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During routine monitoring in Ojo de Liebre Lagoon, Mexico, a juvenile black turtle ( Chelonia mydas ) was captured, physically examined, measured, weighed, sampled, and tagged. The turtle showed no clinical signs suggestive of disease. Eleven months later, this turtle was recaptured in the same area, during which one lesion suggestive of fibropapilloma on the neck was identified and sampled for histopathology and molecular analysis. Histopathology revealed hyperkeratosis, epidermal hyperplasia, acanthosis, papillary differentiation and ballooning degeneration of epidermal cells, increased fibroblasts in the dermis, and angiogenesis, among other things. Hematological values were similar to those reported for clinically healthy black turtles and did not show notable changes between the first capture and the recapture; likewise, clinicopathological evaluation did not show structural or functional damage in the turtle's systems. The chelonid alphaherpesvirus 5 (ChHV5) UL30 gene was amplified and sequenced for phylogeny; Bayesian reconstruction showed a high alignment with the genus Scutavirus of the Eastern Pacific group. This is one of the first reports of ChHV5 in a cutaneous fibropapilloma of a black turtle in the Baja California peninsula.

Stable plastid transformation in Nicotiana tabacum has been achieved by using two different methods, the biolistic method, using a particle gun, and the polyethylene glycol (PEG)-mediated transformation. PEG-mediated plastid transformation involves the treatment of isolated protoplasts (plant cells without cell wall) with PEG in the presence of DNA. We have previously shown that in Nicotiana tabacum both methods are equally efficient. The PEG-mediated transformation efficiencies range between 20 and 50 plastid transformants per experiment (10 6 viable treated protoplasts). One advantage of the PEG method is that no expensive equipment such as a particle gun is required. The only crucial points are the handling and the cultivation of protoplasts. Furthermore, markers for the selection of transformed plastids are required. One of the most often used selection markers is the aadA gene which encodes for spectinomycin and streptomycin resistance. Here we describe a simplified and inexpensive protocol for the transformation of plastids in Nicotiana tabacum using an optimized protoplast culture protocol. PEG-mediated plastid transformation has the potential to be developed into a high-throughput, automated pipeline.

Stable plastid transformation in Nicotiana tabacum has been achieved by using two different methods, the biolistic method, using a particle gun, and the polyethylene glycol (PEG)-mediated transformation. PEG-mediated plastid transformation involves the treatment of isolated protoplasts (plant cells without cell wall) with PEG in the presence of DNA. We have previously shown that in Nicotiana tabacum both methods are equally efficient. The PEG-mediated transformation efficiencies range between 20 and 50 plastid transformants per experiment (10(6) viable treated protoplasts). One advantage of the PEG method is that no expensive equipment such as a particle gun is required. The only crucial points are the handling and the cultivation of protoplasts. Furthermore, markers for the selection of transformed chloroplasts are required. One of the most often used selection markers is the aadA gene which encodes for spectinomycin and streptomycin resistance. Here we describe a simplified and inexpensive protocol for the transformation of chloroplasts in Nicotiana tabacum using an optimized protoplast culture protocol.

Introduction: The importance of cerebral ischemia in clinical practice has driven the development of diverse experimental models of its different forms and different methods of producing it have been described. Here we report our experience with the use of the internal carotid artery (ICA) as an access route for the permanent endoluminal occlusion of the middle cerebral artery (MCA) in Wistar rats, and a detailed description of the procedure., Aims: Our aim at the at the Universidad de Cuenca Research Institute was to establish a standard technique for the constant and repeatable production of a focal cerebral infarct in the MCA territory in Wistar rats., Materials and Methods: Using a cervical approach, the ICA was dissected in 20 Wistar rats and, by means of an arteriotomy, a 25 mm long nylon 3-0 or 4-0 filament was introduced until the MCA was obstructed. The infracted area was delimited with 2,3,5-triphenyltetrazolium chloride and its volume was calculated using computer software., Results: Positive results were obtained in 10 cases, the mean volume of the infarct was 488.84 mm3, and the standard error was found to be 63.91 mm3. Subcortical infarcts occurred on three occasions. Two cases presented subarachnoid haemorrhage and two others showed parenchymatous haemorrhages; there were three deaths., Conclusions: The use of the ICA as an entry point is an effective alternative for the endovascular production of focal cerebral ischemia and the use of a nylon 4-0 filament was more efficient than using a calibre 3-0 filament.