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 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 determine the prevalence of biofilm formation under long-term cell culture conditions in serum samples of dairy cattle, goats, cats, and dogs, and to determine whether there is an association between nanobacteria and biofilm formation. Sample Population-Serum samples of clinically normal animals (313 dairy cattle, 48 goats, 140 dogs, and 44 cats) and animals with various medical conditions (60 dogs and 116 cats). Procedure-Serum was incubated under cell culture conditions and observed for biofilm formation by use of light microscopy, electron microscopy, and spectroscopy. A polymerase chain reaction assay was developed to identify 16S rRNA gene sequences of nanobacteria. Results-Biofilm formation developed in serum samples of 304 of 313 (97%) cattle, 44 of 48 (92%) goats, 44 of 44 (100%) cats, and 126 of 140 (90%) dogs. Prevalence of serum samples with positive results for biofilm formation was not significantly different between cats or dogs with and without medical conditions associated with pathologic extraskeletal calcification processes. Scanning electron microscopy and spectroscopy of biofilm samples revealed small coccoid particles consisting mainly of calcium and phosphate. Polymerase chain reaction assay failed to amplify sequences of nanobacteria. Conclusions and Clinical Relevance-Under longterm cell culture conditions, biofilm made up of aggregates of calcium and phosphate crystals does form in serum samples of clinically normal dairy cattle, goats, cats, and dogs. Disease, however, does not predispose to biofilm formation in serum samples of dogs and cats. Our findings did not support the existence of nanobacteria in serum samples of cattle, goats, cats, and dogs.

Objective-To evaluate the association between airway reactivity and age, sex, body weight, and radiographic findings in cats. Animals-32 mature cats that constituted 2 age groups (17 young cats that were 1 to 2 years old and 15 old cats that were 12 to 13 years old). Procedure-Cats were placed in the chamber of a barometric whole-body plethysmograph (volume, 38 L), and box pressure was measured at baseline and after aerosol administration of increasing concentrations of carbachol. Airway reactivity was assessed by monitoring increases in enhanced pause (PENH), a unitless variable that measures bronchoconstriction as derived from dose-response curves. The endpoint chosen was the provocative concentration of carbachol that increased PENH to 300% of the baseline value (PCPENH300). Results-We did not find a correlation between PCPENH300 and sex, body weight, number of eosinophils, PENH before bronchoconstriction, respiratory frequency, tidal volume, or minute ventilation. Airway reactivity was significantly less in the old cats (mean +/- SD PCPENH300, 0.578 +/- 0.051%), compared with the value for the young cats (0.053 +/- 0.006%). Radiographic patterns differed significantly between groups of cats; a greater proportion of old cats (12/15) had bronchointerstitial patterns, compared with the proportion of young cats (4/17). Conclusion and Clinical Relevance-These data support the notion that age exerts a strong influence on airway reactivity in adult cats, and radiographic differences suggest that structural changes in older cats may contribute to this effect. These findings have important implications for interpretation of results of airway reactivity tests in cats.

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.

Objective-To compare the minimum alveolar concentration (MAC) of isoflurane required to prevent corticocerebral activation, autonomic responses, and purposeful movements after somatic or visceral stimulation in cats anesthetized with isoflurane. Animals-17 healthy spayed female cats. Procedure-Bispectral index (BIS), autonomic parameters, and purposeful movements were monitored before and after somatic or visceral stimuli in cats anesthetized with isoflurane. End-tidal (ET) isoflurane concentration was varied to determine MAC values for cortical arousal (MACBIS), autonomic responsiveness (MACBAR), and purposeful movement (MAC). Bispectral index values greater than 60 were considered to represent corticocerebral activation. Results-Minimum alveolar concentration for purposeful movement was significantly less than MACBIS and MACBAR for both somatic and visceral stimulation. Individual MAC values for somatic stimulation were not significantly different from respective MAC values for visceral stimulation. The percentage of cats that had a BIS response greater than 60 was inversely related to the end-tidal isoflurane concentration. Conclusions and Clinical Relevance-Corticocerebral arousal and subcortical autonomic reflexes occured at isoflurane anesthetic concentrations at which reflexive or purposeful movements were absent. These results suggested that isoflurane had a preferential effect on voluntary motor output at low end-tidal isoflurane concentrations, and that sensory pathways, subcortical sympathetic output, and cortical responsiveness are less susceptible to the anesthetic effects of isoflurane. Bispectral index values obtained after somatic or visceral stimulation were sensitive for the detection of early changes in cortical excitability.

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.