Low temperature and mTOR inhibition favor stem cell maintenance in human keratinocyte cultures.

Adult autologous human epidermal stem cells can be extensively expanded ex vivo for cell and gene therapy. Identifying the mechanisms involved in stem cell maintenance and defining culture conditions to maintain stemness is critical, because an inadequate environment can result in the rapid conversion of stem cells into progenitors/transient amplifying cells (clonal conversion), with deleterious consequences on the quality of the transplants and their ability to engraft. Here, we demonstrate that cultured human epidermal stem cells respond to a small drop in temperature through thermoTRP channels via mTOR signaling. Exposure of cells to rapamycin or a small drop in temperature induces the nuclear translocation of mTOR with an impact on gene expression. We also demonstrate by single-cell analysis that long-term inhibition of mTORC1 reduces clonal conversion and favors the maintenance of stemness. Taken together, our results demonstrate that human keratinocyte stem cells can adapt to environmental changes (e.g., small variations in temperature) through mTOR signaling and constant inhibition of mTORC1 favors stem cell maintenance, a finding of high importance for regenerative medicine applications.

Occurrence of claw asymmetries in fattening pigs and potential impact on the development of sole ulcerations.

BACKGROUND: Claw abnormalities, particularly claw asymmetries, are associated with lameness in pigs and can be a welfare issue. However, the prevalence and development of claw asymmetries in pigs of different age is unknown. Therefore, the aim of this study was to analyze the claw symmetry over the pig lifetime from birth to slaughter as well as the occurrence of sole ulcerations in fattening pigs possibly caused by such asymmetric claws. RESULTS: From third day of life until slaughtering, asymmetric growth of the claws was detected more frequently and more severely with increasing age as determined by three-step scoring. Sole ulcerations were detected in slaughtered pigs only with a prevalence of 64.2% (197/307 examined animals). The risk for a sole ulceration was 3.6-fold higher for pigs with strongly asymmetric claws (≥ 30% size difference of the claw footing area) compared to slightly asymmetric claws (≥ 5-15% size difference of the claw footing area) (odds ratio (OR) = 3.6). It was even higher for pigs showing intermediately asymmetric claws (≥ 15-30% size difference of the claw footing area) (OR = 2.7). CONCLUSIONS: The study showed a significant increase in the prevalence of claw asymmetries over the pigs' lifetime, which can lead to serious pathologic findings with increasing age such as sole ulcerations. Most likely, the unbalanced weight load on single claws in combination with hard flooring can result in claw damages. Moreover, a genetic component cannot be excluded because claw asymmetries were already detected in suckling piglets.

Using Deep Learning to Detect Spinal Cord Diseases on Thoracolumbar Magnetic Resonance Images of Dogs.

Deep Learning based Convolutional Neural Networks (CNNs) are the state-of-the-art machine learning technique with medical image data. They have the ability to process large amounts of data and learn image features directly from the raw data. Based on their training, these networks are ultimately able to classify unknown data and make predictions. Magnetic resonance imaging (MRI) is the imaging modality of choice for many spinal cord disorders. Proper interpretation requires time and expertise from radiologists, so there is great interest in using artificial intelligence to more quickly interpret and diagnose medical imaging data. In this study, a CNN was trained and tested using thoracolumbar MR images from 500 dogs. T1- and T2-weighted MR images in sagittal and transverse planes were used. The network was trained with unremarkable images as well as with images showing the following spinal cord pathologies: intervertebral disc extrusion (IVDE), intervertebral disc protrusion (IVDP), fibrocartilaginous embolism (FCE)/acute non-compressive nucleus pulposus extrusion (ANNPE), syringomyelia and neoplasia. 2,693 MR images from 375 dogs were used for network training. The network was tested using 7,695 MR images from 125 dogs. The network performed best in detecting IVDPs on sagittal T1-weighted images, with a sensitivity of 100% and specificity of 95.1%. The network also performed very well in detecting IVDEs, especially on sagittal T2-weighted images, with a sensitivity of 90.8% and specificity of 98.98%. The network detected FCEs and ANNPEs with a sensitivity of 62.22% and a specificity of 97.90% on sagittal T2-weighted images and with a sensitivity of 91% and a specificity of 90% on transverse T2-weighted images. In detecting neoplasms and syringomyelia, the CNN did not perform well because of insufficient training data or because the network had problems differentiating different hyperintensities on T2-weighted images and thus made incorrect predictions. This study has shown that it is possible to train a CNN in terms of recognizing and differentiating various spinal cord pathologies on canine MR images. CNNs therefore have great potential to act as a "second eye" for imagers in the future, providing a faster focus on the altered image area and thus increasing workflow in radiology.