages of the disease. Urine samples of 38 patients with type 1 diabetes [mean age (+/-SD), 12.8 +/- 4.5 y; diabetes duration, 5.7 +/- 4.3 y; HbA1c, 8.0 +/- 1.6%; urinary albumin excretion, 12.6 +/- 14.4 mg/g creatinine (Cr)] and those of 60 age-matched healthy control subjects were assayed for AGEs, pentosidine and pyrraline, and markers of oxidative stress, 8-hydroxy-2'-deoxyguanosine (8-OHdG) and acrolein-lysine. Of these four markers, urinary concentrations of pentosidine, 8-OHdG, and acrolein-lysine were significantly higher in the patients with diabetes than in the healthy control subjects. For the patient group, pentosidine correlated significantly with 8-OHdG and acrolein-lysine, and pyrraline correlated significantly with acrolein-lysine. Urinary pentosidine, 8-OHdG, and acrolein-lysine but not pyrraline correlated significantly with urinary albumin excretion. Patients with microalbuminuria (> or =15 mg/g Cr) showed significantly higher levels of all four markers than did normoalbuminuric patients and control subjects. The present study indicates that accumulation of AGEs, whose formation is closely linked to oxidative stress, and resultant endothelial dysfunction may start early in the course of type 1 diabetes. This means that the risk of vascular complications may be present at an early age and that the best possible glycemic control should be emphasized from the diagnosis of diabetes.

Self-organizing maps (SOM) method was developed by Kohonen (1997) was first applied to the problems of chemical analysis, such as AES (Auger electron spectroscopy), XPS (X-ray photoelectron spectroscopy), and XRD (X-ray diffraction) data. Using the constructed 2-dimensional SOM, it became clear that the things which are described quantitatively by linguistic expressions can be explained more quantitatively by the position of the spectra data on the SOM as well as with a gray level expression. Also, the composition of the unknown spectra can be determined very precisely by the SOM, which is constructed using the composition of the known spectra.

An extremely high vacuum system has been developed in order to transfer specimens long-distance without any contamination of the clean surface. The syytem is composed of vacuum chambers, vacuum pumps, vacuum gauges, valves and specimen transports. All components exposed to the extremely high vacuum environment of the main chamber use metal sealings for high temperature bakeout and this can reduce outgassing remarkably. A tantalum getter pump and a bakeable cryopump evacuate the main chamber. The main chamber has achieved vacuum of pressure 10 −10 Pa. Magnetic levitation transport system installed in the main chamber can transfer specimens in vacuum of less than 10 −8 Pa without any contamination because it has no sliding part.

A simple and rapid method for the determining arsenate and phosphate in rainwater is proposed. These compounds form molybdoarsenate– and molybdophosphate–Malachite Green aggregates with MoMG reagent (MoMG = a mixture of ammouium molybdate and Malachite Green). The aggregates were first collected on a nitrocellulose membrane filter (pore size 3μm) and then dissolved with the filter in a small amount of 2-methoxyethanol. Absorbance (627 nm), denoted as A(As + P), was found to be proportional to the sum of the arsenate and phosphate concentrations with a molar absorptivity of 2.8 × 105 (mol dm−3)−1 cm−1. Following the addition of thiosulphate, an agent which reduces arsenate, the absorbance A(P) was measured as described above. A(P) corresponded to the concentration of phosphate alone and A(As + P)–A(P), to arsenate concentration only. This method is shown to be a rapid and highly accurate means for determining arsenate and phosphate levels within the 1–150 nmol dm−3 concentration range.

Stimulation and modulation of tumor necrosis factor-alpha (TNF-alpha) production during treatment of the murine macrophage-like cell line J774/JA-4 with 25 oligosaccharides were studied. Direct stimulation of TNF-alpha production by oligosaccharides was measured with a cytotoxic assay using the L929 cell line. Twelve samples showed a significantly higher production (Por = 0.01) of TNF-alpha than the controls. Modulation of TNF-alpha production by treatment with oligosaccharides, followed by stimulation with lipopolysaccharide (LPS) from E. coli, was examined using the L929 bioassay system. In three samples TNF-alpha production increased significantly, but in four samples, production was reduced significantly (Por = 0.01). No samples showed modulation of growth or viability of L929 cells within the first 26 hr. The present results are useful in the application of these oligosaccharides which is potentially applicable in medical and food technology.

Molecular cloning of canine interleukin-8 (IL-8) was performed to establish a basis for its investigation in the canine immune system. From a cDNA pool constructed from LPS-stimulated popliteal lymph node cells, canine IL-8 cDNA covering the whole coding region was amplified by polymerase chain reaction. The nucleotide sequence of a canine IL-8 clone, designated pcIL-8#38, was highly similar to those of human, rabbit and porcine IL-8, and comprised 353 bp with an open reading frame that encoded 101 amino acids. Analysis of the deduced amino acid sequence of insert DNA in pcIL-8#38 showed 76.5, 80.2, and 87.0% similarities with human, rabbit and porcine IL-8 proteins, respectively. Insert DNA of pcIL-8#38 was transferred to a mammalian expression vector, pcDL-SRα296, and transfected into Cos7 cells. The supernatant of the transfectant had neutrophil chemotactic activity when it was examined by the neutrophil migration assay, suggesting that our cloned cDNA was biologically active. The cloned canine IL-8 cDNA will be useful for canine inflammatory disease and comparative immunology research.

Hydroxylysylpyridinoline (HP), an intermolecular cross-linking amino acid, is present only in bone and cartilage collagen. We examined urinary HP levels in order to assess bone collagen metabolism using HPLC. Urinary HP excretion levels (nmol/mmol creatinine) of healthy control groups gradually increased with aging. Urinary HP excretion levels of females were higher than those of age-adjusted males. On the other hand, the urinary excretion levels of HP in osteoporotic patients were significantly higher than those of age and sex-adjusted controls. It is suggested that the determination of urinary HP excretion levels is useful in the clinical evaluation of metabolic bone diseases, especially osteoporosis.

The urinary excretion levels of hydroxylysine glycosides [Hyl-Gs; O-beta-D-Galactopyranosylhydroxylysine (GH) and 2-O-alpha-D-glucopyranosyl-O-beta-D-galactopyranosylhydroxylysine (GGH)], provide a new index of collagen metabolism. The determination of urinary hydroxylysine glycoside excretion levels was applied to 20 osteoporotic patients and 208 healthy control subjects (69 females and 139 males) in order to evaluate the conditions of bone collagen metabolism. Urinary Hyl-Gs were analyzed by the method of pre-column fluorescent derivatized HPLC. Compared to the age and sex-adjusted control, the urinary excretion levels of hydroxylysine glycosides, especially GH, in osteoporotic patients were significantly higher than in the age and sex-adjusted control, and the urinary GGH/GH ratio was lower. Sixteen of the 20 patients whose Hyl-Gs levels we followed exhibited significantly lower values after 200 d compared to the initial levels. These results suggest that clinical therapeutics affected the bone resorption and, therefore, demonstrated the usefulness of determining urinary excretion levels of Hyl-Gs for evaluating the conditions of bone metabolism.

The surface oxide layer of fine iron particles prepared by the vaporization method was investigated by X-ray diffraction (XRD), electron diffraction (ED), Mossbauer spectroscopy, X-ray photoelectron spectroscopy (XPS), infrared (IR) absorption measurement and high resolution transmission electron microscopic (HR-TEM) observation. The thickness of the surface oxide layer was about 5 nm and the component from outside to inner core was characterized as amorphous-FeOOH/γ-Fe2O3 and Fe3O4/Fe. The existence of FeO at the metal/oxide interface was not confirmed.

The possibility that oxidative stress caused by chemotherapy would accelerate formation of pentosidine was investigated. The pentosidine levels of erythrocyte membrane protein, plasma, and urine of pulmonary cancer in patients who had been administered cisplatin were higher than those of unadministered patients. Injection of cisplatin to SD mice resulted in a gradual increase in pentosidine levels. These results suggest that oxidative stress caused by cisplatin accelerates pentosidine formation in vivo.

A method has been developed for rapid analysis of the urinary hydroxylysine glycosides [glucosyl-galactosyl-hydroxylysine (GGH) and galactosyl-hydroxylysine (GH)] as collagen metabolites. The glycosides were fluorometrically derivatized with 5-dimethylaminonaphthalene-1-sulfonyl chloride, and then subjected to reversed-phase high-performance liquid chromatography using a gradient system. Suitable standard hydroxylysine glycosides which were not diastereoisomers were prepared from human urine. Urinary GGH and GH were analyzed within 1 h, including column washing. This method was applied to the urine samples of 68 healthy persons of different ages.

Silicon bipolar 4:1 multiplexer and 1:4 demultiplexer ICs with retiming functions, and a 1/64 prescaler IC with a comparator for a phase lock loop are reported. All clock dividers are static so that the ICs operate at any data rate from DC to over 10 Gb/s. The output data of the multiplexer and the input data of the demultiplexer are retimed by the data-rate clock at a flip-flop that minimizes multiplexer output pattern jitter and enlarges demultiplexer input phase margin. The ICs use a Si bipolar process featuring U-groove isolated double poly-silicon transistors on a SOI substrate, highly accurate poly-silicon resistors, MOS capacitors, and three levels of metal interconnects, including surface gold metal for packaging. Packaged ICs operate above 10.5 Gb/s, 12.0 Gb/s, and 16.0 GHz for the multiplexer, demultiplexer, and prescaler, respectively. >

A digital video recording system using a four-beam magneto-optical disk drive has been developed. In order to realize continuous parallel read/write operation on an ordinary single-spiral disk, periodic single track jumping during one disk rotation has been employed. Digital video signal recording has been carried out on the entire disk.© (1990) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

High track density magneto-optical recording, using a crosstalk canceller which eliminates crosstalk from two adjacent tracks, has been developed. By this recording method, both land area and groove area can be used for data storage tracks, and twice the track density, cornpared with the conventional method, has been achieved. Applying this crosstalk canceller also realizes intersymbol interference compensation.

Work function variation with surface segregation of underlayer elements was studied to enable the utilization of saturation behavior in segregation phenomena for work function control. Surface composition of Nb film/Ti film/Cu substrate was monitored by X-ray photoelectron spectroscopy (XPS) at elevated temperatures. Cu and Ti segregated after a certain period of time; the surface concentration of Cu saturated first followed by that of Ti. Both Cu and Ti were observed only at the surface region less than 1nm thick. Using a Kelvin probe, a monotonous work function decrease with increasing Cu and Ti concentration was observed. The amount of decrease at Cu saturation (Cu: 5 at.%, Ti: 20 at.%) was 0.2 eV and that at Ti saturation (Cu: 5 at.%, Ti: 30 at.%) was 0.3 eV. It was determined that the sample worked as a getter for residual gas and the work function on the surface with saturated segregation was stable.

Si nanocrystals were formed in a SiO2 thin film by irradiation of a high-intensity electron beam at about 900 K inside an ultrahigh vacuum field emission transmission electron microscope (UHV-FE-TEM). The structure of the nanocrystals was investigated by high-resolution transmission electron microscopy (HRTEM) and electron diffraction analysis. Relations among the lattice distances suggest that the structure is not a diamond structure. For the structural model, a beta-tin structure with elongation along the c-axis and shrinkage along the a-axes compared to the diamond structure is proposed.

The purpose of the present study was to conduct a basic work of the tr ace analysis of phospholipids in biological urine and serum samples. Phospholipids were extracted with chloroform using Extrelut column and developed by TLC. Solutions of PL-C and Al-P were sprayed on the TLC plate (silica gel), and released phosphate was detected with molybdate-Malachite Green solution. This method can be easily used for the separation and determination of phospholipid content as low as pmol.

The microwave surface resistance of YBaCuO thick films was measured using a demountable copper cavity operated at 3 GHz in the TE011 mode over a temperature range from 4.2 K to 300 K. The films were deposited onto end plates and cylinder walls of silver substrates or nickel-buffered copper substrates by a low-pressure plasma-spraying technique combined with a melt-reaction method. The area of the end plate was 177 cm2 and that of the cylinder wall was 396 cm2. The surface resistance was obtained from the Q values measured before and after substituting either the end plate or the cylinder wall for one covered with the film. The ratio of the surface resistance of the film to that of the copper was lower than 0.08 at 5.2 K and 0.65 at 77.3 K for the end plate and was 0.15 at 5.1 K and 1.91 at 77.3 K for the cylinder wall.

The adsorption of hydrogen on Ni and NiTiO 2 composite ultrafine particles (UFPs) was studied by temperature-programmed desorption (TPD) experiments. The desorption characteristics of hydrogen on Ni and NiTiO 2 composite UFPs were similar to those of a Ni single crystal or polycrystal. The amount of adsorption of hydrogen on NiTiO 2 composite UFPs was dependent upon the temperature of exposure, and the phenomenon attributed to the spillover of hydrogen from Ni onto TiO 2 . The CO hydrogenation reaction on NiTiO 2 composite UFPs proceeded above 373 K. TPD spectra showed new desorption peaks after the catalytic reaction above 373 K and suggested the formation of new adsorption states.

A free-space optical bus, which consists of cascaded optical switch devices, is proposed. The switch devices have multiple functions, such as data transmission, data detection, and data repetition. Basic interconnection characteristics were measured with vertical-to-surface transmission electrophotonic devices. Optical switching energy as low as 150 FJ and up to 4 MHz and a relatively high data-transmission rate, i.e., more than 20 MHz, were obtained. Connection cascadability and extendibility were confirmed.