Relationship between assays of inflammation and coagulation: A novel interpretation of the canine activated clotting time

The processes of inflammation and coagulation are known to be interconnected through several mechanisms; however, the influence of inflammation on the interpretation of coagulation assays remains unknown. Blood was collected from 87 dogs admitted to a tertiary referral intensive care unit (ICU) and 15 control dogs. The association between 2 markers of inflammation [mature neutrophil count and C-reactive protein (CRP)] and 5 coagulation parameters [activated clotting time (ACT), prothrombin time (PT), activated partial thromboplastin time (aPTT), antithrombin (AT), and platelet count (plt)] were evaluated through correlation analysis. The study population was then divided into 4 groups based on severity of ACT prolongation with comparisons to all other variables assessed through an analysis of variance (ANOVA) test. A strong correlation for a biological system was demonstrated between ACT and CRP (r = 0.66; P < 0.0001). Statistically significant results were also found between aPTT and AT with the markers of inflammation, but the correlations were weaker. Within ACT groups of increasing severity, higher CRP concentrations (P < 0.0001) and lower AT activities (P < 0.0001) were identified. This study provides evidence for an association between assays of inflammation and coagulation and suggests that modification of our traditional interpretations of coagulation assays may be required. As a point-of-care test, ACT is a simple and inexpensive tool that can be used to assess an underlying inflammatory or hemostatic process.     

Characterization of topical antiinflammatory compounds in Rosmarinus officinalis L

The topical antiinflammatory activity of three extracts at increasing polarity (n-hexane, chloroform, and methanol) from the leaves of Rosmarinus officinalis L. (Labiatae) has been tested using the croton oil ear test in mice. Both the n-hexane and the chloroform (CE-1) extracts from the leaves showed a dose-dependent activity, the last one possessing an antiinflammatory potency similar to that of indomethacin, the nonsteroidal antiinflammatory drug used as a reference drug (ID50 = 83 and 93 microg/cm2, respectively). The bioassay-oriented fractionation of CE-1 led to the identification of tritepenes, ursolic acid, oleanolic acid, and micromeric acid as the main antiinflammatory principles. Furthermore, the CE-1 extract obtained from the residue of the steam distillation of the leaves (extract A) showed the same antiinflammatory potency of CE-1, suggesting this waste product as a source of antiinflammatory products.     

Amino acid composition and antioxidant capacity of Spanish honeys

The amino acid composition of 53 honey samples from Spain, consisting of 39 floral, 5 honeydew, and 9 blend honeys, has been determined. Physicochemical characteristics, polyphenolic content, amino acid composition, and estimation of the radical scavenging capacity against the stable free radical DPPH of the honey samples were analyzed. The resulting data have been statistically evaluated. The results showed that pH, acidity, net absorbance, electrical conductivity, and total polyphenolic contents of the honeys showed a strong correlation with the radical scavenging capacity. The correlation between the radical scavenging capacity of honey and amino acid contents was high with 18 of the 20 amino acids detected, with correlation values higher than those obtained for polyphenolic content. These results suggest that the amino acid composition of honey is an indicator of the sample's scavenging capacity.     

Efficiency of DNA typing methods for detection of smoked paprika "pimenton de la vera" adulteration used in the elaboration of dry-cured Iberian pork sausages

The purpose of this work was to develop a PCR method for the identification of smoked paprika "Pimento?n de la Vera" adulteration with paprika elaborated from varieties of pepper foreign to the la Vera region, in central western Spain. Three autochthonous varieties of pepper, Jaranda, Jariza, and Bola, and the varieties Papri Queen, Papri King, Sonora, PS9794, and Papri Ace, foreign to the La Vera region, were used in the study. Analyses of the ITS and 5.8S rDNA, RAPD-PCR, SSR, and ISSR were tested. RAPD-PCR, SSR, and ISSR analyses allowed differentiation among the varieties of paprika analyzed. There was no difference in the sequence of ITS1-5.8S rDNA-ITS2. In addition, with the RAPD-PCR primers S13 and S22, two molecular markers were obtained of 641 and 704 bp, respectively, which allowed all of the smoked paprika varieties to be differentiated from paprikas elaborated with the five foreign varieties. These two molecular markers were investigated as a basis for detecting the adulteration of smoked paprika with paprika elaborated from foreign varieties of pepper.     

Use of probiotic bacteria in the rearing of Senegalese sole (Solea senegalensis) larvae

Three candidate probiotics, which had shown antimicrobial activity in vitro against two fish pathogens, were used in the rearing of Senegalese sole larvae and postlarvae. These probiotics improved the survival of starved sole yolk‐sac larvae. A feeding experiment of sole larvae and postlarvae comprised three treatments: A, B and C. Cultures of a Vibrio sp. (2J18) were added to treatment A, whereas a gram‐positive (J84) and a Shewanella sp. strain (2J27), were added to treatment B, while in a control treatment C no bacteria were added. Addition of bacteria in treatment B increased survival of larvae in the first phase of the experiment [0–20 days after hatching (DAH) (P<0.05)] and decreased the numbers of colony‐forming units (CFU) in larval gut 5 DAH compared with the control treatment (P<0.05). No differences were observed in survival (25–47%) during the second phase of the experiment (20–60 DAH). Nevertheless, the total numbers of CFU in fish gut 40 DAH were significantly lower in treatment B (P<0.05). The colonization rates of the added bacteria were the highest 5 and 11 DAH, and the highest average values reached were 65%, 82% and 17% of the total CFU count for the strains 2J18, 2J27 and J84 respectively.     

Effects of natural phenolic compounds on the antioxidant activity of lactoferrin in liposomes and oil-in-water emulsions

The effect of natural phenolic compounds on the antioxidant and prooxidant activity of lactoferrin was studied in liposomes and oil-in-water emulsions containing iron. The antioxidants tested with lactoferrin were alpha-tocopherol, ferulic acid, coumaric acid, tyrosol, and natural phenolic extracts obtained from three different extra-virgin olive oils and olive mill wastewater. The natural extracts of olive oils and mill wastewaters were composed mainly of polyphenols and simple phenolics, respectively. Lipid oxidation at 30 degrees C was determined by the formation of hydroperoxides and fluorescent compounds resulting from oxidized lipid interactions. All phenolic compounds showed synergistic properties in reinforcing the antioxidant activity of lactoferrin in lipid systems containing iron. The highest synergistic effects were observed for the phenolic extracts rich in polyphenols of extra-virgin olive oils and lactoferrin. This synergistic effect was higher in liposomes than in emulsions.     

Prevalence of malignancy when solitary versus multiple lesions are detected during abdominal ultrasonographic examination of dogs with spontaneous hemoperitoneum: 31 cases (2003–2008)

Objective – To compare the histopathologic diagnosis in dogs with spontaneous hemoperitoneum when abdominal ultrasonographic examination detects a solitary versus multiple lesions.
Design – Retrospective cross-sectional study.
Setting – Private veterinary hospital.
Animals – Client-owned dogs presented with spontaneous hemoperitoneum between March 1, 2003 and June 1, 2008.
Interventions – Dogs were divided into 2 groups based on presence of a solitary or multiple abdominal ultrasonographic lesions. Prevalences were compared between groups for malignancy and specifically hemangiosarcoma.
Measurements and Main Results – Ten of 31 (32%) dogs had a solitary abdominal ultrasonographic lesion and 21 of 31 (68%) had more than 1 lesion. The bleeding tissue was characterized as malignant in 8 of 10 (80%) dogs with solitary lesions and 17 of 21 (81%) dogs with multiple lesions; there was no significant difference ( P =1.0) between groups. In this study no association ( P =0.26) was found between the number of abdominal ultrasonographic lesions observed and subsequent diagnosis of hemangiosarcoma.
Conclusions – Solitary abdominal ultrasonographic lesions in dogs with spontaneous hemoperitoneum do not necessarily indicate a lower prevalence of malignancy.     

Change in Sox9 protein localization through gonad development in Russian sturgeon (Acipenser gueldenstaedtii)

The Russian sturgeon is a highly prized species reared in aquaculture. The process of gonad development in this critically endangered species is poorly understood. The aim of this study was to describe the localization of Sox9 protein during gonad development of the Russian sturgeon from the day of hatching to the 1440 day post hatching (dph). The larvae at age 1, 10, 25 dph and prepared gonads of 300, 720, 1440 dph individuals were immunohistochemistry‐stained for Sox9 detection. Sox9‐positive regions were detected in larvae in primordial germ cells cytoplasm. Analysis of 300 dph sturgeon gonads revealed the presence of the Sox9 protein in cytoplasm of some oocytes in the chromatin nucleus stage. In testes at 720 dph, Sox9 was observed in the cytoplasm of type A and early B spermatogonias. In the ovaries, Sox9 was observed in the cytoplasm of diplotene oocytes and prefollicular cells. In testes of 1440 dph sturgeon, Sox9 was present in the nucleus of the spermatocytes and in types A and B spermatogonias cytoplasm. Analysis of ovaries at 1440 dph reveals multiple diplotene oocytes with a Sox9‐positive cytoplasm. Furthermore, in 720 and 1440 dph, sturgeon presence of intersexual gonads was detected. In intersex gonads, Sox9 was observed in the cytoplasm of diplotene oocytes and type A spermatogonias. This study may be the first attempt to determine Sox9 protein localization during ontogenesis of the Russian sturgeon. Localization of Sox9 protein may become a useful marker of the maturation level in testis of the Russian sturgeon.     

Soil microbiome biomass,activity, composition and CO2 emissions in a long-term organic and conventional farming systems

The implementation of environmentally friendly agricultural policies has increased the need to compare agricultural aspects of conventional (CON) and organic farming (ORG) systems. The objective of the present work was to compare the effects of an organic and conventional long-term experiment on bacterial and fungal biomass and activity, as well as soil CO₂ emission and readily available nitrogen forms in a soil cultivated with Helianthus annuus L. The microbial biomass was more active and abundant in ORG as well as soil CO₂ emission. Despite being less abundant, fungi were more active than bacteria in both ORG and CON experiments. 16S rRNA gene sequencing showed that the ORG treatment had a significantly greater bacterial richness than CON. Cyanobacteria, Actinobacteria and Proteobacteria were the most abundant phyla contributing more than others to the differences between the two systems. Moreover, the soil ${\mathit{NH}}_4^{+}$ and ${\mathit{NO}}_2^{-}$ content was not significantly different between ORG and CON, while ${\mathit{NO}}_3^{-}$ was less in ORG. ORG sunflower yield was significantly less compared with CON. While much remains to be discovered about the effects of these agricultural practices on soil chemical properties and microbial diversity, our findings may contribute to this type of investigation.     

Standardization of canine meningioma grading: Validation of new guidelines for reproducible histopathologic criteria

Canine meningiomas are currently graded using the human grading system. Recently published guidelines have adapted the human grading system for use in dogs. The goal of this study was to validate the new guidelines for canine meningiomas. To evaluate the inter-observer agreement, 5 veterinary surgical pathologists graded 158 canine meningiomas following the human grading system alone or with the new guidelines. The inter-observer agreement for histologic grade and each of the grading criteria (mitotic grade, invasion, spontaneous necrosis, macronucleoli, small cells, hypercellularity, pattern loss and anaplasia) was evaluated using the Fleiss kappa index. The diagnostic accuracy (sensitivity and specificity) was assessed by comparing the diagnoses obtained with the 2 grading systems with a consensus grade (considered the reference classification). The consensus histologic grade was obtained by agreement between 4 experienced veterinary neuropathologists following the guidelines. Compared with the human grading alone, the canine-specific guidelines increased the inter-observer agreement for: histologic grade (κ = 0.52); invasion (κ = 0.67); necrosis (κ = 0.62); small cells (κ = 0.36); pattern loss (κ = 0.49) and anaplasia (κ = 0.55). Mitotic grade agreement remained substantial (κ = 0.63). The guidelines improved the sensitivity in identifying grade 1 (95.6%) and the specificity in identifying grade 2 (96.2%) meningiomas. In conclusion, the new grading guidelines for canine meningiomas are associated with an overall improvement in the inter-observer agreement and higher diagnostic accuracy in diagnosing grade 1 and grade 2 meningiomas.