Objective-To investigate telomere lengths in tissues of domestic shorthair (DSH) cats of various ages, evaluate the relationship between telomere length and age of cats, and investigate telomerase activity in the somatic tissues of cats. Sample Population-Tissues obtained from 2 DSH cats and blood samples obtained from 30 DSH cats. Procedure-DNA isolated from blood cells and somatic tissue samples was subjected to terminal restriction fragment (TRF) analysis to determine mean telomere repeat lengths. Protein samples were subjected to analysis by use of a telomeric repeat-amplification protocol to assess telomerase activity. Results-Mean TRF values of cats ranged from 4.7 to 26.3 kilobase pairs, and there was significant telomeric attrition with increasing age of cat. Telomerase activity was not found in a wide range of normal tissues obtained from 2 cats. Conclusions and Clinical Relevance-Analysis of these results clearly indicates that telomeres are shorter in older cats, compared with young cats; therefore, telomeres are implicated in the aging process. The analysis of telomerase activity in normal somatic tissues of cats reveals a pattern of expression similar to that found in human tissues. Impact for Human Medicine-Fundamental differences in the biological characteristics of telomeres and telomerase exist between humans and the other most widely studied species (ie, mice). The results reported here reveal similarities in telomere and telomerase biologic characteristics between DSH cats and humans. Hence, as well as developing our understanding of aging in cats, these data may be usefully extrapolated to aging in humans.

Objective-To determine whether transitional cell carcinoma (TCC) cells incubated in media containing 5-aminolevulinic acid (ALA) would produce sufficient protoporphyrin IX (PpIX) to cause lethal phototoxic effects when exposed to 635-nm light. Sample Population-Canine TCC cells (K9TCC). Procedure-Cultured K9TCC cells were exposed to graded doses of ALA, and PpIX concentrations were determined. Cells then were exposed to various doses of 635-nm light from a diode laser, and cell viability was assayed. Results-Production of PpIX was dependent on time and dose of ALA. The K9TCC cells incubated with ALA produced sufficient PpIX to cause lethal phototoxic effects when exposed to 635-nm light. Phototoxic effects were dependent on time and dose of ALA. Increasing laser power density and energy density decreased cell survival. Conclusions and Clinical Relevance-ALA is an effective photosensitizer for in vitro photodynamic treatment of K9TCC cells. Further studies are warranted to assess the safety and efficacy of ALA as a photosensitizer for use in treating dogs with TCC. Impact for Human Medicine-On the basis of this study, dogs with TCC may be useful in the development of protocols for ALA-based photodynamic therapy of humans affected with muscle-invasive bladder cancer.

Objective-To use transient and stable transfection of Chinese hamster ovary cells to clone the gene encoding feline erythropoietin (feEPO) protein, characterize the expressed protein, and assess its biological activity. Sample Population-Cultures of Chinese hamster ovary or TF-1 cells. Procedure-The gene encoding feEPO was cloned into a eukaryotic expression plasmid. Chinese hamster ovary cells were transiently or stably transfected with the plasmid. Expressed recombinant feEPO (rfeEPO) protein was purified from transiently transfected cells. The protein was characterized by use of SDS gel electrophoresis and western blot analysis. Biological activity was assessed by measuring thymidine incorporation by TF-1 erythroleukemic cells. Results-Purified rfeEPO from supernatants of transiently transfected cells was determined to be 34 to 40 kilodaltons (kd) by use of SDS gel electrophoresis, whereas the molecular weight predicted from the amino acid sequence was 21.5 kd. The banding pattern and high molecular weight suggested the protein was glycosylated. The rfeEPO proteins derived from transient or stable transfections subsequently were determined to be biologically active in vitro. Conclusions and Clinical Relevance-The gene encoding feEPO can be transfected into eukaryotic cells, and the expressed rfeEPO protein is biologically active in vitro. Cats with chronic renal failure often are anemic as a result of reduced expression of erythropoietin (EPO). Treatment with human-derived EPO stimulates RBCs in anemic cats; however, treatment is often limited by the development of antibodies directed against the recombinant human protein, which can then cross-react with endogenous feEPO. Recombinant feEPO may prove beneficial for use in cats with chronic renal failure.

Objective-To evaluate response of euthyroid cats to administration of recombinant human thyroid-stimulating hormone (rhTSH). Animals-7 healthy cats. Procedure-Each cat received each of 5 doses of rhTSH (0, 0.025, 0.050, 0.100, and 0.200 mg), IV, at 1-week intervals. Serum concentration of total thyroxine (TT4) and free thyroxine (fT4) was measured immediately before each injection (time 0) and 2, 4, 6, and 8 hours after administration of each dose. Results-Overall TT4 response did not differ significantly among cats when administered doses were ≥ 0.025 mg. Serum TT4 concentrations peaked 6 to 8 hours after administration for all doses greater than 0.025 mg. For all doses greater than 0.025 mg, mean +/- SEM TT4 concentration at 0, 6, and 8 hours was 33.9 +/- 1.7, 101.8 +/- 5.9, and 101.5 +/- 5.7 nmol/L, respectively. For all doses greater than 0.025 mg, mean fT4 concentration at 0, 6, and 8 hours was 38.7 +/- 2.9, 104.5 +/- 7.6, and 100.4 +/- 8.0 pmol/L, respectively. At 8 hours, the fT4 response to 0.025 and 0.050 mg was less than the response to 0.100 and 0.200 mg. Adverse reactions after rhTSH administration were not detected. Conclusions and Clinical Relevance-The TSH stimulation test can be performed in cats by IV administration of 0.025 to 0.200 mg of rhTSH and measurement of serum TT4 concentrations at time of injection and 6 or 8 hours later. Clinical validation of the TSH stimulation test would facilitate development of additional tests of thyroid gland function, such as a TSH assay.

The objective of this study was to describe the kinetics of urinary recovery and to evaluate the effects of postmucosal factors on urinary recovery of 5 intravenously administered saccharides. Ten cats received an isotonic sugar solution containing lactulose, rhamnose, xylose, methylglucose, and sucrose intravenously. These sugars were selected because of their prior use for intestinal permeability and mucosal function testing in humans and dogs. Urethral catheterization with a closed collection system was used for collection of cumulative urine samples prior to and 2, 4, 6, 8, 10, 12, and 24 h after administration of the sugar solution. High-pressure anion exchange liquid chromatography with pulsed amperometric detection was used to measure the concentrations of each sugar in the urine and calculate urinary recovery. Twenty-four hour cumulative urinary recovery for each sugar from the cats, was lower than expected compared to dogs and humans. All 5 sugars had the highest percentage of urinary recovery during the first 2 h after administration. Mean sugar elimination rate constants and half-lives ranged from 0.268/h for methylglucose to 0.415/h for lactulose and 1.67 h for lactulose to 2.59 h for methylglucose, respectively. Metabolism and incomplete urine collection are possible reasons for lower cumulative urinary recoveries of these 5 sugars in cats compared with dogs. Although these 5 sugars are not ideal marker molecules, they may still be useful for intestinal permeability and mucosal function testing in cats.