Characterization of cold shock domain proteins and SUMOylation system from Oryza sativa

In the first part of this study, two novel c&barbelow;old s&barbelow;hock domain p&barbelow;roteins from rice (OsCSP) were cloned and subsequently characterized their roles during stress conditions and development. OsCSP1 and OsCSP2 ( Oryza sativa CSD protein) encode putative proteins consisting of an N-terminal CSD and glycine-rich regions that are interspersed by 4 and 2 CX2CX4HX4C (CCHC) retroviral-like zinc fingers, respectively. Using an in vitro DNA binding assay, I demonstrate that OsCSPs exhibit conserved ssDNA binding activity. In vivo functional complementation in a cold-sensitive bacterial strain, that lacks four cold inducible cold shock domain proteins revealed that OsCSPs function as RNA chaperones, similar to their bacterial and winter wheat counterparts. To understand the functions of these genes in rice, I studied the transcriptional regulation in response to abiotic stress conditions. Under cold stress, OsCSP transcript levels are only transiently and marginally increased and the encoded proteins did not accumulate. These transcript and protein data are in sharp contrast with the observed data for winter wheat and Arabidopsis cold shock domain proteins under cold stress. In these species, both transcripts and protein levels of CSPs are increased upon cold stress. Based on these data, it can be hypothesized that the accumulation of cold shock domain proteins may play an important role in determining the cold acclimation capability of the plants. Expression analysis at the protein and RNA levels during development revealed that OsCSPs are highly expressed in the reproductive and meristematic tissues. These results indicate a potential role for rice cold shock domain proteins in plant growth and reproductive development.;In this study, I also characterized the post-translational modification of plant cold shock domain proteins by SUMOylation. Post-translational modifications can impart rapid changes in protein function. SUMOylation involves the reversible attachment of a small protein called SUMO (small ubiquitin-like modifier) to target proteins. The SUMO protein has a similar three dimensional structure as that of ubiquitin and the process of SUMOylation is very similar to that of ubiquitination. However, unlike ubiquitination, SUMOylation is not implicated in protein degradation. SUMO modification can affect the target protein stability, sub-cellular localization protein-protein interactions. Using a computational approach on rice and Arabidopsis cold shock domain proteins, I identified canonical SUMOylation motifs in both rice CSPs and one of the Arabidopsis CSPs. Using in vitro assays, I demonstrated that both OsCSPs can undergo SUMOylation. Using mutational approaches, I identified an important lysine residue for SUMOylation in Arabidopsis AtCSP1. By employing GFP-tagged wild-type and variant AtCSP1 proteins, I demonstrate that SUMOylation appears to affect AtCSP1 protein localization.;In another study, I characterized the entire SUMO conjugation system in rice. The process of SUMOylation involves a cascade of enzymatic reactions involving activation (E1) enzymes, conjugation (E2) enzymes and ligation (E3) enzymes. I compared the protein sequences of all these genes from rice with those from Arabidopsis, yeast and human. This revealed a high amino acid sequence conservation of individual SUMOylation components from yeast to plants and animals. In Arabidopsis, the SUMOylation system has been implicated in plant development and in mediating abiotic stress responses. To understand the role of the rice SUMOylation system during development, I studied the SUMO conjugate profiles and the expression of individual SUMO component genes in various tissues at different stages of plant development. The highest levels of SUMOylated proteins were observed in panicles and meristematic tissues. Expression studies revealed that SUMO component genes are highly expressed in reproductive tissues like developing seeds and panicles. Together, these data implicate an important role for the rice SUMOylation system in plant growth and reproductive development. To understand the role of SUMOylation system in rice, I studied SUMO conjugate profiles and the transcriptional regulation of individual SUMO components during cold, salt and ABA stress conditions. Rice responds to these stresses by accumulating SUMO conjugated proteins, suggesting that protein SUMOylation helps to mediate plant stress responses. Studies on the transcriptional regulation of individual SUMO pathway genes during these stress conditions revealed that most are transcriptionally down-regulated. However, a particular SUMO E3 ligase gene (OsSIZ2) is transiently up-regulated upon exposure to all three stress conditions. Considering the importance of E3 ligases in improving the efficiency and specificity of the SUMO conjugation reactions, OsSIZ2 may mediate accumulation of SUMO conjugates during these stress conditions. Taken together, these data suggest a role for SUMOylation in rice development and stress responses. (Abstract shortened by UMI.)

A Buddhist Monastery and Temple with a Religious Center in New York City

The Buddhist Temple form has remained unchanged in the last 2,500 years. The thesis will become an exploration of contemporary issues of acculturation and transition of Buddhism in the United States. The thesis will examine architecturally, the individual spirit of contemplation and meditation of a monistic religion. This allows the thesis to transform, architecturally and typologically, a Buddhist Temple and Monastery of the Theravada Sect, with a Religious Center

Supported Planar Bilayers for the Formation of Study of Immunological Synapses and Kinapse

Supported planar bilayers are powerful tools that can be used to model the molecular interactions in an immunological synapse. To mimic the interactions between lymphocytes and antigen presenting cells, we use Ni2+-chelating lipids to anchor recombinant cell adhesion and MHC proteins to the upper leaflet of a bilayer with poly-histidine tags. Planar bilayers are generated by preparing lipid, treating the glass surfaces where the bilayer will form, and then forming the bilayer in a specialized chamber containing a flow-cell where the lymphocytes will be added. Then, bilayers are charged with Ni and his-tagged recombinant proteins are added. Finally, lymphocytes are injected into the flow cell and TIRF microscopy can be used to image synapse formation and the mechanisms that control T cell locomotion, sites of receptor sorting, and sites of receptor degradation

Wireless Emitter Location Estimation Based on Linear and Nonlinear Algorithms using TDOA Technique

Low-power devices such as cell phones, and wireless routers are commonly used to control Improvised Explosive Devices (IEDs) and as the communication nodes for the sake of command and control. Quickly locating the source of these signals is ambitious, specifically in a metropolitan environment where buildings and towers may cause intervention. This presents a geolocation system that compounds the attributes of several proven geolocation and error mitigation methods to locate an emitter of interest in an urban environment. The proposed geolocation system uses a Time Difference of Arrival (TDOA) approach to estimate the position of the emitter of interest. Using multiple sensors at known locations, TDOA estimates are achieved by the cross-correlation of the signal received at all the sensors. A Weighted Least Squares (WLS) solution, Linear least Square (LLS) method and maximum likelihood (ML) estimation is used to estimate the emitter's location. If the variance of this location estimate is too high, a sensor is detected and identified as possessing a Non-Line of Sight (NLOS) path from the emitter. This poorly located sensor is then removed from the geolocation system and a new position estimate is computed with the remaining sensor TDOA information. The performance of the TDOA system is determined through modeling and simulations. Test results confirm the feasibility of identifying a NLOS sensor, thereby improving the geolocation system's accurateness in a metropolitan environment

IN VITRO CYTOTOXIC STUDY OF GREEN SYNTHESIZED GOLD AND SILVER NANOPARTICLES USING ECLIPTA PROSTRATA (L.) AGAINST HT-29 CELL LINE

Objective: The objective of this study is to synthesize gold and silver nanoparticles using the extracts of Eclipta prostrata.Methods: The nanoparticles synthesize is carried out using the powdered leaves and mixed with distilled water. The filtered extract was then mixedwith aqueous solution of HAuCl4 (1 mM) and AgNo3 (1 mM), and the reaction volume was made up to 100 ml. Then, the characterization of nanoparticleswas carried out using ultraviolet, infrared, scanning electron microscope, and the cytotoxic activity of the nanoparticles were investigated against HT-29 cancer cell lines.Results: From the study, it was found that the plant extract was able to synthesize nanoparticles, and the synthesized nanoparticles were found to betoxic against the cancer cell line HT-29.Conclusion: In the present study, both silver and gold nanoparticles were synthesized using the plant extract of E. prostrata. The synthesizednanoparticles were found to be effective against HT 29 cancer cells. The green synthesized nanoparticles were found to be cost-effective, simpler andenvironmentally safe. As the nanotechnology is an emerging field in medicine, the biological synthesis of nanoparticles helps in the other way. Fromthe present study, the nanoparticles synthesized were thus proved against various studies novely. Hope this paves way for the better development ofnanoparticle production in the large scale amount.Keywords: Nanoparticles, Eclipta prostrata, Scanning electron microscope, HT-29 cell line. §Both authors are equally contribute

WALNUT PEDUNCULAGIN A PROBABLE SERM FOR BREAST CANCER TREATMENT

Walnuts constituents have been actively used in nutrition in slowing cancer growth by its anti-proliferative and anti-angiogenic mechanisms. Pedunculagin, a natural chemical constituent of walnut were performed to check for its binding with Estrogen Receptor (ER) receptor using Insilco approach. Most of current approach towards breast cancer therapies is aimed at blocking ER signaling pathway using various estrogen antagonists like synthetic estrogen receptor modulators (SERMs) which down regulates the signaling cascade leading to tumor activity. This study is aimed to find the binding interactions between Pedunculagin and ER. Pedunculagin structure was extracted from CHEMSPIDER database and Estrogen Receptor (ER) 1QKN was selected, further ligand and receptor structures were minimized with steepest descent and conjugate gradient with minimized energy levels were found to be -14211.04235 Kcal/mol for ligand molecule and -12518.66882 Kcal/mol for ER. Six interacting binding sites were identified between ligand and ER and docking studies showed second active binding site of receptor showed strong affinity and interacting groups. Pedunculagin may act as SERM in modulating the ER signaling activity and may be a probable therapeutic molecule for treating breast cancer

Edge-of-grid voltage control in distribution networks

As the electric power supply systems are undergoing major changes with the integration of renewables, the issues related to voltage regulation and system protection are arising. In this scenario, advanced voltage regulation technologies that provide voltage control at the grid-edge, that is at the low-voltage secondary side of the distribution circuit, have emerged as a potential solution to address the shortcomings of traditional voltage control practices in distribution systems. In this work, these technologies are modeled and algorithms are developed to strategically deploy them, tune their control parameters, and evaluate their voltage regulation performance. A two-stage optimization framework is proposed for optimal placement and real-time control of the low-voltage static var compensators to minimize the energy losses while maintaining the voltage regulation. Integration of high levels of distributed generation such as photovoltaic (PV) systems impacts the voltage regulation by causing steady-state voltage variations and transient voltage fluctuations. This work further develops a procedure to tune the control parameters of PV smart inverters to mitigate these voltage issues. Furthermore, the PV penetration levels in a distribution network can be increased without creating voltage problems by dynamic controlled reactive power absorption at several strategic buses. This concept is modeled and demonstrated in this work. Furthermore, the high levels of PV generation can interfere with the overcurrent protection schemes prevalent in distribution networks. An analytical approach is proposed in this work to estimate the distribution feeder PV accommodation limits with respect to overcurrent protection issues as the impact criteria, without needing to simulate numerous PV screening scenarios to assess the impactElectrical and Computer Engineerin

Is emergency cesarean section more risky than elective cesarean section in women with previous cesarean section?

Background: Cesarean section is one of the most performed surgical procedures all over the world, but unfortunately cesarean sections are associated with a great deal of maternal morbidity and mortality. In the past the rate of cesarean section has increased for many avoidable and unavoidable indications both in developed and developing countries. The objective of this study was to compare maternal morbidity and mortality in elective repeat cesarean section (El-RCS) and emergency repeat cesarean section Em-RCS.Methods: Prospective data was recorded on management practices, associated complications and morbidity and mortality on 15664 consecutive cases of previous cesarean section reporting at 30 medical colleges/teaching hospitals for delivery.Results: Of the 15664 women with a previous cesarean section, 5399 (34.5%) women underwent elective repeat cesarean section, 7752 (49.5%) women who underwent emergency repeat cesarean section and 2513 (16.0%) had successful trial of labor (S-TOL). There was    failed trial of labor (F-TOL) in 1522 cases and requiring an emergency cesarean section for delivery of baby. Therefore, total no. of 7752 women had an emergency cesarean section. The overall maternal morbidity was 22.5%, 20.7% in Em-RCS and El-RCS respectively. Blood loss was more than 1000ml in 7.2% of Em-RCS where as in El-RCS it was 8.8%, blood transfusion was 7.5% in Em-RCS where as it was 6.5% in El-RCS, dehiscence of scar in Em-RCS was 4.7% as compared to 2.2% in El-RCS, uterine rupture was 1.2% in Em-RCS as compared to 0.7 % in El-RCS found statistically significant. Post-operative complication was 5.9% cases in Em-RCS where as in El-RCS was 5.8% (p=0.79 non-significant). Maternal mortality was reported in 12 (0.2%) cases of Em-RCS as compared to 5 (0.1%) cases in El-RCS (p=0.37) which was not statistically significant.  Conclusions: Maternal morbidity was found more in emergency repeat cesarean section than in elective repeat cesarean section. Complications and referral of women who are likely to undergo cesarean section should be diagnosed at an early stage so that the maternal   morbidity and mortality can be prevented.