To develop a technique for isolation and culture of canine insulinoma cells and assess expression of cellular hexokinases (glucokinase and hexokinase I) and expression and secretion of insulin from these cells in vitro.
Pancreatic insulinomas and normal pancreatic tissue from 4 and 3 dogs, respectively.
Tissues were collected by surgical excision or at necropsy. Insulinoma cells from 2 dogs were cultured for up to 10 weeks with standard techniques; insulin synthesis in vitro was confirmed by immunohistochemical analysis of freshly prepared slides of cultured cells, and insulin secretion was assessed by measurement of insulin concentrations in culture medium with an ultrasensitive mouse insulin ELISA. Expression of cellular hexokinases in insulinomas and adjacent normal (nontumor) pancreatic tissue from the same dog (n = 3) was examined by quantitative reverse transcriptase PCR assay.
Insulinoma cells survived for up to 10 weeks but did not proliferate in culture. Insulin was detected in isolated cells and secreted into culture medium for up to 10 weeks. Both cellular hexokinases were expressed; glucokinase appeared to be overexpressed in insulinomas, compared with normal pancreatic tissue from the same dogs.
CONCLUSIONS AND CLINICAL RELEVANCE
Canine insulinomas expressed hexokinases responsible for glucose responsiveness. Insulinoma cells were successfully maintained in short-term culture; cultured cells remained functional for 10 weeks as evidenced by cellular insulin content and had detectable secretion of insulin into the culture medium for ≥ 5 weeks. Apparent glucokinase overexpression by insulinomas suggested a possible mechanism underlying excessive insulin release by these tumors.
OBJECTIVE To determine the tonicity effects of β-hydroxybutyrate, acetoacetate, and lactate in canine RBCs.
SAMPLE RBCs from approximately 40 dogs.
PROCEDURES 2 in vitro methods were used to conduct 4 experiments. The modified osmotic fragility assay was used to measure the ability of ketoacid salts added to serial sucrose dilutions to protect RBCs from osmotic hemolysis. In a second assay, a handheld cell counting device was used to measure changes in RBC diameter to assess the tonicity effect of solutions of ketoacid and lactate salts.
RESULTS For the modified osmotic fragility assay, all ketoacid salts had an osmoprotective effect, but the effect was determined to be completely attributable to the tonicity effect of added cations (sodium and lithium) and not the ketoacid moieties. However, both the sodium and lithium lactate salts provided osmoprotection attributable to both the cation and lactate anion. For the second assay, RBC diameter was significantly increased with the addition of urea (an ineffective osmole) but did not change with the addition of glucose (an effective osmole), which established the behaviors of ineffective and effective osmoles in this assay. The RBC diameter was significantly increased over that of control samples by the addition of sodium β-hydroxybutyrate, lithium acetoacetate, and lithium lactate but was decreased by the addition of sodium lactate.
CONCLUSIONS AND CLINICAL RELEVANCE For both assays, β-hydroxybutyrate and acetoacetate acted as ineffective osmoles, whereas lactate acted as an effective osmole in 3 of 4 experiments.
OBJECTIVE To evaluate mean corpuscular volume difference (dMCV) as a marker for hypertonicity induced by water deprivation in dogs.
ANIMALS 5 healthy Greyhounds maintained in a research colony.
PROCEDURES Water was withheld for 24 hours. Blood and urine samples were collected before (time 0) and every 6 hours during water deprivation. Serum and urine osmolality were measured on the basis of freezing point depression, and dMCV was calculated from routine hematologic variables.
RESULTS Serum and urine osmolality significantly increased and body weight decreased over time in healthy Greyhounds during water deprivation, although most dogs developed only a slight increase in serum osmolality. The dMCV also increased over time, but the value at 24 hours did not differ significantly from the value at time 0. However, a significant correlation was found between serum osmolality and dMCV. A dMCV ≥ 5 fL yielded 100% specificity for predicting hypertonicity when hypertonicity was defined as serum osmolality ≥ 310 mOsM.
CONCLUSIONS AND CLINICAL RELEVANCE dMCV may be a useful marker for detection of mild hypertonicity in dogs and may have clinical and research applications for use in screening canine populations for hypertonicity.
Procedures—A CGMS was placed and used to obtain calculated glucose measurements before, during, and after anesthesia in each dog. Periodically, CGMS measurements were compared with concurrent measurements of glucose concentration in peripheral venous blood obtained with a portable chemistry analyzer (PCA).
Results—CGMS-calculated glucose measurements were significantly different from PCA blood glucose measurements during most of the anesthetic period. The CGMS values differed from PCA values by > 20% in 54 of 126 (42.9%) paired measurements obtained during the anesthetic period. Hypoglycemia was evident in CGMS measurements 25 of 126 (19.8%) times during anesthesia. By comparison, only 1 incident of hypoglycemia was detected with the PCA during the same period.
Conclusions and Clinical Relevance—Use of the CGMS for routine monitoring of interstitial glucose concentration as an indicator of blood glucose concentration during anesthesia cannot be recommended. Additional investigation is necessary to elucidate the cause of discrepancy between CGMS results and PCA data during anesthesia.
To evaluate and compare regulation of diabetes mellitus (DM) in dogs with cataracts and well-controlled DM that received an ophthalmic preparation of prednisolone acetate versus diclofenac sodium.
22 client-owned dogs with cataracts and well-controlled DM.
A prospective, randomized, double-masked, experimental study was conducted. On days 0 and 32, serum fructosamine concentrations (SFCs), clinical scores, and body weights were determined. Dogs were assigned to receive a topically administered ophthalmic preparation of either prednisolone acetate 1% or diclofenac sodium 0.1% in each eye 4 times daily for 28 days. Data analysis was conducted with generalized linear mixed models.
Findings indicated no meaningful differences in SFCs, clinical scores, or body weights between the treatment groups on days 0 or 32. Clinical score on day 0 was positively associated with SFC, as indicated by the corresponding rate of change such that each 1 -unit increase in clinical score was associated with an approximately 45.6 ± 9.4 μmol/L increase in SFC. In addition, the least squares mean ± SEM SFC was higher in spayed females (539.20 ± 19.23 μmol/L; n = 12) than in castrated males (458.83 ± 23.70 μmol/L; 8) but did not substantially differ between sexually intact males (446.27 ± 49.72 μmol/L; 2) and spayed females or castrated males regardless of the treatment group assigned.
CONCLUSIONS AND CLINICAL RELEVANCE
Findings indicated no evidence for any differential effect on DM regulation (assessed on the basis of SFCs, clinical scores, and body weights) in dogs treated topically with an ophthalmic preparation of prednisolone versus an ophthalmic preparation of diclofenac. Additional research investigating plasma concentrations of topically applied ophthalmic glucocorticoid medications is warranted. (Am J Vet Res 2019;80:1129-1135)