Factors associated with survival to hospital discharge for cats treated surgically for thoracic trauma.

OBJECTIVE To determine the most common types of injuries in cats surgically treated for thoracic trauma, complications associated with surgical treatment, and factors associated with mortality rate and evaluate the effectiveness of the animal trauma triage (ATT) scoring system for predicting outcome. DESIGN Retrospective case series with nested observational study. ANIMALS 23 client-owned cats surgically treated for thoracic trauma at 7 veterinary teaching hospitals between 1990 and 2014. PROCEDURES Medical records were reviewed to collect data on signalment, medical history, clinical signs and physical examination findings at initial evaluation, clinicopathologic findings, initial emergency treatments and diagnostic tests performed, type of trauma sustained, imaging findings, surgery details, postoperative complications, duration of hospitalization, and cause of death, if applicable. All variables were evaluated for associations with survival to hospital discharge. RESULTS Types of trauma that cats had sustained included dog bite or attack (n = 8 [35%]), motor vehicle accident (6 [26%]), other animal attack (2 [9%]), impalement injury or fall (2 [9%]), projectile penetrating trauma (1 [4%]), or unknown origin (4 [17%]). Intrathoracic surgery was required for 65% (15/23) of cats. The overall perioperative mortality rate was 13% (3/23). Mean ± SD ATT scores for surviving and nonsurviving cats were 6.4 ± 2.2 and 10.0 ± 1.7, respectively. Nineteen of 20 cats with no cardiopulmonary arrest survived to discharge, compared with 1 of 3 cats with cardiopulmonary arrest. Only these 2 variables were significantly associated with outcome. CONCLUSIONS AND CLINICAL RELEVANCE The perioperative mortality rate was low in this series of cats with thoracic trauma; however, those with cardiopulmonary arrest were less likely to survive to hospital discharge than other cats. Cats with a low ATT score were more likely to survive than cats with a high ATT score.

Perioperative mortality rate and risk factors for death in dogs undergoing surgery for treatment of thoracic trauma: 157 cases (1990-2014).

OBJECTIVE To determine perioperative mortality rate and identify risk factors associated with outcome in dogs with thoracic trauma that underwent surgical procedures and to evaluate the utility of the animal trauma triage (ATT) score in predicting outcome. DESIGN Retrospective case series. ANIMALS 157 client-owned dogs. PROCEDURES Medical records databases of 7 veterinary teaching hospitals were reviewed. Dogs were included if trauma to the thorax was documented and the patient underwent a surgical procedure. History, signalment, results of physical examination and preoperative laboratory tests, surgical procedure, perioperative complications, duration of hospital stay, and details of follow-up were recorded. Descriptive statistics and ATT scores were calculated, and logistic regression analysis was performed. RESULTS 123 of 157 (78%) patients underwent thoracic surgery, and 134 of 157 (85.4%) survived to discharge. Mean ± SD ATT score for nonsurvivors was 8 ± 2.4. In the multivariable model, female dogs and dogs that did not experience cardiac arrest as a postoperative complication had odds of survival 6 times and 102 times, respectively, those of male dogs and dogs that did experience cardiac arrest as a postoperative complication. Additionally, patients with a mean ATT score < 7 had odds of survival 5 times those of patients with an ATT score ≥ 7. CONCLUSIONS AND CLINICAL RELEVANCE The overall perioperative mortality rate was low for patients with thoracic trauma undergoing surgery in this study. However, male dogs and dogs that experienced cardiac arrest had a lower likelihood of survival to discharge. The ATT score may be a useful adjunct to assist clinical decision-making in veterinary patients with thoracic trauma.

Experimental observations and numerical modeling of lipid-shell microbubbles with calcium-adhering moieties for minimally-invasive treatment of urinary stones.

A novel treatment modality incorporating calcium-adhering microbubbles has recently entered human clinical trials as a new minimally-invasive approach to treat urinary stones. In this treatment method, lipid-shell gas-core microbubbles can be introduced into the urinary tract through a catheter. Lipid moities with calcium-adherance properties incorporated into the lipid shell facilitate binding to stones. The microbubbles can be excited by an extracorporeal source of quasi-collimated ultrasound. Alternatively, the microbubbles can be excited by an intraluminal source, such as a fiber-optic laser. With either excitation technique, calcium-adhering microbubbles can significantly increase rates of erosion, pitting, and fragmentation of stones. We report here on new experiments using high-speed photography to characterize microbubble expansion and collapse. The bubble geometry observed in the experiments was used as one of the initial shapes for the numerical modeling. The modeling showed that the bubble dynamics strongly depends on bubble shape and stand-off distance. For the experimentally observed shape of microbubbles, the numerical modeling showed that the collapse of the microbubbles was associated with pressure increases of some two-to-three orders of magnitude compared to the excitation source pressures. This in-vitro study provides key insights into the use of microbubbles with calcium-adhering moieties in treatment of urinary stones.

The Secreted Protease PrtA Controls Cell Growth, Biofilm Formation and Pathogenicity in Xylella fastidiosa.

Pierce's disease (PD) is a deadly disease of grapevines caused by the Gram-negative bacterium Xylella fastidiosa. Though disease symptoms were formerly attributed to bacteria blocking the plant xylem, this hypothesis is at best overly simplistic. Recently, we used a proteomic approach to characterize the secretome of X. fastidiosa, both in vitro and in planta, and identified LesA as one of the pathogenicity factors of X. fastidiosa in grapevines that leads to leaf scorching and chlorosis. Herein, we characterize another such factor encoded by PD0956, designated as an antivirulence secreted protease "PrtA" that displays a central role in controlling in vitro cell proliferation, length, motility, biofilm formation, and in planta virulence. The mutant in X. fastidiosa exhibited reduced cell length, hypermotility (and subsequent lack of biofilm formation) and hypervirulence in grapevines. These findings are supported by transcriptomic and proteomic analyses with corresponding plant infection data. Of particular interest, is the hypervirulent response in grapevines observed when X. fastidiosa is disrupted for production of PrtA, and that PD-model tobacco plants transformed to express PrtA exhibited decreased symptoms after infection by X. fastidiosa.

Postoperative Respiratory Function and Survival After Pneumonectomy in Dogs and Cats.

OBJECTIVE: To describe indications for, and outcomes after, pneumonectomy in dogs and cats, including assessment of immediate postoperative respiratory function in comparison to dogs undergoing single lung lobectomy. STUDY DESIGN: Retrospective case series. ANIMALS: Dogs (n=16) and cats (n=7) with naturally occurring pulmonary disease. METHODS: Medical records (1990-2014) of dogs and cats undergoing right or left pneumonectomy were reviewed. Data retrieved included signalment, history, preoperative diagnostics, operative descriptions, postoperative data including respiratory function, and postdischarge outcomes. For respiratory function comparisons, medical records of dogs having undergone a single lung lobectomy via median sternotomy (n=15) or intercostal thoracotomy (n=15) were reviewed. RESULTS: Twenty-three cases (16 dogs, 7 cats) were included. Pneumonectomy was performed for congenital (1 dog, 1 cat), neoplastic (8 dogs, 1 cat), and infectious (7 dogs, 5 cats) disease. Postoperative aspiration pneumonia occurred in 2 dogs; 15 of 16 dogs (94%) and 6/7 cats (86%) survived to hospital discharge. After pneumonectomy, dogs had a significantly higher postoperative PaO2 on 21% oxygen (P=.033) and lower postoperative A-a gradient (P=.004) compared to dogs undergoing single lung lobectomy. Survival times (right-censored at last follow-up) for dogs ranged from 2 days to 7 years (estimated median=1,868 days) and for cats from 1-585 days. CONCLUSION: Dogs and cats have acceptable respiratory function immediately postoperatively and most have protracted long-term survival after pneumonectomy for a variety of pulmonary diseases.

Elemental Content of Calcium Oxalate Stones from a Canine Model of Urinary Stone Disease.

One of the most common types of urinary stones formed in humans and some other mammals is composed of calcium oxalate in ordered hydrated crystals. Many studies have reported a range of metals other than calcium in human stones, but few have looked at stones from animal models such as the dog. Therefore, we determined the elemental profile of canine calcium oxalate urinary stones and compared it to reported values from human stones. The content of 19 elements spanning 7-orders of magnitude was quantified in calcium oxalate stones from 53 dogs. The elemental profile of the canine stones was highly overlapping with human stones, indicating similar inorganic composition. Correlation and cluster analysis was then performed on the elemental profile from canine stones to evaluate associations between the elements and test for potential subgrouping based on elemental content. No correlations were observed with the most abundant metal calcium. However, magnesium and sulfur content correlated with the mineral hydration form, while phosphorous and zinc content correlated with the neuter status of the dog. Inter-elemental correlation analysis indicated strong associations between barium, phosphorous, and zinc content. Additionally, cluster analysis revealed subgroups within the stones that were also based primarily on barium, phosphorous, and zinc. These data support the use of the dog as a model to study the effects of trace metal homeostasis in urinary stone disease.

Evaluation of acid-base disorders in dogs and cats presenting to an emergency room. Part 2: comparison of anion gap, strong ion gap, and semiquantitative analysis.

OBJECTIVE: To compare the diagnostic performance of the anion gap (AG) with 2 physicochemical approaches to identify unmeasured anions. DESIGN: Prospective cohort study. SETTING: University teaching hospital. ANIMALS: Eighty-four dogs and 14 cats presenting to a university teaching hospital emergency room. INTERVENTIONS: All dogs and cats in which venous blood samples for acid-base, lactate, and serum biochemical analysis were all collected within 60 minutes of each other, over a 5-month enrollment period. Unmeasured anions were quantified using each of three approaches: the anion gap (AG), strong ion gap (SIG), and a semiquantitative approach (XA). MEASUREMENTS AND MAIN RESULTS: An increased AG metabolic acidosis was evident in 34/98 of cases. The Stewart approach identified an increased SIG acidosis in 49/98 of cases. There was a strong correlation between SIG and AG (r = 0.89; P < 0.001). The semiquantitative approach identified increased unmeasured anions in 68/98 of cases. There was a moderate correlation between AG and XA (r = 0.68; P < 0.001) and a slightly stronger correlation between SIG and XA (r = 0.75; P < 0.001). Plasma lactate concentrations and AG were poorly correlated (r = 0.22; P = 0.029) and there was no correlation between lactate concentrations and BE (r = 0.19; P = 0.069). CONCLUSIONS: Unmeasured anions occurred commonly in this sample of small animal emergency room patients and physiochemical approaches identified more animals with unmeasured anions than the traditional AG calculation. Further studies are needed to determine if the results of the physicochemical approach improves clinical management and warrants the associated increases in cost and complexity.

Evaluation of acid-base disorders in dogs and cats presenting to an emergency room. Part 1: comparison of three methods of acid-base analysis.

OBJECTIVE: To compare the diagnostic performance of the traditional approach to acid-base analysis with the Stewart approach and a semiquantitative approach. DESIGN: Prospective cohort study. SETTING: University teaching hospital. ANIMALS: A total number of 84 dogs and 14 cats presenting to a university teaching hospital emergency room. PROCEDURES: All dogs and cats in which venous blood samples for acid-base, lactate, and serum biochemical analysis were all collected within 60 minutes of each other, over a 5-month enrollment period. Acid-base analysis was performed using the traditional approach, Stewart approach, and a semiquantitative approach. RESULTS: Traditional acid-base analysis identified respiratory acid-base abnormalities in 14/98 animals and metabolic acid-base abnormalities in 67/98. A mixed disorder of metabolic acidosis and respiratory alkalosis was most common occurring in 29/98 patients. The Stewart approach identified metabolic abnormalities in 82/98 patients; strong ion difference abnormalities were evident in 68/98 cases; an increased strong ion gap acidosis was identified in 49/98 cases; and changes in the quantity of weak acids in 25/98 cases. The semiquantitative approach identified abnormalities in all cases evaluated. Of the 14 patients with a primary respiratory acid-base abnormality, the Stewart approach identified metabolic abnormalities in 9 and the semiquantitative approach found abnormalities in all animals. CONCLUSIONS AND CLINICAL RELEVANCE: The physicochemical approaches diagnosed more acid-base abnormalities in this population than the traditional approach although many of the abnormalities identified were small and of unknown clinical relevance. The physicochemical approaches may provide greater insight as to the underlying etiology of abnormalities, which maybe of particular relevance to cases with changes in albumin and/or phosphorus concentration.

The effect of hetastarch 670/0.75 administered in vivo as a constant rate infusion on platelet closure time in the dog.

OBJECTIVE: To evaluate the effects of hetastarch 670/0.75 on canine platelet function and clinical bleeding following its administration as a constant rate infusion (CRI) at 1 mL/kg/h and 2 mL/kg/h for 24 hours. DESIGN: In vivo, prospective, open-label, crossover study. SETTING: Research laboratory at a university veterinary facility. ANIMALS: Eight healthy, adult male research dogs. INTERVENTIONS: Each dog received 1 mL/kg/h hetastarch for 24 hours then 2 mL/kg/h with a washout period of 10 weeks between each experiment. Platelet closure time (CT) was measured using a platelet function analyzer with collagen adenosine diphosphate (ADP) cartridges. CT measurements were performed at baseline and 6, 12, and 24 hours following initiation of hetastarch infusion. MEASUREMENTS AND MAIN RESULTS: At 1 mL/kg/h, mean CT was significantly increased at the 12- and 24-hour time point relative to the baseline value, although mean CT never rose to a value above the reference interval during the 24-hour infusion. At 2 mL/kg/h, median CT was also significantly increased at the 12- and 24-hour time point relative to the baseline value. Administration of 2 mL/kg/h did progressively prolong the median CT value though only exceeded the reference interval at the 24-hour time point. Despite the prolongation of median CT, there was no clinical evidence of spontaneous bleeding in any dog during the 24-hour infusion at either CRI rate. CONCLUSIONS: Hetastarch 670/0.75 when used as a 24-hour CRI at 1 and 2 mL/kg/h prolongs CT in healthy dogs at 6, 12, and 24 hours. Median CT only exceeded the reference interval at 24 hours at 2 mL/kg/h.

A comparison of the clinical utility of several published formulae for estimated osmolality of canine serum.

OBJECTIVE: To determine the normal osmole gap for 18 previously published formulae used to estimate serum osmolality in dogs. DESIGN: Prospective study. SETTING: University veterinary medical teaching hospital. ANIMALS: Two hundred and fifty client-owned dogs. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Serum samples were saved and frozen at -80°C after routine biochemical analysis as ordered by attending clinicians. An Advanced Micro Osmometer 3300 was used to measure serum osmolality. Eighteen distinct formulae previously reported in the medical literature were used to calculate the osmolality from the biochemical analysis results. The calculated osmolality was then subtracted from the measured osmolality to determine the osmole gap. Osmole gaps for azotemic and hyperglycemic dogs were compared to those of dogs without azotemia or hyperglycemia using each formula. The median measured osmolality for all dogs in the study was 302 mOsm/kg (interquartile range 297-307). The osmole gaps varied widely depending on the formula used to calculate osmolality and the presence or absence of hyperglycemia or azotemia. Eleven formulae led to calculated osmolality and osmole gaps that were not statistically different when hyperglycemia or azotemia was present. Four out of these 11 formulae resulted in osmole gaps near zero. CONCLUSIONS AND CLINICAL RELEVANCE: Multiple formulae reported to calculate serum osmolality can be used in the clinical setting, but they result in significantly different normal osmole gaps. Clinicians should be aware of the specific reference interval for the formula being used. The authors recommend the formula 2(Na(+) ) + [glucose/18] + [BUN/2.8] because it is easy to use and is reliable even when hyperglycemia or azotemia are present.

Ventilator waveforms.

Ventilator waveforms are graphic representations of changes in pressure, flow, and volume within a ventilator circuit. The changes in these parameters over time may be displayed individually (scalars) or plotted one against another (pressure-volume and flow-volume loops). There are 6 basic shapes of scalar waveforms, but only 3 are functionally distinct (square, ramp, and sine). The pressure scalar is a particularly valuable tool when constant flow (e.g., volume control) modes are employed and an inspiratory pause is added. In this setting, inspection of the pressure waveform can allow determination of static, quasistatic, and dynamic compliance, as well as relative changes in airway resistance. Inspection of the pressure waveform can also help to identify many important aspects of patient drug responses, dyssynchrony, and air trapping (auto positive end-expiratory pressure [auto-PEEP]). Depending on the ventilation mode employed, the shape of the flow waveform may be set by the ventilator operator or may be dependent on patient effort and lung mechanics. Decelerating flow patterns have several important advantages when this option is available. Inspection of flow waveforms is crucial in the recognition of dyssynchrony, setting optimal inspiratory times, evaluating responses to bronchodilators, and the recognition of auto-PEEP. The volume waveform often contains somewhat less useful information than the other 2 scalars, but plays a crucial role in the identification of leaks in the circuit. Pressure-volume loops are particularly useful in setting PEEP and peak inspiratory pressure ranges. Inspection of these loops also often helps in the evaluation of lung mechanics, in the identification of circuit leaks, and in the assessment of patient triggering effort. Flow-volume loops are extremely useful in the identification of leaks and excessive airway secretions as well as alterations in airway resistance. Lastly, serial waveform inspection is crucial to the identification and resolution of patient-ventilator dyssynchrony in many cases.

Indication, management, and outcome of brachycephalic dogs requiring mechanical ventilation.

OBJECTIVES: To evaluate the frequency, and need for mechanical ventilation (MV) in a population of brachycephalic dogs (BD) compared with non-BD. Also, to describe the pre-MV abnormalities, ventilator settings used, the cardiovascular and pulmonary monitoring results and complications encountered in the same BD population. In addition, we sought to identify factors associated with successful weaning and describe outcomes of BD requiring MV. DESIGN: Retrospective observational study (1990-2008). SETTING: University Small Animal Teaching Hospital. ANIMALS: Fifteen BD managed with MV. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Signalment, indication for MV, ventilator settings, arterial blood gas values, duration of MV, complications, and outcome were recorded for each patient enrolled in study. BD were more likely to receive MV than non-BD (P=0.036). Out of the 15 dogs that fulfilled the inclusion criteria 7 (47%) underwent MV for impending respiratory fatigue, 6 (40%) for hypoxemia and 2 for hypercapnea. The most common underlying disease was aspiration pneumonia. Duration of MV ranged from 2 to 240 hours (median 15 hours). Seven patients were weaned (47%). Seven dogs had a temporary tracheostomy tube and 5 of them (71%) were weaned. Dogs that were weaned had a significantly greater preweaning trial PaO2/FiO2 ratio than those that were not (359 ± 92 versus 210 ± 57 mm Hg, P=0.025). No significant difference for weaning success between dogs with and those without a tracheostomy was detected (P=0.132). The discharge rate was 27% (all from the respiratory fatigue group). CONCLUSION: Among all dogs admitted to ICU, BD were more likely to receive MV than non-BD. Aspiration pneumonia was frequently identified as the underlying cause of respiratory compromise. The survival rate for BD undergoing MV was not markedly different from previous studies. Weaning of BD from MV may be facilitated by employing preemptive strategies such as performing tracheostomy tube placements.

In vitro evaluation of the efficacy of a veterinary dry heat fluid warmer.

OBJECTIVES: To evaluate the efficacy of a veterinary dry heat fluid warmer on ambient and prewarmed crystalloid fluids and refrigerated packed red blood cells (pRBC). DESIGN: Prospective in vitro study. SETTING: University teaching hospital. ANIMALS: None. INTERVENTIONS: Ambient and prewarmed crystalloid fluids and refrigerated pRBC were delivered via a standard fluid administration set at various rates. A thermistor continuously monitored fluid outflow temperature with and without a dry heat veterinary fluid warmer (study device). RESULTS: The outflow temperature was significantly higher with the study device as compared to control conditions for all fluids and rates tested. The maximum outflow temperature of approximately 35°C (95°F) occurred when the study device was applied to either ambient or prewarmed crystalloid fluids at 50 mL/h. In the study device trials, the outflow temperature of ambient crystalloid fluids ranged from 35.1° to 27.3°C (95.2° to 81.1°F) as the fluid rate increased from 50 to 999 mL/h. Control trials of prewarmed crystalloids produced outflow temperatures that rapidly approached ambient temperature. Addition of the study device to prewarmed crystalloids resulted in outflow temperatures that were similar to that of the corresponding ambient crystalloid trials. Control trials of refrigerated pRBC achieved ambient temperature at rates from 10 to 500 mL/h. With the study device, pRBC were maximally warmed to an outflow temperature of 35.8°C (96.4°F) at 100 mL/h. CONCLUSION: Although the study device generated statistically significant increases in outflow temperature of crystalloid fluids and pRBC, the ability of the device to decrease the metabolic cost of fluid administration is limited to specific clinical scenarios. The use of prewarmed crystalloid fluids with or without the study device offers minimal benefit over ambient temperature crystalloids. Substantial warming of pRBC occurs during administration, even without use of the study device.

Airway microbial culture and susceptibility patterns in dogs and cats with respiratory disease of varying severity.

OBJECTIVE: To compare airway microbiological culture and susceptibility results in 2 groups of dogs and cats: 1 with respiratory failure requiring positive pressure ventilation (PPV) and 1 with respiratory disease. DESIGN: Retrospective study. SETTING: University teaching hospital. ANIMALS: Fifty-two dogs and cats requiring PPV that had an airway microbiologic culture submitted from October 1, 2003 to October 31, 2008 were included. One hundred and four airway microbiologic cultures from dogs and cats with respiratory disease not requiring PPV were randomly sampled for comparison. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients with respiratory failure were more likely to have a gram-negative enteric isolate identified (P<0.001), while patients with respiratory disease were more likely to have a gram-negative nonenteric isolate (P<0.001) or anaerobic isolate (P<0.001) identified. Aerobic bacterial isolates from patients with respiratory failure were less likely to be susceptible to ampicillin (P=0.006), amoxicillin/clavulonate (P<0.001), chloramphenicol (P=0.004), enrofloxacin (P<0.001), ticarcillin/clavulonate (P=0.004), and the combination of ampicillin with enrofloxacin (P<0.001) than were aerobic bacterial isolates from patients with respiratory disease. CONCLUSIONS: Canine and feline patients with respiratory failure severe enough to require PPV exhibit a different pattern of bacterial isolates cultured from their airways when compared with isolates from patients with respiratory disease that has not resulted in ventilator dependence. These isolates are more likely to be resistant to commonly used antimicrobials/antimicrobial combinations than patients in the respiratory disease group. These findings suggest that in canine and feline patients with infectious lower respiratory tract disease, consideration of the severity of the pulmonary insult may allow for better prediction of likely isolates and their antimicrobial susceptibilities. Further prospective studies with a standardized collection technique are warranted.

Comparison of regular insulin infusion doses in critically ill diabetic cats: 29 cases (1999-2007).

OBJECTIVE: To compare biochemical parameters, neurologic changes, length of hospital stay, and clinical improvement in 3 groups of cats with diabetic ketosis/diabetic ketoacidosis (DK/DKA) prescribed varied doses of regular insulin as a continuous rate of infusion (CRI). DESIGN: Retrospective study. SETTING: University teaching hospital. ANIMALS: Twenty-nine client-owned cats with DK/DKA prescribed a regular insulin CRI. INTERVENTIONS: Cats were grouped as follows: 7 cats each in Group 1 and 2, (prescribed 1.1 and 2.2 U/kg/d, respectively), and 15 cats in Group 3 (prescribed increasing doses as needed). MEASUREMENTS AND MAIN RESULTS: None of the groups received the total prescribed dose of insulin. The mean actual dose administered/kg/d ranged from 0.30 (0.21) to 0.87 (0.32) U/kg/d in Groups 1, 2, and 3. There was no difference in mean minimum blood glucose (BG) per 4 hours or change in BG from baseline per 4 hours between Groups 1 and 2 (P=0.63, 0.50). There was no difference between groups regarding the time required to reach a BG ≤ 13.9 mmol/L (250 mg/dL), serum phosphorus or potassium concentrations relative to baseline values (P=0.53, 0.90), length of time until urine or serum ketones were no longer detected (P=0.73), the animal commenced eating (P=0.24), or length of hospital stay (P=0.63). Four of the cats had declining mentation during hospitalization; there were no relationships between osmolality at presentation, either prescribed or administered insulin dose, and mentation changes. Three of the 4 cats with declining mentation survived. Twenty-seven of the 29 cats (93%) survived to discharge. CONCLUSIONS: In this study, prescribing the published canine dose (2.2 U/kg/d) of regular insulin to cats with DK/DKA does not appear to increase the frequency of adverse neurologic or biochemical sequelae compared with cats that are prescribed the published cat dose (1.1 U/kg/d). The use of a sliding scale for determination of infusion rates significantly reduces the amount of insulin cats receive in this setting. Determination of whether adverse sequelae would occur more frequently if cats with DK/DKA received the full insulin prescribed doses of 1.1, 2.2, or >2.2 U/kg/d is warranted. Further controlled studies are necessary to determine if higher doses of insulin are associated with beneficial effects on morbidity or mortality.

Airway smooth muscle proliferation and mechanics: effects of AMP kinase agonists.

Obesity is a risk factor for asthma. The purpose of this study was to determine whether metformin, an agent used in the treatment of an obesity-related condition (type II diabetes), might have therapeutic potential for modifying the effects of obesity on airway smooth muscle (ASM) function. Metformin acts via activation of AMP-activated protein kinase (AMPK), a cellular sensor of energy status. In cultured murine ASM cells, metformin (0.2-2 mM) caused a dose-dependent inhibition of cell proliferation induced by PDGF (10(-8) M) and serotonin (10(-4) M). Another AMPK activator, 5-aminoimidazole-4-carboxamide-1-beta-Driboruranoside (AICAR), also inhibited PDGF-induced proliferation. Furthermore, cells treated with metformin or AICAR, also exhibited an attenuation in the rate of cytoskeletal remodeling, as quantified by spontaneous nanoscale motions of microbeads tightly anchored to the cytoskeleton (CSK) of the ASM cell. ASM cells treated with metformin or AICAR, however, exhibited no appreciable differences in stiffness as measured by optical magnetic twisting cytometry (OMTC) or their abilities to stiffen in response to contractile agonist serotonin. Taken together, these findings suggest that metformin, probably through activation of AMPK, reduces the rate of ongoing reorganization of the CSK and inhibits ASM cell proliferation.

Filamin-a and rheological properties of cultured melanoma cells.

Here we report the rheological properties of cultured hsFLNa (filamin-A)-expressing (FIL+) and hsFLNa-deficient (FIL-) melanoma cells. Using magnetic twisting cytometry over a wide range of probing frequencies, and targeting either cortical or deeper cytoskeletal structures, we found that differences in stiffness of FIL+ versus FIL- cells were remarkably small. When probed through deep cytoskeletal structures, FIL+ cells were, at most, 30% stiffer than FIL- cells, whereas when probed through more peripheral cytoskeletal structures FIL- cells were not different except at very high frequencies. The loss tangent, expressed as an effective cytoskeletal temperature, was systematically greater in FIL- than FIL+ cells, but these differences were small and showed that the FIL+ cells were only slightly closer to a solidlike state. To quantify cytoskeletal remodeling, we measured spontaneous motions of beads bound to cortical cytoskeletal structures and found no difference in FIL+ versus FIL- cells. Although mechanical differences between FIL+ and FIL- cells were evident both in cortical and deeper structures, these differences were far smaller than expected based on measurements of the rheology of purified actin-filamin solutions. These findings do not rule out an important contribution of filamin to the mechanical properties of the cortical cytoskeleton, but suggest that effects of filamin in the cortex are not exerted on the length scale of the probe used here. These findings would appear to rule out any important contribution of filamin to the bulk mechanical properties of the cytoplasm, however. Although filamin is present in the cytoplasm, it may be inactive, its mechanical effects may be small compared with other crosslinkers, or mechanical properties of the matrix may be dominated by an overriding role of cytoskeletal prestress.

Oncostatin M causes eotaxin-1 release from airway smooth muscle: synergy with IL-4 and IL-13.

BACKGROUND: Eotaxin is implicated in asthmatic eosinophilia. Oncostatin M (OSM) causes eotaxin release from fibroblasts. OBJECTIVE: We sought to examine the effects and mechanism of action of OSM and other IL-6 family cytokines on eotaxin release from human airway smooth muscle cells. METHODS: Eotaxin 1 release was measured by means of ELISA. Western blotting was used to examine mitogen-activated protein kinase and signal transducer and activator of transcription 3 (STAT-3) phosphorylation. Eotaxin promoter activity was analyzed in cells transfected with wild-type STAT-3, a mutant form of STAT-3 that cannot be phosphorylated, and a constitutively active form of STAT-3. The mRNA and protein expression of IL-4R alpha, the signaling receptor for IL-4 and IL-13, was evaluated by means of real-time PCR and flow cytometry, respectively. RESULTS: OSM increased eotaxin 1 release and augmented IL-4- or IL-13-induced eotaxin release, whereas other IL-6 family cytokines did not. OSM caused a greater increase in STAT-3 phosphorylation and STAT-3-mediated gene transcription than other IL-6 family cytokines. OSM increased eotaxin promoter activity and augmented IL-13- and IL-4-induced increases in promoter activity. The constitutively active form of STAT-3 increased eotaxin promoter activity, whereas the mutant form of STAT-3 that cannot be phosphorylated significantly reduced eotaxin promoter activity induced by OSM or IL-4 plus OSM. OSM increased IL-4R alpha mRNA and protein levels. CONCLUSIONS: OSM induces eotaxin 1 expression in human airway smooth muscle cells by a mechanism involving STAT-3. OSM synergizes with IL-13 and IL-4 to increase eotaxin 1 expression, possibly as a result of effects on IL-4R alpha expression.

Oncostatin M causes VEGF release from human airway smooth muscle: synergy with IL-1beta.

Vascular endothelial growth factor (VEGF), a potent angiogenesis factor, likely contributes to airway remodeling in asthma. We sought to examine the effects and mechanism of action of IL-6 family cytokines on VEGF release from human airway smooth muscle (HASM) cells. Oncostatin M (OSM), but not other IL-6 family cytokines, increased VEGF release, and IL-1beta enhanced OSM-induced VEGF release. OSM increased VEGF mRNA expression and VEGF promoter activity, whereas IL-1beta had no effect. IL-1beta did not augment the effects of OSM on VEGF promoter activity but did augment OSM-induced VEGF mRNA expression and mRNA stability. The STAT3 inhibitor piceatannol decreased both OSM-induced VEGF release and synergy between OSM and IL-1beta, without affecting responses to IL-1beta alone. Piceatannol also inhibited OSM-induced VEGF mRNA expression. In contrast, inhibitors of MAPK pathway had no effect on OSM or OSM plus IL-1beta-induced VEGF release. OSM increased type 1 IL-1 receptor (IL-1R1) mRNA expression, as measured by real-time PCR, and piceatannol attenuated this response. Consistent with the increase in IL-1R1 expression, OSM markedly augmented IL-1beta-induced VEGF, MCP-1, and IL-6 release. In summary, our data indicate OSM causes VEGF expression in HASM cells by a transcriptional mechanism involving STAT3. IL-1beta also synergizes with OSM to increase VEGF release, likely as a result of effects of IL-1beta on VEGF mRNA stability as well as effects of OSM on IL-1R1 expression. This is the first description of a role for OSM on IL-1R1 expression in any cell type. OSM may contribute to airway remodeling observed in chronic airway disease.

Effect of leptin on allergic airway responses in mice.

BACKGROUND: Epidemiologic data indicate that the incidence of asthma is increased in obese patients. OBJECTIVE: Because the serum levels of the satiety hormone and proinflammatory cytokine leptin are increased in obese individuals, we sought to determine whether leptin can augment allergic airway responses. METHODS: We sensitized and challenged BALB/cJ mice with ovalbumin. Alzet micro-osmotic pumps were implanted in the mice to deliver a continuous infusion of either saline or leptin (1.75 mug/g/d). Two days later, the mice were challenged with either aerosolized saline or ovalbumin once per day for 3 days. We measured airway responsiveness, performed bronchoalveolar lavage, and obtained blood to measure serum leptin and IgE 24 or 48 hours after the last challenge. RESULTS: Leptin infusion increased serum leptin concentrations, which were increased further after ovalbumin sensitization and challenge. Ovalbumin challenge increased bronchoalveolar lavage fluid cells and cytokines, serum IgE, lung cytokine mRNA expression, and responses to inhaled, aerosolized methacholine. It is important to note that the changes in methacholine responsiveness and IgE were augmented in leptin- versus saline-infused mice. CONCLUSIONS: These results indicate that serum leptin is increased during allergic reactions in the airways and may play a role in the relationship between obesity and asthma.