Induced pluripotent stem cells from pigs and other ungulate species: an alternative to embryonic stem cells?

Robust embryonic stem cell (ESC) lines from livestock species have been difficult to derive and maintain, and unlike mouse ESC, have not contributed to our ability to understand directed differentiation in vitro. Nor have such cells yet provided a simpler means than pronuclear injection to manipulate the genomes of agriculturally important species, such as cattle, sheep and pigs. Induced pluripotent stem cells (iPSC) generated by reprogramming somatic cells, such as fibroblasts, with a set of stemness genes, most usually but not exclusively POU5F1, SOX2, KLF4 and c-MYC, offer an alternative to ESC in these regards, as they exhibit a pluripotent phenotype resembling that of ESC, yet are readily generated in the laboratory. Accordingly, such cells, in association with cloning technologies, may be useful for introducing complex genetic changes into livestock, although this potential has yet to be demonstrated. Porcine iPSC may be especially valuable because the pig is a prime biomedical model for tissue transplantation. In general, iPSC from livestock, like those from humans, are of the epiblast type and depend upon FGF2 and activin/nodal signalling systems to maintain their pluripotency and growth. Recent experiments, in which newly reprogrammed porcine and bovine cells were selected on a LIF-based medium in presence of specific protein kinase inhibitors, have allowed iPSC cells of the na?ve type, resembling the more amenable blastocyst-derived mouse ESC and iPSC to be isolated. However, hurdles still remain if such cells are to achieve their biotechnological promise.     

Upregulation of TNF-α Production by IFN-γ and LPS in Cultured Canine Keratinocytes: Application to Monosaccharides Effects

Activated keratinocytes play a key role in the cutaneous immune system by their interactions with other cell types through the production of cytokines with both autocrine and paracrine activity. But there is little knowledge about epidermal cytokines in the dog. In this study, cultured canine keratinocytes were stimulated by human recombinant interferon γ (IFN-γ) and lipopolysaccharide (LPS) and cell supernatants were tested for tumour necrosis factor α (TNF-α) concentration using a cell viability assay on a murine cell line. We show that IFN-γ in combination with LPS significantly increases TNF-α secretion by canine keratinocytes. The best stimulations were obtained using confluent cultures and the association of IFN-γ (400 ng/ml) and LPS (40 μg/ml). The experimental protocol we describe represents a new method for studying keratinocyte activation and its modulation in the dog. We provide an example of application of our method: the study of the effects of different monosaccharides on canine keratinocyte activation.     

Dose measurement in small animal radiology: how intense is the influence of distance and position of the investigator in relation to the patient?

Radiation exposure of body parts of persons, needed to restrain small animal during the examination, varies depending on the distance and position in relation to the patient, the region investigated, and the exposure settings applied. The aim of the study was to quantify the effects of these factors on the ambient dose. The dosimeter was positioned with varying distance from the direct beam (from 30 to 150 cm) and on different level above the floor (55 cm, 85 cm, and 150 cm. The study consisted of two parts. In the first part a plastic water tank (thickness: 18 cm) was used as source of scatter radiation. Different exposure settings (77 kVp; 20 - 10 - 5 - 2.5 - 1.25 mAs) were applied. In the second part dose was measured during the examination of the abdomen of a large dog (thickness: 18 cm; 77 kVp, 20 mAs) and of the skull of a cat (thickness: 6 cm; 55 kVp, 20 mAs). At the level of the patient (85 cm above the floor) the dose decline relative to the distance followed a quadratic function. In the series "abdomen-dog" the mean dose values ranged from 51.6 microSv (30 cm distance) to 1.02 microSv (150 cm distance). The corresponding doses for the series "skull-cat" were 0.98 microSv and 0.02 microSv, respectively. Comparably lower doses were measured on the lines along the table, when non-irradiated parts of the body were located between the exposed patient volume and the dosimeter. At 150 cm above the floor higher doses were observed relative to the doses at the level of the table. The following conclusions can be drawn: (1.) Depending on the target volume the ambient dose varies in a wide range. (2.) An increase of the distance of only few centimetres is reducing exposure considerably. Therefore persons should make use of it whenever this is possible. (3.) Persons should stand on the short sides of the table while fixing the patient. (4.) Head and neck are relatively highly exposed. Methods to protect the thyroid gland and the eye lens are indicated. (5.) Since different exposure levels can be applied in digital radiography, the range of dose levels is rather wide. Therefore strict dose discipline (e.g. by use of exposure tables and dose indicators) is necessary to avoid continuing overexposure.