Lipodendriplexes: A promising nanocarrier for enhanced nucleic acid delivery

The main objective of the current study was to developed an optimized system for enhanced nucleic acid delivery with minimum cytotoxicity. For efficient therapeutic gene delivery, PAMAM dendrimer, with ethylenediamine core was chosen due to its biodegradable and non-immunogenic nature. However, the cytotoxicity and unusual biodistribution by this polycationic polymeric system urged the need of protective shielding. Therefore, a non-covalent interaction of dendriplexes with lipids i.e. liposomal modification was chosen as a technique to overcome the associated drawbacks. The methodology section of the thesis deals with the preparation of liposomes, dendriplexes, lipodendriplexes, their subsequent physicochemical characterization and in vitro, in ovo and in vivo studies. In the results section, the characterization of dendriplexes has been discussed, which was the prerequisite of lipodendriplex formation. Dendriplexes formation was confirmed by gel retardation and fluorescence quenching assay. Dynamic light scattering and laser Doppler anemometry also confirmed the stable complex formation, with a desired size range suitable for cellular internalization. Based on highest pDNA transfection efficiency and appropriate cell viability profile, an N/P ratio of 12 has been chosen for complexation with the liposomal formulation. A broad range of liposomal formulations has been investigated and the influence of liposome to PAMAM ratios on surface charge and size of lipodendriplexes are described in detail. Surface morphology of the liposomes and lipodendriplexes has been analysed using atomic force microscopy and their sizes were compared with the results obtained from dynamic light scattering. Transfection studies have been discussed for different lipodendriplexes formulations. It has been observed that the DPPC:CH-PAMAM lipodendriplexes showed a significant improvement in pDNA transfection as compared to other lipodendriplexes formulations and their parent dendriplexes. The higher gene expression was also confirmed using fluorescence microscopy by GFP expression analysis. Cytotoxicity studies including MTT, ROS, lysosomal disruption and DNA damage assays has been discussed in detail. From the results, it has been depicted that the lipid modification of dendriplexes shields the terminal amino group induced toxicity and consequently improves the cell viability. Biocompatibility studies has been performed to check to the compatibility of the complexes in the presence of blood components. Heparin competition and erythrocyte hemolysis assay revealed the biocompatible nature of the lipodendriplexes. Another objective of the study was to deliver the siRNA to investigate the gene silencing effect. Knockdown experiments showed a pronounced downregulation of luciferase, GFP and MDR1 genes by lipodendriplexes compared to dendriplexes or the control siRNA groups. The role of MDR1 in cell migration and colonization of cancer cells has also been described in detail. Downregulation of MDR1 gene by lipodendriplexes exhibited significant inhibition of tumor metastasis and colonization. The knockdown effect of MDR1 gene was also investigated in 3D cell culture environment. Cell migration and ring closure assays were performed using 3D tumor spheroid and ring bioprinting model. A significant reduction in the cell migration was observed in the case of lipodendriplexes treated group. Next step was to investigate the enhanced intracellular accumulation of tyrosine kinase inhibitor (imatinib mesylate) after the downregulation of P-gp (responsible for drug efflux) and subsequent apoptosis in colon carcinoma. Apoptosis assay was done in 2D and 3D cultures using flow cytometry and live dead staining, respectively. The results of 2D culture were in agreement with the data from 3D cell cultures. Cell cycle analysis also confirmed the imatinib mesylate induced apoptosis, indicated by an arrest of Sub-G1 phase depicting an effective downregulation of P-glycoprotein by lipodendriplexes. In order to minimise the unethical use of animals, an alternative in ovo chorioallantoic membrane model (an in vivo like environment) has been used to determine the efficacy and biocompatibility of the complexes. Lipodendriplexes exhibited successful reporter gene (GFP) expression in the CAM with no toxicity observed within the CAM vasculature. After establishing the improved gene transfection and toxicity profile in in vitro conditions, in vivo biodistribution and toxicity assessment of the complexes has performed in female BALB/c mice and discussed in detail. In vivo biodistribution has revealed that encapsulation of dendriplexes with liposomes has essentially increased the cellular uptake of the complexes, which was confirmed by ex vivo imaging of the dissected organs. Acute toxicity studies showed that the lipodendriplexes treated group did not produce any change in body weight, organ to body ratio and in organ tissue histopathology. Serum biomarkers and hematological studies also revealed the biocompatibility in an in vivo environment. From the findings in this thesis, it can be concluded that development of such non-viral nano carrier system could serve for an efficient gene transfection with better safety profile, both for in vitro and in vivo therapeutics. Further in vivo studies using different pre-clinical models for specific targeted delivery against different types of cancer and genetic disorders will help to realise the therapeutic potential of this system

Psychological Evaluation of Sports Persons with Disability

The purpose of the present study was to compare the sensation seeking and anxiety state of sports person with impairment vision problem. The total thirty (15Cricketers and 15 Sprinters) male Open National tournament players were selected for this study. The age of the subjects were ranged between 18 to 25 years. The data on sensation seeking and anxiety state of the subjects were obtained by using a questionnaire developed by Neary and Zuckerman (1976). The t test was used to determine the difference between the mean score of the cricketers and sprinters. Results revealed that there was a significant difference between Cricketers and sprinters in their sensation seeking and anxiety state at 0.05 level of significance with 28 degree of freedom. Study showed that cricketers have higher level of sensation seeking and anxiety state as compared to sprinters. Key Words: Persons with vision impairment, Cricketers, Sprinters, Sensation Seeking and Anxiety State

Psychological Evaluation of Sports Persons with Disability

The purpose of the present study was to compare the sensation seeking and anxiety state of sports person with impairment vision problem. The total thirty (15Cricketers and 15 Sprinters) male Open National tournament players were selected for this study. The age of the subjects were ranged between 18 to 25 years. The data on sensation seeking and anxiety state of the subjects were obtained by using a questionnaire developed by Neary and Zuckerman (1976). The t test was used to determine the difference between the mean score of the cricketers and sprinters. Results revealed that there was a significant difference between Cricketers and sprinters in their sensation seeking and anxiety state at 0.05 level of significance with 28 degree of freedom. Study showed that cricketers have higher level of sensation seeking and anxiety state as compared to sprinters. Key Words: Persons with vision impairment, Cricketers, Sprinters, Sensation Seeking and Anxiety State.

Novel Low Power and Low Transistor Count Flip-Flop Design with High Performance

The paper proposed a new design of static SET flip-flop for low power applications. In this work, comparative analysis of existing architecture for flip-flops along with the proposed design is made. The comparison is done on the basis of power and power delay product, transistor count is also included. Due to continuous increase in integration of transistors and growing needs of portable equipments, low power design is of prime importance. The proposed design has the best power and the second best PDP than the existing architectures. Proposed FF has the least transistor count hence reducing the manufacturing cost and area. All simulations are performed on TSpice using BSIM models in 130 nm process node. The simulation results show that for all supply voltages, proposed FF has the best power consumption, second best PDP and the lowest transistor count. So this design is best suited for low power and high performance portable applications. Keywords: Transmission Gate, Short circuit current, Edge Triggered, Optimizatio

A New Area and Power Efficient Single Edge Triggered Flip-Flop Structure for Low Data Activity and High Frequency Applications

In this work, a new area and power efficient single edge triggered flip-flop has been proposed. The proposed design is compared with six existing flip-flop designs. In the proposed design, the number of transistors is reduced to decrease the area. The number of clocked transistors of the devised flip-flop is also reduced to minimize the power consumption. As compared to the other state of the art single edge triggered flip-flop designs, the newly proposed design is the best energy efficient with the comparable power delay product (PDP) having an improvement of up to 61.53% in view of power consumption. The proposed flip-flop also has the lowest transistor count and the lowest area. The simulation results show that the proposed flip-flop is best suited for low power and low area systems especially for low data activity and high frequency applications. Keywords: PDP, reliability, delay, process node, clock frequenc

Evaluation of Selection Criteria of Sports Persons in Eastern Region of SAI, India

The intent of this study was to evaluate the organizational structure, administrative frame work and facilities related to selection criteria of   Sports Authority of India, Eastern region, Kolkata. The sample of the present study was drawn randomly from the administrator, coaches and players of the SAI, Eastern region. The size of the sample was 300 comprising 200 players, 60 coaches or sports-experts and 40 administrators.  At the outset survey procedure was adopted for collecting data by using extensive questionnaire on administrators, coaches and players of SAI  Eastern region. The data thus collected were tabulated and put to statistical treatment. In a view of the spelled study and nature of the data,‘t’ test was employed to explore the difference among the assorted categories of subjects on various items of organization, administration and facilities. And “Kruskal-Wallis” test was also applied. It is non-parametric technique, which is commonly referred as one-way analysis of various ranks

MORAL ANTECEDENTS OF ATTITUDE FOR THE PURCHASE OF COUNTERFEIT LUXURY PRODUCTS: MEDIATING EFFECT OF PRICE SENSITIVITY

In the recent past demand for counterfeiting has enhanced worldwide and has become serious problem for business and trade of every country. Increasing demand of counterfeit products needs in-depth investigation for stoppage of this crime. The effects of counterfeit purchases are not restricted to branded products but, they also enhance consumer safety risk, reduction in revenues and damage to image of branded products. The aim of this study is to investigate the mediating role of price sensitivity between moral influences and purchase intention of consumer for purchase of counterfeit luxury products. Population for this investigation comprised of non-deceptive purchasers of counterfeit products of district Dera Ismail Khan. For the said purpose one hundred questionnaires were circulated through simple random sampling and out of which 95 completed questionnaires were received and utilized for statistical analysis. The findings of the present study yielded mediation of price sensitivity between the relationship of moral influences and purchase intention. Key Words: Counterfeit Luxury Products, Purchase intention, Moral Influences &nbsp

A COMPARISON OF THE LEGAL AND SOCIAL ATTITUDES TOWARDS ADULTERY IN SAUDI ARABIA AND PAKISTAN

Adultery is considered a major sin in Islam and is prohibited in all Muslim societies. However, there are differences in the legal and social attitudes towards adultery in different Muslim countries. This article aims to compare the legal and social attitudes towards adultery in Saudi Arabia and Pakistan, two Muslim-majority countries with different legal systems and cultural norms. The article begins with an overview of the Islamic perspective on adultery, as outlined in the Quran and Hadith. It then delves into the legal frameworks governing adultery in Saudi Arabia and Pakistan, including the penal codes and the judicial procedures for prosecuting, and punishing adulterers. Furthermore, the article explores the social attitudes towards adultery in these two countries, including the cultural norms and religious beliefs that influence people's perceptions of this issue. It examines the role of family and community in shaping these attitudes, as well as the impact of modernization and globalization on traditional values. The article draws upon a range of sources, including Islamic texts, legal documents, academic literature, and media reports. It provides a comprehensive analysis of the similarities and differences between Saudi Arabia and Pakistan in their legal and social attitudes towards adultery. Overall, this article contributes to the understanding of how Islam shapes the legal and social attitudes towards adultery in different Muslim societies, and the implications of these attitudes for the lives of people living in these societies

Economical Evaluation of Sensation Seeking Among Different Levels Weight Lifters

The purpose of the present study was to compare the sensation seeking trait on different levels of weight lifters. The total hundred (50 State level and 50 All- India intervarsity level weight lifters) males were selected for this study. The age of the subjects were ranged between 18 to 25 years. The data on sensation seeking of the subjects were obtained by using a questionnaire developed by Neary and Zuckerman (1976). The t test was used to determine the difference between the mean score of different levels of weight lifters. Results revealed that there was a significant difference between different levels of weight lifters at 0.05 level of significance with 98 degree of freedom. Study showed that All- India intervarsity level weight lifters have higher level of sensation seeking as compared to State level weight lifters. Key words: weight lifters, Sensation seeking, Thrill and Adventure Seeking, Experience Seeking, Disinhibition, Boredom Susceptibility

Mathematical Modeling and Review of Pine Wilt Disease

Pine Wilt Disease (PWD), caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus, causes significant losses in coniferous forests in eastern Asia, including Japan, China, and South Korea, as well as western Europe, including Portugal. The results of the research papers given at the International Symposium on Pine Wilt Disease (IUFRO Working Party Meeting 4.04.03) in Nanjing, China, in July 2009 are summarized in this article. The basic themes discussed included pine wilt disease (PWD), the pinewood nematode (PWN) Bursaphelenchus xylophilus, and other PWN-associated microorganisms that play a significant role in PWD, such as bacteria (e.g., Pseudomonas fluorescens). The majority of the papers are based on PWD-PWN research in East Asia and Russia. The following are some of the specific topics covered: 1) fundamental concepts of PWD development, 2) pathogenicity, 3) host-parasite relationships, including histopathology of diseased conifers and the role of toxins from bacteria-nematode ecto-symbionts, 4) PWN life cycle and transmission, 5) B. xylophilus dissemination models, 6) associations (with other nematodes), 7) diagnostics, 8) quarantine and control of the PWN and 9) biocontrol of the PWN