Development of a facial expression scale using footrot and mastitis as models of pain in sheep

Management of pain in sheep is limited by the challenges of recognising and accurately quantifying pain in this species. The use of facial expression scoring to assess pain is a well-utilised, practical tool in both humans and non-human animals. The objective of this study was to develop a standardised facial expression pain scale for adult sheep, that could be used reliably and accurately to detect pain associated with naturally occurring painful diseases, such as footrot and mastitis. We also investigated whether the scale could be reliably and accurately utilised by observers after training, enabling the development of an on-farm pain assessment tool. The Sheep Pain Facial Expression Scale (SPFES) was able to correctly identify sheep suffering from disease with a high degree of accuracy (AUC; Footrot: 0.81, Mastitis: 0.80). Diseased sheep scored higher on the scale than controls on the day of disease identification (P<0.05) and diseased sheep showed changes in their facial expression after treatment (P<0.001). The abnormal facial expressions of diseased sheep reduced over time, and at recovery were in line with control sheep. Control sheep did not change their facial expression over time. Five scorers who were trained to use the developed scale also assessed the facial expressions of sheep. The scorers were blind to treatment and session. Scorers reliably (ICC: 0.86) and accurately (α = 0.86) identified changes in the facial expression of sheep with footrot over time (P<0.05), and scored control sheep consistently low over time. The SPFES offers a reliable and effective method of assessing pain in sheep after minimal training.We would like to thank the EU VII Framework Program (FP7-KBBE-2010-4) for funding this study as part of the AWIN project. Boehringer Imgelheim Ltd are thanked for providing the meloxicam used in this study.This is the author accepted manuscript. The final version is available from Elsevier via http://dx.doi.org/10.1016/j.applanim.2016.01.00

Intratumoral patterns of clonal evolution in gliomas

Few studies have explored the patterns of clonal evolution in gliomas. Here, we investigate the cytogenetic patterns of intratumoral clonal evolution of gliomas and their impact on tumor histopathology and patient survival. Cytogenetic analysis of 90 gliomas was performed in individual tumor cells (>200 cells/tumor) using multicolor (N = 16 probes) interphase-FISH. Overall, chromosome gains were more frequent than chromosome losses. Gains of chromosome 7 and/or EGFR amplification were detected in 91% of the cases, whereas del(9p21) (77%) and del(10q23) (78%) were the most frequent chromosome losses. Virtually, all cases (99%) showed >/=2 tumor cell clones, with higher numbers among high- versus low-grade gliomas (p = 0.001). Nine different cytogenetic patterns were found in the ancestral tumor clones. In most gliomas, ancestral clones showed abnormalities of chromosome 7, 9p, and/or 10q and cytogenetic evolution consisted of acquisition of additional abnormalities followed by tetraploidization. Conversely, early tetraploidization was associated with low-grade astrocytomas-2/3 pilocytic and 3/6 grade II diffuse astrocytomas-and combined loss of 1p36/19q13 with oligodendrogliomas, respectively; both aberrations were associated with a better patient outcome (p = 0.03). Overall, our results support the existence of different pathways of intratumoral evolution in gliomas

Amplified and Homozygously Deleted Genes in Glioblastoma: Impact on Gene Expression Levels

BACKGROUND: Glioblastoma multiforme (GBM) displays multiple amplicons and homozygous deletions that involve relevant pathogenic genes and other genes whose role remains unknown. METHODOLOGY: Single-nucleotide polymorphism (SNP)-arrays were used to determine the frequency of recurrent amplicons and homozygous deletions in GBM (n = 46), and to evaluate the impact of copy number alterations (CNA) on mRNA levels of the genes involved. PRINCIPAL FINDINGS: Recurrent amplicons were detected for chromosomes 7 (50%), 12 (22%), 1 (11%), 4 (9%), 11 (4%), and 17 (4%), whereas homozygous deletions involved chromosomes 9p21 (52%) and 10q (22%). Most genes that displayed a high correlation between DNA CNA and mRNA levels were coded in the amplified chromosomes. For some amplicons the impact of DNA CNA on mRNA expression was restricted to a single gene (e.g., EGFR at 7p11.2), while for others it involved multiple genes (e.g., 11 and 5 genes at 12q14.1-q15 and 4q12, respectively). Despite homozygous del(9p21) and del(10q23.31) included multiple genes, association between these DNA CNA and RNA expression was restricted to the MTAP gene. CONCLUSIONS: Overall, our results showed a high frequency of amplicons and homozygous deletions in GBM with variable impact on the expression of the genes involved, and they contributed to the identification of other potentially relevant genes

The Expression of Connexins and SOX2 Reflects the Plasticity of Glioma Stem-Like Cells

Glioblastoma (GBM) is the most malignant primary brain tumor, with an average survival rate of 15 months. GBM is highly refractory to therapy, and such unresponsiveness is due, primarily, but not exclusively, to the glioma stem-like cells (GSCs). This subpopulation express stem-like cell markers and is responsible for the heterogeneity of GBM, generating multiple differentiated cell phenotypes. However, how GBMs maintain the balance between stem and non-stem populations is still poorly understood. We investigated the GBM ability to interconvert between stem and non-stem states through the evaluation of the expression of specific stem cell markers as well as cell communication proteins. We evaluated the molecular and phenotypic characteristics of GSCs derived from differentiated GBM cell lines by comparing their stem-like cell properties and expression of connexins. We showed that non-GSCs as well as GSCs can undergo successive cycles of gain and loss of stem properties, demonstrating a bidirectional cellular plasticity model that is accompanied by changes on connexins expression. Our findings indicate that the interconversion between non-GSCs and GSCs can be modulated by extracellular factors culminating on differential expression of stem-like cell markers and cell-cell communication proteins. Ultimately, we observed that stem markers are mostly expressed on GBMs rather than on low-grade astrocytomas, suggesting that the presence of GSCs is a feature of high-grade gliomas. Together, our data demonstrate the utmost importance of the understanding of stem cell plasticity properties in a way to a step closer to new strategic approaches to potentially eliminate GSCs and, hopefully, prevent tumor recurrence

Match running performance and physical capacity profiles of U8 and U10 soccer players

Aim This study aimed to characterize match running performance of very young soccer players and evaluate the relationship between these data and physical capacities and technical skills. Methods Distances covered at different speed thresholds were measured during 31 official matches using GPS technology in U10 (n = 12; age 10.1 ± 0.1 years) and U8 (n = 15; age 7.9 ± 0.1 years) national soccer players. Counter movement jump performance (CMJ), 20 m shuttle running (20 m-SR), linear sprint performance (10, 20, 30 m), shuttle (SHDT) and slalom dribble tests (SLDT) were performed to determine the players physical capacities and technical skills. Results Physical capacities and technical skills were higher in U10 versus U8 players [P 0.05, ES: 0.74). The U10 players covered more total (TD) and high-intensity running distance (HIRD) than their younger counterparts did (P 0.05, ES: 0.99). TD and HIRD covered across the three 15 min periods of match play did not decline (P > 0.05, ES: 0.02–0.55). Very large magnitude correlations were observed between the U8 and U10 players performances during the 20 m-SR versus TD (r = 0.79; P < 0.01) and HIRD (r = 0.82; P < 0.01) covered during match play. Conclusions Data demonstrate differences in match running performance and physical capacity between U8 and U10 players, and large magnitude relationships between match play measures and physical test performances. These findings could be useful to sports science staff working within the academies

Biallelic mutations in SORD cause a common and potentially treatable hereditary neuropathy with implications for diabetes

Here we report biallelic mutations in the sorbitol dehydrogenase gene (SORD) as the most frequent recessive form of hereditary neuropathy. We identified 45 individuals from 38 families across multiple ancestries carrying the nonsense c.757delG (p.Ala253GlnfsTer27) variant in SORD, in either a homozygous or compound heterozygous state. SORD is an enzyme that converts sorbitol into fructose in the two-step polyol pathway previously implicated in diabetic neuropathy. In patient-derived fibroblasts, we found a complete loss of SORD protein and increased intracellular sorbitol. Furthermore, the serum fasting sorbitol levels in patients were dramatically increased. In Drosophila, loss of SORD orthologs caused synaptic degeneration and progressive motor impairment. Reducing the polyol influx by treatment with aldose reductase inhibitors normalized intracellular sorbitol levels in patient-derived fibroblasts and in Drosophila, and also dramatically ameliorated motor and eye phenotypes. Together, these findings establish a novel and potentially treatable cause of neuropathy and may contribute to a better understanding of the pathophysiology of diabetes