Understanding Hydrogen-Bond Patterns in Proteins using a Novel Statistical Model

Proteins are built from basic structural elements and their systematic characterization is of interest. Searching for recurring patterns in protein contact maps, we found several network motifs, patterns that occur more frequently in experimentally determined protein contact maps than in randomized contact maps with the same properties. Some of these network motifs correspond to sub-structures of alpha helices, including topologies not previously recognized in this context. Other motifs characterize beta-sheets, again some of which appear to be novel. This topological characterization of patterns serves as a tool to characterize proteins, and to reveal a high detailed differences map for comparing protein structures solved by X-ray crystallography, NMR and molecular dynamics (MD) simulations. Both NMR and MD show small but consistent differences from the crystal structures of the same proteins, possibly due to the pair-wise energy functions used. Network motifs analysis can serve as a base for many-body energy statistical energy potential, and suggests a dictionary of basic elements of which protein secondary structure is made

Role and immunomodulatory profile of histamine receptors by H1 and H2 antagonists

The present study was designed to delineate the immunomodulatory role of histamine receptors (H1- and H2-) on induction of antibody response to sheep red blood cells (SRBC), as well as the antibody generation profile, in rabbit system, systemically. The rabbits in two groups received pheniramine (H1-receptor antagonist) and ranitidine (H2-receptor antagonist), respectively, via intramuscular route and were immunized with SRBC intravenously to evaluate suppression or enhancement of antibody responses in sem. A third, control group, received vehicle and were immunized in a similar manner. Histamine released from effector cells (mast cells and basophils) _in vivo_ during inflammatory reactions could influence a detectable antibody response to SRBC as early as day 7-postimmunization (post-I), which lasted until day 28- post-I. Pheniramine-treated rabbits had significantly (*Pa ≤ 0.05 and **Pa ≤ 0.01) more suppressed total serum antibody (IgM + IgG) to SRBC as compared to ranitidine-treated ad cotrol rabbits, while ranitidine-treated rabbits showed different pattern (suppressed or enhanced) during the whole study period. Ranitidine suppressed total antibody level at days 7- and 14- post-I, and enhanced at days 21- and 28- post-I. IgM suppression at day 7- and enhancement at days 14-, 21- and 28- post-I, while IgG suppression during whole study period, as compared to control group was significant (*Pa ≤ 0.05 and **Pa ≤ 0.01) as assessed by direct hemagglutination assay* ad whole SBC-ELISA method**. Here we report that histamine receptor type 2 (H2R)-antagonists have a dominant role on immunosuppression and in immunoregulation of humoral immune responses. Histamine receptor type 2 (H2R)-antagonists are mainly involved in B cell differentiation and proliferation over histamine receptor type 1 (H1R)-antagonists

Continuing teacher professional development in post-Soviet Kyrgyzstan

After the collapse of the USSR, Kyrgyzstan became an independent Central Asian state in 1991. Since its independence, there have been enormous changes in the political, social and economic life of the young state, which strives to become a democratic and aspiring market-oriented economy. Its education system has also begun experiencing changes. However, most of the reforms brought in this area continue to be short-term and ever-changing. Economic hardship does not allow the continuation of the previously state-funded teacher retraining system, which further creates deteriorating quality education in the schools. On one hand, the country continues to follow the Soviet in-service teacher education system and procedures that became hard to accomplish. On the other hand, ‘lip service’ reforms continue to irritate teachers who are left in uncertainty and despair. This paper examines the existing continuing teacher professional development programs in post-Soviet Kyrgyzstan and identifies related issues of quality, access and lack of structure. These all reflect the overall economic decline of the region, which is one of the impoverished Central Asian states. It brings out the necessity for more teacher needs-based programs and the acknowledgement that teacher self-initiated programs/approaches to professional development are necessary

Functional and Mechanical Role of Splice Variant of Mucin4 (MUC4/X) and Trefoil Factors in Pancreatic Cancer Pathogenesis

Pancreatic Cancer (PC) is one of the vicious cancers as it ranks third in the race of leading cause of cancer-related death. Lack of early diagnostic marker, poor understanding of molecular mechanism of the disease and failure to conventional chemotherapy makes this disease dreadful. Mucin 4 (MUC4), a high molecular weight glycoprotein is one of the top differentially expressed molecules in PC while not expressed in normal pancreas. Accumulating evidence from our lab suggested its tumorigenic role in PC by increasing cell proliferation, invasion, chemotherapy resistance, tumor growth, and metastasis. Previously, our lab and other has identified 24 different splice variant of MUC4 among them MUC4/X is devoid of exon 2 and 3 and MUC4/Y is devoid of exon 2. Exon 2 encodes for the largest domain of MUC4 suggesting that MUC4/X is devoid of the largest domain of MUC4 which variable tandem repeat. Though lots of effort has been made to identify its role in PC, there is still a gap on understanding its splice variant in PC as splice variant has an invaluable role in tumor pathogenesis. Recently splice variant has emerged as one of the key players for tumorigenesis and MUC4 is one of the key players for PC pathogenesis, we aim to identify the functional and mechanical role of MUC4/X, a splice variant which is devoid of the largest domain of MUC4 yet contains all other functional domain, in PC pathogenesis. Thus, in this part of dissertation, we sought to identify the role of splice variant MUC4/X, a unique splice variant of wild-type MUC4 which contain all functional domain except largest tandem repeat. First, we identified that, MUC4/X in aberrantly expressed in poorly differentiated PC clinical sample. Then our invitro experimental evidence suggested overexpression of MUC4/X in PC cells is involved in increased cell proliferation, invasion and metastasis. Moreover, our orthotopic transplantation system also corroborated our in-vitro findings which showed increased volume of tumor and metastasis to distant organ. Using inducible tet-on system to overexpress MUC4/X in the presence of WT-MUC4 in CAPAN-1 cells, we identified that MUC4/X has increased cell proliferation and invasion suggesting their role as tumorigenic alone as well as in the presence of WT-MUC4. Our mechanical investigation indicate that overexpression of MUC4/X led to upregulation of integrin β1-FAK-ERK pathway which might be potential mechanism for MUC4/X mediated PC tumorigenesis. Lack of early effective diagnostic marker and resistance to chemotherapy are the major reasons for poor PC patient outcome. There is a pressing need to identify highly specific and sensitive biomarker as well as precise understanding of chemoresistance of PC. Trefoil factors (TFFs) are small secretory molecules mostly associated with mucin. Their primary role is to protect gastrointestinal tract partnering with mucin. Report on aberrant expression, potential as biomarker and role in tumorigenicity has conveyed for many cancers, however, their role in PC is still elusive. Recently they have emerged as a part of gene signature of classical subtype of PC, a subtype which showed gemcitabine resistance towards PC. As it is high time to identify effective biomarker and understanding the role of chemoresistance in PC, in this part of my thesis, we focused to evaluate TFFs diagnostic potential using a training and validation cohort of PC clinical sample. Here, we comprehensively investigated the diagnostic potential of all the member of trefoil family, i.e., TFF1, TFF2, and TFF3 (TFFs) in combination with CA19.9 for detection of PC. In silico analysis of publicly available datasets and expression analysis from human and spontaneous PC mouse model revealed a significantly increased expression of TFFs in precursor lesions and PC cases. Additionally, we performed a comprehensive analysis in the sample set (n= 377) comprising of independent training and validation set using ELISA consisted of benign controls (BC), chronic pancreatitis (CP), and various stages of PC. Our analysis revealed that TFF1 and TFF2 were significantly elevated in early stages of PC in comparison to BC (P Additionally, we also aim to identify the molecular landscape of TFFs role in gemcitabine resistance of PC which integrates analyzing publicly available cancer genome dataset, dissecting transcriptomic and signaling pathways and identification of biochemical interaction. From TCGA database analysis revealed a significant positive correlation between TFF1 and GR predictor of PC (P=0.0001). Our in vitro studies showed that SW1990-TFF1-KD cells induced apoptosis, reduced colony formation capacity and modulated many apoptotic regulators such increase of cleaved caspases and decrease of CIAP in the presence of gemcitabine. Furthermore, TFF1 was observed to be colocalized with MUC5AC, in human and mouse PC tissues suggesting their partnering are critical for PC pathogenesis. Interestingly, our chromatin immunoprecipitation indicates that 16 fold enrichment of GATA-6, an overexpressed transcription factor in classical subtype of PC, was observed on two distinct TFF1 promoter sites and GATA-6-siRNA repressed expression of TFF1. Moreover, protein-protein docking studies revealed the interaction of TFF1 with CXCR4 at Phe-172, Ser-122 and Glu-1 and TFF1 recombinant protein treatment in SW1990 cells increased CXCR4 mediated downstream signaling critical for GR. In this part, our overall data demonstrate that TFF1 may play a crucial role in gemcitabine resistance which is regulated by GATA6 and by interacting with MUC5AC