Canine fetal echocardiography: correlations for the analysis of cardiac dimensions

The aim of this study was to develop regression models for correlation of canine fetal heart development with body size to characterize normal development or suggest cardiac anomalies. Twenty clinically healthy pregnant bitches, either brachycephalic and non-brachycephalic, were examined ultrasonographically. Transabdominal fetal echocardiography was conducted every 4 days from the beginning of cardiac chambers differentiation until parturition. Ten cardiac parameters were measured: length, width and diameter of the heart; heart area; left and right ventricular dimensions; left and right atrial dimensions; and aortic and pulmonary artery diameter. Femoral length, biparietal diameter and abdominal cross-sectional area were also recorded. Regression equations were developed for each parameter of fetal body size, and linear and logarithmic models were compared. The model with the highest correlation coefficient was chosen to produce equations to calculate relative dimensions based on the correlations. Only the left-ventricular chamber differed between the two racial groups. Biparietal diameter was the independent parameter that produced the highest correlation coefficient for the most fetal cardiac dimensions, although good correlations were also observed using femoral length and abdominal cross-sectional area. Heart width and heart diameter were used as surrogates of cardiac development, as these measurements showed the best statistical correlation. Quantitative evaluation of fetal cardiac structures can be used to monitor normal and abnormal cardiac development.     

Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Immunological and pathological investigations in equine experimental uveitis

Background

The pathogenic mechanism of equine recurrent uveitis (ERU) is still poorly defined and many variations between experimental animal models and spontaneous disease exist.

Objectives

The aim of our study was to investigate if Th17 cell-mediated response plays role in the pathogenesis of the used experimental model in horses and to reveal its pathological findings.

Methods

Experimental uveitis was induced in 6 healthy horses. The concentrations of retinal autoantigen CRALBP and IL-17 were measured using ELISA in aqueous humor and vitreous body of the 12 inflamed eyes as well as in 12 control non-inflamed eyes taken from 6 horses in slaughter house. After centrifugation of the two eye media, smears were prepared and cytological investigation was performed. Tissue specimens were taken from all eye globes and were submitted to histopathological investigation.

Results

CRALBP and IL-17 concentrations were significantly elevated in eye media of horses with experimental uveitis in comparison with controls. Cytological and histopathological findings corresponded to the changes characteristic of chronic immune-mediated inflammation with mononuclear cell infiltration of uvea, choroid, retina, and eye media as well as severe retinal destruction.

Conclusions

Our study demonstrated the involvement of the retinal autoantigen CRALBP as well as IL-17 in the pathogenesis of experimental uveitis in horses. These findings suggests that this experimental uveitis in horses may serve as a suitable animal model for investigation of IL-17- mediated immune response during spontaneous autoimmune uveitis in horses as well as in humans.

     

Effect of maternal cells transferred with colostrum on cellular responses to pathogen antigens in neonatal calves

OBJECTIVE: To assess the effect of maternal cells or cellular components on neonatal immune responses to intracellular pathogens in calves. ANIMALS: 15 Holstein calves. PROCEDURES: Calves were fed whole colostrum, frozen colostrum, or cell-free colostrum within 4 hours after birth. Leukocytes were obtained from calves before feeding colostrum and 1, 2, 7, 14, 21, and 28 days after ingestion. Proliferative responses against bovine viral diarrhea virus (BVDV) and mycobacterial purified protein derivatives were evaluated. Dams received a vaccine containing inactivated BVDV, but were not vaccinated against mycobacterial antigens. RESULTS: All calves had essentially no IgG in circulation at birth, but comparable and substantial concentrations by day 1. Calves that received whole colostrum had enhanced responses to BVDV antigen 1 and 2 days after ingestion of colostrum. In contrast, calves that received frozen colostrum or cell-free colostrum did not respond to BVDV. No differences were identified among the 3 groups in response to mycobacterial antigens. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that transfer of live maternal cells from colostrum to neonatal calves enhanced responses to antigens against which the dams had previously responded (BVDV), but not to antigens to which the dams were na?ve (mycobacterial purified protein derivatives). Results suggested that cell-mediated immune transfer to neonates can be enhanced by maternal vaccination.     

Resynchronization of estrus in beef cattle: ovarian function, estrus and fertility following progestin treatment and treatments to synchronize ovarian follicular development and estrus

The objective was to optimize rebreeding of nonpregnant, previously inseminated beef cattle. In Experiment 1, 43 cows received a used intravaginal progesterone-releasing insert (IVPRI; Days 0-7) 12.3 d after ovulation and received concurrently no treatment, 100 microg gonadotropin releasing hormone (GnRH), 1 mg estradiol cypionate (ECP), or 150 mg progesterone. Emergence of a new ovarian follicular wave was most synchronous (P < 0.0001) in the GnRH group. In Experiment 2, 675 heifers were given GnRH or no treatment on Day 0, fed melengestrol acetate (MGA; 0.5 mg/head/d) from Days 0-5 (Day 0 = 13-14 d after timed insemination; TAI), given 0.5 mg ECP or nothing on Day 7, and reinseminated 6-12 h after onset of estrus. Estrus was more synchronous (P < 0.05) in heifers given GnRH versus no treatment on Day 0. In Experiment 3, 317 TAI heifers were resynchronized with either MGA or a used IVPRI with or without ECP on Day 7; estrus was more synchronous (P < 0.05) and pregnancy rates were higher (54.1% versus 39.2%, P < 0.05) in heifers given a used IVPRI than those fed MGA. For resynchronization of heifers, pregnancy rates were not significantly improved with GnRH treatment, but were higher with a used IVPRI than with MGA.     

The second-generation curcumin analogue RL71 elicits G2/M cell cycle arrest and apoptosis in canine osteosarcoma cells

Canine osteosarcoma is an aggressive cancer, comprising 85% of canine bone neoplasms. Current treatment practices of surgery and chemotherapy increase 1-year survival by only 45%. The curcumin analogue RL71, has demonstrated potent in vitro and in vivo efficacy in several models of human breast cancer through increased apoptosis and cell cycle arrest. Thus, the present study aimed to investigate efficacy of curcumin analogues in two canine osteosarcoma cell lines. Osteosarcoma cell viability was assessed using the sulforhodamine B assay and mechanisms of action were determined by analysing the levels of cell cycle and apoptotic regulatory proteins via Western blotting. Further evidence was obtained using flow cytometry to detect cell cycle distribution and the number of apoptotic cells. RL71 was the most potent curcumin analogue with EC₅₀ values of 0.64 ± 0.04 and 0.38 ± 0.009 μM (n = 3) in D-17 (commercial) and Gracie canine osteosarcoma cells, respectively. RL71 significantly increased the ratio of cleaved-caspase 3 to pro-caspase 3 and the level of apoptotic cells at the 2× and 5× EC₅₀ concentration (p < 0.001, n = 3). Furthermore, at the same concentration, RL71 significantly increased the number of cells in the G2/M phase. In conclusion, RL71 has potent cytotoxic activity in canine osteosarcoma cells triggering G2/M arrest and apoptosis at concentrations achievable in vivo. Future research should further investigate molecular mechanisms for these changes in other canine osteosarcoma cell lines prior to in vivo investigation.     

Using midday stem water potential for scheduling deficit irrigation in mid–late maturing peach trees under Mediterranean conditions

Irrigation techniques that reduce water applications are increasingly applied in areas with scarce water resources. In this study, the effect of two regulated deficit irrigation (RDI) strategies on peach [Prunus persica (L.) Batsch cv. “Catherine”] performance was studied over three growing seasons. The experimental site was located in Murcia (SE Spain), a Mediterranean region. Two RDI strategies (restricting water applications at stage II of fruit development and postharvest) based on stem water potential (Ψ_s) thresholds (?1.5 and ?1.8 MPa during fruit growth and ?1.5 and ?2.0 MPa during postharvest) were compared to a fully irrigated control. Soil water content (θ_v), Ψ_s, gas exchange parameters, vegetative growth, crop load, yield and fruit quality were determined. RDI treatments showed significantly lower values of θ_v and Ψ_s than control trees when irrigation water was restricted, causing reductions in stomatal conductance and photosynthesis rates. Vegetative growth was reduced by RDI, as lower shoot lengths and pruning weights were observed under those treatments when compared to control. However, fruit size and yield were unaffected, and fruit quality was slightly improved by RDI. Water savings from 43 to 65 % were achieved depending on the year and the RDI strategy, and no negative carryover effect was detected during the study period. In conclusion, RDI strategies using Ψ_s thresholds for scheduling irrigation in mid–late maturing peach trees under Mediterranean conditions are viable options to save water without compromising yield and even improving fruit quality.     

Measured and estimated water use and crop coefficients of grapevines trained to overhead trellis systems in California’s San Joaquin Valley

The use of overhead trellis systems for the production of dry-on-vine (DOV) raisins and table grapes in California is expanding. Studies were conducted from 2006 to 2009 using Thompson Seedless grapevines grown in a weighing lysimeter trained to an overhead arbor trellis and farmed as DOV raisins for the first two years and for use as table grapes thereafter. Maximum canopy coverage for the two lysimeter vines across years was in excess of 80 %. Seasonal (15 March–31 October) evapotranspiration for the lysimeter vines (ET_Lys) was 952 mm in 2007 (farmed as DOV raisins) and 943 and 952 mm (when farmed as table grapes). The maximum crop coefficient (K _cLys) across all 4 years ranged from 1.3 to 1.4. These maximum values were similar to those estimated using the relationship where K _c is a function of the amount of shaded area measured beneath the canopy at solar noon (K _c = 0.017 × percent shaded area). Covering the lysimeter’s soil surface with plastic (and then removing it) numerous times during the 2009 growing season (1 June–14 September) reduced ET_Lys from an average of 6.4 to 5.6 mm day^?1 and the K _c from 1.07 to 0.93. A seasonal basal K _c (K _cb) was calculated for grapevines using an overhead trellis system with a 13 % reduction in the K _cLys across the growing season.