The isolation and characterization of bovine adult derived adipose stem cells for the use in nuclear transfer

Since the cloning of Dolly there has been little change in the efficiency of nuclear transfer (NT). Research is beginning to investigate the characteristics of donor cells. Adiposetissue is an abundant source of adult-derived cells that have displayed “stemness” in-vitro(Gimble et al., 2003). The overall goal of this research was to define the in-vitro characteristicsof bovine adipose-derived adult stem cells (ADAS) for the use in NT. Isolation methods weredetermined by a 3 x 3 factorial design. 1 g of subcutaneous fat was collected and subjected to0.10%, 0.25% or 0.50% collagenase type I solution for 1, 2 and 3 h. Nucleated cells werecounted using heochst stain. There was no significant difference (P\u3e0.05) in number of nucleated cells released during the incubation period or collagenase concentrations. Viable cells were determined by those that remained adherent 24 h post plating. Incubation in 0.25% collagenase for 2 h had the consistently highest percentage of viable cells (45%). The lifespan and growth characteristics were determined by in a 2 x 2 factorial experiment of DMEM or DMEM:F12 supplemented or not supplemented with growth factors. DMEM with growth factors supplementation was significantly shorter lifespan (P\u3e0.05) than DMEM:F12. The averagelifespan was ~30 population doublings (PDs), with 1 cell cycle every two days until passage 8(P8). Two bovine ADAS cell lines were differentiated into adipocytes, chondrocytes and osteoblasts at middle and late passages along side of adult derived skin fibroblasts. Differentiation was confirmed by histological staining resulting in early passage ADAS cells staining more intensely compared to late passage ADAS cells and skin fibroblasts. Global levels of DNA methylation and histone acetylation were analyzed from P1 to P6 in ADAS and skin fibroblasts from three animals. There was no significant difference (P\u3e0.05) between cell types for DNA methylation or histone acetylation. The percentage of cleaved and developing blastocyst embryos from the ADAS cells (62% and 8%) and skin fibroblasts cells (42% and 8%)were not different (P\u3e0.05). Interspecies nuclear transfer utilized eland ADAS cells into enucleated bovine oocytes. A total of 3 interspecies embryos (1%) developed to blastocyst

Child–Adult Differences in Using Dual-Task Paradigms to Measure Listening Effort

The purpose of the project was to investigate the effects modifying the secondary task in a dual-task paradigm to measure objective listening effort. To be specific, the complexity and depth of processing were increased relative to a simple secondary task

Child–Adult Differences in Using Dual-Task Paradigms to Measure Listening Effort

The purpose of the project was to investigate the effects modifying the secondary task in a dual-task paradigm to measure objective listening effort. To be specific, the complexity and depth of processing were increased relative to a simple secondary task

Monitoring Insulin Aggregation via Capillary Electrophoresis

Early stages of insulin aggregation, which involve the transient formation of oligomeric aggregates, are an important aspect in the progression of Type II diabetes and in the quality control of pharmaceutical insulin production. This study is the first to utilize capillary electrophoresis (CE) with ultraviolet (UV) detection to monitor insulin oligomer formation at pH 8.0 and physiological ionic strength. The lag time to formation of the first detected species in the aggregation process was evaluated by UV-CE and thioflavin T (ThT) binding for salt concentrations from 100 mM to 250 mM. UV-CE had a significantly shorter (5–8 h) lag time than ThT binding (15–19 h). In addition, the lag time to detection of the first aggregated species via UV-CE was unaffected by salt concentration, while a trend toward an increased lag time with increased salt concentration was observed with ThT binding. This result indicates that solution ionic strength impacts early stages of aggregation and β-sheet aggregate formation differently. To observe whether CE may be applied for the analysis of biological samples containing low insulin concentrations, the limit of detection using UV and laser induced fluorescence (LIF) detection modes was determined. The limit of detection using LIF-CE, 48.4 pM, was lower than the physiological insulin concentration, verifying the utility of this technique for monitoring biological samples. LIF-CE was subsequently used to analyze the time course for fluorescein isothiocyanate (FITC)-labeled insulin oligomer formation. This study is the first to report that the FITC label prevented incorporation of insulin into oligomers, cautioning against the use of this fluorescent label as a tag for following early stages of insulin aggregation

Health-related quality of life and mental health in the medium-term aftermath of the Prestige oil spill in Galiza (Spain): a cross-sectional study

<p>Abstract</p> <p>Background</p> <p>In 2002 the oil-tanker <it>Prestige </it>sank off the Galician coast. This study analyzes the effect of this accident on health-related quality of life (HRQoL) and mental health in the affected population.</p> <p>Methods</p> <p>Using random sampling stratified by age and sex, 2700 residents were selected from 7 coastal and 7 inland Galician towns. Two exposure criteria were considered: a) residential exposure, i.e., coast versus interior; and b) individual exposure-unaffected, slightly affected, or seriously affected-according to degree of personal affectation. SF-36, GHQ-28, HADS and GADS questionnaires were used to assess HRQoL and mental health. Association of exposure with suboptimal scores was summarized using adjusted odds ratios (OR) obtained from logistic regression.</p> <p>Results</p> <p>For residential exposure, the SF-36 showed coastal residents as having a lower likelihood of registering suboptimal HRQoL values in physical functioning (OR:0.69; 95%CI:0.54–0.89) and bodily pain (OR:0.74; 95%CI:0.62–0.91), and a higher frequency of suboptimal scores in mental health (OR:1.28; 95%CI:1.02–1.58). None of the dimensions of the other questionnaires displayed statistically significant differences.</p> <p>For individual exposure, no substantial differences were observed, though the SF-36 physical functioning dimension rose (showed better scores) with level of exposure (91.51 unaffected, 93.86 slightly affected, 95.28 seriously affected, p < 0.001).</p> <p>Conclusion</p> <p>Almost one and a half years after the accident, worse HRQoL and mental health levels were not in evidence among subjects exposed to the oil-spill. Nevertheless, some of the scales suggest the possibility of slight impact on the mental health of residents in the affected areas.</p

Genome-Wide Search Reveals the Existence of a Limited Number of Thyroid Hormone Receptor Alpha Target Genes in Cerebellar Neurons

Thyroid hormone (T3) has a major influence on cerebellum post-natal development. The major phenotypic landmark of exposure to low levels of T3 during development (hypothyroidism) in the cerebellum is the retarded inward migration of the most numerous cell type, granular neurons. In order to identify the direct genetic regulation exerted by T3 on cerebellar neurons and their precursors, we used microarray RNA hybridization to perform a time course analysis of T3 induced gene expression in primary cultures of cerebellar neuronal cell. These experiments suggest that we identified a small set of genes which are directly regulated, both in vivo and in vitro, during cerebellum post-natal development. These modest changes suggest that T3 does not acts directly on granular neurons and mainly indirectly influences the cellular interactions taking place during development

Unraveling the Early Events of Amyloid-β Protein (Aβ) Aggregation: Techniques for the Determination of Aβ Aggregate Size

The aggregation of proteins into insoluble amyloid fibrils coincides with the onset of numerous diseases. An array of techniques is available to study the different stages of the amyloid aggregation process. Recently, emphasis has been placed upon the analysis of oligomeric amyloid species, which have been hypothesized to play a key role in disease progression. This paper reviews techniques utilized to study aggregation of the amyloid-β protein (Aβ) associated with Alzheimer’s disease. In particular, the review focuses on techniques that provide information about the size or quantity of oligomeric Aβ species formed during the early stages of aggregation, including native-PAGE, SDS-PAGE, Western blotting, capillary electrophoresis, mass spectrometry, fluorescence correlation spectroscopy, light scattering, size exclusion chromatography, centrifugation, enzyme-linked immunosorbent assay, and dot blotting

Deepwater Horizon oil spill exposure and child health: a longitudinal analysis

The BP Deepwater Horizon oil spill (DHOS) created widespread concern about threats to health among residents of the Louisiana Gulf Coast. This study uses data from the Resilient Children, Youth, and Communities study—a longitudinal cohort survey of households with children in DHOS-affected areas of South Louisiana—to consider the effect of DHOS exposure on health trajectories of children, an especially vulnerable population subgroup. Results from latent linear growth curve models show that family DHOS exposure via physical contact and job/income loss both negatively influenced initial child health. However, the effects of physical exposure dissipated over time while the effects of job/income loss persisted. This pattern holds for both general child health and the number of recent physical health problems children had experienced. These findings help to bridge the literature on disaster impacts and resilience/vulnerability, with the literature on socioeconomic status as a fundamental cause of health outcomes over the life course