Oligodendroglial vacuolar degeneration in the bilateral motor cortices and astrocytosis in epileptic beagle dogs

We performed a pathologic examination of the brains of three dogs in an epileptic beagle colony. Histologically, all the cases had diffuse astrocytosis in the cerebral cortex and basal ganglia as well as the hippocampus, whereas they showed acute nerve cell change in the hippocampus and some other areas of the cerebrum. One of these animals showed laminar myelin pallor associated with the presence of many vacuoles in the IV to VI layers of the bilateral motor cortices. Most of the vacuoles contained fine granules stained with luxol-fast-blue stain. Ultrastructural examination revealed that some oligodendrocytes and perineuronal satellite oligodendrocytes in the bilateral cerebral motor cortices of the two affected dogs had many vacuoles surrounded by myelin-like lamellar structures. These findings suggest a possibility that astrocytosis in the cerebrum and vacuolar degeneration of oligodendrocytes in the cerebral motor cortex may be, at least in part, related to the occurrence or development of seizures.     

Iron uptake in a bicarbonate-resistant cell line of tobacco

Abstract

In alkaline soils, plant growth is impaired mainly by high pH and high concentration of bicarbonates. The bicarbonate concentration increases the pH value, and causes deficiency of iron. A bicarbonate-resistant cell line (BR line) of tobacco (Nicotiana tabacum L. cv. Burley21) was selected by adding excess bicarbonate ions (20 mmol L^?1) to the culture medium. The pH of the medium was buffered 8.0 to 8.3. Under these conditions, about 80% of iron in the medium became insoluble. However, under such conditions, the BR line grew well. In this report, we examined some characteristics of the growth and iron uptake in the BR line under iron-deficient (i.e. high pH or no-iron) condition. At pH 5.8, the Fe³⁺ reduction activity was not significantly different between the non-selected line and the BR line. At pH 8.0, however, the Fe³⁺ reduction activity of the BR line was higher than that of the non-selected line. In no-iron condition, the growth of the non-selected line was markedly reduced after 2 weeks, while the BR line was not affected. The content of malic acid in both lines increased with the medium pH, and the content in the BR line was higher than that in the non-selected line. The BR line was able to adapt to the conditions, which restricted iron uptake, such as high bicarbonate concentration, high pH, and low iron conditions. The high ability of Fe³⁺ reduction was maintained at even high pH conditions. Further, the BR line may be able to improve the utilization of iron in the cells.     

Development of a PCR test for the identification of Staphylococcus intermedius based on the 16S rDNA sequence

The development of a PCR assay based on the 16S ribosomal RNA gene (rDNA) sequence was carried out for the identification of Staphylococcus intermedius. Sixty-six strains of S. intermedius, 70 of Staphylococcus aureus and 2 of Staphylococcus hyicus were examined for the assay. The 16S rDNA, of which the PCR target fragment makes up 901 bp corresponding to the sequence data of the gene, was detected in all strains of S. intermedius, but it was not detected in any strains of either S. aureus or S. hyicus. These results suggest that the PCR allows a simple and precise identification of S. intermedius.     

Isolation of Neospora caninum from the brain of a naturally infected adult dairy cow

Neospora caninum was isolated from the brain of a 2-year-old dairy cow that had aborted confirmed N. caninum-infected fetuses on two occasions. The cow had an indirect fluorescent antibody titer of 1:1600 to N. caninum. The cow was killed 24 days after its second abortion and the brain was bioassayed for N. caninum in nude mice. Multifocal areas of perivascular cuffing and glial nodules were observed in the cerebrum and mesencephalon of the cow, but N. caninum was not identified in histological sections of the brain. All three nude mice inoculated with brain homogenate of the cow, developed emaciation and paralysis. Microscopical examination of the nude mice revealed systemic N. caninum infection with demonstrable tachyzoites in various organs. The parasites isolated from fresh mouse brain were transferred successfully into Vero cell cultures. PCR procedure on the purified tachyzoites obtained from the Vero cell cultures amplified the specific DNA sequence for N. caninum.     

Polymer distribution in the cell wall of a wood-polyethylenimine composite

Summary A light microscopic (LM) visual technique to determine the polyethylenimine (PEI) distribution in a wood-PEI composite was studied by the adsorption of acid red or the circulation of Cu(II) ion with PEI in the composite. The amount of the adsorbed dye or Cu(II) on the composite increased linearly with an increase in the PEI content of the wood-PEI. The dyed composites were observed with LM under monochromatic light, corresponding to their absorption maxima at 560 nm for acid red and at 640 nm for copper chelate. The microscopic image was photographically recorded and the negative was subjected to densitometric analysis. At a polymer content lower than 4% in the composite, about 70% of total polymer was located in the middle lamella region, leaving only 30% in the secondary wall. With increasing PEI content of the composite, the concentration in the secondary wall increased, and about 80% of the polymer was located in the secondary wall at a PEI content of 27%. There were no noticeable differences between the amount of polymer detected by dyeing with acid red and by chelating with Cu(II). From the results of the line analysis of the intensity of the characteristic X-ray of uranium, the distribution of the PEI in the cell wall agreed closely with that determined by the LM method. It was concluded that the LM visual technique to determine the PEI location in the wood-PEI composite by dyeing with acid red or by chelating with Cu(II) is a simple and reliable method.     

Condensation reactions of phenolic resins III: self-condensations of 2,4-dihydroxymethylphenol and 2,4,6-trihydroxymethylphenol (1)

This study reexamined the kinetics of the condensation reactions of hydroxymethylphenols with the purpose of elucidating the reaction mechanisms. This report discusses experimental results on the self-condensations of 2,4-dihydroxymethylphenols (2,4-DHMP) and 2,4,6-trihydroxymethylphenol (THMP), focusing on the order of reaction. The relations between the initial rates of reaction and the initial concentrations of reactants were investigated. Results quite different from those of previous reports were obtained. The order of reaction of the selfcondensation of 2,4-DHMP was found to be 1.1, which did not change with the alkali/2,4-DHMP molar ratio. The order of reaction of the self-condensation of THMP was found to vary with both the concentration of THMP and the alkali/THMP molar ratio. In the region of THMP concentrations above 1.5mol/l, the order of reaction was confirmed to be 2.0, which did not change with the alkali/THMP molar ratio. In the region of THMP concentrations below 1.0mol/l, the order of reaction varied with the alkali/THMP molar ratio, showing fractional numbers of 1.2–1.6. These results indicate that unimolecular reaction(s) and bimolecular reaction(s) take place simultaneously as the ratedetermining step in the condensation reactions of 2,4-DHMP and THMP and that the reaction mechanism changes with the species of reactant and, in some cases, with the reaction conditions.Part of this study was presented at the Wood Adhesives 2000 symposium, South Lake Tahoe, Nevada, USA, June 2000     

Blackening of Diospyros genus xylem in connection with boron content

The amount of boron contained in the xylem of Japanese persimmon (Diospyros kaki Thunb.) and ebony (Diospyros ebenum Koen) was determined by inductively coupled plasma (ICP) emission spectrometry and a modified curcumin–acetic acid method. The boron content was compared between the heartwood and sapwood of ebony as well as between a blackened portion (“kurogaki”) and normal portion of Japanese persimmon. The kurogaki contained a higher level of boron than the normal portion of the same individual, although the boron content varied among individuals. Moreover, the boron content of the heartwood of ebony was much higher than that of the sapwood. These results suggest the participation of boron in the blackening of Japanese persimmon. Because both kurogaki and heartwood of ebony are durable to fungal attack, the blackening of Diospyros genus appears to be related to the formation of defensive substances in which boron seems to take part. The convenient curcumin–acetic acid method is an alternative to the ICP method with comparable accuracy.     

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.     

Distribution of a fish pathogen Listonella anguillarum in the Japanese flounder Paralichthys olivaceus hatchery

The present study was undertaken to investigate the distribution of Listonella anguillarum in a Japanese flounder (Paralichthys olivaceus) hatchery. A total of 2704 isolates were obtained from the developing fish, live diets and artificial feeds of Japanese flounder and their rearing water, 439 of which were identified as L. anguillarum by the combining incubation on thiosulfate-citrate-bile salt-sucrose (TCBS) agar at 35 °C overnight with polymerase chain reaction (PCR) detection for the VAH1 hemolysin gene. L. anguillarum was detected in all seven rotifer samples, with densities of 2.5 × 10³ to 4.6 × 10⁶ colony forming units (CFU) g^− 1. Both the analyzed samples of Nannochloropsis oculata contained this bacterium at densities of 1.6 × 10⁴ to 1.4 × 10⁵ CFU g^− 1. L. anguillarum was detected in only one of four samples of Artemia nauplii with a density of 4.8 × 10⁵ CFU g^− 1 (35%) and it was not detected in the two analyzed artificial feed samples. L. anguillarum was detected in 11 of 18 specimens of larval and juvenile Japanese flounder at densities of 5.0 × 10¹ to 7.4 × 10⁵ CFU g^− 1, while it was not detected in the two analyzed egg specimens of Japanese flounder. These results indicate that L. anguillarum associated with the developing Japanese flounder is likely derived from rearing water and live diets such as rotifers. Further, it is strongly suggested that L. anguillarum is a transient bacterium of the intestinal microflora for the Japanese flounder but is a permanently indigenous one for the Japanese flounder hatcheries.