Effect of hypoxia on generation of neurospheres from adipose tissue-derived canine mesenchymal stromal cells

Adipose tissue-derived mesenchymal stromal cells (AT-MSCs) are good candidates for cell therapy due to the accessibility of fat tissue and the abundance of AT-MSCs therein. Neurospheres are free-floating spherical condensations of cells with neural stem/progenitor cell (NSPC) characteristics that can be derived from AT-MSCs. The aims of this study were to examine the influence of oxygen (O₂) tension on generation of neurospheres from canine AT-MSCs (AT-cMSCs) and to develop a hypoxic cell culture system to enhance the survival and therapeutic benefit of generated neurospheres.AT-cMSCs were cultured under varying oxygen tensions (1%, 5% and 21%) in a neurosphere culture system. Neurosphere number and area were evaluated and NSPC markers were quantified using real-time quantitative PCR (qPCR). Effects of oxygen on neurosphere expression of hypoxia inducible factor 1, α subunit (HIF1A) and its target genes, erythropoietin receptor (EPOR), chemokine (C-X-C motif) receptor 4 (CXCR4) and vascular endothelial growth factor (VEGF), were quantified by qPCR. Neural differentiation potential was evaluated in 21% O₂ by cell morphology and qPCR.Neurospheres were successfully generated from AT-cMSCs at all O₂ tensions. Expression of nestin mRNA (NES) was significantly increased after neurosphere culture and was significantly higher in 1% O₂ compared to 5% and 21% O₂. Neurospheres cultured in 1% O₂ had significantly increased levels of VEGF and EPOR. There was a significant increase in CXCR4 expression in neurospheres generated at all O₂ tensions. Neurosphere culture under hypoxia had no negative effect on subsequent neural differentiation. This study suggests that generation of neurospheres under hypoxia could be beneficial when considering these cells for neurological cell therapies.     

Preservation of manipulated yeast diet

Manipulated yeast diet can be usedfor seed production of aquacultural organisms.Various methods for preserving the yeast dietduring the periods of circulation in marketwere tested, and the preservation of the yeastdiet by freeze-drying was the best. With thispreservation method, the manipulated yeastswere maintained fairly well (up to 71%) whenstored for three weeks under refrigeratedcondition (4 °C), while more than 80% ofthe yeast protoplasts disappeared in 5 days incontrol. Preservation temperature had an effecton retaining the yeast diet, but adding 3.5%NaCl concentration in the culture medium at thetime of yeast cultivation did not.     

Lethal Effects of Pulsed High-Voltage Discharge on Marine Plankton and Escherichia coli

Ballast seawater is considered globally as a major vector for invasions of non-indigenous organisms. Several technologies have been tested for their ability to remove organisms from ballast water. In the present study, we constructed a novel pulsed high-voltage discharge (PHVD) system that could operate in either high current mode with several hundred amperes or shockwave generating mode with relatively lower current in seawater. In laboratory-scale experiments, the PHVD system with shockwave-generating mode was found to be more effective in killing zooplankton (1.9- to 4.0-fold) and phytoplankton (3.3-fold) than high current mode at discharge with 300–500 pulses at 7.1 kV. Further experiments were carried out at different voltages and pulse-numbers to examine effects of the shockwave-generating PHVD system on viabilities of one zooplankton larva, two phytoplankton species, and an indicator bacterium suspended in seawater in a static chamber. For zooplankton, live cells were not detected at discharge with 400 pulses at 13 kV. For phytoplankton, the initial live cells of a dinoflagellate was decreased by 77?±?0.5%, and the initial chl a concentration of a diatom was decreased by 76?±?6% at discharge with 700 pulses at 13 kV. For an indicator bacterium Escherichia coli, live cells were not detected at discharges with 200 or 700 pulses at 13 kV. Measurements of ATP content of organisms showed congruent results with those obtained by the above methods, suggesting it may be a rapid method for evaluating treatment efficiency. Though further scale-up studies are necessary, these results suggest that the PHVD system have a high potential for applying to ballast seawater treatment.     

Quality Effects of Rye (Secale cerealeL.) Chromosome Arm 1RL Transferred to Wheat (Triticum aestivumL.)

Quality effects of rye (Secale cerealeL.) chromosome arm 1RL transferred to wheat (Triticum aestivumL.) were characterised by comparison of a group 1R(1B) substitution lines and 1BL.1RS translocation lines. The experimental materials were sister lines derived from the mating Mironovskaya10/NE7060//NE80413. 1R(1B) substitution lines were identified by the presence of ryeomega- andgamma-secalins among 70% ethanol soluble proteins, combined with the presence of high molecular weight (HMW) secalin proteins in total grain protein extracts. Genes on 1RL reduced grain weight, grain hardness, Mixograph time, Mixograph tolerance and SDS sedimentation volumes. 1RL had no effect on flour yield or grain and flour protein concentrations. The HMW secalin proteins encoded by genes on 1RL most probably caused the decline in dough strength seen in 1R(1B) lines relative to that of 1BL.1RS sister lines. Reduced grain hardness might also be related to the presence of HMW secalins, although a role for additional, unidentified genes on 1RL could not be discounted.     

Detection and characterization of bovine-like coronaviruses from four species of zoo ruminants

Five coronaviruses (CoVs) were detected in diarrheal feces from four zoo ruminant species: one wisent (Bison bonasus), two Himalayan tahr (Hemitragus jemlahicus), one sitatunga (Tragelaphus spekii), and one nyala (Tragelaphus angasii). We sequenced and analyzed the spike (S) and hemagglutinin/esterase (HE) genes of these viruses and compared the nucleotide (nt) and deduced amino acid (aa) sequences with those of other bovine CoV (BcoV) strains. Comparison of the entire deduced aa sequences of the S and HE glycoproteins revealed no specific differences that would account for discrimination between bovine-like CoV strains from zoo ruminants and BcoVs strains. In addition, the 99.9% aa identity among the five CoV strains revealed that the ruminants were infected by the same strain. Phylogenetically, bovine-like CoVs belong to group 2a CoVs, which are related most closely to the BcoV strains recently isolated in Korea. These data suggest that cattle are potential reservoirs for CoVs that are capable of infecting zoo ruminants.     

Biaxial Extensional Viscosity of Sheeted Noodle Dough

The flows encountered during the sheeting process contain a more extensional component and more stretching than simple shear. Evaluating the properties of the extensional rheological flow of wheat flour dough is essential to better understand the sheeting process and the final quality of the product. Our results show that the curves of biaxial extensional viscosity versus extensional strain rate for dough from different wheat cultivars including Dark Northern Spring (DNS), Hard Red Winter (HRW), and Western White Wheat (WW) with protein contents of 7.81–18.09% and water contents of 32–40% could be discriminated. During a lubricated compression test, the sheeted dough displayed a region of extensional thickening followed by a region of mild extensional thinning, giving rise to an s-shaped extensional stress-strain rate curve. The higher degree of extensional thinning was exhibited for the sheeted dough prepared from the flour of DNS and HRW wheat mixture with an extensional thinning index of n = 0.65–0.74; while the lower degree of extensional thinning was displayed for the DNS wheat with an extensional thinning index of n = 0.89–0.96. The data of biaxial extensional viscosity (η_Bmax) versus water content and protein content could be described by power relationships. The results suggest that biaxial extensional viscosity can be a useful modeling and process design parameter that objectively represents the rheological properties during dough sheeting.     

Genetic analysis and PCR-based identification of major <Emphasis Type="Italic">Fusarium</Emphasis> species causing head blight on wheat in Japan

Identifying the Fusarium species cause Fusarium head blight (FHB) and produces mycotoxins in wheat and other cereal is difficult and time consuming because of confusing phenotypic classification systems. In Japan, the F. graminearum complex, F. culmorum, F. avenaceum, and Microdochium nivale predominantly cause FHB. The internal transcribed spacer (ITS) and 5.8S of rDNA, a partial sequence of β-tubulin and mitochondrial cytochrome b (cytb) genes of the four species were PCR-amplified and analyzed. On the basis of the ITS, β-tubulin and cytb sequences, F. avenaceum and M. nivale are distinct from the F. graminearum complex and F. culmorum, whereas the F. graminearum complex is closely related to F. culmorum. Moreover, thiophanate–methyl-resistant isolates of the F. graminearum complex and F. culmorum did not have an amino acid substitution at amino acid codon 198 or 200 of β-tubulin. In contrast, very highly or highly thiophanate–methyl-resistant isolates of M. nivale had Glu (GAG) substituted with Ala (GCG) or Lys (AAG) at codon 198, respectively. The allele-specific PCR assay was used to identify the F. graminearum complex and F. culmorum, and these Fusarium species could be distinguished rapidly.     

Dairy-cattle health in Gyeongnam, Korea.

An animal-health monitoring system in the Gyeongnam area was started in 1997 to develop statistically valid data for use in estimating disease frequencies in dairy cattle, and the associated costs. The objectives of this study were to: (1) describe what was done to implement and maintain the system in Gyeongnam; (2) present selected disease frequencies; (3) discuss the epidemiological consideration of what was done and implications for results obtained.Veterinary medical officers (VMOs), comprising professors and graduate students from Gyeongsang National University, faculty of Gyeongnam Livestock Promotion Institute and clinic veterinarians, served as data collectors. After training on current disease and management problems of dairy cattle, interview techniques, sampling methods and data-collection instruments, the VMOs participated in selection of the sample herds and data gathering. Forty (n=40) of 167 dairy herds were selected randomly using a computer-generated list of random numbers and the VMOs visited farms once in a month for 12 months to collect data about management, disease, inventory, production, preventive treatment, financial and other relevant data. Strict data-quality control devices were used. Specific feed-back was developed for the producers and data collectors.The six disorders found most frequently in cows (from the highest to the lowest) were breeding problems, clinical mastitis, birth problems, gastrointestinal problems, metabolic problems and lameness. In young stock, respiratory, multiple system, breeding and gastrointestinal problems were predominant, whereas in calves, gastrointestinal, respiratory and integumental problems predominated.     

Evidence that Cache Valley virus induces congenital malformations in sheep

An outbreak of congenital abnormalities occurred in sheep at San Angelo, Texas, between December 1986 and February 1987. Of 360 lambs born, 19.2% had arthrogryposis or other musculo-skeletal problems and hydranencephaly (AGH), and the total neonatal loss was 25.6%. In 1987, all ewes that were tested with AGH lambs had antibody to Cache Valley virus (CVV), whereas 62% of the ewes with normal lambs had CVV-specific antibody. Pre-colostral serum samples from AGH lambs had neutralizing antibody to CVV. An increase in prevalence of CVV-specific antibody, from 5% during the spring of 1986 to 63.4% during the winter of 1987, occurred during a time that included the gestation of these affected lambs, as well as a period of increased rainfall. The isolation of a CVV-related strain from a sentinel sheep in October 1987 confirmed the continued presence of this virus in the pasture where this outbreak occurred and provided a recent field strain for future studies.     

EFFECT OF METHYLCELLULOSE ON UPPER GASTROINTESTINAL QUALITY IN DOGS

This study was performed to evaluate and optimize a small bowel contrast technique using barium and methylcellulose in dogs. Ten healthy dogs underwent both a conventional upper gastrointestinal study that used only barium and a modified study that used barium and methylcellulose. The control group received 10 mL/kg of 40% barium suspension. Experimental groups were divided into three subgroups given 15 mL/kg of different viscosities (low, moderate, and high viscosity) of 0.5% methylcellulose after 4 mL/kg of 40% barium suspension. Compared with the control group, dogs receiving methylcellulose had higher-quality upper gastrointestinal studies. Moderate viscosity of methylcellulose was superior to the other methylcellulose groups. In conclusion, the use of methylcellulose is a simple and effective method for improving the image quality in an upper GI examination.