In vivo genetic engineering of murine pancreatic beta cells mediated by single-stranded adeno-associated viral vectors of serotypes 6, 8 and 9.

AIMS/HYPOTHESIS: The genetic engineering of pancreatic beta cells could be a powerful tool for examining the role of key genes in the cause and treatment of diabetes. Here we performed a comparative study of the ability of single-stranded (ss) adeno-associated viral vectors (AAV) of serotypes 6, 8 and 9 to transduce the pancreas in vivo. METHODS: AAV6, AAV8 and AAV9 vectors encoding marker genes were delivered to the pancreas via intraductal or systemic administration. Transduced cells were analysed by immunostaining. AAV9 vectors encoding hepatocyte growth factor (HGF) were delivered intraductally to a transgenic mouse model of type 1 diabetes and glycaemia was monitored. RESULTS: AAV6, AAV8 and AAV9 mediated efficient and long-term transduction of beta cells, with AAV6 and AAV8 showing the highest efficiency. However, alpha cells were poorly transduced. Acinar cells were transduced by the three serotypes tested and ductal cells only by AAV6. In addition, intraductal delivery resulted in higher AAV-mediated transduction of the pancreas than did systemic administration. As proof of concept, intraductal delivery of AAV9 vectors encoding for the beta cell anti-apoptotic and mitogenic HGF preserved beta cell mass, diminished lymphocytic infiltration of the islets and protected mice from autoimmune diabetes. CONCLUSIONS/INTERPRETATION: Intraductal administration of AAV6, AAV8 and AAV9 is an efficient way to genetically manipulate the pancreas in vivo. This technology may prove useful in the study of islet physiopathology and in assessment of new gene therapy approaches designed to regenerate beta cell mass during diabetes.

Successful medical management of pseudomembranous cystitis in three cats with lower urinary tract obstruction.

CASE REPORT: The present case series describes the clinical course and outcome of three cats diagnosed with pseudomembranous cystitis. This is an uncommon presentation of lower urinary tract obstruction but can be easily be identified by ultrasonography, revealing severe bladder wall thickening and thin hyperechoic luminal strips. The condition can be secondary to severe bacterial urinary tract infection. All cats were successfully treated with medical management only, mainly based on antimicrobials and individualised supportive therapy. CONCLUSION: Further evaluation of this condition is necessary in order to determine potential underlying aetiologies, pathophysiological mechanisms and the most appropriate standardised treatment.

Early Levallois technology and the Lower to Middle Paleolithic transition in the Southern Caucasus.

The Lower to Middle Paleolithic transition (~400,000 to 200,000 years ago) is marked by technical, behavioral, and anatomical changes among hominin populations throughout Africa and Eurasia. The replacement of bifacial stone tools, such as handaxes, by tools made on flakes detached from Levallois cores documents the most important conceptual shift in stone tool production strategies since the advent of bifacial technology more than one million years earlier and has been argued to result from the expansion of archaic Homo sapiens out of Africa. Our data from Nor Geghi 1, Armenia, record the earliest synchronic use of bifacial and Levallois technology outside Africa and are consistent with the hypothesis that this transition occurred independently within geographically dispersed, technologically precocious hominin populations with a shared technological ancestry.

The TD6 level lithic industry from Gran Dolina, Atapuerca (Burgos, Spain): production and use.

Technological analysis of lithic artefacts recovered at the Aurora stratum of Atapuerca-TD6 shows that this Lower Pleistocene assemblage is similar to Mode I Technology (=Oldowan tradition) documented at many African sites. Diachronic comparison of the different levels of Gran Dolina allows us to conclude that this particular form of early European technology lacks the production of big flakes to manufacture large tools such as bifaces and cleavers. Rather, it is characterized by the presence of small artefacts, including flakes, denticulates, notches, and side-scrapers, many of which bear use-wear traces of butchery and woodworking. The dominant production technique is orthogonal, which is also reflected in the core recovered at the slightly older level of TD4. The raw materials also found in the Middle Pleistocene occupations at Atapuerca, though with significant proportion differences, have a local origin and include varieties of flint, quartzite and sandstone as well as limestone and quartz. TD6 small artefacts were made from most of these, although the retouched pieces seem to have been preferentially made of the best quality flint, i.e., Cretaceous flint, pointing to the existence of differential use of lithic material, and therefore, some degree of planned knapping behaviour. Most of the "chaînes opératoires" or reduction sequences took place inside the cave, although some artefacts, elaborated on Cretaceous flint, seem to have been retouched off site, possibly near the supply sources.