Comparative Analysis of mRNA Expression of Surface Antigens between Histiocytic and Nonhistiocytic Sarcoma in Dogs

Background

Definitive diagnosis of histiocytic sarcoma (HS) in dogs is relatively difficult by conventional histopathological examination because objective features of HS are not well defined.

Hypothesis

Quantitative analysis of mRNA expression of selected cellular surface antigens (SAs) specific to HS in dogs can facilitate objective and rapid diagnosis.

Animals

Dogs with HS (n = 30) and dogs without HS (n = 36), including those with other forms of lymphoma (n = 4), inflammatory diseases (n = 6), and other malignant neoplasias (n = 26).

Methods

Retrospective clinical observational study. Specimens were collected by excisional biopsy, needle core biopsy, or fine needle aspiration. To determine HS detection efficacy, mRNA expression levels of selected SAs specific to HS in dogs, including MHC class IIα, CD11b, CD11c, and CD86, were quantitatively analyzed using real‐time quantitative polymerase chain reaction.

Results

Each SA mRNA expression level was significantly higher in HS dogs than in non‐HS dogs (P = .0082). Cutoff values for discriminating between HS and non‐HS dogs based on these expression levels were calculated on the basis of receiver‐operating characteristic analysis. Accuracy of the cutoff values, including MHC class IIα, CD11b, CD11c, and CD86, was 87.9, 86.4, 86.4, and 84.8%, respectively.

Conclusions and Clinical Importance

Our results suggest that quantitative analysis of mRNA expression of the selected SAs could be an adjunctive diagnostic technique with high diagnostic accuracy for HS in dogs. Substantial investigation is required for exclusion of diseases with similar cell types of origin to lymphoma.     

COMPARISON BETWEEN ULTRASONOGRAPHIC AND CLINICAL FINDINGS IN 43 DOGS WITH GALLBLADDER MUCOCELES

Cholecystectomy is the current standard recommended treatment for dogs with gallbladder mucoceles. However, medical management with monitoring has also been recommended for asymptomatic dogs. The purpose of this retrospective study was to compare ultrasonographic patterns of gallbladder mucoceles with clinical disease status in a group of dogs. For each included dog, the ultrasonographic pattern of the mucocele was classified into one of six types: type 1, immobile echogenic bile; type 2, incomplete stellate pattern; type 3, typical stellate pattern; type 4, kiwi like pattern and stellate combination; type 5, kiwi like pattern with residual central echogenic bile; and type 6, kiwi like pattern. A total of 43 dogs were included. Twenty‐four dogs, including 11 dogs with gallbladder rupture, were symptomatic. Nineteen dogs were asymptomatic. Cholecystectomy (n = 19), medical therapy (n = 17), or monitoring (n = 6) treatments were applied according to clinical signs and owners’ requests. One dog suspected of having gallbladder rupture was euthanized. Frequencies of gallbladder mucocele patterns were as follows: type 1 = 10 (23%), type 2 = 13 (30%), type 3 = 5 (12%), type 4 = 11 (26%), type 5 = 4 (9%), and type 6 = 0. In dogs with gallbladder rupture, type 2 (8/13) was the most common. No significant correlations were found between ultrasonographic patterns of gallbladder mucoceles and clinical disease status or gallbladder rupture. Findings indicated that ultrasonographic patterns of gallbladder mucoceles may not be valid bases for treatment recommendations in dogs.     

Ultrasonographic evaluation of renal dimension and resistive index in clinically healthy Korean domestic short-hair cats

Renal length, height, width, resistive index (RI), size of cortex, and medulla were determined by renal ultrasonography in 50 healthy Korean domestic short-hair cats. In the sagittal plane, the renal length was 3.83 ± 0.51 cm (mean ± SD) in the left kidney and 3.96 ± 0.48 cm in the right kidney, whereas the renal height was 2.42 ± 0.27 cm in the left kidney and 2.36 ± 0.28 cm in the right kidney. In the transverse plane, the renal height was 2.42 ± 0.28 cm in the left kidney and 2.38 ± 0.27 cm in the right kidney, whereas the renal width was: 2.65 ± 0.35 cm in the left kidney and 2.63 ± 0.31 cm in the right kidney. In the dorsal plane, the renal length was 3.84 ± 0.53 cm in the left kidney and 3.97 ± 0.54 cm in the right kidney, whereas the renal width was 2.65 ± 0.34 cm in the left kidney and 2.66 ± 0.33 cm in the right kidney. There were no significant differences (p > 0.05) among the same structure sizes measured in different planes. In the sagittal plane, the size of the renal cortex was 0.47 ± 0.08 cm in the left kidney and 0.47 ± 0.08 cm in the right kidney, whereas of the size of the renal medulla was 0.55 ± 0.30 cm in the left kidney and 0.50 ± 0.07 cm in the right kidney. RI evaluated by pulsed wave Doppler sonography was 0.52 ± 0.05 in the left kidney and 0.55 ± 0.05 in the right kidney. The actual renal dimensions determined by gross examination were not statistically different from those determined by ultrasonography. Furthermore the renal dimensions and RI were statistically correlated to the body weight of cats.     

Genetic parameters for various random regression models to describe total sperm cells per ejaculate over the reproductive lifetime of boars

The objective of this study was to model the variances and covariances of total sperm cells per ejaculate (TSC) over the reproductive lifetime of AI boars. Data from boars (n = 834) selected for AI were provided by Smithfield Premium Genetics. The total numbers of records and animals were 19,629 and 1,736, respectively. Parameters were estimated for TSC by age of boar classification with a random regression model using the Simplex method and DxMRR procedures. The model included breed, collector, and year-season as fixed effects. Random effects were additive genetic, permanent environmental effect of boar, and residual. Observations were removed when the number of data at a given age of boar classification was < 10 records. Preliminary evaluations showed the best fit with fifth-order polynomials, indicating that the best model would have fifth-order fixed regression and fifth-order random regressions for animal and permanent environmental effects. Random regression models were fitted to evaluate all combinations of first- through seventh-order polynomial covariance functions. Goodness of fit for the models was tested using Akaike's Information Criterion and the Schwarz Criterion. The maximum log likelihood value was observed for sixth-, fifth-, and seventh-order polynomials for fixed, additive genetic, and permanent environmental effects, respectively. However, the best fit as determined by Akaike's Information Criterion and the Schwarz Criterion was by fitting sixth-, fourth-, and seventh-order polynomials; and fourth-, second-, and seventh-order polynomials for fixed, additive genetic, and permanent environmental effects, respectively. Heritability estimates for TSC ranged from 0.27 to 0.48 across age of boar classifications. In addition, heritability for TSC tended to increase with age of boar classification.     

Molecular characterization of Korean rabies virus isolates

The nucleoprotein (N) and glycoprotein (G) of 11 Korean rabies virus (RABV) isolates collected from animals diagnosed with rabies between 2008 and 2009 were subjected to molecular and phylogenetic analyses. Six isolates originated from domestic animals (cattle and dogs) and five were obtained from wild free-ranging raccoon dogs. The similarities in the nucleotide sequences of the N gene among all Korean isolates ranged from 98.1 to 99.8%, while those of the G gene ranged from 97.9 to 99.3%. Based on the nucleotide analysis of the N and G genes, the Korean RABV isolates were confirmed as genotype I of Lyssavirus and classified into four distinct subgroups with high similarity. Phylogenetic analysis showed that the Korean isolates were most closely related to the non-Korean NeiMeng1025B and 857r strains, which were isolated from rabid raccoon dogs in Eastern China and Russia, respectively. These findings suggest that the Korean RABV isolates originated from a rabid raccoon dog in Northeastern Asia. Genetic analysis of the Korean RABV isolates revealed no substitutions at several antigenic sites, indicating that the isolates circulating in Korea may be pathogenic in several hosts.     

Occurrence of Honeysuckle Yellow Vein Virus (HYVV) containing a monopartite DNA-A genome in Korea

Two newly emerged begomoviruses were isolated from naturally infected tomato (Solanum lycopersicum) plants grown in greenhouses at Jeju Island and Dangjin in Korea and their genomes were characterized. These viruses-infected plants had very small leaves that curled upward, yellow margins and a leathery appearance, and a bushy and stunted appearance with short internodes. Nucleotide (nt) sequence analysis of their genomes showed that they have a DNA-A component of a monopartite begomovirus. Their genomes comprised 2763 and 2764 nucleotides with six open reading frames. The results of nt sequence similarity analysis of DNA-A genome between the two Korean isolates and isolates of Tobacco leaf curl Japan virus (TbLCJV), Honeysuckle yellow vein virus (HYVV), Honeysuckle yellow vein mosaic virus (HYVMV), and Eupatorium yellow vein virus in Japan (EpYVV) showed that they are likely similar to HYVV-[Masuda] (89.4–92.8% nt identity). Consequently, we tentatively propose the two isolates’ names as HYVV-Jeju and -DJ according to the ICTV geminivirus rules. Phylogenetic relationship analysis of 33 DNA-A genome sequences using PAUP* 4.0b10 and MrBayes revealed that HYVV-Jeju and -DJ belong to the Far East Asian begomovirus species complex. Within the Far East Asian begomovirus species complex, HYVV-Jeju and -DJ are distantly related to EpYVV, HYVMV, and TbLCJV groups. Based on the presence of a recombination fragment spanning the C3 ORF, a recombinant origin was suggested for both HYVV-Jeju and –DJ, with parents close to Japanese isolates HYVMV-[SP1:00] and Eupatorium yellow vein virus (EpYVV)-[Suya]. In addition, the presence of a further recombination fragment spanning the IR suggested the parents of HYVV-DJ were close to HYVV-Jeju and EpYVV-[Suya].     

Isolation and Characterization of Efficient Active Compounds Using High-Performance Centrifugal Partition Chromatography (CPC) from Anti-Inflammatory Activity Fraction of Ecklonia maxima in South Africa

Ecklonia maxima is a brown seaweed, which is abundantly distributed in South Africa. This study investigated an efficient approach using high-performance centrifugal partition chromatography (HPCPC), which has been successfully developed for the isolation and purification of phlorotannins, eckmaxol, and dieckol from the ethyl acetate fraction of E. maxima (EEM). We evaluated EEM for its inhibitory effect against lipopolysaccharide (LPS)-induced inflammatory responses in zebrafish embryos. The separation of eckmaxol and dieckol from samples of EEM using HPCPC was found to be of high purity and yield under an optimal solvent system composed of n-hexane:ethyl acetate:methanol:water (2:7:3:7, v/v/v/v). To evaluate the anti-inflammatory efficacy of EEM containing active compounds, zebrafish embryos exposed to LPS were compared with and without EEM treatment for nitric oxide (NO) production, reactive oxygen species (ROS) generation, and cell death two days after fertilization. These evaluations indicate that EEM alleviated inflammation by inhibiting cell death, ROS, and NO generation induced by LPS treatment. According to these results, eckmaxol and dieckol isolated from brown seaweed E. maxima could be considered effective anti-inflammatory agents as pharmaceutical and functional food ingredients.