Canine model of ischemic stroke with permanent middle cerebral artery occlusion: clinical and histopathological findings

The aim of the present study was to assess the clinical and histopathological findings in a canine model of ischemic stroke. Cerebral ischemic stroke was induced by middle cerebral artery occlusion in four healthy beagle dogs using silicone plugs. They showed neurological signs of forebrain dysfunction such as reduced responsiveness, head turning, circling, postural reaction deficits, perceptual deficits, and hemianopsia. These signs gradually regressed within 4 weeks without therapy. On magnetic resonance imaging, T2 hyperintensity and T1 hypointensity were found in the cerebral cortex and basal ganglia. These lesions were well-defined and sharply demarcated from adjacent brain parenchyma with a homogenous appearance. No abnormalities of the cerebrospinal fluid were observed. At necropsy, atrophic and necrotic lesions were observed in the cerebral cortex. The cerebral cortex, basal ganglia, and thalamus were partially unstained with triphenyl-tetrazolium chloride. Histopathologically, typical features of infarction were identified in cortical and thalamic lesions. This study demonstrates that our canine model resembles the conditions of real stroke patients.     

Enhancement of protective immune responses by oral vaccination with Saccharomyces cerevisiae expressing recombinant Actinobacillus pleuropneumoniae ApxIA or ApxIIA in mice

We previously induced protective immune response by oral immunization with yeast expressing the ApxIIA antigen. The ApxI antigen is also an important factor in the protection against Actinobacillus pleuropneumoniae serotype 5 infection; therefore, the protective immunity in mice following oral immunization with Saccharomyces cerevisiae expressing either ApxIA (group C) or ApxIIA (group D) alone or both (group E) was compared with that in two control groups (group A and B). The immunogenicity of the rApxIA antigen derived from the yeast was confirmed by a high survival rate and an ApxIA-specific IgG antibody response (p < 0.01). The highest systemic (IgG) and local (IgA) humoral immune responses to ApxIA and ApxIIA were detected in group E after the third immunization (p < 0.05). The levels of IL-1β and IL-6 after challenge with an A. pleuropneumoniae field isolate did not change significantly in the vaccinated groups. The level of TNF-α increased in a time-dependent manner in group E but was not significantly different after the challenge. After the challenge, the mice in group E had a significantly lower infectious burden and a higher level of protection than the mice in the other groups (p < 0.05). The survival rate in each group was closely correlated to the immune response and histopathological observations in the lung following the challenge. These results suggested that immunity to the ApxIA antigen is required for optimal protection.     

Guided bone regeneration with beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone membrane in partial defects of canine humerus

This study was performed to evaluate the effect of beta-tricalcium phosphate and poly L-lactide-co-glycolide-co-epsilon-caprolactone (TCP/PLGC) membrane in the repair of partial bone defects in canine proximal humerus. Three adult mixed-breed dogs were used during the experimental period. The length of the defect was quarter of the full length of humerus, and width of the defect was quarter of middle diameter of the lateral aspect of humerus. The humeri of each dog were divided into treatment (TCP/PLGC) and control groups. The defect was covered with TCP/PLGC membrane in treatment group. To evaluate regeneration of the bone, computerized tomography (CT) and histopathologic examination were performed. The radiopaque lines were appeared at the original defect sites in TCP/PLGC group but below the original site in control at 4th week. Radiopacity and thickness of the defect sites, and radiopaque lines were more increased at 8th week than those of 4th week. Histopathologic findings revealed fibrous connective tissue migration into the defect and the migration inhibited the structure of new cortex to be placed in the original level in control whereas new cortex growth was found in the level of original line in TCP/PLGC group. However, the new cortical bone in the TCP/PLGC group was thinner and less organized than the adjacent intact cortex, and the amount of new cancellous bones were also scanty. The result suggested that TCP/PLGC membrane is a good guided bone regeneration material to restore the original morphology of humerus in partial defect.     

Tapered conduits can buffer hydraulic conductance from path-length effects

The model of West, Brown and Enquist showed that total hydraulic resistance in trees can be independent of path length, provided that vascular conduits taper sufficiently. This model assumes that the tree branch network is volume-filling, so that segment lengths increase exponentially from tree top to base. We show that partial buffering of hydraulic resistance from path-length effects can occur even for moderate tapering, and that this effect is stronger when segment lengths are fixed. Still needed are measurements of tracheary size and hydraulic resistance designed to test this model, which shows how hydraulic limitation of tree height growth may be mitigated.     

Study on half-scale model test for light-frame shear wall

A half-scale model of a light-frame shear wall was developed to evaluate the racking performance of a fullscale shear wall (prototype). The effect of nail size on the performance of the shear wall was also investigated using models constructed with three types of nail. Materials for the model were determined through experimental methods, which included nail-head push-through, stud-to-sheathing nail connection, and static bending tests. Materials with which the model was made to be in similarity to the prototype were three-layer 4.8-mm plywood, 39.72-mm long nails, and 1 × 2 lumber cut from 2 × 4 studs. In accordance with ASTM E 72 and ASTM E 564, racking resistance tests were conducted on 20 shear walls. The results showed that the maximum load capacities of the prototype walls could be evaluated by the model without significantly different failure modes. Tests on the effect of nail size revealed that increasing the nail head diameter may improve the performance of shear walls.Parts of this paper were presented at the International Conference on Effective Utilization of Plantation Timber (ICEUPT'99), Chi-Tou, Taiwan, May 1999; and the World Conference on Timber Engineering (WCTE2000), Whistler, Canada, July–August 2000     

Observations on the Quality Characteristics of Waxy (Amylose‐Free) Winter Wheats

Previous investigations have suggested waxy (amylose‐free) wheats (Triticum aestivum L.) possess weak gluten properties and may not be suitable for commercial gluten extraction. This limitation could prevent the use of waxy wheat as a source of unique starch, because gluten is a by‐product of the wheat starch purification process. Fifty waxy wheat lines were used to determine the extent to which gluten protein and other grain quality related traits might vary and, consequently, allow the development of waxy wheat with acceptable gluten properties. Among the waxy lines, significant variation was observed for all measured quality traits with the exception of flour protein concentration. No waxy entries statistically equaled the highest ranking nonwaxy entry for grain volume weight, falling number, flour yield, or mixograph mix time. No waxy lines numerically exceeded or equaled the mean of the nonwaxy controls for falling number, flour yield, or mixograph mix time. For grain and flour protein related variables, however, many waxy lines were identified well within the range of acceptability, relative to the nonwaxy controls used in this study. Approximately 50% of the waxy lines did not differ from the highest ranking nonwaxy cultivar for grain and flour protein concentrations. Forty‐three (86%) of the tested waxy lines were not sig‐nificantly different from the nonwaxy line with the highest mixograph mixing tolerance, 22/50 (44%) of the waxy wheat lines did not differ from the highest ranking nonwaxy line in gluten index scores, and 17/50 (34%) did not differ from the highest ranking nonwaxy line in extracted wet gluten. All waxy experimental lines produced gluten via Glutomatic washing. The quality of the gluten, as measured both by mixograph and gluten index, varied widely among the waxy lines tested. These observations suggest that weak gluten is not a natural consequence of the waxy trait, and waxy cultivars with acceptable gluten properties can be developed.     

Prediction of bending properties for structural glulam using optimized distributions of knot characteristics and laminar MOE

This study established a prediction model for bending properties of glued-laminated timber (glulam) using optimized knot and modulus of elasticity (MOE) distributions of lumber laminate as the main input variables. For this purpose, knot and MOE data were investigated for all pieces of lumber that were prepared for glulam manufacturing, and statistical distributions of knot size, knot number in one lumber, and MOE of each laminate were optimized as distribution functions. These knot and MOE data were used as input variables in the prediction model for bending properties, and were also used in generating virtual glulam using the inverse transform method. Prediction of bending properties for glulam was carried out using the transformed section method, which is partially provided in ASTM D 3737 (Annex A4). Predicted values were compared with those from full-scale four-point bending tests for 60 six-layered glulams with 10 different laminar combinations. Finally, the allowable bending properties of glulam for each specific laminate combination were determined by calculating the fifth percentile of the modulus of rupture and the average modulus of elasticity from virtual test results of more than 1000 virtual glulams. From the results of this study, predicted bending properties for glulam and their distributions could be used for structural design in both allowable stress design and limit state design.     

Vitamin D Status in Different Stages of Disease Severity in Dogs with Chronic Valvular Heart Disease

Background

In humans with heart disease, vitamin D deficiency is associated with disease progression and a poor prognosis. A recent study showed that serum 25‐hydroxyvitamin D [25(OH)D] concentration, the hallmark of vitamin D status, was lower in dogs with heart failure than in normal dogs, and a low concentration was associated with poor outcome in dogs with heart failure.

Objectives

To elucidate the vitamin D status of dogs with chronic valvular heart disease (CVHD) at different stages of disease severity.

Animals

Forty‐three client‐owned dogs with CVHD.

Methods

In this cross‐sectional study, dogs were divided into 3 groups (14 dogs in Stage B1, 17 dogs in Stage B2, and 12 dogs in Stage C/D) according to ACVIM guidelines. Dogs underwent clinical examination including echocardiography. Serum 25(OH)D concentrations were measured in each dog.

Results

Serum 25(OH)D concentration was significantly lower in Stage B2 (median, 33.2 nmol/L; range, 4.9–171.7 nmol/L) and C/D (13.1 nmol/L; 4.9–58.1 nmol/L) than in Stage B1 (52.5 nmol/L; 33.5–178.0 nmol/L) and was not significantly different between Stage B2 and Stage C/D. Among clinical variables, there were significant negative correlations between 25(OH)D concentration and both left atrial‐to‐aortic root ratio and left ventricular end‐diastolic diameter normalized for body weight.

Conclusions and Clinical Importance

These results indicate that vitamin D status is associated with the degree of cardiac remodeling, and the serum 25(OH)D concentration begins to decrease before the onset of heart failure in dogs with CVHD.     

Cell-free extract from porcine induced pluripotent stem cells can affect porcine somatic cell nuclear reprogramming

Pretreatment of somatic cells with undifferentiated cell extracts, such as embryonic stem cells and mammalian oocytes, is an attractive alternative method for reprogramming control. The properties of induced pluripotent stem cells (iPSCs) are similar to those of embryonic stem cells; however, no studies have reported somatic cell nuclear reprogramming using iPSC extracts. Therefore, this study aimed to evaluate the effects of porcine iPSC extracts treatment on porcine ear fibroblasts and early development of porcine cloned embryos produced from porcine ear skin fibroblasts pretreated with the porcine iPSC extracts. The Chariot^TM reagent system was used to deliver the iPSC extracts into cultured porcine ear skin fibroblasts. The iPSC extracts-treated cells (iPSC-treated cells) were cultured for 3 days and used for analyzing histone modification and somatic cell nuclear transfer. Compared to the results for nontreated cells, the trimethylation status of histone H3 lysine residue 9 (H3K9) in the iPSC-treated cells significantly decreased. The expression of Jmjd2b, the H3K9 trimethylation-specific demethylase gene, significantly increased in the iPSC-treated cells; conversely, the expression of the proapoptotic genes, Bax and p53, significantly decreased. When the iPSC-treated cells were transferred into enucleated porcine oocytes, no differences were observed in blastocyst development and total cell number in blastocysts compared with the results for control cells. However, H3K9 trimethylation of pronuclear-stage-cloned embryos significantly decreased in the iPSC-treated cells. Additionally, Bax and p53 gene expression in the blastocysts was significantly lower in iPSC-treated cells than in control cells. To our knowledge, this study is the first to show that an extracts of porcine iPSCs can affect histone modification and gene expression in porcine ear skin fibroblasts and cloned embryos.     

Protective effect of butylated hydroxylanisole against hydrogen peroxide-induced apoptosis in primary cultured mouse hepatocytes

Butylated hydroxyanisole (BHA) is a synthetic phenolic compound consisting of a mixture of two isomeric organic compounds: 2-tert-butyl-4-hydroxyanisole and 3-tert-butyl-4-hydroxyanisole. We examined the effect of BHA against hydrogen peroxide (H₂O₂)-induced apoptosis in primary cultured mouse hepatocytes. Cell viability was significantly decreased by H₂O₂ in a dose-dependent manner. Additionally, H₂O₂ treatment increased Bax, decreased Bcl-2, and promoted PARP-1 cleavage in a dose-dependent manner. Pretreatment with BHA before exposure to H₂O₂ significantly attenuated the H₂O₂-induced decrease of cell viability. H₂O₂ exposure resulted in an increase of intracellular reactive oxygen species (ROS) generation that was significantly inhibited by pretreatment with BHA or N-acetyl-cysteine (NAC, an ROS scavenger). H₂O₂-induced decrease of cell viability was also attenuated by pretreatment with BHA and NAC. Furthermore, H₂O₂-induced increase of Bax, decrease of Bcl-2, and PARP-1 cleavage was also inhibited by BHA. Taken together, results of this investigation demonstrated that BHA protects primary cultured mouse hepatocytes against H₂O₂-induced apoptosis by inhibiting ROS generation.