Six reagent waters, eleven diets, and two culture sizes were evaluated for culturing C. dubia. Different filtration techniques were used to prepare the reagent waters. The eleven diets were comprised of two algae augmented with eight supplements. Reproduction and growth were assessed to discern differences among C. dubia raised in mass cultures and cultured in individual cups, during which, bacterial population densities, lipid, protein, and carbohydrate concentrations of the diets were measured. Results showed that a glass-distilled, carbon filtered, deionized reagent water and a Selenastrum capricornutum- Cerophyl® diet were optimum for culturing. Mass culturing supported the highest reproduction and growth, while no correlation was found between nutritional measurements and production.

Both in-situ and ex-situ giant pandas (Ailuropoda melanoleuca), display shifts in bamboo species and part preference throughout the year. The effects of this shifting preference on gastrointestinal (GIT) microbiota were observed using traditional culturing methods to characterize normal GIT microflora from fecal samples and behavioral feeding data of adult male and female pandas over a fourteen-month period. Linear and quadratic fits were used to determine any significant relationships between the time of year and part preference on the GIT microflora (P<0.05). Significant values for time of year were observed with the linear fit in total aerobes (P-value=0.0368), streptococci (P-value=0.0120), and lactobacilli (P-value=0.0166) and quadratic fits in streptococci (P-value=0.0382) and Bacteroides spp. (P-value=0.0134) at á=0.05. Significant linear relationships were observed with part preference and lactobacilli and Bacteroides spp., P-values of 0.0028 and 0.0030, respectively, indicating that part preference and time of year may affect the flux of panda GIT microflora.

<p>One main hurdle to xenotransplantation, i.e. transplantation between different species, is the immunological barrier that the organ meets in the recipient. The aim of this thesis was to characterise xenogeneic rejection mechanisms by using the concordant mouse-to-rat heart transplantation model.</p><p>Graft-infiltrating immune cells could be isolated from both rejecting and non-rejecting grafts using ex vivo propagation, a technique based on incubation of graft biopsies in culture medium for 48 hours. The numbers of recovered T lymphocytes were considerably higher in grafts undergoing cell-mediated rejection than in grafts undergoing acute vascular rejection (AVR) or in non-rejecting transplants. Thus, ex vivo propagation should be a valuable tool for further studies of cell-mediated rejection.</p><p>Cytokine patterns in the grafts, as measured by a quantitative real-time RT-PCR method, showed that AVR and cell-mediated rejection are associated with an increase of both pro-inflammatory cytokines (IL-1β and TNF-α) and more specific cytokines (IL-2, IL-10, IL-12p40 and IFN-γ). These data differed considerably from the patterns seen in the spleens of the recipients. Cell-mediated xenograft rejection was also found to be associated with a local accumulation of hyaluronan.</p><p>Oral administration of xenogeneic cells stimulated a production of antibodies that could induce hyperacute rejection of cardiac xenografts when passively transferred to graft recipients. This is in contrast to several models for autoimmune diseases and allogeneic transplantation where oral administration of antigens is an effective way to induce unresponsiveness. Hence, future attempts to induce oral tolerance in xenotransplantation should be done with caution.</p>

One main hurdle to xenotransplantation, i.e. transplantation between different species, is the immunological barrier that the organ meets in the recipient. The aim of this thesis was to characterise xenogeneic rejection mechanisms by using the concordant mouse-to-rat heart transplantation model. Graft-infiltrating immune cells could be isolated from both rejecting and non-rejecting grafts using ex vivo propagation, a technique based on incubation of graft biopsies in culture medium for 48 hours. The numbers of recovered T lymphocytes were considerably higher in grafts undergoing cell-mediated rejection than in grafts undergoing acute vascular rejection (AVR) or in non-rejecting transplants. Thus, ex vivo propagation should be a valuable tool for further studies of cell-mediated rejection. Cytokine patterns in the grafts, as measured by a quantitative real-time RT-PCR method, showed that AVR and cell-mediated rejection are associated with an increase of both pro-inflammatory cytokines (IL-1β and TNF-α) and more specific cytokines (IL-2, IL-10, IL-12p40 and IFN-γ). These data differed considerably from the patterns seen in the spleens of the recipients. Cell-mediated xenograft rejection was also found to be associated with a local accumulation of hyaluronan. Oral administration of xenogeneic cells stimulated a production of antibodies that could induce hyperacute rejection of cardiac xenografts when passively transferred to graft recipients. This is in contrast to several models for autoimmune diseases and allogeneic transplantation where oral administration of antigens is an effective way to induce unresponsiveness. Hence, future attempts to induce oral tolerance in xenotransplantation should be done with caution.

From 2006-2011, four population reintroduction techniques were applied to three sites within a reach of the upper Clinch River in Virginia designated suitable for population restoration of the federally endangered oyster mussel (Epioblasma capsaeformis). These techniques were: 1) translocation of adults (Site 1), 2) release of laboratory-propagated sub-adults (Site 1), 3) release of 8-week old laboratory-propagated juveniles (Site 2), and 4) release of stream-side infested host fishes (Site 3). Demographic data were collected in 2011 and 2012 by systematic quadrat and capture-mark-recapture sampling to assess reintroduction success, evaluate reintroduction techniques, and compare survey approaches for monitoring freshwater mussels. Estimates of abundance and density of translocated adults ranged from 450-577 individuals and 0.09-0.11/m2 in 2011, and 371-645 individuals and 0.07-0.13/m2 in 2012. Estimates of abundance and density of laboratory-propagated sub-adults ranged from 1,678-1,943 individuals and 0.33-0.38/m2 in 2011, and 1,389-1,700 individuals and 0.27-0.33/m2 in 2012. Additionally, three recruits were collected at Site 1. No E. capsaeformis were collected at Sites 2 and 3. Capture-mark-recapture sampling produced similar mean point estimates as systematic quadrat sampling, but with typically more precision. My results indicated that the release of larger individuals (>10 mm) is the most effective technique for restoring populations of E. capsaeformis, and that systematic quadrat and capture-mark-recapture sampling have useful applications in population monitoring that are dependent on project objectives. Systematic quadrat sampling is recommended when the objective is to simply estimate and detect trends in population size for species of moderate to larger densities (>0.2/m2). Capture-mark-recapture sampling should be used when objectives include assessing a reintroduced population of endangered species or at low density, obtaining precise estimates of population demographic parameters, or estimating population size for established species of low to moderate density (0.1-0.2/m2). The ability to grow endangered juveniles to larger sizes in captivity requires improving grow-out culture methods of laboratory-propagated individuals. A laboratory experiment was conducted to investigate the effects of temperature (20-28 C) on growth and survival of laboratory-propagated juveniles of the Cumberlandian combshell (Epioblasma brevidens), E. capsaeformis, and the wavyrayed lampmussel (Lampsilis fasciola) in captivity. Results indicated that 26 C is the optimum temperature to maximize growth of laboratory-propagated juveniles in small water-recirculating aquaculture systems. Growing endangered juveniles to larger sizes will improve survival in captivity and after release into the wild. As a result, hatcheries can reduce the time that juveniles spend in captivity and thus increase their overall production and enhance the likelihood of success of mussel population recovery efforts by federal and state agencies, and other partners. / Master of Science

No description available.

The mesenchymal stromal cell (MSC) is of great interest for its immunomodulatory and regenerative properties. However, to research and use these MSCs it is essential to identify and characterize them as such. They need to fulfill the MSCs' minimal criteria which assess the differentiation potential, cell surface markers, and adherence. In this study, cells donated from human bone marrow were identified as MSC according to the minimal criteria. Methods used were flow cytometry, immunofluorescent staining, and ELISA. Furthermore, the population was cultured in three different media (DMEM-LG with either 10% FBS, 2% FBS, or 10% FBS supplemented with 10% conditioned media from human urinary bladder carcinoma cells (T24)) for 21 days whereupon tested for the mesenchymal characteristics, cells were counted and size measured at every passage. All cultures maintained their mesenchymal character, however, cells grown in 2% FBS became a considerably more heterogenous population regarding cell size and granularity, perhaps because of senescence. Additionally, these cells somewhat decreased in proliferation and resulted in 1 x 106 cells after 21 days, however, this was not a significant decrease when compared to the 10% FBS culture which had 2.16 x 106 cells after 21 days (p=0.061). On the contrary, the culture supplemented with T24 conditioned media resulted in a significantly higher cell count with 4.75 x 106 cells (p=0.008). Further studies could investigate which components in the conditioned media contributed to the proliferation. Moreover, the cell population in this study could not be characterized as MSC with certainty as additional cell surface markers should be tested.

No description available.

No description available.

Endometrial adenocarcinoma (EAC) is the most frequently diagnosed gynecological cancer of the female genital tract in the Western world. Research studies in EC is difficult to conduct on human tumor samples due to the complex nature of tumor arousal and genetic heterogeneousness in the human population. Therefore, inbred animal models can be promising tools to use in EC research due to similar histopathology and pathogenesis as humans. Studies performed on MAP2K1 and CCND1 has shown that their altered expression play a crucial role in carcinogenesis. CCND1 has been demonstrated to have oncogenic properties when overexpressed in human neoplasias. The aim of this study is to investigate gene expression levels of MAP2K1 and CCND1 in BDII rat model of endometrial adenocarcinoma cells. Quantitative real-time PCR was used to analyze expression levels of MAP2K1 and CCND1 genes in BDII/Han rat model of endometrial cancer cells using TaqMan approach. The differences in gene expression levels of MAP2K1 and CCND1 between pathologically EAC malignant and nonmalignant cells showed an upregulation of MAP2K1 and CCND1 in EAC malignant cells. The analyzed data presented observable mean differences between MAP2K1 and CCND1 in several endometrial cell lines that were examined. Although no statistical significance was reached, an alteration in gene expression levels in malignant and nonmalignant endometrial cells could be observed. Furthermore, this present study shows observable upregulation of MAP2K1 and CCND1 in endometrial carcinoma cells vs. nonmalignant endometrium cells and encourages further investigation of the role of CCND1 and MAP2K genes in endometrial carcinogenesis.