DIFFERENTIAL ABILITIES OF THE CHICKEN PIT1 ISOFORMS TO REGULATE THE CHICKEN GROWTH HORMONE PROMOTER

Pit1, a pituitary-specific transcription factor, regulates differentiation of cells of the PIT1 lineage in the anterior pituitary. PIT1 also regulates the synthesis of peptide hormones from these cell types, including growth hormone (GH). A founding member of the POU-homeodomain family of transcription factors, PIT1 is characterized by a serine-threonine rich N-terminal transactivation domain and a C-terminal POU-domain. Alternative forms of PIT1, differing from each other in the N-terminal domain have been reported in several species, but the functional implication of having multiple isoforms is not known. Several Pit1 isoform mRNAs exist in chickens which have not been characterized. The main aim of this study was to determine which, if any, of the chicken PIT1 isoforms regulated the chicken Gh (cGh) promoter. PIT1β2, a novel isoform of chicken PIT1 was discovered, and known and novel isoforms (PIT1α, PIT1β1, PIT1β2 and PIT1γ) were characterized. A luciferase reporter construct containing 1775bp of the cGh promoter driving expression of firefly luciferase was used to determine the ability of the isoforms to regulate the target gene promoter activity in chicken LMH cells. We showed that three of the isoforms, PIT1α, PIT1β1 and PIT1β2, expressed from recombinant plasmids, regulated the cGh promoter, while PIT1γ did not. All the isoforms localized to the nucleus in both non-pituitary and pituitary cells. Results from gel-shift assays show that PIT1γ did not bind the proximal PIT1-binding site of the cGh promoter as well as the other isoforms, suggesting a possible mechanism behind the inactivity. Our result did not suggest a negative regulatory role for this isoform. In contrast, we found a functional advantage for having multiple isoforms. PIT1β1, the isoform that activated the promoter most strongly, when co-transfected with other activating isoforms, such as PIT1α and PIT1β2, induced significantly higher level of activation than one isoform alone. Whether this increased activation required, or was facilitated by, heterodimerization of two isoforms is not known. Nevertheless, identification of isoforms with specific functions will facilitate identification of their respective interacting partners, which are essential for GH gene expression

Showcasing New Tourism Destination by Using Gis: a Study of Sikkim

Purpose: The main purpose of this research is to find out new tourist destinations and suggest developmental strategies through scientific ways for solving overtourism-related problems.   Theoretical framework: In this research paper, the researchers emphasized finding new tourist spots and analyzing their potential both in touristic and infrastructural value was assessed by the researcher with the help of a most modern tool like Geographic Information System. The researchers have explained a scientific process of exploring new tourist destinations or tourist spots by using GIS and have shown how to develop proper infrastructure around those spots.   Design/methodology/approach: A “Multicriteria Spatial Decision Support System modeling” was used to find out and validate the existing and new tourist spots. In the pair-wise comparison method, a criterion versus criteria was created to compare each pair of criteria and assign relative ratings using the scale of pair-wise comparison. In this research, the Simple Additive Weighting (SAW) method is used because it gives the “most acceptable results for majority of single-dimensional problems”. The researcher has created this map to support the ‘Destination Fetching Model’, and it can also be used for promoting sustainable tourism in the area. If the entire land cover is visible to the planners with an aerial view, they can plan for tourism infrastructure and superstructures from anywhere. GIS-based layer data has created an "all covered" aerial map with the help of advanced GIS technology, which not helped the planners to understand the current land condition, but also showed the open area which can be utilized for new tourism set-up. Manmade and natural resources are also classified through this LULC model.   Findings: These new tourist spots will help planners and stakeholders to distribute excessive crowds to comparatively new destinations. It is observed that most of the Indian tourism destinations suffer from “over Tourism” in peak season. This research will help spread the flow of tourists to the nearest newly added destinations.   Research, Practical & Social implications: Scientific tourism planning through GIS will also help tourism service providers and planners to add infrastructure to the new destinations, and would also help local people to generate income through tourism. Originality/value: This research will solve the “over tourism” problem of any destination or state, and help generate more visitor’s day at the destination which is proportionate to extra revenue generation for the local, regional and national tourism stakeholders

Partogram: an important tool in managing labour!

Background: Various types and designs of partographs are being used at various centers. WHO introduced simplified version of partogram, for the use by skilled birth attendant. Preprinted paper versions of the partograph are availableMethods: 100 antenatal women were selected for study. Patients with vertex presentation and singleton pregnancy were taken. Patients who came late in labour and those with cephalopelvic disproportion were not included in the study. Cervical Dilatation in cms was assessed by per vaginal examination every 2 hourly, fetal Heart Rate every ½ hourly, uterine contractions and maternal pulse measured every ½ hourly, maternal BP and temperature were measured every 4th hourly.Results: Using WHO simplified partogram, characteristics of labour and neonatal outcome was evaluated. Among the 100 women included in the study, 78 required augmentation of labour, here 6 of them crossed the alert line and underwent LSCS. Of the 22 women who did not require augmentation, 4 crossed the alert line and underwent LSCS.We didn’t find any difference in monitoring of labour using simplified version of WHO partogram compared with other partograms, apparently it’s more simpler to plot and easy to understand.Conclusions: An alert line on partogram should be based on lower 10th centile rate of cervical dilatation of the local population. We found this rate as 1cm/hr, which corresponds to the slope of alert lineon standard partogram. Based on this we conclude, simplified partogram is good enough for monitoring labour progress

Cooperativity of Rb, Brca1, and p53 in malignant breast cancer evolution

Breast cancers that are triple-negative for the clinical markers ESR1, PGR, and HER2 typically belong to the Basal-like molecular subtype. Defective Rb, p53, and Brca1 pathways are each associated with triple-negative and Basal-like subtypes. Our mouse genetic studies demonstrate that the combined inactivation of Rb and p53 pathways is sufficient to suppress the physiological cell death of mammary involution. Furthermore, concomitant inactivation of all three pathways in mammary epithelium has an additive effect on tumor latency and predisposes highly penetrant, metastatic adenocarcinomas. The tumors are poorly differentiated and have histologic features that are common among human Brca1-mutated tumors, including heterogeneous morphology, metaplasia, and necrosis. Gene expression analyses demonstrate that the tumors share attributes of both Basal-like and Claudin-low signatures, two molecular subtypes encompassed by the broader, triple-negative class defined by clinical markers

Chimeric Antigen Receptor Signaling Domains Differentially Regulate Proliferation and Native T Cell Receptor Function in Virus-Specific T Cells

The efficacy of T cells expressing chimeric antigen receptors (CARs) for solid tumors has been limited by insufficient CAR T cell expansion and persistence. The use of virus-specific T cells (VSTs) as carriers for CARs may overcome this limitation since CAR-VSTs can be boosted by viral vaccines or oncolytic viruses. However, there is limited understanding of the optimal combination of endodomains and their influence on the native T cell receptor (TCR) in VSTs. We therefore compared the function of GD2.CARs expressing the TCR zeta chain (ζ) alone or combined with endodomains from CD28 and 4-1BB in varicella zoster virus-specific (VZV) T cells. VZVSTs expressing GD2-CARs recognized VZV-derived peptides and killed GD2-expressing tumor cells. However, after repeated stimulation through their native TCR, the expansion of GD2-CAR.CD28ζ-VZVSTs was 3.3-fold greater (p < 0.001) than non-transduced VZVSTs, whereas GD2-CARζ- and GD2-CAR.41BBζ inhibited VZVST expansion (p < 0.01). Compared to control VZVSTs, GD2-CAR.ζ VZVSTs showed a greater frequency of apoptotic (p < 0.01) T cells, whereas prolonged downregulation of the native αβ TCR was observed in GD2-CAR.41BBζ VZVSTs (p < 0.001). We confirmed that CD28ζ can best maintain TCR function by expressing GD2.CARs in Epstein-Barr virus-specific T cells and CD19-CARs in VZVSTs. In response to CAR stimulation VSTs with CD28ζ endodomains also showed the greatest expansion (6 fold > GD2-CAR.41BBζ VZVSTs (p < 0.001), however anti-tumor efficacy was superior in GD2-CAR.41BBζ-VZVSTs. These findings demonstrate that CAR signaling domains can enhance or diminish the function of the native TCR and indicate that only CD28ζ may preserve the function of the native TCR in tonically signaling CAR-VSTs

Development and Characterization of Mouse Models for Breast Cancer

While previous studies using genetically engineered mice (GEM) have indicated potential effects of several aberrations observed in human breast cancer, the combined role of loss of RBf, P53 and BRCA1 has not been assessed before. As these events frequently occur together in human breast cancer, we use GEMs to show that these pathways have a synergistic tumor suppressor role in the mammary gland. We show that loss of Rbf alone is not enough to promote mammary tumors, but combined loss of Rbf and p53 can lead to mammary adenocarcinoma with a reduction in apoptosis and low latency. In addition to p53 we studied the role of Brca1 loss either in conjunction with loss of Rbf or with both Rbf and p53. We found that these three important tumor suppressors have a synergistic effect in mammary tumor progression and combined loss of all three further reduces mammary tumor latency and leads to progression to distant metastasis. Genomic analysis suggests that the combined loss of Rbf, p53 and Brca1 results in increased genetic instability, overexpression of metastasis promoting genes and an altered micro RNA profile that may also contribute to increased malignancy. So far analysis of genetic lesions in human cancers has focused on the accumulation of multiple events within the epithelial cell. However it is still unresolved as to why human BRCA1 mutated cancers have their characteristic "basal" like features. We show here for the first time, that the target cell of origin of the Brca1 mutation determines the nature of the tumor that evolves and hence the right combination of genetic mutations and cell of origin is important in modeling human cancers. All of our mouse models generated in this study can be used as preclinical animal models, which both genetically and biologically model the initiation and progression of human breast cancer.Doctor of Philosoph

Notch Signaling in Kidney Development, Maintenance, and Disease

Kidney development involves formation of nephrons intricately aligned with the vasculature and connected to a branched network of collecting ducts. Notch signaling plays multiple roles during kidney development involving the formation of nephrons composed of diverse epithelial cell types arranged into tubular segments, all the while maintaining a nephron progenitor niche. Here, we review the roles of Notch signaling identified from rodent kidney development and injury studies, while discussing human kidney diseases associated with aberrant Notch signaling. We also review Notch signaling requirement in maintenance of mature kidney epithelial cell states and speculate that Notch activity regulation mediates certain renal physiologic adaptations