Echinococcus Granulosus in the Endangered Patagonian Huemul (Hippocamelus bisulcus).

Cystic echinococcosis (CE) is a zoonotic parasitic disease associated with Echinococcus granulosus. The parasite is maintained by domestic and wild canids as definitive hosts with several ungulate species as intermediate hosts in domestic and peridomestic transmission cycles. In Chile, CE is endemic, and the role of livestock and dogs (Canis lupus familiaris) in the cycle and the accidental infection of humans are widely documented at rural sites. However, the role of wild herbivores in wild cycles or the potential transmission of CE from livestock is still unknown in Chile and the rest of South America. We used molecular techniques to describe CE infecting a Patagonian huemul (Hippocamelus bisulcus) in Cerro Castillo National Reserve (Aysén region, Chile). We make inferences about the risk of disease spillover from sympatric domestic and wild species. The DNA-based molecular analysis revealed that the huemul was infected with E. granulosus G1 genotype, sharing haplotypes with other G1 samples collected from sheep (Ovis aries) and cattle (Bos taurus) worldwide. Geographic overlap between sheep and huemul populations in the reserve likely facilitates parasite spillover into wild deer populations, with shepherd or stray dogs and wild foxes (Lycalopex culpaeus) potentially acting as bridging hosts between livestock and the endangered huemul. Further studies are warranted to understand the implications of E. granulosus for huemul conservation throughout the Chilean Patagonia.

Corynebacterium pseudotuberculosis Infection in Patagonian Huemul ( Hippocamelus bisulcus ).

Corynebacterium pseudotuberculosis is an intracellular bacteria and the etiologic agent of caseous lymphadenitis in domestic and wildlife species. We report C. pseudotuberculosis infection in Patagonian huemul ( Hippocamelus bisulcus ) from the Cerro Castillo National Reserve, Region of Aysen, Chile. Subcutaneous abscesses in the abdominal and pectoral regions from two animals were sampled and bacteriologic isolation was performed. In both cases, we isolated a C. pseudotuberculosis strain belonging to the ovine genotype. In addition, one isolate was resistant to ciprofloxacin and streptomycin. We report that H. bisulcus is a susceptible species to this bacterium, which is transmitted by direct or indirect contact with domestic sheep ( Ovis aries ) and which represents a potential conservation threat to populations of H. bisulcus . Additional research and prevention efforts should be addressed.

Human embryoid body-derived stem cells in tissue engineering-enhanced migration in co-culture with bladder smooth muscle and urothelium.

OBJECTIVES: To evaluate the affinity between human stem cells and human bladder cells by analyzing their migration and proliferation patterns in co-culture. Human stem cells have great potential for tissue engineering purposes. Co-culture of stem cells with mature cells may promote differentiation. METHODS: Equal numbers of green fluorescent protein-labeled human embryonic germ cell derivates (SDECs) were plated, either alone or in the presence of red fluorescence-labeled (PKH 26) human bladder smooth muscle cells (SMCs) or urothelial cells (UROs). The co-cultures shared the same media (EGM2MV). The migration patterns of the different cell lines were measured daily, using an integrated grid, for 8 days with fluorescence microscopy. RESULTS: SDECs, grown alone, had a robust basal migration rate of between 0.3 and 0.7 mm/day compared with SMCs, which had a rate of 0.1 to 0.3 mm/day and UROs with a rate of 0.1 to 0.2 mm/day. Stem cell migration was enhanced in co-culture with SMCs or UROs to 0.5 to 1.0 mm/day. Migration of SDECs was more linear, directed toward SMCs or UROs, compared with the circumferential growth when plated alone. SMCs, more than UROs, migrated more rapidly in the presence of stem cells. CONCLUSIONS: Human stem cells showed improved migration in the presence of mature human bladder cells and were attracted to them, as shown by the altered direction of growth. Thus, co-culture of human stem cells with host SMCs can enhance seeding of matrices due to positive chemotaxis. Identifying the responsible factors may help to augment chemotaxis between desired cell types and optimize tissue regeneration.

GABAergic lineage differentiation of AF5 neural progenitor cells in vitro.

We have previously described an immortal rat central-nervous-system progenitor cell line, AF5, which is able to exit the cell cycle and assume a differentiated state with neuronal properties. The phenotypic specification of differentiated AF5 cells, however, is not known. In the present study, when induced to differentiate by serum starvation in Neurobasal medium, AF5 cells down-regulate glial fibrillary acidic protein and up-regulate expression of beta-III-tubulin, medium-molecular-weight neurofilament protein, and neuronal growth-associated protein 43. Expression of the gamma-aminobutyric acid (GABA) lineage marker, glutamic acid decarboxylase 67 (GAD67), increases during differentiation, suggesting that AF5 cells adopt a GABAergic lineage. Time-course analysis of the GABAergic neuron specification transcription factor, Pitx2, by reverse transcription/polymerase chain reaction, has shown an increase in the Pitx2 transcript 48 h after initiation of differentiation. In differentiated AF5 cells, expression of the Pitx2 target gene products GAD65 and GABA transporter-1 increases. Cellular GABA levels in differentiated AF5 cells increase by about 26-fold, and GABA release into the medium is 150-fold higher compared with that of undifferentiated cells. Therefore, AF5 cells can be induced to differentiate to a neuronal phenotype with a GABAergic lineage.

Human embryoid body-derived stem cells in bladder regeneration using rodent model.

OBJECTIVES: To evaluate the capability of a human embryonic germ (hEG) cell-derived cell line (SDEC), previously characterized in our laboratory, seeded on porcine small intestinal submucosa (SIS) to regenerate the injured rat bladder. METHODS: Fluorescent-labeled SDEC cells seeded on SIS for 8 days in vitro were used as bladder grafts in rats. A total of 30 congenitally athymic rats (six groups of 5 rats each), underwent partial cystectomy and replacement with plain SIS (groups 1 to 3) or cell-seeded SIS (groups 4 to 6). The rats were sacrificed after 7 (groups 1 and 4), 14 (groups 2 and 5), and 28 (groups 3 and 6) days. The bladders were analyzed by histopathologic examination and fluorescence microscopy. RESULTS: No graft rejection or diminution in bladder capacity occurred. Plain SIS implants had multiple calcareous deposits, not seen with the cell-seeded implants. Macroscopically, at 7 days, the grafts were healed with a cellular lining on the luminal aspect in groups 4 to 6. Microscopically, the rat bladder was completely regenerated 28 days after stem cell-seeded SIS implantation. Labeled stem cells were identified throughout the graft and contributed significantly to bladder regeneration. CONCLUSIONS: The results of this study have demonstrated the successful replacement of a bladder defect in a rat model using hEG cell-derived cells seeded on SIS grafts. Longer term analysis of these bladder grafts will allow evaluation of function, cell migration, and differentiation processes of human stem cells.

AF5, a CNS cell line immortalized with an N-terminal fragment of SV40 large T: growth, differentiation, genetic stability, and gene expression.

Central nervous system progenitor cells that are self-renewing in culture and also differentiate under controlled conditions are potentially useful for developmental studies and for cell-based therapies. We characterized growth and plasticity properties and gene expression in a rat mesencephalic cell line, AF5, that was immortalized with an N-terminal fragment of SV40 large T (T155g). For over 150 population doublings in culture, the growth rate of AF5 cells remained steady, the cells remained responsive to bFGF, and telomerase activity and telomere lengths were unchanged. While karyotype analyses revealed some chromosomal abnormalities, these were also unchanged over time; additionally, no mutations in p53 gene sequences were found, and wild-type p53 activation was normal. AF5 cells produced PDGF, TGFbeta1, TGFbeta2, and bFGF. Similar to primary progenitor cells, AF5 cells retained their plasticity in culture; they could be propagated in an undifferentiated state as "neurospheres" in serum-free media or as adherent cultures in serum-containing media, and they differentiated when allowed to become confluent. Adherent subconfluent actively growing cultures expressed a marker for immature neurons, nestin, while few cells expressed the mature neuronal cell marker betaIII-tubulin. Confluent cultures ceased growing, developed differentiated morphologies, contained few or no nestin-expressing cells, and acquired betaIII-tubulin expression. Global gene expression was examined using a 15,000 gene microarray, comparing exponential growth with and without bFGF stimulation, and the differentiated state. The AF5 cell line exhibited stable genetic and growth properties over extended periods of time, while retaining the ability to differentiate in vitro. These data suggest that the AF5 cell line may be useful as an in vitro model system for studies of neural differentiation.

Administración de servicios de enfermería / Management of nursing services

Conceptualiza la administración como un proceso dinámico que constituye un instrumento para brindar una atención de calidad en enfermería, tanto en el manejo de la atención del usuario de salud a nivel individual, familiar o de comunidad; el manejo de las unidades y de los recursos humanos