Urinary transforming growth factor-beta1 in feline chronic renal failure

Transforming growth factor-beta1 (TGF-beta1), an inflammatory cytokine, plays a role in tissue fibrosis, such as glomerular sclerosis and tubulointerstitial fibrosis of the kidneys. In the present study, the urinary TGF-beta1 level of cats diagnosed with chronic renal failure (CRF) was measured to investigate its relationship to the pathogenesis of feline CRF. Urinary TGF-beta1 levels (TGF-beta1/creatinine ratio) were significantly increased compared with healthy controls, whereas serum levels of TGF-beta1 were not. These results indicate that TGF-beta1 is expressed in the kidneys of CRF cats, and that it was reflected in the urinary TGF-beta1 level. Therefore, TGF-beta1 may play a role in feline CRF, and urinary TGF-beta1 could be used as a clinical marker for renal fibrosis.     

Establishment of a chicken monoclonal antibody panel against mammalian prion protein

A panel of chicken monoclonal antibodies (mAbs) was developed against prion protein (PrP), the sequence of which is a highly conserved molecule among mammals. A portion of the splenocytes from chickens immunized with recombinant mouse PrP was fused with the chicken B cell line, MuH1. The remaining splenocytes were used to generate the recombinant mAbs by phage display. A total of 36 anti-PrP mAbs, 2 from cell fusion and 34 from phage display were established. The specificity of these mAbs was determined by Western blot and ELISA using various PrP antigens including recombinant PrPs, synthetic PrP peptides and PrPs from brains or scrapie-infected neuroblastoma cell line. These mAbs were classified into three main groups, protease K (PK)-sensitive (Group I), PK cleavage site proximal (Group II) and PK-resistant (Group III), based on their abilities to recognize PrP following PK-treatment. Some mAbs were found to selectively recognize different glycoforms of PrP as well as the metabolic fragments of PrP. Furthermore, we found that PrP recognition by chickens differed from that by PrP-knockout mouse. These results indicate that these newly generated PrP antibodies from chickens will help to research the PrP and to establish the diagnosis of prion disease.     

Inhibition of p16 tumor suppressor gene expression via promoter hypermethylation in canine lymphoid tumor cells

To investigate the epigenetic regulation of the p16 gene in canine lymphoid tumor cells, its methylation status was examined in four canine lymphoid tumor cell lines. In three canine lymphoid tumor cell lines (CLBL-1, GL-1, and UL-1) with low-level p16 mRNA expression, 20 CpG sites in the promoter region of p16 gene were consistently methylated although all of the CpG sites were not methylated in another cell line (CL-1) and normal lymph node cells. The expression level of p16 mRNA in these three cell lines was restored after cultivation in the presence of a methylation inhibitor, 5-Aza-2′-deoxycitidine, indicating inactivation of p16 gene via hypermethylation. This study revealed the inactivation of p16 gene through hypermethylation of its CpG island in a fraction of canine lymphoid tumor cells.     

Total plant sterols,steryl ferulates and steryl glycosides in milling fractions of wheat and rye

The total plant sterol, steryl ferulate and steryl glycoside contents in wheat and rye milling fractions show that there are significant differences in both sterol content and composition between various milling fractions collected from a commercial mill. Total sterols were analysed by gas chromatography after acid and alkaline hydrolyses. The steryl conjugates were first extracted with acetone, fractionated on solid phase extraction (SPE) cartridges and then analysed individually (steryl glycosides by gas chromatography and steryl ferulates by reverse phase-high performance liquid chromatography with UV detection. Differences in sterol contents of the wheat samples were greater than in the rye samples. The highest total sterol content was found in wheat germ, but surprisingly high sterol contents, that were comparable to bran, were found in some flour fractions. Contents of steryl ferulates were high in the bran fractions contributing up to 17% of total sterols. The variation in the content of steryl glycosides in the samples was lower and contributed less than 10% of total sterols. These results show that much of the bioactive components may be lost when certain flour fractions produced in common flour milling procedures are discarded. However, some of these fractions with significantly high sterol contents could possibly be introduced into milling products used in breadmaking and the food industry without greatly compromising consumer acceptability.     

Time–course monitoring of serum amyloid A in a cat with pancreatitis

Abstract: Time‐course changes in the concentration of serum amyloid A (SAA), a major acute phase protein, were measured in a cat with pancreatitis over an 831‐day period and compared with changes in WBC count and feline trypsin‐like immunoreactivity (fTLI). SAA concentration was increased at the onset of the disease and gradually decreased over 5 days of treatment with an improvement in the clinical condition. In contrast, fTLI concentration and WBC count were not increased at the onset of the disease but increased gradually during the 5 days of treatment. Long‐term monitoring from days 68 to 831 revealed a good correlation between SAA concentration and the reoccurrence of clinical signs in the cat; however, WBC count did not increase even with the exacerbation of disease. These findings suggest that the SAA concentration may be a useful marker for evaluating response to treatment and disease exacerbation in feline pancreatitis.