Comparison of artificial intelligence to the veterinary radiologist's diagnosis of canine cardiogenic pulmonary edema.

Application of artificial intelligence (AI) to improve clinical diagnosis is a burgeoning field in human and veterinary medicine. The objective of this prospective, diagnostic accuracy study was to determine the accuracy, sensitivity, and specificity of an AI-based software for diagnosing canine cardiogenic pulmonary edema from thoracic radiographs, using an American College of Veterinary Radiology-certified veterinary radiologist's interpretation as the reference standard. Five hundred consecutive canine thoracic radiographs made after-hours by a veterinary Emergency Department were retrieved. A total of 481 of 500 cases were technically analyzable. Based on the radiologist's assessment, 46 (10.4%) of these 481 dogs were diagnosed with cardiogenic pulmonary edema (CPE+). Of these cases, the AI software designated 42 of 46 as CPE+ and four of 46 as cardiogenic pulmonary edema negative (CPE-). Accuracy, sensitivity, and specificity of the AI-based software compared to radiologist diagnosis were 92.3%, 91.3%, and 92.4%, respectively (positive predictive value, 56%; negative predictive value, 99%). Findings supported using AI software screening for thoracic radiographs of dogs with suspected cardiogenic pulmonary edema to assist with short-term decision-making when a radiologist is unavailable.

Differential Desensitization Observed at Multiple Effectors of Somatic µ-Opioid Receptors Underlies Sustained Agonist-Mediated Inhibition of Proopiomelanocortin Neuron Activity.

Activation of somatic µ-opioid receptors (MORs) in hypothalamic proopiomelanocortin (POMC) neurons leads to the activation of G-protein-coupled inward rectifier potassium (GIRK) channels and hyperpolarization, but in response to continued signaling MORs undergo acute desensitization resulting in robust reduction in the peak GIRK current after minutes of agonist exposure. We hypothesized that the attenuation of the GIRK current would lead to a recovery of neuronal excitability whereby desensitization of the receptor would lead to a new steady state of POMC neuron activity reflecting the sustained GIRK current observed after the initial decline from peak with continued agonist exposure. However, electrophysiologic recordings and GCaMP6f Ca2+ imaging in POMC neurons in mouse brain slices indicate that maximal inhibition of cellular activity by these measures can be maintained after the GIRK current declines. Blockade of the GIRK current by Ba2+ or Tertiapin-Q did not disrupt the sustained inhibition of Ca2+ transients in the continued presence of agonist, indicating the activation of an effector other than GIRK channels. Use of an irreversible MOR antagonist and Furchgott analysis revealed a low receptor reserve for the activation of GIRK channels but a >90% receptor reserve for the inhibition of Ca2+ events. Altogether, the data show that somatodendritic MORs in POMC neurons inhibit neuronal activity through at least two effectors with distinct levels of receptor reserve and that differentially reflect receptor desensitization. Thus, in POMC cells, the decline in the GIRK current during prolonged MOR agonist exposure does not reflect an increase in cellular activity as expected.SIGNIFICANCE STATEMENT Desensitization of the µ-opioid receptor (MOR) is thought to underlie the development of cellular tolerance to opiate therapy. The present studies focused on MOR desensitization in hypothalamic proopiomelanocortin (POMC) neurons as these neurons produce the endogenous opioid ß-endorphin and are heavily regulated by opioids. Prolonged activation of somatic MORs in POMC neurons robustly inhibited action potential firing and Ca2+ activity despite desensitization of the MOR and reduced activation of a potassium current over the same time course. The data show that somatic MORs in POMC neurons couple to multiple effectors that have differential sensitivity to desensitization of the receptor. Thus, in these cells, the cellular consequence of MOR desensitization cannot be defined by the activity of a single effector system.

Induction of stable ER-plasma-membrane junctions by Kv2.1 potassium channels.

Junctions between cortical endoplasmic reticulum (cER) and the plasma membrane are a subtle but ubiquitous feature in mammalian cells; however, very little is known about the functions and molecular interactions that are associated with neuronal ER-plasma-membrane junctions. Here, we report that Kv2.1 (also known as KCNB1), the primary delayed-rectifier K(+) channel in the mammalian brain, induces the formation of ER-plasma-membrane junctions. Kv2.1 localizes to dense, cell-surface clusters that contain non-conducting channels, indicating that they have a function that is unrelated to membrane-potential regulation. Accordingly, Kv2.1 clusters function as membrane-trafficking hubs, providing platforms for delivery and retrieval of multiple membrane proteins. Using both total internal reflection fluorescence and electron microscopy we demonstrate that the clustered Kv2.1 plays a direct structural role in the induction of stable ER-plasma-membrane junctions in both transfected HEK 293 cells and cultured hippocampal neurons. Glutamate exposure results in a loss of Kv2.1 clusters in neurons and subsequent retraction of the cER from the plasma membrane. We propose Kv2.1-induced ER-plasma-membrane junctions represent a new macromolecular plasma-membrane complex that is sensitive to excitotoxic insult and functions as a scaffolding site for both membrane trafficking and Ca(2+) signaling.

Regulation of Kv2.1 K(+) conductance by cell surface channel density.

The Kv2.1 voltage-gated K(+) channel is found both freely diffusing over the plasma membrane and concentrated in micron-sized clusters localized to the soma, proximal dendrites, and axon initial segment of hippocampal neurons. In transfected HEK cells, Kv2.1 channels within cluster microdomains are nonconducting. Using total internal reflection fluorescence microscopy, the number of GFP-tagged Kv2.1 channels on the HEK cell surface was compared with K(+) channel conductance measured by whole-cell voltage clamp of the same cell. This approach indicated that, as channel density increases, nonclustered channels cease conducting. At the highest density observed, only 4% of all channels were conducting. Mutant Kv2.1 channels that fail to cluster also possessed the nonconducting state with 17% conducting K(+) at higher surface densities. The nonconducting state was specific to Kv2.1 as Kv1.4 was always conducting regardless of the cell-surface expression level. Anti-Kv2.1 immunofluorescence intensity, standardized to Kv2.1 surface density in transfected HEK cells, was used to determine the expression levels of endogenous Kv2.1 in cultured rat hippocampal neurons. Endogenous Kv2.1 levels were compared with the number of conducting channels determined by whole-cell voltage clamp. Only 13 and 27% of the endogenous Kv2.1 was conducting in neurons cultured for 14 and 20 d, respectively. Together, these data indicate that the nonconducting state depends primarily on surface density as opposed to cluster location and that this nonconducting state also exists for native Kv2.1 found in cultured hippocampal neurons. This excess of Kv2.1 protein relative to K(+) conductance further supports a nonconducting role for Kv2.1 in excitable tissues.

Corrigendum: Classification of feline hypertrophic cardiomyopathy-associated gene variants according to the American College of Medical Genetics and Genomics guidelines.

[This corrects the article DOI: 10.3389/fvets.2024.1327081.].

Classification of feline hypertrophic cardiomyopathy-associated gene variants according to the American College of Medical Genetics and Genomics guidelines.

Introduction: The correct labeling of a genetic variant as pathogenic is important as breeding decisions based on incorrect DNA tests can lead to the unwarranted exclusion of animals, potentially compromising the long-term health of a population. In human medicine, the American college of Medical Genetics (ACMG) guidelines provide a framework for variant classification. This study aims to apply these guidelines to six genetic variants associated with hypertrophic cardiomyopathy (HCM) in certain cat breeds and to propose a modified criterion for variant classification. Methods: Genetic samples were sourced from five cat breeds: Maine Coon, Sphynx, Ragdoll, Devon Rex, and British Short- and Longhair. Allele frequencies were determined, and in the subset with phenotypes available, odds ratios to determine the association with HCM were calculated. In silico evaluation followed with joint evidence and data from other publications assisting in the classification of each variant. Results: Two variants, MYBPC3:c.91G > C [A31P] and MYBPC3:c.2453C > T [R818W], were designated as pathogenic. One variant, MYH7:c.5647G > A [E1883K], was found likely pathogenic, while the remaining three were labeled as variants of unknown significance. Discussion: Routine genetic testing is advised solely for the MYBPC3:c.91G > C [A31P] in the Maine Coon and MYBPC3:c.2453C > T [R818W] in the Ragdoll breed. The human ACMG guidelines serve as a suitable foundational tool to ascertain which variants to include; however, refining them for application in veterinary medicine might be beneficial.

Use of a percutaneously controlled urethral hydraulic occluder for treatment of refractory urinary incontinence in 18 female dogs.

OBJECTIVE: To evaluate the efficacy and safety of a percutaneously controlled urethral hydraulic occluder (HO) device for the treatment of refractory urinary incontinence (RUI) in female dogs with various urogenital anomalies. STUDY DESIGN: Case series. ANIMALS: Female dogs with RUI (n = 18). METHODS: Retrospective evaluation of dogs after a silicone ring (HO) was surgically placed around the proximal urethra. The ring was connected to a subcutaneous injection port with actuating tubing. Residual incontinence was treated with percutaneous infusion of sterile saline into the device to provide extraluminal urethral compression. Dogs were assessed for continence (owner-assessed 10-point continence scale) and complications at standard time points. RESULTS: All 18 dogs had significantly improved continence scores (P < .001) after HO placement (median and mean score pre-HO = 2.8 and 3.3; post-HO = 10 and 8.9, respectively) with a median follow-up time of 32 months. "Functional" continence (score ≥ 9) was achieved in 67% of dogs after HO placement, though only 13/18 clients were compliant with inflations. Of dogs belonging to compliant owners, 12 (92%) had a functional continence score. Six dogs (33%) did not require inflation to achieve continence. Urethral obstruction occurred as a complication in 3 dogs. CONCLUSIONS: Use of an HO device was an effective long-term treatment for RUI when traditional options failed. The technique was associated with some complications, and these risks should be considered before use.

Size of cell-surface Kv2.1 domains is governed by growth fluctuations.

The Kv2.1 voltage-gated potassium channel forms stable clusters on the surface of different mammalian cells. Even though these cell-surface structures have been observed for almost a decade, little is known about the mechanism by which cells maintain them. We measure the distribution of domain sizes to study the kinetics of their growth. Using a Fokker-Planck formalism, we find no evidence for a feedback mechanism present to maintain specific domain radii. Instead, the size of Kv2.1 clusters is consistent with a model where domain size is established by fluctuations in the trafficking machinery. These results are further validated using likelihood and Akaike weights to select the best model for the kinetics of domain growth consistent with our experimental data.

Investigating transmission of SARS-CoV-2 using novel face mask sampling: a protocol for an observational prospective study of index cases and their contacts in a congregate setting.

INTRODUCTION: This study aims to measure how transmission of SARS-CoV-2 occurs in communities and to identify conditions that lend to increased transmission focusing on congregate situations. We will measure SARS-CoV-2 in exhaled breath of asymptomatic and symptomatic persons using face mask sampling-a non-invasive method for SARS-CoV-2 detection in exhaled air. We aim to detect transmission clusters and identify risk factors for SARS-CoV-2 transmission in presymptomatic, asymptomatic and symptomatic individuals. METHODS AND ANALYSIS: In this observational prospective study with daily follow-up, index cases and their respective contacts are identified at each participating institution. Contact definitions are based on Centers for Disease Control and Prevention and local health department guidelines. Participants will wear masks with polyvinyl alcohol test strips adhered to the inside for 2 hours daily. The strips are applied to all masks used over at least 7 days. In addition, self-administered nasal swabs and (optional) finger prick blood samples are performed by participants. Samples are tested by standard PCR protocols and by novel antigen tests. ETHICS AND DISSEMINATION: This study was approved by the Colorado Multiple Institutional Review Board and the WHO Ethics Review Committee. From the data generated, we will analyse transmission clusters and risk factors for transmission of SARS-CoV-2 in congregate settings. The kinetics of asymptomatic transmission and the evaluation of non-invasive tools for detection of transmissibility are of crucial importance for the development of more targeted control interventions-and ultimately to assist with keeping congregate settings open that are essential for our social fabric. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov (#NCT05145803).

Immunofluorescent Localization of Plakoglobin Is Altered in Endomyocardial Biopsy Samples from Dogs with Clinically Relevant Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC).

Diagnosing the early stages of canine Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC) is complicated by day-to-day arrhythmia variability, and absence of reliable, transthoracic echocardiographic features. Definitive diagnosis requires histopathologic identification of transmural fibrofatty replacement of the right ventricle. Reduction of immunofluorescent signal for plakoglobin (PG) at the intercalated disc (ID) is reported in ARVC-affected humans and boxers. Our objective was to determine whether reduced immunofluorescent signal for PG in endomyocardial biopsy samples (EMBs) correspond with a histopathologic diagnosis of ARVC. Here, 49 dogs were evaluated: 43 with advanced cardiac disease and 6 non-clinical boxers with mild to moderate ventricular arrhythmia (VA) burden. EMBs were obtained from all dogs; samples were prepared with antibodies recognizing cadherin (PC) and PG and evaluated with confocal microscopy. Investigators were blinded to breed and clinical status. ARVC was histopathologically diagnosed in 8 out of 49 dogs. Of these, three out of eight had clinical signs consistent with ARVC (two boxers, one English bulldog) and reduced PG signal at ID; five out of eight were non-clinical boxers with moderate VA and no reduction in PG. A total of 41 out of 49 dogs were histopathologically diagnosed with non-ARVC cardiac disease; 1 out of 41 showed reduction of PG at ID, while 40 out of 41 had no PG reduction. These results suggest that EMB PG signal is reduced in dogs with advanced ARVC, but not in the occult phase of the disease. Additionally, presence of PG at ID supports a diagnosis of non-ARVC cardiac disease in dogs with clinical signs. These results may offer an additional test that helps differentiate advanced ARVC from other myocardial diseases.

ACVIM consensus statement guidelines for the classification, diagnosis, and management of cardiomyopathies in cats.

Cardiomyopathies are a heterogeneous group of myocardial disorders of mostly unknown etiology, and they occur commonly in cats. In some cats, they are well-tolerated and are associated with normal life expectancy, but in other cats they can result in congestive heart failure, arterial thromboembolism or sudden death. Cardiomyopathy classification in cats can be challenging, and in this consensus statement we outline a classification system based on cardiac structure and function (phenotype). We also introduce a staging system for cardiomyopathy that includes subdivision of cats with subclinical cardiomyopathy into those at low risk of life-threatening complications and those at higher risk. Based on the available literature, we offer recommendations for the approach to diagnosis and staging of cardiomyopathies, as well as for management at each stage.

Temporal changes in clinical and radiographic variables in dogs with preclinical myxomatous mitral valve disease: The EPIC study.

BACKGROUND: The Evaluation of pimobendan in dogs with cardiomegaly caused by preclinical myxomatous mitral valve disease (EPIC) study monitored dogs with myxomatous mitral valve disease (MMVD) as they developed congestive heart failure (CHF). OBJECTIVES: To describe the changes in clinical and radiographic variables occurring as dogs with MMVD and cardiomegaly develop CHF, compared to similar dogs that do not develop CHF. ANIMALS: One hundred and thirty-five, and 73 dogs that did or did not develop CHF, respectively. MATERIALS AND METHODS: The following variables were evaluated in 2 groups of dogs (dogs that did or did not develop CHF): Heart rate (HR), clinic respiratory rate (RR), home-measured resting respiratory rate (RRR), rectal temperature (RT), body weight (BW), and vertebral heart sum (VHS). Absolute value and rate of change of each variable were calculated for each day a dog was in study. Daily means were calculated and plotted against time. The onset of CHF or last visit before leaving the study were set as reference time points. RESULTS: The most extreme values and rate of change occurred in variables immediately before onset of CHF. Vertebral heart sum increased earliest. Heart rate, RR, and RRR also increased. Rectal temperature and BW decreased. Increases in RR and RRR were most extreme and occurred immediately before CHF. CONCLUSIONS AND CLINICAL IMPORTANCE: Dogs with MMVD and cardiomegaly experience increases in HR, RR, RRR, and VHS, and decreases in BW and RT as they develop CHF. The variables with highest absolute change and rate of change were RR and RRR. These findings reinforce the value of RR and RRR as indicators of impending or incipient CHF.

Bacteremia associated with toothbrushing and dental extraction.

BACKGROUND: Antibiotic prophylaxis recommendations for the prevention of infective endocarditis are based in part on studies of bacteremia from dental procedures, but toothbrushing may pose a greater threat. The purpose of this study was to compare the incidence, duration, nature, and magnitude of endocarditis-related bacteremia from single-tooth extraction and toothbrushing and to determine the impact of amoxicillin prophylaxis on single-tooth extraction. METHODS AND RESULTS: In this double-blind, placebo-controlled study, 290 subjects were randomized to (1) toothbrushing, (2) single-tooth extraction with amoxicillin prophylaxis, or (3) single-tooth extraction with identical placebo. Blood was drawn for bacterial culturing and identification at 6 time points before, during, and after these interventions. The focus of our analysis was on bacterial species reported to cause infective endocarditis. We identified 98 bacterial species, 32 of which are reported to cause endocarditis. Cumulative incidence of endocarditis-related bacteria from all 6 blood draws was 23%, 33%, and 60% for the toothbrushing, extraction-amoxicillin, and extraction-placebo groups, respectively (P<0.0001). Significant differences were identified among the 3 groups at draws 2, 3, 4, and 5 (all P<0.05). Amoxicillin resulted in a significant decrease in positive cultures (P<0.0001). CONCLUSIONS: Although amoxicillin has a significant impact on bacteremia resulting from a single-tooth extraction, given the greater frequency for oral hygiene, toothbrushing may be a greater threat for individuals at risk for infective endocarditis.

Primary Sjögren's Syndrome: health experiences and predictors of health quality among patients in the United States.

OBJECTIVE: To assess the health related quality of life of patients with primary Sjögren's Syndrome (PSS) in a large US sample. METHODS: Questionnaires were mailed to 547 patients with a confirmed diagnosis of PSS (PhysR-PSS) and all active members of the Sjögren's Syndrome Foundation USA (SSF-PSS), half of whom identified a friend without PSS to also complete the survey. RESULTS: 277 PhysR-PSS patients were compared to 606 controls. The mean age was 62 years in the PhysR-PSS group and 61 years in the control group. 90% in both groups were women. Time from first symptom to diagnosis of PSS was a mean of 7 years. Sicca related morbidity, fatigue severity, depression and pain (assessed by validated questionnaires, PROFAD-SSI, FACIT-F, CES-D, BPI) were significantly greater, and all eight SF-36 domains were significantly diminished, in patients compared to controls. Somatic fatigue was the dominant predictor of physical function and of general health. Depression was the dominant predictor of emotional well being. Health care utilization was higher in patients than controls, including out of pocket dental expenses (mean: PhysR-PSS = $1473.3, controls = $503.6), dental visits (mean: PhysR-PSS = 4.0, controls = 2.3), current treatments (mean: PhysR-PSS = 6.6, controls = 2.5), and hospitalizations (53% PhysR-PSS, vs. 40% controls). CONCLUSION: Diminished health quality and excess health costs are prevalent among PSS patients. Health experiences and functional impact of PSS is similar among US and European patients. Delayed diagnosis, sicca related morbidity, fatigue, pain and depression are substantial suggesting unmet health needs and the importance of earlier recognition of PSS.

Assessment of serum N-terminal pro-B-type natriuretic peptide concentration for differentiation of congestive heart failure from primary respiratory tract disease as the cause of respiratory signs in dogs.

OBJECTIVE: To determine whether serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentration is useful in discriminating between cardiac and noncardiac (ie, primary respiratory tract disease) causes of respiratory signs (ie, coughing, stertor, stridor, excessive panting, increased respiratory effort, tachypnea, or overt respiratory distress) in dogs. DESIGN: Multicenter cross-sectional study. ANIMALS: P 115 dogs with respiratory signs. PROCEDURES: Dogs with respiratory signs were solicited for study. Physical examination, thoracic radiography, and echocardiography were used to determine whether respiratory signs were the result of cardiac (ie, congestive heart failure) or noncardiac (ie, primary respiratory tract disease) causes. Serum samples for NT-proBNP assay were obtained at time of admission for each dog. Receiver-operating characteristic curves were constructed to determine the ability of serum NT-proBNP concentration to discriminate between cardiac and noncardiac causes of respiratory signs. RESULTS: Serum NT-proBNP concentration was significantly higher in dogs with cardiac versus noncardiac causes of respiratory signs. In dogs with primary respiratory tract disease, serum NT-proBNP concentration was significantly higher in those with concurrent pulmonary hypertension than in those without. A serum NT-proBNP cutoff concentration > 1,158 pmol/L discriminated between dogs with congestive heart failure and dogs with primary respiratory tract disease with a sensitivity of 85.5% and a specificity of 81.3%. CONCLUSIONS AND CLINICAL RELEVANCE: Measuring serum NT-proBNP concentration in dogs with respiratory signs helps to differentiate between congestive heart failure and primary respiratory tract disease as an underlying cause.

Effect of Targeted Pulsed Electromagnetic Field Therapy on Canine Postoperative Hemilaminectomy: A Double-Blind, Randomized, Placebo-Controlled Clinical Trial.

Intervertebral disc disease is one of the leading causes of paralysis in dogs. Pulsed electromagnetic field (PEMF) therapy has been advocated for improving wound healing and pain reduction; however, robust clinical trials are lacking. The present prospective, double-blind, placebo-controlled trial evaluated targeted PEMF therapy administered to 53 client-owned dogs who underwent hemilaminectomy for naturally occurring disk extrusion intervertebral disc disease. The dogs were randomized to receive either targeted PEMF (n = 27) or placebo treatment (n = 28). Wound healing, evaluated by visual analog score and wound evaluation scale, was significantly improved at 6 wk postoperatively in the treatment compared with the control group (P = .010 and .023, respectively). Pain medications were administered less frequently in dogs receiving PEMF treatment during the 7 day postoperative period compared with the control treatment group (P = .010) with codeine administered 1.8 times more frequently in the control group. No untoward effects were recorded in either treatment group. More frequent evaluation of outcome measures with larger patient numbers, as well as histologic samples, may be useful in future studies. Dogs receiving PEMF therapy following postoperative hemilaminectomy demonstrated improved wound scores at 6 wk and reduced mean number of owner-administered pain medications compared with the control group therapy.

Evaluation of serum 25-hydroxyvitamin D concentrations in a heterogeneous canine ICU population.

OBJECTIVE: To identify the prevalence of low serum 25-hydroxyvitamin D (25-OH-D) concentrations in critically ill dogs and determine whether vitamin D concentration measured at admission correlates with illness severity and patient outcome. DESIGN: Prospective observational study from December 2013 to July 2014. SETTING: Private referral teaching hospital ICU. ANIMALS: One hundred eleven critically ill dogs admitted to the ICU, and 28 apparently healthy control dogs. INTERVENTIONS: Measurement of 25-OH-D concentration and assessment of physiological parameters required to calculate illness severity scores. MEASUREMENTS AND MAIN RESULTS: Blood was sampled to measure serum 25-OH-D concentration, point-of-care laboratory data (packed cell volume, total plasma protein, venous blood gas, electrolytes, plasma lactate, and blood glucose concentration), platelet count, and serum albumin within 12 hours of admission. Primary disease etiologies were determined. Mentation score, Acute Patient Physiologic and Laboratory Evaluation (APPLE) fast score, length of ICU hospitalization, and survival to discharge were recorded. Low serum 25-OH-D concentrations were detected in 25% (28/111) of the ICU cohort; 25-OH-D concentrations (median; IQR) (89.2; 26.3 ng/mL [222; 140.0 nmol/L]) were significantly lower in hospitalized dogs compared with the healthy cohort (127.5; 44.5 ng/mL [318; 111.0 nmol/L]) (P < 0.0001). Serum 25-OH-D concentrations had a weak positive correlation with albumin (r = 0.34, P = 0.0003), but not with any other blood analyte evaluated. Serum concentration of 25-OH-D was also weakly correlated with APPLEfast score (r = 0.31, P = 0.001). 25-OH-D concentrations (median; IQR) were significantly higher in ICU dogs that survived to discharge (95.5; 55 ng/mL [238; 137 nmol/L]) compared with nonsurvivors (60.0; 54 ng/mL [149; 135 nmol/L]; P < 0.001). CONCLUSIONS: Low serum 25-OH-D was recorded in one-quarter of critically ill dogs, and was significantly lower in nonsurvivors in comparison to survivors. There was weak correlation between serum 25-OH-D and illness severity. Further studies are required to clarify relationships between vitamin D status and outcome.

Comparison of closed root planing with versus without concurrent doxycycline hyclate or clindamycin hydrochloride gel application for the treatment of periodontal disease in dogs.

OBJECTIVE To compare improvements in values for periodontal disease indices in dogs following treatment with closed root planing (CRP) alone, CRP with concurrent 8.5% doxycycline hyclate gel application, and CRP with concurrent 2% clindamycin hydrochloride reverse-polymer hydrogel application. DESIGN Randomized, blinded, controlled clinical trial. ANIMALS 34 client-owned dogs with periodontal pockets measuring 3.5 to 5.5 mm deep. PROCEDURES Dogs were randomly assigned to receive 1 of 3 treatments: CRP alone (n = 10) or CRP plus 8.5% doxycycline hyclate (12) or 2% clindamycin hydrochloride (12) gel applied within the periodontal pockets. Indices of periodontal disease severity were recorded before and 12 weeks after treatment, and outcomes were compared among treatment groups. RESULTS Except for gingivitis index, no significant differences were identified among the 3 treatment groups in the amount of improvement observed in values for periodontal disease indices following treatment. A minor but clinically unimportant improvement in mean gingivitis index values was identified for dogs treated with CRP plus doxycycline gel, which differed significantly from improvements in the other 2 groups. Teeth that were initially more severely affected (pocket depths, 5.0 to 5.5 mm) had the greatest amount of improvement, whereas teeth with only mildly high initial pocket depths (3.5 to 4.0 mm) had less improvement. CONCLUSIONS AND CLINICAL RELEVANCE Overall, addition of doxycycline or clindamycin gel application to CRP for the treatment of periodontal disease in dogs yielded no clinically relevant benefit over CRP during the 12-week follow-up period.

Utility of admission lactate concentration, lactate variables, and shock index in outcome assessment in dogs diagnosed with shock.

OBJECTIVE: To determine whether admission venous plasma lactate concentration, calculated lactate variables, or shock index (SI) could discriminate hospital survivors from nonsurvivors in dogs admitted with shock. DESIGN: Prospective investigation performed over a 19-month period. SETTING: Large urban private teaching hospital. ANIMALS: Twenty-three dogs consecutively admitted to the ICU from January 2008 to July 2009 with initial peripheral venous plasma lactate concentration >2 mmol/L (18.0 mg/dL) and clinical and hemodynamic parameters consistent with shock. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Heart rate, systolic blood pressure, and venous plasma lactate concentrations were serially recorded at predefined time points and used to calculate SI (SI = heart rate/systolic blood pressure) and lactate variables, including lactime (time lactate > 2.0 mmol/L), lactate clearance ([lactateinitial - lactatedelayed ]/lactateinitial × 100), and LACAREA (area under the lactate concentration versus time curve). Primary outcome was survival to discharge. Overall survival rate was 61%. Admission venous plasma lactate concentration did not differ between groups (P = 0.2). Lactime was shorter in survivors versus nonsurvivors (P = 0.02). Lactate clearance at 1, 10, 16, 24, and 36 hours, and final lactate clearance were greater in survivors versus nonsurvivors (P < 0.05). LACAREA at time intervals 0-1, 1-4, 4-10, 10-16, 16-24, 24-30, and 30-36 hours was larger in nonsurvivors versus survivors (P < 0.05). Total LACAREA did not differ between groups (P = 0.09). Admission SI and time to normalize SI (SI < 0.9) were not different between survivors and nonsurvivors (P > 0.05). CONCLUSIONS: While admission venous plasma lactate concentration could not discriminate between hospital survivors and nonsurvivors, lactate variables showed clinical utility to predict outcome in dogs with shock. Further studies are needed to determine SI reference ranges and optimal SI cut-off values to improve its prognostic ability in sick dogs.

A genetically encoded probe for imaging nascent and mature HA-tagged proteins in vivo.

To expand the toolbox of imaging in living cells, we have engineered a single-chain variable fragment binding the linear HA epitope with high affinity and specificity in vivo. The resulting probe, called the HA frankenbody, can light up in multiple colors HA-tagged nuclear, cytoplasmic, membrane, and mitochondrial proteins in diverse cell types. The HA frankenbody also enables state-of-the-art single-molecule experiments in living cells, which we demonstrate by tracking single HA-tagged histones in U2OS cells and single mRNA translation dynamics in both U2OS cells and neurons. Together with the SunTag, we also track two mRNA species simultaneously to demonstrate comparative single-molecule studies of translation can now be done with genetically encoded tools alone. Finally, we use the HA frankenbody to precisely quantify the expression of HA-tagged proteins in developing zebrafish embryos. The versatility of the HA frankenbody makes it a powerful tool for imaging protein dynamics in vivo.