Arterial blood gas analysis in 53 racehorses with a diagnosis of Small Airway Inflammatory Disease (SAID)

In 53 racehorses with a mean age of 4.5 years old presented for poor performance, Small Airway Inflammatory Disease (SAID) was diagnosed by bronchoalveolar lavage (BAL). Thirty of these horses (58.5%) had arterial pCO₂ above normal range (> 46 mmHg), while pO₂ was within normal range (> 80 mmHg) in both hypercapnic (group A) and normocapnic (group B) horses although pO₂ was significantly lower in group A horses. The horses were subsequently subdivided into two groups according to the duration of symptoms (group 1: less than 4 weeks; group 2: longer than 4 weeks). Horses from group 2 had significantly higher values of pCO₂ (p < 0.01) , HCO₃- (p < 0.01) and TCO₂ (p < 0.05) when compared to horses from group 1. It was concluded that the duration of the inflammatory process may play a role in the alteration of blood/alveolar gas exchanges and acid-base status in SAID affected racehorses.     

Comparison of serum amyloid A and C-reactive protein as diagnostic markers of systemic inflammation in dogs

The diagnostic performance of canine serum amyloid A (SAA) was compared with that of C-reactive protein (CRP) in the detection of systemic inflammation in dogs. Sera from 500 dogs were retrospectively included in the study. C-reactive protein and SAA were measured using validated automated assays. The overlap performance, clinical decision limits, overall diagnostic performance, correlations, and agreement in the clinical classification between these 2 diagnostic markers were compared. Significantly higher concentrations of both proteins were detected in dogs with systemic inflammation (SAA range: 48.75 to > 2700 mg/L; CRP range: 0.4 to 907.4 mg/L) compared to dogs without systemic inflammation (SAA range: 1.06 to 56.4 mg/L; CRP range: 0.07 to 24.7 mg/L). Both proteins were shown to be sensitive and specific markers of systemic inflammation in dogs. Significant correlations and excellent diagnostic agreement were observed between the 2 markers. However, SAA showed a wider range of concentrations and a significantly superior overall diagnostic performance compared with CRP.     

Development and validation of a spectrophotometric method for quantification of total glucosinolates in cruciferous vegetables

Given their putative role in chemoprevention, validated methods are needed for quantification of total glucosinolates. Based on the colorimetric reaction of ferricyanide with 1-thioglucose, released by alkaline hydrolysis of glucosinolates, we developed a simple and sensitive method for spectrophotometric quantification of total glucosinolates in cruciferous vegetables. Lyophilized and ground vegetables are extracted with 80% boiling methanol. Extracted glucosinolates are isolated using a strong anion exchange column and then hydrolyzed with 2 N NaOH to release 1-thioglucose. Ferricyanide is added, and the decrease in absorbance is measured at 420 nm, with final values adjusted for background. Recovery of internal standard (sinigrin) was 107%. Intra- and interassay coefficients of variation were 5.4% and 15.8%, respectively. Dose response was linear with sinigrin and amount of plant material extracted (R(2) ≥ 0.99). Using sinigrin, the lower limit of quantification was 0.6 mg. This straightforward method may be an alternative to time-consuming and costly chromatographic methods.     

Main odorants in Jura flor-sherry wines. Relative contributions of sotolon, abhexon, and theaspirane-derived compounds

The aromatic profile of Jura flor-sherry wines (also called "yellow wines") has been little studied. Only acetaldehyde, diethoxy-1,1-ethane, and sotolon have been described as key odorants. In the present work, three wines (vintages 2002 and 2003) were investigated by gas chromatography-mass spectrometry and gas chromatography-olfactometry (GC-O) aroma extract dilution analysis. The goal was to assess the relative impact of varietal, fermentation, and oak-barrel compounds by using two complementary extraction procedures. No grape terpenoids were found after the long barrel aging (6 years and 3 months). On the other hand, two candy/fruity esters issued from yeast exhibited high flavor dilution factor (FD) values: ethyl isobutyrate (64-1024) and ethyl isovalerate (128-1024). As expected, many oak-related odorants were found in the XAD 2 flavor extracts, mainly homofuraneol [2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone] (cotton candy, FD = 16-256) and cis-β-methyloctalactone (butter, woody, FD = 256). Most probably issued from oxidation of the grape constituent theaspirane, an exceptional grenadine odor was perceived by GC-O up to dilution 64-1024. Chemical oxidation experiments and GC-high-resolution mass spectrometry (HRMS) allowed us to identify it as 4-hydroxy-7,8-dihydro-β-ionone (RI(CPsil5CB) = 1373), a hydrolysis-derived product of dihydrodehydro-β-ionone. With an extraction dedicated to hydrophilic compounds, the key role of sotolon was confirmed (112-387 μg/kg; FD = 256-1024). This procedure enabled us to also evidence its ethyl analogue, abhexon (31-74 μg/kg; FD = 64-256).     

Characterization of Palytoxin Binding to HaCaT Cells Using a Monoclonal Anti-Palytoxin Antibody

Palytoxin (PLTX) is the reference compound for a group of potent marine biotoxins, for which the molecular target is Na⁺/K⁺-ATPase. Indeed, ouabain (OUA), a potent blocker of the pump, is used to inhibit some PLTX effects in vitro. However, in an effort to explain incomplete inhibition of PLTX cytotoxicity, some studies suggest the possibility of two different binding sites on Na⁺/K⁺-ATPase. Hence, this study was performed to characterize PLTX binding to intact HaCaT keratinocytes and to investigate the ability of OUA to compete for this binding. PLTX binding to HaCaT cells was demonstrated by immunocytochemical analysis after 10 min exposure. An anti-PLTX monoclonal antibody-based ELISA showed that the binding was saturable and reversible, with a K_d of 3 × 10⁻¹⁰ M. However, kinetic experiments revealed that PLTX binding dissociation was incomplete, suggesting an additional, OUA-insensitive, PLTX binding site. Competitive experiments suggested that OUA acts as a negative allosteric modulator against high PLTX concentrations (0.3–1.0 × 10⁻⁷ M) and possibly as a non-competitive antagonist against low PLTX concentrations (0.1–3.0 × 10⁻⁹ M). Antagonism was supported by PLTX cytotoxicity inhibition at OUA concentrations that displaced PLTX binding (1 × 10⁻⁵ M). However, this inhibition was incomplete, supporting the existence of both OUA-sensitive and -insensitive PLTX binding sites.     

Familial canine pituitary-dependent hyperadrenocorticism in wirehaired Dachshunds

Canine pituitary hyperadrenocorticism (Cushing's disease) caused by neoplasia of the corticotrope cells is one of the most common endocrine diseases especially in smaller dog breeds. Cushing's disease was diagnosed in eleven wire-haired Dachshunds and for further six wire-haired Dachshunds Cushing's disease was suspected on the basis of clinical signs. A joined pedigree could be ascertained for all these 17 dogs. Eleven of these dogs were so closely related to each other, that they were summarized in four nucleus families. Two fullsiblings were examined by means of clinical, laboratory diagnostic and morphological methods. The main lesions consisted of atrophic dermatosis with alopecia, increase of activity of liver enzymes in plasma and bilateral adrenocortical hyperplasia and therefore corresponded to the typical signs of a secondary hyperadrenocorticism. A rather unusual finding was the pituitary carcinoma in one of these dogs. Similarly to human patients affected by hyperadrenocorticism, real-time PCR analysis showed a 2.9-fold increase of expression of the canine MDR1 gene in the liver of one affected wirehaired Dachshund. This study documents the first familial occurrence of pituitary-dependent hyperadrenocorticism in wirehaired Dachshunds, the overexpression of the MDR1 gene in the dog and the third case of familial hyperadrenocorticism in dogs ever described.     

In vitro study to assess the efficacy of CDK4/6 inhibitor Palbociclib (PD‐0332991) for treating canine mammary tumours

Therapy of canine mammary tumours (CMTs) with classical antitumour drugs is problematic, so better therapeutic options are needed. Palbociclib (PD‐0332991) is an innovative and effective anticancer drug for the treatment of breast cancer in women. Palbociclib is an inhibitor of cyclin‐dependent kinase 4 (CDK4) and CDK6, which are key regulators of the cell cycle machinery and thus cell proliferation. In the present in vitro study, we investigated whether Palbociclib also represents a candidate drug to combat CMT. For this purpose, the effect of Palbociclib was analysed in P114 and CF41 cells, two CMT cell lines with an endogenous CDK4/6 co‐expression. Incubation of P114 and CF41 cells with Palbociclib resulted in a dose‐ and time‐dependent loss of phosphorylated retinoblastoma protein (pRb), a classical CDK4/6 substrate within the cell cycle machinery. Moreover, treatment of CMT cells with Palbociclib‐induced cell cycle arrest affected cell viability, prevented colony formation and impaired cell migration activity. Palbociclib also inhibited the growth of P114 and CF41 cell spheroids. Immunohistochemical analysis of canine patient samples revealed a consistent expression of CDK6 in different canine mammary carcinoma types, but an individual and tumour‐specific expression pattern of phosphorylated pRb independent of the tumour grade. Together, our findings let us suggest that Palbociclib has antitumour effects on CMT cells and that canine patients may represent potential candidates for treatment with this CDK4/6 inhibitor.     

Emphysematous pyelitis and cystitis associated with vesicoureteral reflux in a diabetic dog

A 12-year-old female dog with a 3-month history of poor response to diabetes treatment had an acute worsening of symptoms, including weakness and blindness. The dog had elevated blood glucose, alkaline phosphatase and urea concentration, hyposthenuria, glycosuria, hematuria, and pyuria. Escherichia coli was isolated from the urine. Radiographs and ultrasound examination showed that the dog had unilateral emphysematous pyelitis and concurrent cystitis associated with vesicoureteral reflux.