The Prostate Cancer Active Lifestyle Study (PALS): A randomized controlled trial of diet and exercise in overweight and obese men on active surveillance.

BACKGROUND: Active surveillance (AS) is increasingly used to monitor patients with lower risk prostate cancer (PCa). The Prostate Cancer Active Lifestyle Study (PALS) was a randomized controlled trial to determine whether weight loss improves obesity biomarkers on the causal pathway to progression in patients with PCa on AS. METHODS: Overweight/obese men (body mass index >25 kg/m2) diagnosed with PCa who elected AS were recruited. The intervention was a 6-month, individually delivered, structured diet and exercise program adapted from the Diabetes Prevention Program with a 7% weight loss goal from baseline. Control participants attended one session reviewing the US Dietary and Physical Activity Guidelines. The primary outcome was change in glucose regulation from baseline to the end of the 6-month intervention, which was measured by fasting plasma glucose, C-peptide, insulin, insulin-like growth factor 1, insulin-like growth factor binding protein-3, adiponectin, and homeostatic model assessment for insulin resistance. RESULTS: Among 117 men who were randomized, 100 completed the trial. The mean percentage weight loss was 7.1% and 1.8% in the intervention and control arms, respectively (adjusted between-group mean difference, -6.0 kg; 95% confidence interval, -8.0, -4.0). Mean percentage changes from baseline for insulin, C-peptide, and homeostatic model assessment for insulin resistance in the intervention arm were -23%, -16%, and -25%, respectively, compared with +6.9%, +7.5%, and +6.4%, respectively, in the control arm (all p for intervention effects ≤ .003). No significant between-arm differences were detected for the other biomarkers. CONCLUSIONS: Overweight/obese men with PCa undergoing AS who participated in a lifestyle-based weight loss intervention successfully met weight loss goals with this reproducible lifestyle intervention and experienced improvements in glucose-regulation biomarkers associated with PCa progression.

No Effect of High Eating Frequency Compared with Low Eating Frequency on Appetite and Inflammation Biomarkers: Results from a Randomized Crossover Clinical Trial.

BACKGROUND: Eating frequency (EF) focuses on the total number of eating occasions per day and may influence metabolic health. OBJECTIVES: We sought to examine the effect of high compared with low EF on appetite regulation and inflammatory biomarkers among healthy adults. METHODS: Data are from a randomized, crossover trial (the Frequency of Eating and Satiety Hormones study). Participants (n = 50) completed 2 isocaloric 21-d study periods of low EF (3 eating occasions/d) and high EF (6 eating occasions/d) in random order with a 14-d washout period in between. Participants were free-living and consumed their own food, using study-directed, structured meal plans with identical foods and total energy in both study periods. On days 1 and 21 of each EF period, fasting blood was collected during in-person clinic visits to assess plasma concentrations of ghrelin, leptin, adiponectin, and high-sensitivity C-reactive protein (hs-CRP). Linear mixed models with EF, diet sequence, and period as fixed effects and participant as random effect were used to estimate the intervention effect. Interaction effects between EF and body fat percentage were examined. RESULTS: Among the 50 participants who completed the trial, 39 (78%) were women, 30 (60%) were Non-Hispanic White, and 40 (80%) had a body mass index of <25 kg/m2, and the mean age was 32.1 y. The differences between high and low EF in fasting ghrelin (geometric mean difference: 17.76 ng/mL; P = 0.60), leptin (geometric mean difference: 2.09 ng/mL; P = 0.14), adiponectin (geometric mean difference: 381.7 ng/mL; P = 0.32), and hs-CRP (geometric mean difference: -0.018 mg/dL; P = 0.08) were not statistically significant. No significant interaction was observed between EF and body fat percentage on appetite regulation and inflammatory biomarkers. CONCLUSIONS: No differences was observed in fasting ghrelin, leptin, adiponectin, and hs-CRP comparing high and low EF. Future studies are needed to understand the physiology of EF and appetite as they relate to metabolic health. This trial was registered at clinicaltrials.gov as NCT02392897.

The discovery of multiple active mys-related LTR-retroelements within the Neotominae subfamily of cricetid rodents.

Retrotransposon families in the rodent family Cricetidae have been understudied in contrast to Muridae, both taxa classified within the superfamily Muroidea. Therefore, we carried out a study to advance our knowledge of the unique mys LTR-retroelement identified in Peromyscus leucopus, by incorporating intra-ORF PCR, quantitative dot blots, DNA and protein library screens, the generation of molecular phylogenies, and analyses of orthologous LTR-retroelement loci. These analyses led to the discovery of three additional related families of LTR-retroelements, which include a 2900 bp full-length element of mys-related sequences (mysRS), an 8000 bp element containing the mys ORF1 sequence (mORF1) with ERV-related sequences downstream in the reverse orientation, as well as an 1800 bp element primarily consisting of mys ORF2 (mORF2) related sequences flanked by LTRs. Our data revealed only a few full-length mys elements among genera of the Neotominae subfamily of cricetid rodents, most existing as partial copies. The mysRS and mORF1 elements are also limited to the genomes of the Neotominae subfamily, whereas mORF2 appears to be restricted to the Peromyscus genus. Molecular phylogenies demonstrating concerted evolution along with an assessment of orthologous loci in Peromyscus for the presence or absence of elements are consistent with activity of these novel LTR-retroelement families within this genus. Together with known activity of various families of non-LTR retroelements in Peromyscus species, we propose that retrotransposons have been continually contributing to the dynamics of Peromyscus genomes promoting genomic diversity and may be correlated with the evolution of more than 50 identified Peromyscus species.

Fluid Analysis in the Equine Patient: Cerebrospinal, Synovial, and Peritoneal Fluids.

Fluid analysis is an important part of the diagnostic work-up of equine patients presenting with disorders of the nervous system, musculoskeletal system, and abdominal cavity. Proper specimen handling and processing are paramount for complete and accurate interpretation of fluid samples. Normal cerebrospinal fluid is a low-cellularity, low-protein fluid requiring specific sample handling to ensure accurate results. Joint and abdominal fluid analyses are completed in practice or submitted for analysis to a reference laboratory. This article discusses fluid sample handling and processing considerations for the equine practitioner and reviews cytologic evaluation of normal and abnormal cerebrospinal, synovial, and peritoneal fluid samples.

RENAL NEOPLASIA FROM A SINGLE POPULATION OF PTEROPODID BATS.

Neoplastic diseases have rarely been reported in the family Pteropodidae, and primary malignant renal neoplasms are generally uncommon across animal species. This case series describes four cases of primary renal neoplasia: three renal cell carcinomas and one nephroblastoma in three species of pteropodid bats, specifically large flying foxes (Pteropus vampyrus, n = 2), straw-colored fruit bat (Eidolon helvum, n = 1), and a little golden-mantled flying fox (Pteropus pumilus, n = 1). Two of the cases were diagnosed antemortem using ultrasonography, computed tomography, and cytology; and one of these bats with a renal cell carcinoma was treated successfully with a unilateral nephrectomy. The remaining two cases were diagnosed at necropsy.

Comparison of 3 corneal cytology collection methods for evaluating equine ulcerative keratitis: Cytobrush, kimura platinum spatula, and handle edge of scalpel blade.

PURPOSE: To compare corneal cytology samples from three common sampling techniques: cytobrush (CB), Kimura platinum spatula (KS), and the handle edge of a scalpel blade (SB). METHODS: Equine patients presenting to the University of Florida College of Veterinary Medicine with ulcerative keratitis were included. Following diagnosis of corneal ulcer and sampling for microbial culture, two cytology samples per technique were collected with sterile CB, KS, and SB in a randomized order. Cytologic evaluation was performed by two observers masked to collection method. Objective measures of sample cellularity, quality, distribution, and identification of infectious organisms were recorded per 10 monolayer cell populations using 50× magnification with oil immersion which were compared to culture results. Variables were compared using ANOVA with Student's t test when appropriate and Cohen's kappa (k) to evaluate inter- and intra-observer agreement (IOA) between observers and techniques. RESULTS: Twenty equine patients (120 samples) were included. The IOA between observers was substantial (k = 0.75 ± 0.06) for cytological parameters. SB provided the most cellular samples (P < .01). There was a trend toward agreement (k = 0.12 ± 0.16) in technique for sample quality (P = .08). CB and SB had significantly poorer cell distribution than KS (P < .05). Infection was confirmed in 12 of 20 patients with SB and CB techniques having a significantly higher diagnostic yield than KS (P < .05) and was most consistent with infection confirmed on culture. CONCLUSIONS: The SB provided the most diagnostic samples but all three techniques are clinically useful in evaluating equine ulcerative keratitis.

Mutations in the gene encoding GlyT2 (SLC6A5) define a presynaptic component of human startle disease.

Hyperekplexia is a human neurological disorder characterized by an excessive startle response and is typically caused by missense and nonsense mutations in the gene encoding the inhibitory glycine receptor (GlyR) alpha1 subunit (GLRA1). Genetic heterogeneity has been confirmed in rare sporadic cases, with mutations affecting other postsynaptic glycinergic proteins including the GlyR beta subunit (GLRB), gephyrin (GPHN) and RhoGEF collybistin (ARHGEF9). However, many individuals diagnosed with sporadic hyperekplexia do not carry mutations in these genes. Here we show that missense, nonsense and frameshift mutations in SLC6A5 (ref. 8), encoding the presynaptic glycine transporter 2 (GlyT2), also cause hyperekplexia. Individuals with mutations in SLC6A5 present with hypertonia, an exaggerated startle response to tactile or acoustic stimuli, and life-threatening neonatal apnea episodes. SLC6A5 mutations result in defective subcellular GlyT2 localization, decreased glycine uptake or both, with selected mutations affecting predicted glycine and Na+ binding sites.

Silica and barium: Comparison of microscopic appearance and characterization of effects of silica on cytomorphology.

BACKGROUND: Silica from plastic red top sample collection tubes and barium cause recognized artifacts in slide preparations for microscopic examination. OBJECTIVES: The objectives of this study were to evaluate and directly compare the microscopic appearance of silica and barium particles and various slide preparation techniques (e.g., use of coverslips, oil immersion, and different stains). A secondary objective of this study was to evaluate the effects of silica particles on cellular morphology after mechanical trauma with cytocentrifugation. METHODS: Fluid samples (deionized water, pleural effusion, peritoneal effusion, cerebrospinal fluid, and urine) were collected and evaluated in silica- and non-silica-containing tubes. Barium was added to silica and non-silica samples. Direct and cytocentrifuge preparations were compared to evaluate the effect of silica particles on cellular morphology. Preparations were stained with Wright-Giemsa, rhodizonic acid disodium salt, Alizarin Red, Grocott's methenamine silver, and Prussian blue. RESULTS: Silica and barium particles were identifiable via light microscopy with and without polarized light, although silica particles diminished with immersion oil. Barium particles retained their structure and diminished less under oil. Cytoseal mounting medium for coverslip placement resulted in diminished refractility of silica and some barium particles. Silica particles with mechanical interaction during cytocentrifugation resulted in disrupted cellular morphology with many lysed cells. Silica and barium particles were negative for all special stains tested. CONCLUSIONS: Silica from plastic red top tubes adversely affects cell morphology in cytocentrifuge preparations, potentially affecting manual differential cell counts and compromising diagnostic interpretation. Samples intended for microscopic evaluation should not be collected in silica-containing tubes.

Successful multimodal treatment of extreme hypertriglyceridemia in a juvenile diabetic dog.

OBJECTIVE: To describe the therapeutic protocol used to normalize severe hypertriglyceridemia in a dog. CASE SUMMARY: A 7-month-old, 1.2-kg female Pomeranian presented with acute polyuria, polydipsia, and ocular discoloration. Diagnoses included diabetic ketosis, severe hypertriglyceridemia (>225 mmol/L [>20,000 mg/dl]), lipemia retinalis, and bilateral uveitis. The triglyceride concentration was near normal within 2 days of initiating treatment with fenofibrate, regular insulin constant rate infusion (CRI), manual therapeutic plasma exchange (TPE), and a low-fat diet. All clinical signs resolved. The dog has had no relapse of hypertriglyceridemia at the time of writing the manuscript, 6 months later, with continued treatment of diabetes mellitus. NEW OR UNIQUE INFORMATION PROVIDED: This is the first case report documenting the combination of fenofibrate, insulin CRI, and manual TPE for treatment of severe hyperlipidemia in a dog. Detailed protocols for manual TPE and a novel insulin CRI are provided. A discussion of multiple spurious biochemical and hematologic errors associated with the severe hypertriglyceridemia is also provided.

Cochlear implantation performance outcomes in patients over 80 years old.

Objective: To examine device datalogging characteristics and postoperative performance in the very elderly CI users. Methods: Retrospective chart review of postoperative speech recognition outcomes and datalogging characteristics in patients older than 80 years old who underwent cochlear implantation and remain active users with longer than 6 months post-activation follow-up. Resultss: The mean age at implantation was 84.8 ± 3.8 years old. Pre- and post-operative AzBio sentences in quiet scores were 12.4 ± 16.4% and 53.0 ± 23.5%, respectively (p < .001). Pre- and post-operative consonant-nucleus-consonant (CNC) word scores were 9.4 ± 12.8% and 40.5 ± 20.7%, respectively (p < .001). A nonsignificant negative correlation was noted between the age of implantation and postoperative CNC words and AzBio sentence performance. Significantly decreased performance was noted in the subpopulation with a preoperative diagnosis of dementia. Mean daily use was 10.9 h per day. When available the mean daily usage distribution was 16% speech in noise, 19.1% speech, 51% quiet, 3.8% music, and 9.6% noise. A significant positive correlation was noted between daily use and AzBio sentence and CNC word performance. Conclusions: Very elderly patients have significant postoperative auditory performance benefits after CI. Mean daily use is comparable to previously published results in the younger population. Age of implantation does not play a significant role in overall performance. Elderly patients who are medically cleared for implantation receive significant postoperative benefits.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Comparison of biochemical and hematologic values obtained via jugular venipuncture and peripheral intravenous catheters in dogs.

BACKGROUND: Sampling from a peripheral intravenous catheter (PIVC) might be a more efficient and less traumatic collection of blood for serum biochemistry (SB) or CBC than direct venipuncture (DV). Agreement between results of samples obtained by these methods has not been evaluated in dogs. OBJECTIVES: The primary objectives were to determine whether sampling from PIVC could be used in place of DV for dogs. We hypothesized DV and PIVC samples would have clinically equivalent SB and CBC results. ANIMALS: Sixty-one client-owned dogs were included in each study arm. METHODS: This was a partially randomized method-comparison study. Paired DV and PIVC samples obtained within 1 to 2 minutes after, or approximately 24 hours after, placement of a PIVC in a cephalic vein were evaluated for agreement and bias using percentage difference plots (with a priori application of consensus total allowable error), Bland-Altman analysis, Passing-Bablok regression analysis, Wilcoxon signed rank test, and McNemar's test. RESULTS: There was statistically and clinically acceptable agreement and no bias between sampling methods for the majority of results. Analytes with the most frequent disagreement were aspartate aminotransferase, total bilirubin, potassium, bicarbonate, and leukocyte differential counts, as well as red blood cell count, hemoglobin, hematocrit, and packed cell volume in the hospitalized PIVC sampling group. Few observed differences would change clinical decision making. CONCLUSIONS AND CLINICAL IMPORTANCE: PIVC sampling can provide generally acceptable SB and CBC results for most dogs, but clinicians should be aware of a few values for which disparate results might occasionally be obtained.

Cytologic, histopathologic, and clinical features of granulomatous colitis in a French Bulldog.

A young French Bulldog was presented with clinical signs of chronic gastrointestinal disease, unresponsive to medical therapies. Parasite screenings and abdominal ultrasound failed to identify the etiology of the clinical signs. Cytologic evaluation of a rectal scraping sample diagnosed presumptive granulomatous colitis (GC) based on the presence of numerous macrophages with characteristic abundant, pink, granular cytoplasm, which showed an intense pink color when stained with periodic acid-Schiff. Tissue biopsy samples and Escherichia coli fluorescence in situ hybridization analysis confirmed the cytologic diagnosis. The cytologic, histopathologic, and clinical features and staining properties of GC in a French Bulldog are reported. Rectal scraping should be considered a part of the diagnostic evaluation in patients with suspected GC.

Frequency and classification of errors in laboratory medicine at a veterinary teaching hospital in the United States.

BACKGROUND: The accuracy of laboratory data is of utmost importance. Data regarding laboratory error in human laboratories are often extrapolated into veterinary settings. One study investigated the rate and type of errors in a European commercial veterinary laboratory, but that data might not directly apply to an educational setting. OBJECTIVES: This study determined the frequency and type of errors in laboratory medicine at a veterinary teaching hospital. METHODS: Errors associated with clinical pathology samples were recorded over two 60-day periods. The first period included a time when new students and house officers started at the veterinary school. The second time period was 6 months later. The errors were assigned to categories, and the frequency of each was calculated. Sample hemolysis, icterus, and lipemia were evaluated separately using an automated index, as these conditions could be pathologic or the result of error. Frequencies of error and hemolysis, icterus, and lipemia were assessed between the groups. RESULTS: Total error rates were 4.7% and 3.5% for the first and second periods, respectively. The frequency of each error subclassification was similar to those observed in the veterinary and human literature, with preanalytic error predominating. Statistically significant differences in the overall error rate and percentage of preanalytic errors that occurred outside of and within the laboratory were observed comparing differences between the two periods. CONCLUSIONS: The overall error rate in this veterinary teaching hospital was slightly higher than that previously reported in other settings, although a proportion of errors was as expected. Areas needing improvement were identified, and strategies to reduce error could be developed.

Primary Hyperparathyroidism in a Quarter Horse Mare Associated With a Chief Cell Adenoma.

Primary hyperparathyroidism is rare in large animal species, and little is known regarding its pathophysiology, endocrine and electrolyte derangements, diagnosis, medical management, and prognosis. This report describes the clinicopathologic diagnosis of a parathyroid (PT) gland chief cell adenoma in a 12-year-old Quarter Horse mare, including PT hormone (PTH) and electrolyte disarrangements associated with the neoplasia, the surgical removal of the adenoma, and medical management of the case. This report also describes for the first time the use PTH immunohistochemistry to confirm the nature of this neoplasia in a horse.

Agreement of 2 electrolyte analyzers for identifying electrolyte and acid-base disorders in sick horses.

BACKGROUND: Use of different analyzers to measure electrolytes in the same horse can lead to different interpretation of acid-base balance when using the simplified strong ion difference (sSID) approach. OBJECTIVE: Investigate the level of agreement between 2 analyzers in determining electrolytes concentrations, sSID variables, and acid-base disorders in sick horses. ANIMALS: One hundred twenty-four hospitalized horses. METHODS: Retrospective study using paired samples. Electrolytes were measured using a Beckman Coulter AU480 Chemistry analyzer (PBMA) and a Nova Biomedical Stat Profile (WBGA), respectively. Calculated sSID variables included strong ion difference, SID4 ; unmeasured strong ions, USI; and total nonvolatile buffer ion concentration in plasma (Atot ). Agreement between analyzers was explored using Passing-Bablok regression and Bland-Altman analysis. Kappa (κ) test evaluated the level of agreement between analyzers in detecting acid-base disorders. RESULTS: Methodologic differences were identified in measured Na+ and Cl- and calculated values of SID4 and USI. Mean bias (95% limits of agreement) for Na+ , Cl- , SID4 , and USI were: -1.2 mmol/L (-9.2 to 6.8), 4.4 mmol/L (-4.4 to 13), -5.4 mmol/L (-13 to 2), and -6.2 mmol/L (-14 to 1.7), respectively. The intraclass correlation coefficient for SID4 and USI was .55 (95%CI: -0.2 to 0.8) and .2 (95%CI: -0.15 to 0.48), respectively. There was a poor agreement between analyzers for detection of SID4 (κ = 0.20, 95%CI, 0.1 to 0.31) or USI abnormalities (κ = -0.04, 95%CI, -0.11 to 0.02). CONCLUSIONS AND CLINICAL IMPORTANCE: Differences between analyzer methodology in measuring electrolytes led to a poor agreement between the diagnosis of acid-base disorders in sick horses when using the sSID approach.

Cytologic identification of fungal arthritis in a Labrador Retriever with disseminated Talaromyces helicus infection.

An 8-year-old, neutered male Labrador Retriever presented with acute forelimb lameness. Clinical signs progressed over one week. On physical examination, right cubital joint effusion and bilateral axillary lymphadenomegaly were noted, and severe internal lymphadenomegaly was observed ultrasonographically. Granulomatous lymphadenitis with intralesional fungi was noted cytologically, and the dog was ultimately diagnosed with disseminated Talaromyces helicus infection via PCR of a pure isolate. Extensive medical therapy was pursued, and months later, an arthrocentesis was performed due to continued lameness and severe cubital joint effusion. The synovial fluid contained increased numbers of neutrophils, macrophages, and multinucleated giant cells. Frequent fungal hyphae were found both intracellularly and extracellularly. These basophilic organisms were 2-4 µm in width with internal eosinophilic granules, roughly parallel walls, and occasional to frequent septa. Round to oval yeast-like forms with thin, clear halos were also occasionally identified. Due to the severity of clinical signs, the right thoracic limb was amputated. Histologic examination of the cubital joint revealed marked granulomatous synovitis, fasciitis, panniculitis, and osteomyelitis, all with intralesional fungi. Talaromyces helicus is a very rare cause of disease, reported only in one other dog. Granulomatous lymphadenitis appears to be a feature of this disease, but this report is the first to describe a significant synovial component.

Rationale and design of the frequency of eating and Satiety Hormones (FRESH) study: A randomized cross-over clinical trial.

BACKGROUND: The goal of the Frequency of Eating and Satiety Hormones (FRESH) Study is to understand the relationship between eating frequency (EF) and biomarkers of appetite and disease risk. This report gives the study rationale and design. METHODS: The FRESH study was conducted in n = 50 overweight and obese, but otherwise healthy, male and female adults aged 18-50 years. The protocol included four in-person clinic visits for protocol instruction, blood draws, anthropometry, and meal testing; all other activities were done at home. Participants completed two 21-day phases in random order with a two-week washout between phases. One phase was high EF (6 eating occasions/day) and the other was low EF (3 eating occasions/day). Each phase specified time of day for each eating occasion. Participants prepared their own meals throughout the study using study-provided individualized, structured meal plans ensuring that calories, macronutrients and micronutrients were identical during both study phases. Fasting blood was collected before and after each phase to test intervention effects on the biomarkers. At the end of each phase participants also completed extended appetite testing with meals prepared by the study clinic. RESULTS: Participants were recruited using print, radio, and digital ads. 60 participants consented to enroll; 10 dropped out due to work or school scheduling conflicts and 50 (target sample size) completed the study. Compliance was assessed by completion of daily on-line meal plan checklists. CONCLUSIONS: The FRESH study will provide data on whether higher vs. lower daily EF in the context of constant energy and nutrient intake may be harmful or beneficial based on intervention effects on biomarkers of health and disease risk.

Randomized trial evaluating the role of weight loss in overweight and obese men with early stage prostate Cancer on active surveillance: Rationale and design of the Prostate Cancer Active Lifestyle Study (PALS).

Active surveillance (AS) is increasingly used to monitor patients with low-risk prostate cancer; however, approximately 50% of AS patients experience disease reclassification requiring definitive treatment and little is known about patient characteristics that modify the risk of reclassification. Obesity may be one of the major contributing factors. The Prostate Cancer Active Lifestyle Study (PALS) is a clinical trial evaluating the impact of weight loss among overweight/obese (Body Mass Index (BMI) ≥ 25 kg/m2) men with clinically localized prostate cancer on AS. Two hundred participants will be randomized to either the PALS intervention, a 6-month structured diet and exercise program adapted from the Diabetes Prevention Program followed by 6 months of maintenance, or control (general diet and physical activity guidelines delivered in a single session). The PALS intervention involves one-on-one instruction with a registered dietitian and exercise physiologist to achieve the study goal of loss of 7% of baseline weight. Participation is coordinated so that the 6-month time point coincides with the participants' standard-of-care AS prostate biopsy. Primary outcomes will evaluate the intervention effects on circulating and tissue markers of glucose and insulin regulation, health-related quality of life and pathologic upgrading on follow-up prostate biopsies. Additional analyses will determine whether changes in weight and glucose regulation can be sustained for 6 months after the end of instruction. Findings from this trial may have wide reaching implications for men diagnosed with clinically-localized prostate cancer by providing an active lifestyle-based approach to improve prostate cancer patient outcomes.