Ventilatory failure and successful management for a dog with severe cervical meningioma

A 12-year-old intact male mongrel dog with a weight of 22 kg was referred with a complaint of progressive tetraparesis. Cervical myelography revealed an intradural-extramedullary mass at the second cervical vertebra. After computed tomography (CT) under general anesthesia, the patient showed dyspnea and cyanosis caused by insufficient movement of the chest wall. Positive pressure ventilation was therefore initiated. Hemilaminectomy and partial mass removal were performed 12 hr after the CT. The mass was histopathologically diagnosed as meningioma. Gradual weaning from the mechanical ventilation lasted for 80 hr after the operation. The patient eventually recovered from the ventilatory failure and the tetraparesis at approximately 6 and 14 days after the operation, respectively.     

Cloning,expression, and tissue distribution of bovine interleukin-21

Bovine interleukin-21 (IL-21) cDNA was cloned and sequenced from bovine peripheral blood lymphocytes (PBLs) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin (PHA), and 50 ng/ml phorbol 12-myristate 13-acetate (PMA) for 48 h. The open reading frame of the bovine IL-21 cDNA is 459 bp in length and encodes 152 amino acids. The predicted amino acid sequence is 78.2 and 58.5% homologous to the human and murine IL-21 amino acid sequences, respectively. Recombinant bovine IL-21 was expressed by a baculovirus expression system. The bovine IL-21 was processed to the mature form in insect cells and secreted to the supernatant confirmed by N-terminal amino acid sequencing. The recombinant bovine mature IL-21 induced the proliferation of human IL-2-dependent cells, ILT-MAT. The mRNA expression for bovine IL-21 was observed in the spleen, but not in the brain, heart, lung, liver, and kidney. The bovine IL-21 identified in this study may provide new methods for the enhancement of innate immunity in cows.     

Large-scale production of porcine mature interleukin-18 (IL-18) in silkworms using a hybrid baculovirus expression system

In this report, a hybrid baculovirus expression system, which means a hybrid virus of the Autographa californica nuclear polyhedrosis virus and the Bombyx mori nuclear polyhedrosis virus, was used for the large-scale production of porcine mature interleukin-18 (IL-18) in silkworms. Two recombinant hybrid baculoviruses containing cDNA of the porcine precursor IL-18 and the porcine caspase-1 were constructed and were used to infect silkworm larvae. After the co-infection of the two viruses, porcine mature IL-18 was efficiently produced in the haemolymph. The concentration of IL-18 in the haemolymph was 80-100 microg/ml, as determined by porcine IL-18 specific ELISA. This yield was twenty-times more than that of the insect cell expression system described previously. The porcine mature IL-18 produced by the silkworms strongly induced interferon-gamma (IFN-gamma) production from porcine PBMC. An insect factory system for the large-scale production of useful cytokines for livestock animals will be available in the near future.     

Use of a Nitrate-Nonutilizing Mutant and Selective Media to Examine Population Dynamics of Fusarium oxysporum f. sp. spinaciae in Soil

ABSTRACT Determining the population density of the spinach wilt pathogen Fusarium oxysporum f. sp. spinaciae in soil with conventional Fusarium-selective media is quite difficult because nonpathogenic strains of F. oxysporum also grow on those media and are indistinguishable from the pathogen. Therefore, a nitrate-nonutilizing (nit) mutant of the pathogen and corresponding selective media were tested in an experimental approach to determine the population density of the pathogen. Colony forming units of the pathogen were countable after soil-dilution plating onto nit mutant-selective media MMCPA, CMP, and CGMBP. Colony forming units of wild-type Fusarium spp. were countable using a wildtype Fusarium-selective medium, GMBP. By combining nit mutant- and wild-type-selective media, the population densities of pathogenic and nonpathogenic F. oxysporum in the same soil could be measured selectively. This method was useful in studying population dynamics of the pathogen after different soil treatments. Soil disinfested with hot water or chloropicrin was amended with the nit mutant pathogen, and subsequent changes in population densities of the pathogen were compared with those in nontreated field soil. The pathogen rapidly proliferated in disinfested soil and wilt developed faster than in nontreated soil. When a nonpathogenic isolate of F. oxysporum was added at high density to sterilized soil prior to the pathogen, growth of the pathogen was greatly suppressed. Nonpathogenic F. oxysporum could not, however, reduce the density of preexisting pathogen.     

Expression of proinflammatory cytokine mRNA in the lymphatic organs of adult and neonatal pigs

Inflammatory cytokine mRNA expression in the lymphatic organs of neonatal, 1-month-old and adult pigs was compared. The mRNA expression of interleukin (IL)-1β, IL-6, IL-18 and tumor necrosis factor (TNF)- in the spleen, thymus, tonsil and popliteal and mesenteric lymph nodes was investigated by semi-quantitative RT-PCR. Stronger IL-1β mRNA expression was observed in the 1-day-old and 1-month-old piglets than in the adult pigs. In thymus, tonsil and mesenteric lymph node, IL-1β mRNA expression in 1-day-old piglets was stronger than in 1-month-old pigs. The expression of IL-6 mRNA in the 1-day-old and 1-month-old tonsil tended to be stronger than in the adult pigs. IL-18 and TNF- mRNA expression was constant in all the samples examined. The expression of IL-1β and IL-6 mRNA may reflect an inflammatory reaction against the exo- and endogenous foreign bodies occurring in the lymphatic organs, especially in the tonsil, of neonatal piglets.     

Identifying novel polymorphisms in the canine tryptophan hydroxylase 2 gene

Tryptophan hydroxylase (TPH) serves as the rate-limiting enzyme in the biosynthesis of serotonin, and two forms of TPH genes, TPH1 and TPH2, have been reported with specific nucleotide sequences and expression patterns. We determined the sequences of these two genes and identified polymorphisms within their coding regions. We identified five novel single nucleotide polymorphisms in the TPH2 gene, one of which resulted in a non-synonymous mutation and the remaining four in synonymous mutations. Comparison of the genotype frequencies of the polymorphisms among five popular breeds with different behavioral traits revealed statistically significant interbreed differences for three polymorphisms. These results suggest that these polymorphisms would be useful genetic markers when performing behavior studies in dogs.     

Sequences of canine glutamate decarboxylase (GAD) 1 and GAD2 genes, and variation of their genetic polymorphisms among five dog breeds

Glutamate decarboxylase (GAD) is the primary enzyme in the brain that catalyzes the synthesis of gamma-aminobutyric acid (GABA), the main inhibitory neurotransmitter. There are two isoforms named according to their molecular weights, GAD67 and GAD65, which are encoded by GAD1 and GAD2, respectively. To investigate the association between GAD genes and temperament in domestic dogs, Canis familiaris, we sequenced the full lengths of the coding regions of these genes and identified three single nucleotide polymorphisms (SNPs) in GAD1 and one in GAD2. When comparing genotype and allele frequencies of SNPs among five breeds with different behavioral traits, statistically significant interbreed differences were observed for three SNPs in GAD1. These results suggest that GAD1 SNPs may be useful for behavioral genetic studies in dogs.     

Estimation of water levels in a main drainage canal in a flat low-lying agricultural area using artificial neural network models

The Chiyoda basin is located in the Saga Prefecture of the Kyushu Island, Japan, and lies next to the tidal compartment of the Chikugo River, into which excess water in the basin is drained away. This basin has a total area of approximately 1100 ha and is a typical flat and low-lying agricultural area. The estimation of the water levels at the gates and along the main drainage canal is a crucial issue that has recently been the subject of much research. At these locations farmers and managers need to control the operation of the irrigation and drainage systems during periods of cultivation. An attempt has been made to apply a feed-forward artificial neural network (FFANN) to model and estimate the water levels in the main drainage canal. The study indicated that the artificial neural network (ANN) could successfully model the complex relationship between rainfall and water levels in this flat and low-lying agricultural area. Input variables and the model structure were selected and optimized by trial and error, and the accuracy of the model was then evaluated by comparing the simulated water levels with the observed ones during an irrigation period in July 2007. The water levels at two locations, located upstream and downstream of a main drainage canal, were investigated by using a time series at intervals of 20, 30, and 60 min. At these intervals, rainfall and tide water levels in the Chikugo River were measured, and the backward time-step numbers of the input variables of rainfall and tide water level were searched. For the upstream location, the optimal combination yielding good agreement between the observed and estimated water levels was obtained when the interval of the time series was 60 min. The number of backward time-steps of the input variables of rainfall and tide water level were 5 and 4, respectively. In contrast to the downstream location, the optimal combination was obtained for the interval time series of 20 min with 4 backward time-steps for both the input variables of rainfall and tide water level. The present study could provide farmers and managers with a useful tool for controlling water distribution in the drainage basin, and reduce the cost of installing water level observation points at many locations in the main drainage canal.