Gene array profiling of large hypothalamic CNS regions in lactating and randomly cycling virgin mice

A dramatic example of neuronal and physiological plasticity in adult mammals occurs during the transition from a non-maternal to a maternal, lactating state. In this study we compared gene expression within a large continuous region of the CNS involved in maternal behaviors (hypothalamus, preoptic regions, and nucleus accumbens) between lactating (L) (postpartum Day 7) and randomly cycling virgin (V) outbred mice. Using high density oligonucleotide arrays representing 11,904 genes, two statistical algorithms were used to identify significant differences in gene expression: robust multi array (p \u3c 0.001) (n = 92 genes) and significance analysis of microarrays using a 10% false discover rate (n = 114 genes). 27 common genes were identified as significant using both techniques. A subset of genes (n = 5) were selected and examined by real-time PCR. Our findings were consistent with previous published work. For example, neuropeptide Y (NPY) and proenkephalin were elevated in L mice, whereas POMC was decreased. Increased levels of NPY Y2 receptor and polo-like kinase and decreased levels of endothelin receptor type b in L mice are examples of novel gene expression changes not previously identified. Expression differences occurred in broad classes. Together, our findings provide possible new material on gene expression changes that may support maternal behaviors. The advantages and drawbacks of sampling large CNS regions using arrays are discussed

The Forum: Spring 2003

Spring 2003 journal of the Honors Program at the University of North Dakota. The issue includes stories, poems, essays and art by undergraduate students.https://commons.und.edu/und-books/1051/thumbnail.jp

Transcriptome Analysis of Epithelial and Stromal Contributions to Mammogenesis in Three Week Prepartum Cows

Transcriptome analysis of bovine mammary development has provided insight into regulation of mammogenesis. However, previous studies primarily examined expression of epithelial and stromal tissues combined, and consequently did not account for tissue specific contribution to mammary development. Our objective was to identify differences in gene expression in epithelial and intralobular stromal compartments. Tissue was biopsied from non-lactating dairy cows 3 weeks prepartum, cut into explants and incubated for 2 hr with insulin and hydrocortisone. Epithelial and intralobular stromal tissues were isolated with laser capture microdissection. Global gene expression was measured with Bovine Affymetrix GeneChips, and data were preprocessed using RMA method. Moderated t-tests from gene-specific linear model analysis with cell type as a fixed effect showed more than 3,000 genes were differentially expressed between tissues (P<0.05; FDR<0.17). Analysis of epithelial and stromal transcriptomes using Database for Annotation, Visualization and Integrated Discovery (DAVID) and Ingenuity Pathways Analysis (IPA) showed that epithelial and stromal cells contributed distinct molecular signatures. Epithelial signatures were enriched with gene sets for protein synthesis, metabolism and secretion. Stromal signatures were enriched with genes that encoded molecules important to signaling, extracellular matrix composition and remodeling. Transcriptome differences also showed evidence for paracrine interactions between tissues in stimulation of IGF1 signaling pathway, stromal reaction, angiogenesis, neurogenesis, and immune response. Molecular signatures point to the dynamic role the stroma plays in prepartum mammogenesis and highlight the importance of examining the roles of cell types within the mammary gland when targeting therapies and studying mechanisms that affect milk production

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Health professionals’ perceptions of weight loss programmes and recommendations for future implementation: a qualitative study

Objective Obesity is an increasing public health challenge and most weight loss programmes are still inadequate to support sustainable weight loss. One reason for the continued lack of success might be the dominant biomedical, individualised approach to weight loss. Holistic approaches that focus on overall health and well-being in addition to weight loss are increasingly recommended. In Denmark, health professionals in the municipalities are responsible for developing and conducting weight loss programmes. The objective of this study was to explore what health professional’s perceived as an ideal, holistic weight loss programme that could be feasibly implemented in the municipalities.Design A phenomenological-hermeneutical qualitative study was performed using semistructured interviews.Setting Thirty-two Danish municipalities were weight loss programmes are developed and conducted.Participants Thirty-five health professionals with experience conducting weight loss programmes.Results Three themes emerged from the analysis: Support from the social network are important both during and after a weight loss, Changing the self-belief by positive discussions and doing activities, Maintaining changes through daily life.Conclusion Future municipal weight loss programmes should emphasise overall health and well-being instead of weight loss and adopt a holistic approach including a focus on social relationships, meaningful activities and successes as part of a balanced daily life

Gene array profiling of large hypothalamic CNS regions in lactating and randomly cycling virgin mice

A dramatic example of neuronal and physiological plasticity in adult mammals occurs during the transition from a non-maternal to a maternal, lactating state. In this study we compared gene expression within a large continuous region of the CNS involved in maternal behaviors (hypothalamus, preoptic regions, and nucleus accumbens) between lactating (L) (postpartum Day 7) and randomly cycling virgin (V) outbred mice. Using high density oligonucleotide arrays representing 11,904 genes, two statistical algorithms were used to identify significant differences in gene expression: robust multi array (p This is a manuscript of an article from Molecular Brain Research 139 (2005): 201, doi: 10.1016/j.molbrainres.2005.05.011. Posted with permission.</p

Early historical forest clearance caused major degradation of water quality at Lake Vaeng, Denmark

Although humans have impacted their environment over millennia, details of these impacts, especially on aquatic systems, is still surprisingly scarce despite potential disturbance by early land use. This study examined a high-resolution radiocarbon-dated Holocene record from the Danish Lake Vaeng, using geochemical and biological proxies, and related the observed impacts to other lake records with catchment disturbance. The results indicate a lengthy and varying history of aquatic eutrophication linked to human activity. Modest impacts on the lake coincided with the first signs of landscape disturbance during the Neolithic (c. 4500 cal. yrs BP). Observed impacts intensified in the Late Bronze and Pre-Roman Iron Age. Viking Age/Medieval deforestation and erosional inputs to the lake associated with new ploughing technology (1200 cal. yrs BP), however, led to a major reorganisation of the aquatic ecosystem. Filamentous bloom-forming cyanobacteria, common today in heavily culturally impacted lakes, reached a historical maxima. The lake ecosystem subsequently recovered somewhat but remains eutrophic to date. The erosion record from Lake Vaeng shows a striking similarity with other Danish lake records, especially the notable increase in Medieval Period catchment inputs, which are observed in other European lacustrine records. Numerous European lowland lakes may have shifted into a degraded ecological state millennia ago, but degradation intensified during the onset of the Medieval Period. Hence, assuming preindustrial conditions as relatively pristine reference baselines for more recent cultural eutrophication could be flawed in landscapes intensively used by humans for millennia.Peer reviewe