Inorganic Phosphate Effect in a Hydrogen Peroxide-based Bleaching Agent: Physicochemical, Mechanical, and Morphological Properties of Dental Enamel.

OBJECTIVES: This in vitro study aimed to assess the impact of incorporating calcium glycerophosphate (CaGP) and sodium fluoride (NaF) in addition to 35% hydrogen peroxide concerning the enamel mechanical and morphological properties. METHODS: Specimens of bovine enamel were chosen based on their initial surface hardness (SHi) and subsequently divided into five gel groups (n=12): 1) 35% Hydrogen Peroxide (HP) Gel; 2) HP + 0.1% NaF Gel (HP/NaF); 3) HP + 0.25% CaGP Gel (HP/CaGP); 4) HP + 0.1% NaF + 0.25% CaGP Gel (HP/NaF/CaGP) and 5) HP Blue 35% Gel (HP Blue). The bleaching gels were applied thrice, for 40 min, at intervals of 7 days each. After 21 days, the final surface hardness (SHf), integrated hardness (IH), Polydispersity Index (PdI) and Zeta Potential (Zp), surface roughness (Ra, after and before), and surface/structural analysis by Scanning Electron Microscopy (SEM) were determined. The data were submitted to ANOVA (one-way and two-way) followed by the Student-Newman-Keuls test (α=0.05). RESULTS: The addition of NaF to HP reduced demineralization by 11.5% in relation to HP (p<0.05). The NaF/CaGP association reduction is 22.8 and 20% higher in comparison to HP/NaF/CaGP and HP Blue, respectively. The IH when the PH/NaF/CaGP bleaching gel was applied, was 14% higher compared to HP and HP Blue groups. CONCLUSIONS: It can be concluded that the association of NaF and CaGP with the 35% hydrogen peroxide gel (HP/NaF/CaGP) significantly changed tooth enamel demineralization in terms of surface, depth, roughness, and enamel morphology.

Evaluation of plasma nucleosome concentrations in dogs with a variety of common cancers and in healthy dogs.

BACKGROUND: Cell free DNA, in the form of nucleosomes, is released into circulation during apoptosis and necrosis in a variety of diseases. They are small fragments of chromosomes that are composed of DNA wrapped around a histone core made of four duplicate histone proteins forming an octamer. The nucleosome compartment is a relatively uninvestigated area of circulating tumor biomarkers in dogs. The objectives of this study were to quantify and better characterize nucleosome concentrations in 528 dogs with various common malignancies and compare them to 134 healthy dogs. RESULTS: The sensitivity of increased circulating nucleosome concentrations for the detection of cancer in all dogs was 49.8% with a specificity of 97% with an area under the curve of 68.74%. The top 4 malignancies detected by the test included lymphoma, hemangiosarcoma, histiocytic sarcoma and malignant melanoma. The malignancies least likely to be detected were soft tissue sarcomas, osteosarcoma and mast cell tumors. CONCLUSIONS: A variety of tumor types may cause increased nucleosome concentrations in dogs. Tumors of hematopoietic origin are most likely to cause elevations and local tumors such as soft tissue sarcomas are least likely to cause elevations in plasma nucleosome concentrations.

Atypical patterns of STAT3 phosphorylation in subpopulations B cells in patients with common variable immunodeficiency.

BACKGROUND: Common Variable Immunodeficiency (CVID) is a heterogeneous disorder characterized by defective B cell differentiation and antibody production. Interleukin (IL)-21 activates STAT3, a potent regulator of B cell differentiation into plasma cells. We have studied the phosphorylation of STAT3 in CVID patients and its contribution to B cells subsets. METHODS: We studied 23 CVID patients and 14 healthy donors (HD), determining pSTAT3 in naïve and memory B cells, stimulated with IL-21 at 15 and 60 min. RESULTS: pSTAT3 was increased in total (p = 0.044), naïve (p = 0.023), and memory (p = 0.001) B cells at 60 min in CVID patients compared with HD. We classified patients by the percentage of isotype-switched memory B cells. We observed an increase in pSTAT3 at 60 min in memory B cells in both CVID groups of patients (p = 0.026, p = 0.007, respectively). Interestingly, the analysis of each group individually; demonstrated that patients with decreased memory B cells exhibited an increase in pSTAT3 at 60 min (p = 0.023), while HD had an expected decrease in pSTAT3 (p = 0.045). CONCLUSION: CVID patients showed an increased atypical of pSTAT3, which could affect the differentiation of B cells. Further studies in the IL-21 pathway are necessary to understand how this alteration could promote differentiation defects in patient B cells.

Neoadjuvant chemotherapy and radiation therapy in veterinary cancer treatment: a review.

Multi-modality treatment strategies are more becoming commonplace in veterinary oncology practice yet the evidence base is far inferior to what has been generated in people. Surgery is unquestionably the cornerstone of most solid tumour treatment plans but certain scenarios dictate combining surgery with systemic chemotherapy and radiation therapy as an adjunct. By using these in the neoadjuvant setting, one can leverage certain effects of the treatment to improve local disease control, improve overall survival, gain insight into drug efficacy, reduce surgical morbidity and reduce long-term complications. An unintended consequence of combining therapies is an increased flow of information between members of the care team upfront that in almost all cases leads to improved patient outcomes albeit a difficult metric to quantify. This review sets out to explore some of the principles of neoadjuvant therapies and discuss potential opportunities to expand the evidence base in veterinary medicine.

Development, analytical validation, and initial clinical evaluation of a radioimmunoassay for the measurement of soluble CD25 concentrations in canine serum.

During immune activation, CD25 is expressed by T cells, and its soluble form (sCD25) is released into the extracellular matrix and the bloodstream. In humans, serum sCD25 concentrations are used as a surrogate marker for autoimmune diseases, malignancies, and transplant rejection. However, a canine-specific assay for the measurement of sCD25 in dog serum has not previously been described. Therefore, the aims of this study were to develop and analytically validate a radioimmunoassay to measure sCD25 in canine serum, to establish a reference interval for canine sCD25, and to test the clinical utility of this assay with serum samples for dogs with various diseases. A competitive radioimmunoassay (RIA) was developed and analytically validated. Analytical validation consisted of lower limit of detection (LLOD), dilutional parallelism, spiking recovery, and intra- and inter-assay variability using pooled surplus canine serum samples. A reference interval was established in healthy dogs and serum samples from dogs with various types of neoplasia, IBD, liver disease, suspected pancreatitis, or suspected small intestinal disease and serum samples with an increased C-reactive protein concentration (CRP) were analyzed to test the clinical utility of the assay. LLOD was calculated to be 0.5 ng/mL. The mean (±SD) observed-to-expected ratio (O/E) for serial dilutions was 101.7 ±â€¯14.0%, and the mean (± SD) O/E for spiking recovery was 93.2 ±â€¯4.2%. Coefficients of variation (CVs) for intra-assay variability were ≤12.5% (mean ±â€¯SD: 7.5 ±â€¯4.2%), and inter-assay CVs were ≤15.7% (mean ±â€¯SD: 11 ±â€¯4.4%). A reference interval (RI) for canine sCD25 of 1.2-4.2 ng/mL was established from a population of 112 clinically healthy dogs. Dogs with neoplasia and dogs with suspected small intestinal disease had decreased concentrations of serum sCD25 when compared to healthy dogs (p < 0.0001, respectively). However, the majority of clinical samples used in this study were within the reference interval. Median concentrations of serum sCD25 were 1.9 ng/mL for healthy dogs. Dogs with cancer, IBD, liver disease, suspected pancreatitis, or suspected small intestinal disease, as well as sera with an increased serum CRP concentration, had median serum sCD25 concentrations of 1.6 ng/mL, 2.1 ng/mL, 2.2 ng/mL, 1.7 ng/mL, 1.5 ng/mL, and 1.8 ng/mL, respectively. Thus, the RIA described here is linear, accurate, precise, and reproducible for measuring sCD25 in canine serum. However, this assay shows little clinical utility of sCD25 as a biomarker for dogs with inflammatory, autoimmune, and/or neoplastic conditions.

Residual pollution and vegetation distribution in amended soils 20 years after a pyrite mine tailings spill (Aznalcóllar, Spain).

The present work assesses the residual pollution in the Guadiamar Green Corridor (SW, Spain) after a long-term aging process (18 years) since the accident of the Aznalcóllar pyrite mine. We have focused on the study of trace elements (Cu, Zn, Cd, As and Pb) in soils, their fractionation and the transference to the surrounding vegetation. The residual polluted areas are characterized by scattered plots with absence of vegetation, presenting high concentrations of trace elements, acidic pH and low organic carbon content. Surrounding these polluted plots, two vegetation gradient belts are clearly identified by changes in plant cover and richness. The inhibition of plant growth in the bare soils is related to the highest mobility of soluble and exchangeable Cu, Zn and Cd forms, which significantly decrease with the distance to the polluted plots. Plant richness and cover show differences between belts; bioaccumulation of trace elements in plants also differs, with a preferential accumulation in roots. Despite the low bioavailability of As and Pb in soils, bioaccumulation factors in plants for these elements are significantly higher in belt 1 in relation to belt 2. High Cu and Cd potential toxic concentrations in aerial parts of vegetation are found, posing a risk for livestock and a potential entrance to the food-chain. On the other hand, Lamarckia aurea (L.) Moench (in belt1) and Trifolium campestre Schreb. (in belt2) were the most dominant species in severely polluted soils. Elevated concentrations of trace elements in the vegetation growing in the area indicate plant adaptation mechanisms to live in these severely polluted soils, which can be used as a good bioindicator of pollution in similar polluted areas.

Geographical differences in survival of dogs with non-Hodgkin lymphoma treated with a CHOP based chemotherapy protocol.

BACKGROUND: In humans geographical differences in the incidence and presentation of various cancers have been reported. However, much of this information has not been collected in veterinary oncology. AIM: The purpose of this study was to determine if a geographic difference in progression free survival exists for dogs with lymphoma treated within the US. MATERIALS AND METHODS: Medical records of 775 cases of canine lymphoma from 3 US regions (west, south and north), treated with CHOP chemotherapy, were retrospectively evaluated. Cases were collected from referral institutions and were required to have received at least one doxorubicin treatment and have follow up information regarding time to progression. RESULTS: Significant differences in sex (p = 0.05), weight (p = 0.049), stage (p < 0.001), immunophenotype (p = <0.001), and number of doxorubicin doses (p = 0.001) were seen between regions. Upon univariate analysis, progression free survival (PFS) differed by region (p = 0.006), stage (p = 0.009), sub-stage (p = 0.0005), and immunophenotype (p = 0.001). A multivariable Cox regression model showed that dogs in the western region had a significantly shorter PFS when compared to the south and east. CONCLUSION: PFS was significantly affected by stage, sub-stage and phenotype.

Cortactin deficiency causes increased RhoA/ROCK1-dependent actomyosin contractility, intestinal epithelial barrier dysfunction, and disproportionately severe DSS-induced colitis.

The intestinal epithelium constitutes a first line of defense of the innate immune system. Epithelial dysfunction is a hallmark of intestinal disorders such as inflammatory bowel diseases (IBDs). The actin cytoskeleton controls epithelial barrier integrity but the function of actin regulators such as cortactin is poorly understood. Given that cortactin controls endothelial permeability, we hypothesized that cortactin is also important for epithelial barrier regulation. We found increased permeability in the colon of cortactin-KO mice that was accompanied by reduced levels of ZO-1, claudin-1, and E-cadherin. By contrast, claudin-2 was upregulated. Cortactin deficiency increased RhoA/ROCK1-dependent actomyosin contractility, and inhibition of ROCK1 rescued the barrier defect. Interestingly, cortactin deficiency caused increased epithelial proliferation without affecting apoptosis. KO mice did not develop spontaneous colitis, but were more susceptible to dextran sulfate sodium colitis and showed severe colon tissue damage and edema formation. KO mice with colitis displayed strong mucus deposition and goblet cell depletion. In healthy human colon tissues, cortactin co-localized with ZO-1 at epithelial cell contacts. In IBDs patients, we observed decreased cortactin levels and loss of co-localization with ZO-1. Thus, cortactin is a master regulator of intestinal epithelial barrier integrity in vivo and could serve as a suitable target for pharmacological intervention in IBDs.

Role of adenylyl cyclase in reduced ß-adrenoceptor-mediated vasorelaxation during maturation.

Beta-adrenergic receptor (ßAR)-dependent blood vessel relaxation is impaired in older animals and G protein activation has been suggested as the causative mechanism. Here, we investigated the role of ßAR subtypes (ß1AR, ß2AR, and ß3AR) and cAMP in maturation-dependent vasorelaxation impairment. Aortic rings from 15 Sprague-Dawley male rats (3 or 9 weeks old) were harvested and left intact or denuded of the endothelium. Vascular relaxation in aortic rings from younger and older groups was compared in the presence of ßAR subtype agonists and antagonists along with cAMP and cGMP antagonists. Isolated aortic rings were used to evaluate relaxation responses, protein expression was evaluated by western blot or real time PCR, and metabolites were measured by ELISA. Expression of ßAR subtypes and adenylyl cyclase was assessed, and cAMP activity was measured in vascular tissue from both groups. Isoproterenol- and BRL744-dependent relaxation in aortic rings with and without endothelium from 9-week-old rats was impaired compared with younger rats. The ß1AR antagonist CGP20712A (10-7 M) did not affect isoproterenol or BRL744-dependent relaxation in arteries from either group. The ß2AR antagonist ICI-118,551 (10-7 M) inhibited isoproterenol-dependent aortic relaxation in both groups. The ß3AR antagonist SR59230A (10-7 M) inhibited isoproterenol- and BRL744-dependent aortic ring relaxation in younger but not in older rats. All ßAR subtypes were expressed in both groups, although ß3AR expression was lower in the older group. Adenylyl cyclase (SQ 22536) or protein kinase A (H89) inhibitors prevented isoproterenol-induced relaxation in younger but not in older rats. Production of cAMP was reduced in the older group. Adenylyl cyclase III and RyR3 protein expression was higher in the younger group. In conclusion, altered expression of ß3AR and adenylyl cyclase III may be responsible for reduced cAMP production in the older group.

Role of adenylyl cyclase in reduced ß-adrenoceptor-mediated vasorelaxation during maturation

Beta-adrenergic receptor (βAR)-dependent blood vessel relaxation is impaired in older animals and G protein activation has been suggested as the causative mechanism. Here, we investigated the role of βAR subtypes (β1AR, β2AR, and β3AR) and cAMP in maturation-dependent vasorelaxation impairment. Aortic rings from 15 Sprague-Dawley male rats (3 or 9 weeks old) were harvested and left intact or denuded of the endothelium. Vascular relaxation in aortic rings from younger and older groups was compared in the presence of βAR subtype agonists and antagonists along with cAMP and cGMP antagonists. Isolated aortic rings were used to evaluate relaxation responses, protein expression was evaluated by western blot or real time PCR, and metabolites were measured by ELISA. Expression of βAR subtypes and adenylyl cyclase was assessed, and cAMP activity was measured in vascular tissue from both groups. Isoproterenol- and BRL744-dependent relaxation in aortic rings with and without endothelium from 9-week-old rats was impaired compared with younger rats. The β1AR antagonist CGP20712A (10-7 M) did not affect isoproterenol or BRL744-dependent relaxation in arteries from either group. The β2AR antagonist ICI-118,551 (10-7 M) inhibited isoproterenol-dependent aortic relaxation in both groups. The β3AR antagonist SR59230A (10-7 M) inhibited isoproterenol- and BRL744-dependent aortic ring relaxation in younger but not in older rats. All βAR subtypes were expressed in both groups, although β3AR expression was lower in the older group. Adenylyl cyclase (SQ 22536) or protein kinase A (H89) inhibitors prevented isoproterenol-induced relaxation in younger but not in older rats. Production of cAMP was reduced in the older group. Adenylyl cyclase III and RyR3 protein expression was higher in the younger group. In conclusion, altered expression of β3AR and adenylyl cyclase III may be responsible for reduced cAMP production in the older group.

Circulating Adipokine Levels in Non-diabetic Subjects with High Normal Blood Pressure.

OBJECTIVE: This study aimed to evaluate the levels of the adipokines, resistin and adiponectin in normotensive and high normal blood pressure patients. METHODS: Circulating levels of the adipokines, resistin and adiponectin were measured by enzyme-linked immunosorbent assay (ELISA; R'D Systems, Minneapolis) in 20 high normal blood pressure patients and in 20 age-matched normotensive non-diabetic subjects. Statistical analysis was performed with analysis of variance (ANOVA). RESULTS: The control group showed non-significantly decreased levels of resistin when compared with patients with high normal blood pressure [systolic 130-139 mmHg; diastolic 85-89 mmHg] (12.25 vs 14.38 pg/mL, p = 0.40). There were significantly higher levels of adiponectin in the control group when compared with high normal blood pressure patients (11.3 vs 7.51 µg/mL, p = 0.028). CONCLUSIONS: High normal blood pressure patients have increased levels of resistin and lower values of adiponectin when compared with age-matched non-diabetic normotensive subjects. This may explain why those patients showed more progression to hypertension, atherosclerosis and cardiovascular risk than normotensive subjects.

Identification and evaluation of putative tumour-initiating cells in canine malignant melanoma cell lines.

Tumour-initiating cells (TICs) have been identified in many solid human tumours, including malignant melanoma. In this study, an enriched TIC population was identified in two canine malignant melanoma cell lines (CML1 and CML6M) using cell surface markers and functional assays, including the sphere forming assay, aldehyde dehydrogenase (ALDH) assay, reverse transcriptase-polymerase chain reaction and γH2AX staining for double-stranded DNA (dsDNA)break identification and repair. The CD34(-) population of cells in both cell lines expressed stem cell genes, such as Oct4, Nanog and Ptch1, were more efficient at making spheres in adherence-free media conditions and were able to repair dsDNA breaks faster than the CD34(+) population. A subpopulation of cells with high expression of ALDH was identified in both cell lines by flow cytometry. The findings indicate the presence of TICs in two canine malignant melanoma cell lines.

Onicomadesis y granulomas piogénicos tras inmovilización posquirúrgica de la extremidad superior / Onychomadesis and Pyogenic Granulomas After Postoperative Upper-Limb Immobilization

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PERMANENT GENETIC RESOURCES: Isolation and characterization of 12 microsatellite markers in the middle-spotted woodpecker (Dendrocopos medius).

Twelve polymorphic microsatellite loci were developed for Dendrocopos medius. Polymorphism was assessed for 27 individuals from the southwesternmost population of this woodpecker species. The number of alleles per locus ranged from three to seven, with observed heterozygosity values from 0.444 to 0.852. Genotypic frequencies conformed to Hardy-Weinberg equilibrium, and no evidence for linkage disequilibrium was observed. Multilocus genotypes resulting from this set of markers will be useful to determine genetic diversity and differentiation within and among habitat patches inhabited by D. medius. Three of the loci were polymorphic for Picoides articus.

Sarna neonatal / Neonatal scabies

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[Follicular keratosis and multiple myeloma]. / Queratosis folicular y mieloma múltiple.

Follicular keratosis may be a rare paraneoplastic disease having unknown etiopathogeny that is sometimes associated to myeloma whose diagnosis and evolution follow a parallel course. We describe the case of a 57-year-old female patient diagnosed of multiple myeloma 8 years ago. She developed generalized follicular hyperkeratotic spicules coinciding with leukemization of the myeloma.