Objective—To investigate the in vitro effect of the combination of lignan enterolactone (ENL) or lignan enterodiol (END) with melatonin on steroid hormone secretion and cellular aromatase content in human adrenal carcinoma cells. Sample—Human adrenocortical carcinoma cells.
Procedures—Melatonin plus ENL or END was added to cell culture medium along with cAMP (100μM); control cells received cAMP alone. Medium and cell lysates were collected after 24 and 48 hours of cultivation. Samples of medium were analyzed for progesterone, 17-hydroxyprogesterone, androstenedione, aldosterone, estradiol, and cortisol concentration by use of radioimmunoassays. Cell lysates were used for western blot analysis of aromatase content.
Results—The addition of ENL or END with melatonin to cAMP-stimulated cells (treated cells) resulted in significant decreases in estradiol, androstenedione, and cortisol concentrations at 24 and 48 hours, compared with concentrations in cells stimulated with cAMP alone (cAMP control cells). The addition of these compounds to cAMP-stimulated cells also resulted in higher progesterone and 17-hydroxyprogesterone concentrations than in cAMP control cells; aldosterone concentration was not affected by treatments. Compared with the content in cAMP control cells, aromatase content in treated cells was significantly lower.
Conclusions and Clinical Relevance—The combination of lignan and melatonin affected steroid hormone secretion by acting directly on adrenal tumor cells. Results supported the concept that this combination may yield similar effects on steroid hormone secretion by the adrenal glands in dogs with typical and atypical hyperadrenocorticism.
Objective—To evaluate the effects of IM administration of recombinant human thyroid-stimulating hormone (rhTSH) on plasma total thyroxine (T4) concentrations in euthyroid ferrets.
Animals—25 healthy neutered ferrets (14 female and 11 male) of various ages from 2 populations (laboratory ferrets from Georgia and pet ferrets from Pennsylvania).
Procedures—Each ferret underwent a physical examination and standard hematologic testing to ensure it was healthy and had clinically normal thyroid function. Once determined to be euthyroid, ferrets received a single IM injection of 100 μg of rhTSH. Blood samples were collected into plasma-separator tubes immediately before the rhTSH injection (time 0) and 4 hours after injection to measure T4 concentrations.
Results—Males did not differ from females in regard to prestimulation or poststimulation plasma T4 concentrations; however, prestimulation and poststimulation T4 concentrations were significantly different between the 2 groups of ferrets. A significant difference was also identified between prestimulation T4 concentration (mean ± SD, 21.3 ± 6.1 nmol/L) and poststimulation T4 concentration (29.9 ± 8.2 nmol/L). All 25 ferrets had high poststimulation T4 concentrations (median difference, 7. 5 nmol/L; 10% to 90% interval, 3.26 to 17.70 nmol/L [0.25 to 1.38 μg/dL]; range, 2.50 to 20.70 nmol/L [0.19 to 1.61 μg/dL]); this represented a median increase in T4 concentration of 35% (10% to 90% interval, 18% to 81%; range, 8% to 126%).
Conclusions and Clinical Relevance—Results suggested that rhTSH can be used for thyrotropin stimulation testing in ferrets when administered IM. According to the findings, a euthyroid ferret should have an increase of approximately 30% in plasma T4 concentration 4 hours after rhTSH injection.
Objective—To evaluate the effects of administration of recombinant human (rh) thyroid-stimulating hormone (TSH) for evaluation of thyroid function in euthyroid guinea pigs (Cavia porcellus).
Design—Prospective, experimental study.
Animals—10 healthy, sexually intact, pet guinea pigs (approx 1 year of age).
Procedures—Guinea pigs were given rhTSH (100 μg, IM); plasma thyroxine concentrations were determined prior to and 3 and 4 hours after rhTSH injection. The animals were housed in 2 groups on the basis of sex and fed different commercial maintenance diets according to their husbandry.
Results—There was no significant difference in thyroxine concentrations between males and females before or after rhTSH injection. There was also no difference between thyroxine concentrations at 3 versus 4 hours after rhTSH injection. There was a significant difference between thyroxine concentrations before (median, 9.05 nmol/L [0.70 μg/dL]; 10% to 90% range, 7.39 to 16.99 nmol/L [0.57 to 1.32 μg/dL]) and after (mean ± SD, 23.95 ± 4.2 nmol/L) rhTSH injection. Euthyroid guinea pigs had plasma thyroxine concentrations of at least 7.30 nmol/L (0.57 μg/dL) and an increase of at least 2.6 times prestimulation thyroxine concentrations at 3 or 4 hours after stimulation.
Conclusions and Clinical Relevance—The results suggested that rhTSH administered IM can be used for the TSH stimulation testing in guinea pigs. We suggest thyroxine concentration in a euthyroid guinea pig should at least double 3 to 4 hours after rhTSH injection.
OBJECTIVE To investigate the precision of an ELISA for measurement of serum cortisol concentration (SCC) in dogs, assess agreement between this ELISA and 2 validated chemiluminescence assays (CLAs), and evaluate the clinical implications of any bias associated with this ELISA when measuring SCC in dogs.
DESIGN Evaluation study.
SAMPLE 75 stored, frozen serum samples from client-owned dogs.
PROCEDURES Enzyme-linked immunosorbent assay precision was evaluated by measuring SCC of pooled serum samples. Agreement with standard methods was evaluated with Spearman rank correlation, Passing-Bablok regression, and Bland-Altman analysis to compare SCCs obtained with the ELISA and the 2 CLAs. An error grid was used to evaluate identified bias.
RESULTS Within-laboratory coefficients of variation for pooled serum samples with low, medium, and high SCCs were 21.4%, 28.9%, and 13.0%, respectively. There was a high correlation between ELISA results (for all samples combined) and results of the 2 CLAs (CLA 1, r = 0.96; CLA 2, r = 0.97), but constant and proportional biases between the ELISA and CLAs were present at all concentrations. Clinically important disagreement between ELISA results and CLA results occurred in 16 of 63 (25%) samples, particularly with low and high SCCs.
CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that the rate of clinical disagreement between the ELISA and CLAs was sufficiently high to recommend that equivocal results obtained with the ELISA be confirmed by a reference laboratory. Further evaluation of analytic performance of the ELISA should focus on samples with very high and very low SCCs.
Objective—To characterize the physiologic response
to IV bolus injection of glucose and insulin for development
of a combined glucose-insulin test (CGIT) in
Animals—6 healthy mares and 1 mare each with
pituitary adenoma and urolithiasis.
Procedure—Horses were given a CGIT (glucose,
150 mg/kg; insulin, 0.1 U/kg); results were compared
with a singular IV glucose tolerance test (GTT; 150
mg/kg) and a singular IV insulin sensitivity test (IST;
0.1 U/kg). Healthy horses were also given a CGIT after
receiving xylazine and undergoing stress.
Results—Physiologically, the CGIT resulted in a 2-phase curve with positive (hyperglycemic) and negative
(hypoglycemic) portions; the positive phase came
first (250% of baseline at 1 minute). The descending
segment declined linearly to baseline by approximately
30 minutes and to a nadir at 58% of baseline
by 75 minutes. After a 35-minute valley, a linear
ascent to baseline began. Addition of insulin in the
CGIT increased glucose utilization by approximately
4.5 times during the positive phase but not during the
negative phase. The diseases' effects and experimental
inhibition of insulin secretion with xylazine and
stress were detectable by use of the 2 phases of the
CGIT. Only a single positive phase resulted from the
GTT and a single negative phase from the IST.
Conclusions and Clinical Relevance—The CGIT
resulted in a consistent, well-defined glycemia profile,
which can be disrupted experimentally or by a disease
process. The CGIT has clinical potential because it
provides integrated information and more information
than either the singular GTT or IST. (Am J Vet Res
Objective—To evaluate the effects of 4.7-mg deslorelin acetate implants on egg production and plasma concentrations of 17β-estradiol and androstenedione in Japanese quail (Coturnix coturnix japonica) over 180 days and assess safety of the implants in quail via gross and histologic examination.
Animals—20 female Japanese quail.
Procedures—Following a 7-day period of consistent egg laying, quail were anesthetized and received a 4.7-mg deslorelin implant (treatment group; n = 10) or identical placebo implant (control group; 10) SC between the scapulae. Egg production was monitored daily. Plasma concentrations of 17β-estradiol and androstenedione were measured on days 0 (immediately prior to implant injection), 14, 29, 62, 90, 120, 150, and 180 via radioimmunoassay. Birds were weighed periodically and euthanized at day 180 for complete necropsy.
Results—Egg production was significantly decreased in the treatment group, compared with the control group, from 2 to 12 weeks after implant injection. Egg production ceased in 6 of 10 quail in the treatment group (mean duration of cessation, 70 days). Plasma androstenedione and 17β-estradiol concentrations were significantly lower on day 29 in the treatment group than in the control group. Plama androstenedione and 17β-estradiol concentrations were significantly lower on day 29 in the treatment group then in the control group.
Conclusions and Clinical Relevance—4.7-mg deslorelin acetate implants reversibly decreased egg laying for approximately 70 days in most of the Japanese quail evaluated. Further studies evaluating implants containing different concentrations of the drug are needed in quail and other avian species.
Objective—To characterize signalment, clinical features, clinicopathologic variables, hepatic ultrasonographic characteristics, endocrinologic profiles, treatment response, and age at death of Scottish Terriers with progressive vacuolar hepatopathy (VH) with or without hepatocellular carcinoma (HCC).
Design—Retrospective case series.
Animals—114 Scottish Terriers with progressive VH.
Procedures—Electronic databases from 1980 to 2013 were searched for adult (age > 1 year) Scottish Terriers with histopathologic diagnoses of diffuse glycogen-like VH. Available sections of liver specimens were histologically reevaluated to confirm diffuse VH with or without HCC; 8 dogs with HCC only had neoplastic tissue available. Physical examination, clinicopathologic, treatment, and survival data were obtained.
Results—39 of 114 (34%) dogs with VH had HCC detected at surgery or necropsy or by abdominal ultrasonography. Histologic findings indicated that HCC was seemingly preceded by dysplastic hepatocellular foci. No significant differences were found in clinicopathologic variables or age at death between VH-affected dogs with or without HCC. Fifteen of 26 (58%) dogs with high hepatic copper concentrations had histologic features consistent with copper-associated hepatopathy. Although signs consistent with hyperadrenocorticism were observed in 40% (46/114) of dogs, definitive diagnosis was inconsistently confirmed. Assessment of adrenal sex hormone concentrations before and after ACTH administration identified high progesterone and androstenedione concentrations in 88% (22/25) and 80% (20/25) of tested dogs, respectively.
Conclusions and Clinical Relevance—Results suggested that VH in Scottish Terriers may be linked to adrenal steroidogenesis and a predisposition to HCC. In dogs with VH, frequent serum biochemical analysis and ultrasonographic surveillance for early tumor detection are recommended.