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.     

Chlorantraniliprole application differentially affects adult emergence of Sympetrum dragonflies in rice paddy fields

Paddy and Water Environment - Rice paddy fields are important habitat for many dragonfly species. In Japan, populations of dragonflies inhabiting rice paddies, in particular Sympetrum (Odonata:...     

Pathological changes of tracheal mucosa in chickens infected with infectious laryngotracheitis virus

Six-week-old chickens were inoculated via the posterior thoracic air sac with infectious laryngotracheitis virus. Chickens were sacrificed on various days through day 16 postinoculation (PI), and the trachea was examined by scanning electron microscopy (SEM) and light microscopy (LM). The pathological changes observed on day 1 PI were hypertrophy and hyperplasia of goblet cells. From day 3 PI, the epithelial cells protruded collectively and fused to form syncytia, which contained many intranuclear inclusion bodies. Subsequently, epithelial syncytia desquamated, one after another, and connective tissues were exposed in places. Serofibrinous exudate and detritus were abundant on the surface of the exposed connective tissues and seemed to form a pseudomembrane. On day 5 PI, the remaining epithelial cells began to repair the devastated mucosa just under the pseudomembrane. On day 6 PI, microvillus-rich regenerating epithelial cells were arranged like paving stones. On day 8 PI, the epithelial cells proliferated extensively and formed folds with cyst-like structures. By day 16 PI, the tracheal epithelium was covered with cilia and regained its normal histologic appearance.     

Evaluation of 5 Types of Fishing Material, 2 Sterilization Methods, and a Crimp-Clamp System for Extra-Articular Stabilization of the Canine Stifle Joint

OBJECTIVE: To evaluate the mechanical properties of 5 types of fishing material, 2 sterilization methods, and a commercially designed crimp-clamp system for the extra-articular repair of the canine stifle joint. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Animals were not used in this study. METHODS: Two brands of monofilament nylon fishing line and 3 brands of monofilament nylon leader line were used to determine the effect of steam and ethylene oxide sterilization on strength and elongation of the material. A strand of 36-kg test monofilament nylon fishing material was wrapped around 2 rods or knotted to form a loop around 2 rods on a materials-testing machine. Ten trials of each brand of unsterilized, steam-sterilized, and ethylene oxide-sterilized fishing material were tested. A strand of each material was elongated to failure at a constant displacement of 1,000 mm/min to determine strength. A strand of each material was cycled 10 times to a load of 50 N to determine percent elongation. The brand of fishing material with the greatest strength and least elongation was crimped to form a loop around 2 rods on a materials-testing machine and tested as described above. ANOVA was used to determine the effect of sterilization method, brand of material, knot, wrap, and crimp on strength and elongation of the material, and a post-hoc t test was used when significant differences were found. A Student t test was used to compare fixation techniques (wrap, knot, and crimp). RESULTS: Sterilization by steam or ethylene oxide had no significant effect on the strength of the nylon fishing material. Steam sterilization resulted in significant increases (2- to 4-fold) in elongation of most nylon fishing material when compared with unsterilized material. Ethylene oxide sterilization had minimal effect on elongation of the fishing material. Mason leader line showed no significant change in strength or elongation regardless of sterilization method. Significantly less strength and significantly less elongation were demonstrated in Mason leader line that was crimped as compared with Mason leader line that was knotted. CONCLUSION: Ethlylene oxide was the preferred method of sterilization to preserve strength and minimize elongation of the fishing material. Of the materials tested, Mason leader line had the least elongation and the greatest preservation of strength when ethylene oxide was used as the sterilization method. Mason leader line and Sufix fishing line were comparable choices when steam was used as the sterilization method. Significantly less elongation was demonstrated in crimped Mason leader line as compared with knotted Mason leader line. CLINICAL RELEVANCE: Of the materials tested, Mason leader line and Sufix fishing line had the best mechanical properties for extracapsular stabilization of the canine stifle joint. Crimping is an attractive alternative to knotting and results in a reduction in elongation of the nylon fishing material.     

An outbreak of bovine ephemeral fever in Nagasaki Prefecture in 1988

In October, 1988, bovine ephemeral fever (BEF) occurred in Nagasaki Prefecture and throughout Kyushu island, with the exception Miyazaki Prefecture. The first outbreak occurred in Hirado-shi on October 17. The total number of diseased cattle was 24 in 24 farms in Nagasaki Prefecture. The clinical findings were mainly sudden fever, anorexia, and instability in standing. The serum neutralizing antibodies against BEF virus (BEFV) rose in all infected cattle. Twelve strains of the virus were isolated in HmLu-1 cell cultures made directly from the heparinized blood of 17 infected cattle. The buffy coat was mainly collected from the samples and washed three times with phosphate buffered saline. These isolates were all neutralized by an antiserum against BEFV (Yamaguchi strain). With the aid of an electron microscope, a representative of isolates named Hirado-9 with a length of 150 nm was seen in the sample of infected HmLu-1 cell cultures. Both Hirado-9 and Yamaguchi strains reacted with antisera. The outbreak of BEF in 1988 was the first since 1971 in Nagasaki Prefecture. The result proved that BEFV can be easily isolated in HmLu-1 cell culture from the washed blood cells of infected cattle.     

Restriction fragment length polymorphism for the Yc subunit gene of rat liver glutathione S-transferase

An altered expression of the Yc subunit gene of rat glutathione S-transferase (GST) in the liver of the LEC rat, which is a mutant strain with spontaneous hereditary hepatitis associated with severe jaundice, has been reported. To provide further information concerning the structure of the Yc subunit gene, we carried out the Southern blot hybridization analysis of DNA samples from rats of eight different inbred strains including LEC with cDNA complementary to mRNA specific for the Yc subunit of rat liver GST as a probe. The hybridization patterns of the DNA samples from rats belonging to the different inbred strains showed interstrain variation in the length of restriction fragments with four restriction endonucleases. Since the DNA samples prepared from several rats of one inbred strain gave an identical hybridization pattern, the restriction fragment patterns for the Yc gene could be used as markers for genetic monitoring of inbred rat strains. Although the altered expression of Yc-Yc activity of GST has been observed in the liver of the LEC rat, the characteristic changes in the gene structure of the Yc subunit of LEC rat were not detected in the present hybridization analysis.     

Muscle architecture of the elongated nose in the Asian elephant (Elephas maximus).

The architecture of the M. caninus in the elongated nose was examined in the Asian elephant (Elephas maximus). The following complicated musculature of the M. caninus was observed in the proximal and distal regions of the nose: (1) Proximal region: In the superficial layer, the longitudinal bundles are confirmed in the dorsal part, and the obliquely-oriented ones in the ventral part. In the middle layer, some bundles run ventro-distally, while other ones represent longitudinally-oriented running. The deep layer consists of complicated architecture of many bundles. Some muscle bundles run medio-laterally, while the others extend proximo-distally in this space. (2) Distal region: In the dorsal part of the M. caninus, the bundles run at deep-superficial direction, while in the ventral part the bundles are longitudinally arranged. The bundles run at lateral direction near the septum of the nasal conduits. The N. facialis and N. infraorbitalis send many branches in the lateral area of the M. caninus in the trunk. This muscle architecture of multi-oriented bundles and well-developed innervation to them suggest that they enable the elongated nose to act as a refined manipulator in the Asian elephant.     

Simplified culture method of detached ears and its application to vernalization in wheat

Summary The effects of the addition of sulfurous acid into culture solution and of cold treatment of the solution were examined to simplify the culture of detached wheat ears. In the simplified method, detached ears could be cultured at room temperature on the liquid medium containing 100 g/l sucrose and 0.075% sulfurous acid without any sterilization. The immature seeds in detached ears cultured by this method were treated with low temperature or with chemicals known to have vernalizing effect. The chemical treatment did not affect the chilling requirement of immature embryos, although photoperiodic response and narrow-sense earliness were reduced by kinetin and trypsin. The low temperature treatment drastically affected the chilling requirement, and fully vernalized mature seeds having normal germinability were obtained by treating the detached ears in culture with low temperature from 10 days after anthesis.     

DNA fingerprinting in soybean (Glycine max (L.) Merrill) with oligonucleotide probes for simple repetitive sequences

Summary Soybean DNA fingerprints were analyzed by digoxigenin-labeled oligonucleotide probes complementary to simple repetitive sequences. The clearest and most polymorphic patterns were obtained with (AAT)₆ as a probe, with which all 47 soybean cultivars tested could be distinguished. However, DNA fingerprints of individuals within cultivars showed the same pattern. Using (CT)₈, (GAA)₅ or (AAGG)₄ as probes, clear polymorphic patterns among cultivars and accessions in the subgenus Soja (Glycine max and Glycine soja) were not observed, while quite different patterns were found in accessions in the subgenus Glycine. The results suggest that G. max and G. soja are closer in their genome structure. DNA fingerprints of reciprocal crosses between cultivars and accessions in the subgenus Soja were similar, and contained bands of both parents. In an F₂ population from these crosses, such bands segregated in a Mendelian fashion.