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.     

Adhesion of Corynebacterium pilosum by pili to epithelial cells of bovine vulva

Piliated (P+) and nonpiliated (P-) clones of Corynebacterium pilosum were selected, and their adhesion to the epithelial cells of the bovine vulva and vaginal vestibule was examined. The number of P+ bacteria of C pilosum that adhered to vulval epithelial cells was greater than that of P- bacteria. The adhesion of P+ bacteria, but not P- bacteria, to the epithelial cells was inhibited by the antipilus antiserum; therefore, the adhesion of C pilosum to the epithelial cells of the vulva was primarily dependent on the pili. The number of C pilosum that adhered to the epithelial cells of the bovine vulva and of the vaginal vestibule increased by decreasing the pH.     

A method for isolation of milled-wood lignin involving solvent swelling prior to enzyme treatment

Summary A new lignin isolation method has been developed. Wood and pulp were subjected to ball milling, swelled in an organic solvent, and then treated with a cellulase. The enzyme digestion time could be shortened to 1 day with this method. The lignin obtained has been named Swelled Enzyme Lignin (SEL). Swelling and enzyme digestion conditions and their effects on lignins were investigated. The SEL's from wood could be directly washed with water, while those from pulp had to be washed with aqueous acetic acid because they were water soluble. The purification of crude SEL's was accomplished by extracting them with dioxane-water, and then precipitating and washing with ethyl ether. Lignin yields were 24–67% based on the total amount of lignin present. The characteristics of the SEL's were further investigated by gel-permeation chromatography (GPC), infrared and ¹³C nuclear magnetic resonance (NMR) spectroscopy.Symbols SUKP Softwood Unbleached Kraft Pulp - BUKP Birch Unbleached Kraft Pulp - TD Todomatsu (Abies sachalinensis) wood - MWL Milled Wood Lignin - MWLS (BK, TD) Milled Wood Lignin of SUKP (BUKP, TD) - SEL-C Swelled Enzyme Lignin (crude) - SELS-C SEL-C of SUKP, other materials named in the same way - SELS-90 SEL-C of SUKP and extracted with 90% dioxane-water - SELS-96 SEL-C of SUKP and extracted with 96% dioxane-water, other materials named in the same way     

Probabilistic evaluation of the final moisture content of kiln-dried lumber using the bootstrap method

This paper presents a probabilistic method of evaluating the final moisture content (MC) of lumber obtained at the end of the kiln-drying process. The final MC data of three different drying tests conducted in past studies were analyzed using the bootstrap method. Target MC was tentatively set below 20 % in the analysis. Two characteristic parameters representing the final MC were estimated with bootstrap confidence intervals. These parameters were the standard deviation (SD) and the percentage of the population that met the MC requirement of less than 20 % (P ₂₀). The histograms of the final MC and the subsequent goodness-of-fit tests revealed that the final MC data of two drying tests did not follow any classical probability distributions, including Normal, Log-Normal, Weibull, and Gamma distributions, thus indicating the need for nonparametric statistics. The uncertainty of the final MC could be evaluated with the estimated SD and P ₂₀. After deriving the relationships between P ₂₀ and the corresponding probability that P ₂₀ is not achieved, we demonstrated how such relationships could provide a kiln operator with information to facilitate better decision-making in optimizing a drying schedule.     

Effect of moisture content of members on mechanical properties of timber joints

To investigate the effect of moisture content (MC) of members on the mechanical properties of timber joints, bending tests of precut joints and shear tests of dowel-type joints were carried out using timbers of Japanese cedar (Cryptomeria japonica D. Don) with three moisture conditions: green, kiln-dried with a MC target of 15%, and over-kiln-dried with a MC target of 5%. For the bending test, timbers were processed with a precut processing machine into “koshikake-ari” (a kind of dovetail joint) and “koshikake-kama” (a kind of mortise and tenon joint). A pair of members was jointed together without mechanical fasteners. Bolts (diameter = 12 mm) and nails (diameter = 2.45 mm) were used as dowels in the shear test. Bolted joints were constructed with one bolt and two metal side plates. Two nails and two metal side plates were used for the nailed joint. For precut joints, no clear effect of MC was recognized on maximum moment and initial stiffness. The maximum strength of mechanical joints assembled with kiln-dried wood was changed by the degree of drying. Stiffness of the joints assembled with kiln-dried specimens was larger than that of the joints assembled with green specimens.Part of this study was presented at the 7th International IUFRO Wood Drying Conference, Tsukuba, July 2001     

Combination of serum phosphorylated neurofilament heavy subunit and hyperintensity of intramedullary T2W on magnetic resonance imaging provides better prognostic value of canine thoracolumbar intervertebral disc herniation

The aim of this study was to evaluate the prognostic value of concurrent measurement of serum phosphorylated neurofilament heavy subunit (pNF-H) concentration and intramedullary T2W hyperintensity in paraplegic to paraplegic dogs. Our hypothesis was that concurrent measurement of these would provide a more accurate prediction of functional outcome in dogs with thoracolumbar intervertebral disc herniation (IVDH). A prospective case-control clinical study was designed using 94 dogs with acute onset of thoracolumbar IVDH. The association of serum pNF-H concentration, T2W hyperintensity on sagittal MRI (T2H/L2), deep pain perception and surgical outcome were evaluated with logistic regression analysis after three months for all 94 surgically treated dogs. Sensitivity to predict non-ambulatory outcome was compared among pNF-H and T2H/L2 and combination of both. Logistic regression analysis indicated that serum pNF-H concentration and T2H/L2 were significantly correlated with surgical outcome (P<0.05); however, deep pain perception was not (P=0.41). The results of logistic regression analysis indicated that the odds ratios of unsuccessful long-term outcome were 2.6 for serum pNF-H concentration, 1.9 for T2H/L2 and 2.3 for deep pain sensation. The sensitivity and specificity to predict non-ambulatory outcome for using serum parameter pNF-H>2.6 ng/ml, using T2H/L2 value of>0.84 and using both serum pNF-H and T2H/L2, were 95% and 75.7%, 65% and 86.5%, and 90.0% and 97.5%, respectively. Therefore, combined measurements of serum pNF-H and T2H/L2 might be useful for predicting long-term outcome in dogs with thoracolumbar IVDH.     

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.