Serum cobalamin and methylmalonic acid concentrations in juvenile dogs with parvoviral enteritis or other acute enteropathies.

BACKGROUND: Low serum cobalamin concentrations have been associated with ileal malabsorption in dogs with chronic enteropathy. Increased serum methylmalonic acid (MMA) concentrations indicate cobalamin deficiency on a cellular level. Few studies have evaluated serum cobalamin concentrations or methylmalonic acid concentrations in juvenile dogs with parvoviral enteritis or nonparvoviral acute enteropathies. OBJECTIVES: Evaluate serum cobalamin and methylmalonic acid concentrations in juvenile dogs (6 weeks to 10 months old) with parvoviral enteritis or nonparvoviral acute enteropathy. ANIMALS: Thirty-one juvenile dogs with parvoviral enteritis, 29 dogs with nonparvoviral acute diarrhea (NPVAD), and 40 healthy juvenile control dogs. METHODS: Single-center, prospective, observational, cross-sectional study. Serum cobalamin and, when sufficient serum was available, MMA concentrations were measured. RESULTS: Most serum cobalamin concentrations were within the adult reference interval. Serum cobalamin concentrations in healthy dogs (median, 848 ng/L; range, 293-1912 ng/L) were significantly higher than in dogs with parvoviral enteritis (P = .0002; median, 463 ng/L; range, <150-10 000 ng/L) or dogs with NPVAD (P = .02; median, 528 ng/L; range, 160-8998 ng/L). Serum MMA concentrations were not significantly different between groups (healthy dogs: median, 796 nmol/L; range, 427-1933 nmol/L; parvoviral enteritis: median, 858 nmol/L; range, 554-3424 nmol/L; NPVAD: median, 764 nmol/L; range, 392-1222 nmol/L; P = .1). CONCLUSIONS AND CLINICAL IMPORTANCE: Juvenile dogs with parvoviral enteritis or NPVAD had lower serum cobalamin concentrations than healthy juvenile dogs. However, based on serum MMA concentrations cellular cobalamin deficiency was not apparent.

Comparison of assessment of coagulation in healthy dogs by the TEG 6s and TEG 5000 viscoelastic analyzers.

The TEG 6s (Haemonetics) point-of-care viscoelastic analyzer is portable, compact, simple to use, and has the potential for rapid viscoelastic analysis that can guide the treatment of veterinary patients at the site of care. Although approved for use in people, the TEG 6s has yet to be evaluated for hemostatic analysis in veterinary medicine. Citrated whole blood (CWB) was collected from 27 healthy dogs. An aliquot of CWB from each dog was diluted by 33% with an isotonic crystalloid, representing an in vitro model of hemodilution. Unaltered and diluted CWB samples were analyzed using 2 TEG 6s and 6 TEG 5000 (Haemonetics) analyzers. The 6 TEG 5000 analyzers ran duplicate analyses of either unaltered or diluted samples using 1 of 3 reagents (Haemonetics): Kaolin TEG, RapidTEG, or TEG Functional Fibrinogen. Duplicate TEG 5000 analyses were averaged and compared with a single TEG 6s analysis. Lin concordance correlation coefficient and Bland-Altman plots were used to evaluate agreement of reaction time, kinetic time, alpha angle, maximum amplitude (MA), and G value (G) for samples activated with Kaolin TEG, and agreement of MA for samples activated with RapidTEG between the 2 machines. Overall, agreement between the TEG 6s and TEG 5000 analyzers was poor. Viscoelastic measurements by the TEG 6s and TEG 5000 in healthy dogs were not all interchangeable. Agreement was satisfactory only for MA and G measurements of diluted blood samples activated with Kaolin TEG, and MA measurements for both unaltered and diluted blood samples activated with RapidTEG.

Evaluation of two benchtop blood gas analyzers for measurement of electrolyte concentrations in venous blood samples from dogs.

OBJECTIVE: To assess agreement between 2 benchtop blood gas analyzers developed by 1 manufacturer (BGA 1 and BGA 2 [a newer model with reduced maintenance requirements]) and a reference chemistry analyzer for measurement of electrolyte (sodium, chloride, and potassium) in blood samples from dogs. ANIMALS: 17 healthy staff- and student-owned dogs and 23 client-owned dogs admitted to an emergency and intensive care service. PROCEDURES: Blood collected by venipuncture was placed in lithium heparin-containing tubes. Aliquots were analyzed immediately with each BGA. Samples were centrifuged, and plasma was analyzed with the reference analyzer. Results for each BGA were compared with results for the reference analyzer by Passing-Bablok regression analysis. Percentage differences between BGA and reference analyzer results were compared with published guidelines for total allowable error. RESULTS: Proportional bias was detected for measurement of chloride concentration (slope, 0.7; 95% CI, 0.7 to 0.8), and constant positive bias was detected for measurement of chloride (y-intercept, 34, mmol/L; 95% CI, 16.9 to 38 mmol/L) and potassium (y-intercept, 0.1 mmol/L; 95% CI, 0.1 to 0.2 mmol/L) concentrations with BGA 1. There was no significant bias for measurement of potassium or chloride concentration with BGA 2 or sodium concentration with either BGA. Differences from the reference analyzer result exceeded total allowable error guidelines for ≥ 1 sample/analyte/BGA, but median observed measurement differences between each BGA and the reference analyzer did not. CONCLUSIONS AND CLINICAL RELEVANCE: Good agreement with reference analyzer results was found for measurement of the selected electrolyte concentrations in canine blood samples with each BGA.

Transient Megaesophagus Following Coral Snake Envenomation in Three Dogs (2013-2018).

A 12 yr old dachshund, a 7 yr old English springer spaniel, and a 1.5 yr old French bulldog presented following envenomation by a coral snake. Each patient displayed evidence of varying degrees of lower motor neuron dysfunction, but all three developed transient megaesophagus. Two patients developed secondary aspiration pneumonia, with one requiring mechanical ventilation, which the owners declined, resulting in euthanasia. The third developed hypoventilation without aspiration pneumonia, was mechanically ventilated, and was successfully weaned. In the two surviving patients, the megaesophagus resolved by time of discharge. Coral snake envenomation is an uncommon occurrence, and these are the first documented cases of transient megaesophagus secondary to a North American species.

Resolvin D1 promotes efferocytosis in aging by limiting senescent cell-induced MerTK cleavage.

Inflammation-resolution is mediated by the balance between specialized pro-resolving mediators (SPMs) like resolvin D1 (RvD1) and pro-inflammatory factors, like leukotriene B4 (LTB4). A key cellular process of inflammation-resolution is efferocytosis. Aging is associated with defective inflammation-resolution and the accumulation of pro-inflammatory senescent cells (SCs). Therefore, understanding mechanism(s) that underpin this impairment is a critical gap. Here, using a model of hind limb ischemia-reperfusion (I/R) remote lung injury, we present evidence that aging is associated with heightened inflammation, impaired SPM:LT ratio, defective efferocytosis, and a decrease in MerTK levels in injured lungs. Treatment with RvD1 mitigated I/R lung injury in aging, promoted efferocytosis, and prevented the decrease of MerTK in injured lungs from old mice. Old MerTK cleavage-resistant mice (MerTKCR) exhibited less neutrophils or polymorpho nuclear cells infiltration and had improved efferocytosis compared with old WT controls. Mechanistically, macrophages that were treated with conditioned media (CM) from senescent cells had increased MerTK cleavage, impaired efferocytosis, and a defective RvD1:LTB4 ratio. Macrophages from MerTKCR mice were resistant to CM-induced efferocytosis defects and had an improved RvD1:LTB4 ratio. RvD1-stimulated macrophages prevented CM-induced MerTK cleavage and promoted efferocytosis. Together, these data suggest a new mechanism and a potential therapy to promote inflammation-resolution and efferocytosis in aging.

Resolvin D1 promotes the targeting and clearance of necroptotic cells.

Inflammation-resolution is a protective response that is mediated by specialized pro-resolving mediators (SPMs). The clearance of dead cells or efferocytosis is a critical cellular program of inflammation-resolution. Impaired efferocytosis can lead to tissue damage in prevalent human diseases, like atherosclerosis. Therefore understanding mechanisms associated with swift clearance of dead cells is of utmost clinical importance. Recently, the accumulation of necroptotic cells (NCs) was observed in human plaques and we postulated that this is due to defective clearance programs. Here we present evidence that NCs are inefficiently taken up by macrophages because they have increased surface expression of a well-known "don't eat me" signal called CD47. High levels of CD47 on NCs stimulated RhoA-pMLC signaling in macrophages that promoted "nibbling", rather than whole-cell engulfment of NCs. Anti-CD47 blocking antibodies limited RhoA-p-MLC signaling and promoted whole-cell NC engulfment. Treatment with anti-CD47 blocking antibodies to Ldlr-/- mice with established atherosclerosis decreased necrotic cores, limited the accumulation of plaque NCs and increased lesional SPMs, including Resolvin D1 (RvD1) compared with IgG controls. Mechanistically, RvD1 promoted whole-cell engulfment of NCs by decreasing RhoA signaling and activating CDC42. RvD1 specifically targeted NCs for engulfment by facilitating the release of the well-known "eat me signal" called calreticulin from macrophages in a CDC42 dependent manner. Lastly, RvD1 enhanced the clearance of NCs in advanced murine plaques. Together, these results suggest new molecules and signaling associated with the clearance of NCs, provide a new paradigm for the regulation of inflammation-resolution, and offer a potential treatment strategy for diseases where NCs underpin the pathology.

Pro-resolution therapeutics for cardiovascular diseases.

Studies over the last couple of decades suggest that failed resolution of a chronic inflammatory response is an important driving force in the progression of atherosclerosis. Resolution of inflammation is mediated in part by lipid-derived specialized pro-resolving mediators (SPMs) such as lipoxins, resolvins, protectins and maresins. The major functions of SPMs are to quell inflammation and repair tissue damage in a manner that does not compromise host defense. An imbalance between SPMs and pro-inflammatory mediators like leukotriene B4 (LTB4) are associated with several prevalent human diseases, including atherosclerosis. Because atherosclerosis is marked by persistent, unresolved inflammation and arterial tissue injury, SPMs have garnered immense interest as a potential treatment strategy. This mini review will highlight recent advances in the application of SPMs in atherosclerosis as well as the ability of SPMs to control several of the risk factors associated with cardiovascular diseases.