Expression of the Native Cholera Toxin B Subunit Gene and Assembly as Functional Oligomers in Transgenic Tobacco Chloroplasts

The B subunits of enterotoxigenic Escherichia coli (LTB) and cholera toxin of Vibrio cholerae (CTB) are candidate vaccine antigens. Integration of an unmodified CTB-coding sequence into chloroplast genomes (up to 10,000 copies per cell), resulted in the accumulation of up to 4.1% of total soluble tobacco leaf protein as functional oligomers (410-fold higher expression levels than that of the unmodified LTB gene expressed via the nuclear genome). However, expresssion levels reported are an underestimation of actual accumulation of CTB in transgenic chloroplasts, due to aggregation of the oligomeric forms in unboiled samples similar to the aggregation observed for purified bacterial antigen. PCR and Southern blot analyses confirmed stable integration of the CTB gene into the chloroplast genome. Western blot analysis showed that the chloroplast-synthesized CTB assembled into oligomers and were antigenically identical with purified native CTB. Also, binding assays confirmed that chloroplast- synthesized CTB binds to the intestinal membrane GM1-ganglioside receptor, indicating correct folding and disulfide bond formation of CTB pentamers within transgenic chloroplasts. In contrast to stunted nuclear transgenic plants, chloroplast transgenic plants were morphologically indistinguishable from untransformed plants, when CTB was constitutively expressed in chloroplasts. Introduced genes were inherited stably in subsequent generations, as confirmed by PCR and Southern blot analyses. Increased production of an efficient transmucosal carrier molecule and delivery system, like CTB, in transgenic chloroplasts makes plant-based oral vaccines and fusion proteins with CTB needing oral administration commercially feasible. Successful expression of foreign genes in transgenic chromoplasts and availability of marker-free chloroplast transformation techniques augurs well for development of vaccines in edible parts of transgenic plants. Furthermore, since the quaternary structure of many proteins is essential for their function, this investigation demonstrates the potential for other foreign multimeric proteins to be properly expressed and assembled in transgenic chloroplasts

Smartphone Addiction and Health Issues among Young Adults in India: A Cross Sectional Study

Purpose: Since smartphone usage across the world has increased in the recent years, the present study aims to explore smartphone addiction in young college going adults and common factors associated with it. Method: 306 female college students volunteered to be part of this study. Demographic information including smartphone usage and sleeping hours per day were collected using a self-administered questionnaire. Smartphone Addiction Scale (SAS) was used to evaluate level of addiction to smartphone. Results and discussion: SAS score of participants was 137.64, which shows higher rate of addiction to smartphone. 27.1% of the participants complained of pain and other health related symptoms, 66.3% reported awareness of symptoms occurring due to mobile overuse, 34.6% reported changes in sleeping pattern and 43.5% in sleeping hours. Conclusion: A higher rate of addiction to smartphone is associated with negative effects on health, social life and might present difficulties to performance in education

Vibrational Medicine: Survey of Tribal Sound healing Practices

This thesis is being archived as a Digitized Shelf Copy for campus access to current students and staff only. We currently cannot provide this open access without the author's permission. If you are the author of this work and desire to provide it open access or wish access removed, please contact the Wahlstrom Library to discuss permission.The objective of my thesis is to understand the scientific basis of ancient shamanic and yogic sound healing techniques. The basic principle of Naturopathic philosophy is that healing happens from top down and in order of spiritual health, mental, emotional and physical health. The healing power of sounds vibrations affect energy centers, chakras also called points of higher consciousness which are associated with particular organ systems. This can explain how sound can be used to cause deep healing within our consciousness which reflects in improved physical and emotional health . Important findings from literature reviews that sound healing techniques can stimulate auricular branches of vagus nerve, synchronization of Right and left hemispheres and cause deep brain relaxation. This explains neurological basis of sound healing

Vibrational Medicine: Survey of Tribal Sound healing Practices

This thesis is being archived as a Digitized Shelf Copy for campus access to current students and staff only. We currently cannot provide this open access without the author's permission. If you are the author of this work and desire to provide it open access or wish access removed, please contact the Wahlstrom Library to discuss permission.The objective of my thesis is to understand the scientific basis of ancient shamanic and yogic sound healing techniques. The basic principle of Naturopathic philosophy is that healing happens from top down and in order of spiritual health, mental, emotional and physical health. The healing power of sounds vibrations affect energy centers, chakras also called points of higher consciousness which are associated with particular organ systems. This can explain how sound can be used to cause deep healing within our consciousness which reflects in improved physical and emotional health . Important findings from literature reviews that sound healing techniques can stimulate auricular branches of vagus nerve, synchronization of Right and left hemispheres and cause deep brain relaxation. This explains neurological basis of sound healing

Comparison of glycosylation patterns of phytase from Aspergillus niger (A. ficuum) NRRL 3135 and recombinant phytase

The glycosylation patterns of phytase (E C 3.1.3.8) from Aspergillus niger NRRL 3135 with recombinant phytase from A. niger van Tieghem were compared. The following characteristics were studied: size of the whole molecule, type of linkage (N-linked or O-linked oligosaccharide), profile (number of different types of oligosaccharides present), monosaccharide composition, their order, and configuration. The molecular weights of both glycoproteins, after deglycosylation, was approximately 55 kDa as determined by SDS-PAGE. Both glycoproteins were about 35.29% glycosylated (calculations were made on the basis of 85 kDa molecular weight before deglycosylation). Only N-linked oligosaccharides were present. When N-linked oligosaccharides were released and labeled, the same profile was obtained for both glycoproteins. Mannose residues were detected after digestion by combinations of various exoglycosidases. N-acetylneuraminic acid, galactose, and N-acetylglucosamine residues were not detected. Released mannose residues were (α1-2,3,6) linked. The trisaccharide core structure was non-fucosylated for all the oligosaccharides released from both glycoproteins. The only major difference found between the two glycoproteins was the migration pattern of oligosaccharide bands on gel after digestion with α-mannosidases. The structure of N-linked oligosaccharides ranged from (Man)5(GlcNAc)2 to (Man)10 (GlcNAC)2

Expression Of The Native Cholera Toxin B Subunit Gene And Assembly As Functional Oligomers In Transgenic Tobacco Chloroplasts

The B subunits of enterotoxigenic Escherichia coli (LTB) and cholera toxin of Vibrio cholerae (CTB) are candidate vaccine antigens. Integration of an unmodified CTB-coding sequence into chloroplast genomes (up to 10, 000 copies per cell), resulted in the accumulation of up to 4.1% of total soluble tobacco leaf protein as functional oligomers (410-fold higher expression levels than that of the unmodified LTB gene expressed via the nuclear genome). However, expression levels reported are an underestimation of actual accumulation of CTB in transgenic chloroplasts, due to aggregation of the oligomeric forms in unboiled samples similar to the aggregation observed for purified bacterial antigen. PCR and Southern blot analyses confirmed stable integration of the CTB gene into the chloroplast genome. Western blot analysis showed that the chloroplast-synthesized CTB assembled into oligomers and were antigenically identical with purified native CTB. Also, binding assays confirmed that chloroplast-synthesized CTB binds to the intestinal membrane GM1-ganglioside receptor, indicating correct folding and disulfide bond formation of CTB pentamers within transgenic chloroplasts. In contrast to stunted nuclear transgenic plants, chloroplast transgenic plants were morphologically indistinguishable from untransformed plants, when CTB was constitutively expressed in chloroplasts. Introduced genes were inherited stably in subsequent generations, as confirmed by PCR and Southern blot analyses. Increased production of an efficient transmucosal carrier molecule and delivery system, like CTB, in transgenic chloroplasts makes plant-based oral vaccines and fusion proteins with CTB needing oral administration commercially feasible. Successful expression of foreign genes in transgenic chromoplasts and availability of marker-free chloroplast transformation techniques augurs well for development of vaccines in edible parts of transgenic plants. Furthermore, since the quaternary structure of many proteins is essential for their function, this investigation demonstrates the potential for other foreign multimeric proteins to be properly expressed and assembled in transgenic chloroplasts. © 2001 Academic Press

A study of human resource digital transformation (HRDT): a phenomenon of innovation capability led by digital and individual factors

Surviving in a digitally disrupted world required organizations to continuously innovate and digitally transform their work practices, especially HR processes. Scholarly failure to define and conceptualize the construct of human resource digital transformation (HRDT) suggests the lack of an integrated approach to understanding HRDT as a discipline. This study presents an overarching and integrated conceptual framework of HRDT, grounded in robust qualitative research, to describe factors in successfully implementing HRDT. Drawing insights from 20 senior HR professionals in multinational organizations, this study defines HRDT as a multidimensional construct resulting from successful integration of digital and individual factors into the innovation capability of organizations. Expanding the dynamic-capabilities perspective, this study proposes that in uncertain and turbulent times, such as COVID-19, businesses must constantly upgrade organizational capabilities, manifested in the innovation capability with enablers such as- digital infrastructure, architecture, and individual capability and creativity. This research further elaborates on HRDT indicators

Fisetin in Cancer: Attributes, Developmental Aspects, and Nanotherapeutics

Cancer is one of the major causes of mortality, globally. Cancerous cells invade normal cells and metastasize to distant sites with the help of the lymphatic system. There are several mechanisms involved in the development and progression of cancer. Several treatment strategies including the use of phytoconstituents have evolved and been practiced for better therapeutic outcomes against cancer. Fisetin is one such naturally derived flavone that offers numerous pharmacological benefits, i.e., antioxidant, anti-inflammatory, antiangiogenic, and anticancer properties. It inhibits the rapid growth, invasiveness, and metastasis of tumors by hindering the multiplication of cancer cells, and prompts apoptosis by avoiding cell division related to actuation of caspase-9 and caspase-8. However, its poor bioavailability associated with its extreme hydrophobicity hampers its clinical utility. The issues related to fisetin delivery can be addressed by adapting to the developmental aspects of nanomedicines, such as formulating it into lipid or polymer-based systems, including nanocochleates and liposomes. This review aims to provide in-depth information regarding fisetin as a potential candidate for anticancer therapy, its properties and various formulation strategies