Hepatic AA amyloidosis in a cat: cytologic and histologic identification of AA amyloid in macrophages

A 3‐year‐old, spayed female, Domestic Shorthair cat presented with anorexia, lethargy, vomiting, probable hemoabdomen, and multiple masses on the right lateral liver lobe. Clinicopathologic and imaging abnormalities included anemia, azotemia, icterus, and hepatomegaly with hypoechoic masses. On cytologic evaluation of a fine‐needle aspiration of a liver mass there was abundant extracellular pink‐ to purple‐colored material between hepatocytes. The amorphous material was stained with direct fast scarlet (DFS), and green birefringent areas were observed under polarized light, confirming the presence of amyloid. A unique finding on the cytologic smear were macrophages containing amorphous and fibrillar amyloid‐like protein. Histopathologic examination using H&E and Congo red staining confirmed amyloid deposits within the space of Disse, along the sinusoids, portal tracts, blood vessel walls, and within the cytoplasm of macrophages. Immunohistochemical staining with anti‐AA amyloid antibodies further confirmed the presence of AA amyloid. To the author's knowledge, this is the first report of the cytologic finding of AA amyloid protein within macrophages and DFS stain detection of amyloid on a cytologic smear.     

Effects of Bedding Material on the Lying Behavior in Stabled Horses

The objective of this study was to investigate the effect of straw, sawdust, coconut husk (husk), and coconut fiber (fiber) on the welfare of stable horses by observing their resting behavior. Twenty horses with ages ranging from 3 to 21 years were used at the Equine Research Institute of the Japan Racing Association, Utsunomiya, Japan. Five horses were allocated to each bedding condition. The behavior of each horse was recorded by video camera for 3 days and was continuously sampled from 17:00 to 05:00. The total duration, the number of bouts, and the mean and the maximum duration of bouts in standing rest, sternal lying, and lateral lying were calculated and analysed by the Kruskal-Wallis test and post hoc Steel-Dwass test. There was no difference in the standing rest and the sternal lying among beddings. Significant differences were observed in these values in the lateral lying among the different beddings (P<0.05). The values of the means of the total duration, the number of bouts, and the mean and the maximum duration of bout in the lateral lying were greater when husk was used as the bedding material than when sawdust were used (P<0.05). The results of the observations show that the new bedding materials would be as usable as straw. However, lateral lying was observed less frequently when sawdust were used as bedding; this indicates that use of sawdust as bedding material will decrease the welfare of stabled horses.     

Induction of Soil Suppressiveness Against Rhizoctonia solani by Incorporation of Dried Plant Residues into Soil

ABSTRACT Suppressive effects of soil amendment with residues of 12 cultivars of Brassica rapa on damping-off of sugar beet were evaluated in soils infested with Rhizoctonia solani. Residues of clover and peanut were tested as noncruciferous controls. The incidence of damping-off was significantly and consistently suppressed in the soils amended with residues of clover, peanut, and B. rapa subsp. rapifera 'Saori', but only the volatile substance produced from water-imbibed residue of cv. Saori exhibited a distinct inhibitory effect on mycelial growth of R. solani. Nonetheless, disease suppression in such residue-amended soils was diminished or nullified when antibacterial antibiotics were applied to the soils, suggesting that proliferation of antagonistic bacteria resident to the soils were responsible for disease suppression. When the seed (pericarps) colonized by R. solani in the infested soil without residues were replanted into the soils amended with such residues, damping-off was suppressed in all cases. In contrast, when seed that had been colonized by microorganisms in the soils containing the residues were replanted into the infested soil, damping-off was not suppressed. The evidence indicates that the laimosphere, but not the spermosphere, is the site for the antagonistic microbial interaction, which is the chief principle of soil suppressiveness against Rhizoctonia damping-off.     

Whole-Cell Fatty Acid Composition to Characterize and Differentiate Isolates of Rhizoctonia Species Associated with Turfgrass Diseases in Japan

Cellular fatty acids were analyzed to characterize and differentiate 34 isolates of Rhizoctonia species representing binucleate Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB, AG 2-2 LP, R. circinata var. circinata and var. oryzae associated with turfgrass diseases in Japan. Myristic, pentadecanoic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acids were consistently present in varying quantities in all isolates. Heptadecanoic and 9-heptadecenoic acids were present in isolates of Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB and AG 2-2 LP but not in isolates of R. circinata var. circinata and var. oryzae. Palmitic, oleic and linoleic acids were the major fatty acids found, constituting 88.30-98.37% of the whole-cell fatty acid content. The remaining fatty acids were present in smaller amounts. Isolates within a single group were closely clustered, whereas isolates from different groups were clearly distinguishable based on average linkage cluster analysis of cellular fatty acids. Principal component analysis, based on all fatty acids detected, confirmed the distinct separation of isolates representing the six groups of Rhizoctonia species obtained from turfgrasses. These results suggested that fatty acid analysis is useful for the characterization and differentiation of isolates of Rhizoctonia species associated with turfgrass diseases. Received 21 May 2001/ Accepted in revised form 28 September 2001     

Responses of leaf photosynthesis and plant growth to altered source–sink balance in a Japanese and a Dutch tomato cultivar

We investigated the effects of altered source/sink ratio by leaf or fruit pruning on leaf photosynthetic characteristics and whole-plant growth of ‘Momotaro York’, a Japanese cultivar, and ‘Dundee’, a Dutch cultivar and verified a hypothesis for sink-limitation of plant growth proposed by Tanaka and Fujita (1974). Plants were grown hydroponically with a high-wire system in a greenhouse for 11 weeks. Light-saturated photosynthesis of young, fully expanded leaves measured at atmospheric CO₂ partial pressure of 37 Pa and at an intercellular CO₂ partial pressure of 20 Pa was not influenced by alteration of source/sink ratio for either cultivar. Although soluble sugars were accumulated in leaves under high source/sink conditions, the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase, a rate-limiting factor for CO₂-limited photosynthesis, was not affected by the altered source–sink balance. Net assimilation rate of a whole plant increased with decreasing source/sink ratio, but this can be accounted for by the changes in light interception per unit leaf area, without taking the source–sink relationship into consideration. It was concluded that the altered source/sink ratio did not change leaf photosynthetic capacity and the sink-limitation hypothesis cannot be applied to either cultivar under the conditions of the present study.     

Growth characteristic and sink strength of fruit at different CO2 concentrations in a Japanese and a Dutch tomato cultivar

The aim of the present study was to elucidate how fruit growth was limited by the source and sink capacities in a Japanese (‘Momotaro York’) and a Dutch (‘Dundee’) tomato cultivar. The two cultivars were grown hydroponically with a high-wire system in greenhouses for 25 weeks, and the growth characteristics and sink strength of fruit were determined. Fruits were pruned to four (4F) or one (1F) per truss. The latter were used to determine potential fruit growth, an indicator of fruit sink strength. Growth was also determined under normal (LC) and enriched (HC, 700 μmol mol⁻¹) CO₂ concentrations to examine the effect of source enhancement on fruit production. In both cultivars under normal CO₂, the growth rate of fruit pruned to 4F per truss was lower than that in 1F, indicating that maximum potential fruit growth was not achieved. Under HC conditions, fruit growth rate of ‘Dundee’ achieved in 4F trusses was lower than that in 1F. In ‘Momotaro York’ in HC, fruit growth in 4F trusses was close to potential. This implies that fruit growth was source-limited irrespective of CO₂ concentrations in ‘Dundee’ cultivar while fruit growth in ‘Momotaro York’ under normal and enriched CO₂ conditions was limited by source and sink strengths, respectively. Adjustments of cultural practices including increasing fruit number per truss and/or genetic approaches to enhancing fruit sink strength by breeding may improve fruit yields of Japanese cultivars under high source/sink conditions.     

Postoperative improvement of megaesophagus and esophageal motility in a 19-day-old puppy with gastroesophageal intussusception

Canine gastroesophageal intussusception (GEI) is a rare and life-threatening condition that requires prompt diagnosis and treatment. A 19-day-old Siberian Husky with a 4-day history of regurgitation was diagnosed with GEI based on the findings of computed tomography (CT) performed without anesthesia. Endoscopic reduction of intussusception was impossible; thus, surgical reduction by traction of the duodenum was performed. CT revealed improvement of megaesophagus 82 days postoperatively. Eleven months postoperatively, fluoroscopy showed recovery to nearly normal esophageal motility. Two years postoperatively, no clinical signs were reported. CT is useful to diagnose GEI in neonate puppies with poor abdominal fat and to assess the gastric edema and the anatomical association of stomach with other organs. Fluoroscopy is helpful for evaluating postoperative esophageal motility.     

Development and characterization of monoclonal antibodies against canine trypsin

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.     

Correlation between villous height and the disaccharidase activity in the small intestine of piglets from nursing to growing

Early weaning induces villous atrophy in the small intestine. Reduction in villous height in the small intestine after weaning is associated with reductions in brush‐border enzyme activity. Body weight gain after weaning is, therefore, correlated with villous height. This evidence suggested that the maintenance of small intestinal structure and function after weaning is important for the growth of young pigs. On the other hand, the relationship between villous height and the activity of the digestive enzymes in the small intestine has not been studied with piglets from the suckling to the growing period. Five suckling piglets, four piglets in the proximal stage of weaning, four pigs in the distal stage of weaning and four growing pigs were used. The activities of lactase (LA), sucrase (SA) and maltase (MA) were determined. LA showed a positive correlation with villous height in weaning. SA and MA were positively correlated with villous height from suckling to growing. In a previous study, non‐infectious dyspeptic diarrhea was frequently observed in growing piglets on Japanese swine farms. The maintenance of villous height to retain disaccharidase activity may prevent dyspepsic diarrhea in this stage.     

Identification of the core motif of the BRCA2 C-terminal RAD51-binding domain by comparing canine and human BRCA2

Mammary tumors are the most common tumors in women and non-spayed female dogs. One of the reasons for mammary tumors is mutations of the tumor suppressor gene, BRCA2. BRCA2 participates in homologous recombination repair by interacting with the RAD51 recombinase. BRCA2 has two RAD51-binding domains, consisting of BRC repeats and the C-terminal RAD51-binding domain, respectively. Although several studies have addressed the function of the C-terminal RAD51-binding domain of human BRCA2, the amino acid sequences required for the RAD51-interaction activity remain unclear. In this study, the C-terminal RAD51-binding domains of canine and human BRCA2 were compared; the canine domain displayed a weaker interaction with RAD51. This difference was attributed to the C-terminal portion of the domain via a comparison between canine and human domains. Furthermore, peptides shorter than those previously reported displayed RAD51-interacting activity, and a core motif of this domain consisting of 25 amino acids was identified. Since a mutation (S3323N) was reported in the core motif of this domain, the effect of this mutation was evaluated. The mutant exhibited similar RAD51-binding activity as that of the wild-type protein, suggesting that the mutation was functionally neutral. These data suggested that the C-terminal portion of the BRCA2 C-terminal RAD51-binding domain influenced its RAD51-interaction activity, and a minimum core motif of 25 amino acids was identified in this domain. These data may help clarify BRCA2 function, as well as the tumorigenic effects of BRCA2 mutation.