Cold agglutinin activity in 2 dogs

A 5‐year‐old neutered male Mastiff and an 8‐year‐old spayed female Labrador Retriever were presented to the University of Minnesota Veterinary Medical Center. The Mastiff was presented for evaluation of lameness and pyoderma one month prior in Missouri, where he tested positive for Ehrlichia canis by serum ELISA test, treated with doxycycline. PCR for Ehrlichia sp, Anaplasma sp, Babesia sp, and Bartonella sp, and PCR for antigen receptor rearrangement were negative, serum protein electrophoresis (SPE) revealed polyclonal gammopathy, and mildly reactive lymphoid cells were seen cytologically. The Labrador presented with a proliferative rostral mandibular gingival mass and lipomas for further presurgical evaluation of cold agglutinin activity documented by a commercial laboratory 2 years earlier prior to removal of a grade II mast cell tumor. This dog had a negative SNAP4Dx, normal SPE, and persistently increased serum ALP activity and polyuria/polydipsia suggestive for hyperadrenocorticism. Both dogs had markedly agglutinated RBC in the EDTA samples that dispersed with warming, and normal plasma color. Cold agglutinin activity was demonstrated by direct saline agglutination testing using whole blood and washed erythrocytes demonstrating agglutination at 30°C, 25°C, 15°C, and 4°C, but not at 37°C . CBC results (ADVIA 2120i) from the Mastiff revealed no significant differences in the RBC results obtained at room temperature (RT) and at 37°C; however, the RT run demonstrated negative bias in neutrophil and platelet concentrations attributed to rapid RBC settling. This uncommon hematologic condition may cause artifacts on the automated leukogram and platelet count, and may be subclinical for long periods.     

Ovine and murine T cell epitopes from the non-structural protein 1 (NS1) of bluetongue virus serotype 8 (BTV-8) are shared among viral serotypes

Bluetongue virus (BTV) is a non-enveloped dsRNA virus that causes a haemorrhagic disease mainly in sheep. It is an economically important Orbivirus of the Reoviridae family. In order to estimate the importance of T cell responses during BTV infection, it is essential to identify the epitopes targeted by the immune system. In the present work, we selected potential T cell epitopes (3 MHC-class II-binding and 8 MHC-class I binding peptides) for the C57BL/6 mouse strain from the BTV-8 non-structural protein NS1, using H2^b-binding predictive algorithms. Peptide binding assays confirmed all MHC-class I predicted peptides bound MHC-class I molecules. The immunogenicity of these 11 predicted peptides was then determined using splenocytes from BTV-8-inoculated C57BL/6 mice. Four MHC-class I binding peptides elicited specific IFN-γ production and generated cytotoxic T lymphocytes (CTL) in BTV-8 infected mice. CTL specific for 2 of these peptides were also able to recognise target cells infected with different BTV serotypes. Similarly, using a combination of IFN-γ ELISPOT, intracellular cytokine staining and proliferation assays, two MHC-class II peptides were identified as CD4⁺ T cell epitopes in BTV-8 infected mice. Importantly, two peptides were also consistently immunogenic in sheep infected with BTV-8 using IFN-γ ELISPOT assays. Both of these peptides stimulated CD4⁺ T cells that cross-reacted with other BTV serotypes. The characterisation of these T cell epitopes can help develop vaccines protecting against a broad spectrum of BTV serotypes and differentiate infected from vaccinated animals.     

The Endometrium of Cycling Cows Contains Populations of Putative Mesenchymal Progenitor Cells

Endometrial stem cells have been identified in humans, mice and pigs. This study was designed to determine whether the uterine endometrium of cycling cows contains such cells, to identify markers of stemness and ultimately to isolate putative stem/progenitor cell and evaluate their capability to differentiate into mesodermal derivatives. Uteri from healthy cows in the early (days 1–5) and late luteal phases (days 13–18) of the oestrous cycle were collected. Total RNA and proteins were isolated and searched for gene markers of embryonic (OCT4, NANOG, SOX2) and mesenchymal (CD44, STAT3, CD‐117) stem cells and for protein markers (Oct4, Sox2, Cd44) in Western blots or immunostaining of paraffin‐embedded tissue. Primary cell cultures were isolated; characterized in terms of morphology, colony formation and gene/protein expression; and induced osteogenic and chondrogenic differentiation. We identified expression of embryonic (OCT4 and SOX2, but not NANOG) and mesenchymal (STAT3, CD44 and c‐KIT) gene markers in the endometrium of cycling cows and the encoded proteins (Oct4, Sox2 and Cd44) in both stages of the oestrous cycle. Derived cell lines displayed essentially the same gene expression pattern; however, at the protein level, Oct4 was not detected. No clear influence of the stage of the oestrous cycle was found. Cell lines from late luteal phase displayed osteogenic and chondrogenic differentiation potential upon chemical stimulation. In this research, we demonstrated the presence of mesenchymal progenitor cell populations of apparently mesenchymal origin in the endometrium of cycling cows, in both the early and late phases of the oestrous cycle. The cells isolated from the late luteal phase were more acquiescent to differentiate into mesodermal derivatives than cells in the early luteal phase. Our findings might have implications for the understanding of uterine stem cell biology in cows and other farm animal species.     

MULTIDETECTOR ROW COMPUTED TOMOGRAPHY AND ULTRASOUND CHARACTERISTICS OF CAUDAL VENA CAVA DUPLICATION IN DOGS

Caudal vena cava duplication has been rarely reported in small animals. The purpose of this retrospective study was to describe characteristics of duplicated caudal vena cava in a large group of dogs. Computed tomography (CT) and ultrasound databases from two hospitals were searched for canine reports having the diagnosis “double caudal vena cava.” One observer reviewed CT images for 71 dogs and two observers reviewed ultrasound images for 21 dogs. In all CT cases, the duplication comprised two vessels that were bilaterally symmetrical and approximately the same calibre (similar to Type I complete duplication in humans). In all ultrasound cases, the duplicated caudal vena cava appeared as a distinct vessel running on the left side of the abdominal segment of the descending aorta and extending from the left common iliac vein to the left renal vein. The prevalence of caudal vena cava duplication was 0.46% for canine ultrasound studies and 2.08% for canine CT studies performed at these hospitals. Median body weight for affected dogs was significantly lower than that of unaffected dogs (P < 0.0001). Breeds with increased risk for duplicated caudal vena cava were Yorkshire Terrier (odds ratio [OR] = 6.41), Poodle (OR = 7.46), West Highland White Terrier (OR = 6.33), and Maltese (OR = 3.87). Presence of a duplicated caudal vena cava was significantly associated with presence of extrahepatic portosystemic shunt(s) (P < 0.004). While uncommon in dogs, caudal vena cava duplication should be differentiated from other vascular anomalies when planning surgeries and for avoiding misdiagnoses.     

A new protocol for assessing the conservation priority of groundwater‐dependent ecosystems

1. Species of conservation concern are usually considered important elements in site prioritization for biodiversity conservation. To overcome the lack of information on species conservation status, multidimensional measures of species rarity can be used as proxies of species vulnerability.
2. Under this assumption, a two‐step protocol for site prioritization of aquatic groundwater‐dependent ecosystems is proposed using invertebrate vulnerability estimated from species' traits. In the first step, each species occurring in the sites of interest are scored according to their vulnerability. In the second step, sites are prioritized using species' scores.
3. Species vulnerability scores are based on five dimensions, for which various traits are scored: (i) geography, (ii) ecology, (iii) biology, (iv) population, and (v) evolutionary history. For each species, the scores of the various traits belonging to the same dimension are multiplied to obtain a synthetic score. These scores are then ranked into four classes and, for each dimension, each species receives a new score that reflects its rank. The sum of these scores represents the species' overall score.
4. Site conservation priorities are assessed by combining species scores into three indices: Sum of Species Scores, Biodiversity Conservation Concern (which relates the sum of species scores with the local species richness) and Groundwater Biodiversity Concern (which is the average of the former two). The protocol is illustrated using case studies in Italy and it is fully implemented in the software AQUALIFE which is freely available at: http://app.aqualifeproject.eu by registered users.
5. Sensitivity analyses showed that the protocol is robust against the lack of information on species biology or sampling limitations. However, trait scoring rests with the user, who must be familiar with the study group.
6. This approach can be applied at any spatial scale and to different types of aquatic groundwater‐dependent ecosystems.

     

Wheat spike fertility: inheritance and relationship with spike yield components in early generations

In wheat, grain number is considered as the product of spike dry weight (SDW) and the number of grains per unit of SDW, that is an indicator of spike fertility (SF). The aim of this study was to determine the heritability of SF and the effect of early selection for high SF on its relationship with other spike yield components. Two field experiments were conducted in the south‐eastern Pampas (Argentina) with 400 F₂ and F_2:3 families obtained from two crosses between varieties with contrasting SF (PIG/SSN and B10/KCJ). Heritability estimates in PIG/SSN and B10/KCJ were, respectively, 0.60 and 0.51 by variance component analysis, 0.43 and 0.43 by F₂ : F₃ parent–offspring regression and 0.30 and 0.28 by realized heritability analysis. The existence of transgressive segregation (i.e. the occurrence of families with SF values that were more extreme than those of the parents) was observed. The top 25% F₃ families with the highest SF had 12% more grains per spike, despite a 13% and 5% decrease in SDW per spike and weight per grain, respectively, than the remaining families. These results give support to the application of early selection for high SF.     

Between a rock and a hard place: the impacts of climate change and housing development on breeding birds in California

Although the effects of climate change on species distributions have received considerable attention, land-use change continues to threaten wildlife by contributing to habitat loss and degradation. We compared projected spatial impacts of climate change and housing development across a range of housing densities on California’s birds to evaluate the relative potential impacts of each. We used species-distribution models in concert with current and future climate projections and spatially explicit housing-development density projections in California. We compared their potential influence on the distributions of 64 focal bird species representing six major vegetation communities. Averaged across GCMs, species responding positively to climate change were projected to gain 253,890 km² and species responding negatively were projected to lose 335,640 km². Development accounted for 32 % of the overall reductions in projected species distributions. In terms of land area, suburban and exurban development accounted for the largest portion of land-use impacts on species’ distributions. Areas in which climatic suitability and housing density were both projected to increase were concentrated along the foothills of the Sierra Nevada and areas of the north coast. Areas of decreasing climatic suitability and increasing housing density were largely concentrated within the Central Valley. Our analyses suggest that the cumulative effects of future housing development and climate change will be large for many bird species, and that some species projected to expand their distributions with climate change may actually lose ground to development. This suggests that a key climate change adaptation strategy will be to minimize the impacts of housing development. To do this effectively, comprehensive policies to guide land use decisions are needed at the broader scales of climate change.     

Hydrolytic and oxidative stability of L-(+)-ascorbic acid supported in pectin films: influence of the macromolecular structure and calcium presence

The hydrolytic and oxidative stability of L-(+)-ascorbic acid (AA) into plasticized pectin films were separately studied in view of preserving vitamin C activity and/or to achieve localized antioxidant activity at pharmaceutical and food interfaces. Films were made with each one of the enzymatically tailored pectins (50%, 70%, and 80% DM; Cameron et al. Carbohydr. Polym.2008, 71, 287-299) or commercial high methoxyl pectin (HMP; 72% DM). Since AA stability was dependent on water availability in the network, pectin nanostructure affected the AA kinetics. Higher AA retention and lower browning rates were achieved in HMP films, and calcium presence in them stabilized AA because of higher water immobilization. Air storage did not change AA decay and browning rates in HMP films, but they significantly increased in Ca-HMP films. It was concluded that the ability of the polymeric network to immobilize water seems to be the main factor to consider in order to succeed in retaining AA into film materials.     

Unsaturated fatty alcohol derivatives of olive oil phenolic compounds with potential low-density lipoprotein (LDL) antioxidant and antiobesity properties

A new route for the synthesis of fatty alcohol derivatives of hydroxytyrosol and other olive oil phenolic compounds was developed to allow the preparation of unsaturated derivatives. The biological activity of synthesized compounds was evaluated. Most of the compounds presented a significant antioxidant activity on low-density lipoprotein (LDL) particles. The activity of the tested products was significantly influenced by the number and position of unsaturations as well as modifications on the polar head of the synthesized compounds. Some of them presented modulation of food intake in rats and, due to their molecular similarity with CB(1) endogenous ligands, the endocannabinoid system and PPAR-α were also evaluated as potential targets. The pharmacodynamics could not be totally explained by CB(1) and PPAR-α receptor interactions because only two of the four compounds with biological activity showed a CB(1) activity and all of them presented low PPAR-α affinity, not justifying its whole in vivo activity. The hydroxytyrosol linoleylether (7) increased LDL resistance to oxidation with a capacity similar to that of hydroxytyrosol and was the most active in vivo compound with a hypophagic effect comparable to that of oleoylethanolamine. We consider that this compound could be a good lead compound for future drug development in obesity treatments.     

An aneuploid mouse strain carrying human chromosome 21 with Down syndrome phenotypes

Aneuploidies are common chromosomal defects that result in growth and developmental deficits and high levels of lethality in humans. To gain insight into the biology of aneuploidies, we manipulated mouse embryonic stem cells and generated a trans-species aneuploid mouse line that stably transmits a freely segregating, almost complete human chromosome 21 (Hsa21). This "transchromosomic" mouse line, Tc1, is a model of trisomy 21, which manifests as Down syndrome (DS) in humans, and has phenotypic alterations in behavior, synaptic plasticity, cerebellar neuronal number, heart development, and mandible size that relate to human DS. Transchromosomic mouse lines such as Tc1 may represent useful genetic tools for dissecting other human aneuploidies.