eChapters

Introduction

This section of the manual contains instructions for preparing stock solutions that are commonly used in laboratory protocols for the analysis of DNA, RNA and proteins and visualization of gene or protein expression. Each of these stock solutions is provided as either the most convenient concentration or the most frequently used stock concentrations to be used in the protocols of an individual laboratory.

All chemicals must be reagent grade or molecular biology grade, and the water used in the preparation of all solutions must be the highest quality available. Use sterile-glass distilled deionised water whenever possible. Unless otherwise stated, most solutions required sterilization either by filtration or autoclaving.

Molecular biology labo-ratories are becoming very common in every University and research institutes and a good laboratory manners is extremely important as many of the equipments are very expensive and sensitive. Hence familiarity with equipments becomes very crucial before starting any experiment.

Summary

Lac is a resinous exudation from the body of female scale insect Laccifer lacca Kerr. belongs to family Lacciferidae of phylum Arthropoda, class  Insecta and order Hemiptera. The lac insect lives on native trees in India, China,Thiland, Burma/Myanmar and Malaysia. In India it is chiefly grown on trees like Kusum, “Palas”, and “Ber. It is also purposely cultured on shoots of these tree species. Lac cultivation is an alternative agricultural profession and provides subsidiary source of income for a large number of farmers mainly in Jharkhand, Chhattisgarh, West Bengal, Odisha, North-East states and other parts of the country. India, which is the highest producer of lac, contributes around 55% of the total world requirement followed by China and Thiland. The country exports around 80-90% of its production.

Summary

Nanotechnology is an interdisciplinary areas of science and technology which refers broadly to a field of applied science and technology whose unifying theme of control of matter is on the molecular scale that integrates material science and biology. Due to their advantages over non-biological systems, several research groups have explored the use of biological systems for the synthesis of nanoparticles. Among the different microbes used for the synthesis of nanoparticles, fungi are efficient candidates for fabrication of metal nanoparticles both intra-and extracellulary. The nanoparticles synthesized using fungi present good polydispersity, dimensions and stability. The potential applications of nanotechnology and nanoparticles in different fields have revolutionized the health care, textile and agricultural industries and  they are described in present chapter.

Summary

Molecular evolution is the process of evolution at the scale of DNA, RNA, and Protein. Molecular evolution emerged as a scientific field in the 1960s as researchers from molecular biology, evolutionary biology, and population genetics sought to understand recent discoveries on the structure and function of nucleic acids and protein. It covers two broad areas of study : the evolution of macromolecules, and reconstruction of the evolutionary history of genes and organisms. The evolution of macromolecules refer to the characterization of the changes in the genetic material (DNA or RNA sequences) and its products (Proteins of RNA molecules) during evolutionary process, and to the rates and patterns with which such changes occur, and also the mechanism responsible for such changes. The second area also known as “molecular phylogenetics”, deals with the evolutionary history of organisms and macromolecules as inferred from molecular data and the methodology of tree reconstruction.

Summary

One of the most seductive lures of the genomic revolution is the promise of personalized medicine. The rapid identification of tens of thousands of human genes and hundreds of thousands of DNA variations that might influence disease susceptibility has spawned a new field Pharmacogenetics and Pharmacogenomics deal with the genetic basis underlying variable drug response in individual patients. The traditional pharmacogenetic approach relies on studying sequence variations in candidate genes suspected of affecting drug response i.e. single or few genes. On the other hand, pharmacogenomic studies encompass the sum of all genes, i.e., the genome. Numerous genes may play a role in drug response and toxicity, introducing a daunting level of complexity into the search for candidate genes. The high speed and specificity associated with newly emerging genomic technologies enable the search for relevant genes and their variants to include the entire genome.

Summary

Substance that acts as a catalyst in living organisms, regulating the rate at which life’s chemical reactions proceed without being altered in the process. Enzymes reduce the activation energy needed to start these reactions, without them, most such reactions would not take place at a useful rate. Because enzymes are not consumed, only tiny amounts of them are needed. Enzymes catalyze all aspects of cell metabolism, including the digestion of food, in which large nutrient molecules (including proteins, carbohydrates, and fats) are broken down into smaller molecules; the conservation and transformation of chemical energy and the construction of cellular materials and components. Almost all enzymes are proteins; many depend on a nonprotein cofactor, either a loosely associated organic compound (e.g., a vitamin) or a tightly bound metal ion (e.g., iron, zinc) or organic (often metal-containing) group.

Summary

The process of administration of a drug into the human body to cure a disease is regarded as drug delivery. The common routes of administration of the drugs include oral, nasal, inhalational, transdermal, dermal, ocular, rectal and vaginal. In the current chapter, an attempt has been made to discuss about the various methodologies which are being used to deliver the drug to the human patients.

Summary

Human being is dependent on nature and natural resources to combat diseases from time immemorial. Ayurveda describes the use of traditional herbs in medicinal preparations in ancient India. Such types of evidences are there in classical Chinese and Greek records also. Nowadays, modern tools are used to identify and purify the active principle from the herbal composition having the therapeutic effect. Bioinformatics tools are often used to screen the drug efficacy of natural products. Using computational and Bioinformatics tools, the efficacy of lead molecules can be screened. Moreover, drug targets can be identified using Bioinformatics and proteomics approach. QSAR and molecular docking tools are used to study the probable activity and mode of action of drug molecule. This approach has minimized the cost and time involved in the drug discovery process.

Summary

The present era is witness of understanding the science of human diseases at molecular level. The advances of science in terms of Genomics, Proteomics, Metaboliomics, Lipidomics and bioinformatics have drawn an excellent pathway to discover more promising drugs. It’s still a challenging task to reduce the time line to discover new drug, which is approximately 10-15 years. A drug discovery program requires billions of dollars but without the promise of success. After screening of thousands of compounds only one becomes successful in clinical trials and reaches to the market. Although there are lot many ways to approach drug discovery, nature has gifted many interesting compounds to fight the diseases. On the other hand isolation of pure compounds from natural products like bacteria found in soil or different parts of plants is itself a challenging area. The modern fashion of natural product isolation is guided by bioactivity. Hence a feasible assay to access the bioactivity has got a great importance.

Summary

Plants produce over 200,000 metabolites, which are important source of drugs and industrial materials. They have been traditionally classified as primary and secondary metabolites. The primary metabolites like sugars, amino acids and fatty acids that are needed for general growth and physiological development of plants are widely distributed in nature and are also utilized as food by man. Secondary metabolites are natural products, the absence of which does not result in immediate death, but in long term their impairment may lead to malfunction and reduced survivability. These plant secondary metabolites can be divided into groups that share the same core substructure, originated from the same biosynthetic pathways. Secondary metabolites play a major role in the adaptation of plants to the changing environment and in overcoming stress constraints. In this chapter we discussed the structure, properties, classification and functions of some of the important groups of plant secondary metabolites such as phenols, flavonoids, terpenoids and alkaloids. A brief overview of some of the important fungal secondary metabolites, their structures and importance has also been discussed. Biotechnological approaches which can be adopted to engineer plants for production of secondary metabolites have also been discussed.

Summary

RNA interference has revolutionized the concepts in regulation of gene expression. RNA interference is the mechanism that controls sequence specific gene silencing by double stranded RNA. These non-coding RNAs are gene specific regulators critical in the transformation of normal cells to a malignant stage. Alterations in the non-coding RNAs have been linked to specific cancers and can serve as diagnostic markers. The ability of exogenously introduced siRNAs to specifically bind its complementary mRNA and down regulate its expression has various applications. It has markedly influenced the way the role of certain genes is investigated in tumorigenesis. The siRNA technology has been effectively functional in identification of new components and blocking the expression of critical mediators in tumorigenesis.

Summary

Researchers face a series of similar-yet unique problems to isolate proteins. To solve these they must have dip knowledge to select an appropriate technique. The mechanism, to study the way in which a living system works, is necessary to dismantle the machine and to isolate the component parts so that they may be studied, separately and in their interaction with other parts. The knowledge that is gained in this way may be put to practical use, for example, in the design of medicines, diagnostics, pesticides, or industrial processes. In every case a pure protein is desirable as impurities may either be misleading, dangerous or unproductive, respectively. Protein isolation is, therefore, a very common, almost central, procedure in biotechnology. The purpose of this chapter is thus to fill the beginners with techniques relevant to protein isolation, hopefully to improve their knowledge.

Summary

Nanotechnology, the science of nanoscale structures has been applied in various fields including in biomedical sciences. Nanotechnology is being increasingly used to create materials and devices so as to interact with the body at sub-cellular scales with a high degree of specificity. Nanotechnology is used in the fight against cancer, cardiac and neurodegenerative disorders, infection and other diseases. This chapter provides an overview of some of the applications of nanotechnology in the field of biomedical sciences particularly with a focus on tissue engineering and medical diagnostics.

Summary

The indiscriminate use of pesticides and their persistence in the environment enhances the residue levels in meat, dairy products, eggs and food crops, consequently adversely affecting the human health (Akhtar, 1985). In India millions of tons of pesticides are applied to the agricultural fields to increase the productivity for compensation of the nutritional requirements of increasing population from the same land resources. It was the advent of green revolution that really spurred the growth of pesticide industry. Several reports published reveal that a large amount of organochlorine pesticides is present in vegetables, fruits and soft drinks besides other pesticides compared to World Health Organization (WHO) standards (Martin and Duggan, 1968; Corneliussen, 1969; Ulmann, 1972; Hildebrandt et al., 1986). Organochlorine pesticide persistence, biomagnification and toxicity adversely affect the ecological systems (Khanna et al., 2002). There are a large number of different types of pesticides which differ in their mechanisms of action. Some pesticides are very acutely toxic in humans, while others have low acute toxicity but may cause long term health effects in chronically exposed individuals. Each pesticide class has to be considered individually when predicting effects. Therefore, considering organochlorine pesticides induced disruption of body functions and possibly the involvement of free radicals is cardinal to present study.

Summary

Stem cells are undifferentiated cells which have the potential to divide and multiply indefinitely and can be differentiated to any type of cell of the body of an organism. Due to its peculiar characteristics stem cells are being experimentally used to cure certain dreaded diseases of the humans including different haemoglobinopathies. The stem cells isolated from bone marrow have been cultured, modified and differentiated to get various blood cells like erythrocytes, leukocytes, and thrombocytes etc. Recently haematopoietic stem cells have been transplanted to the sickle cell and beta thalassaemia patients successfully. Gene therapy using the embryonic and cord blood stem cells have been experimentally tried successfully to cure transgenic sickle as well as thalassaemic mice. Induced Pluripotent stem cells (iPS) generated from autologous skin has also been used successfully for treatment of transgenic sickle cell anaemia mouse.

Summary

Multicellular organisms are made up of diverse set of cells. Cells respond to the changes in the particular microenvironment by means of cellular communication. This cell-cell communication is extremely important for the development, maintenance and survival of the organism. This communication is equally important for the unicellular organisms. Cells communicate to one another and respond to the environmental changes using diverse chemical languages, as discussed in this chapter. However, in many cases like multicellular organisms, cell communication is primarily mediated by specific receptor(s) expressed on the cell membranes. These receptors bind to the extracellular signaling molecules (called ligands) and activate specific signaling pathways to bring about particular cellular responses. These different receptors, ligands and signaling pathways are briefly discussed here.

Summary

Animal tissue culture is a biological technique in which a piece of tissue is taken from an animal specimen and cells from this tissue are grown and maintained in an in vitro or artificial environment (Harrison, 1907). Aim of cell / tissue culture is actually to make cells as happy in the in vitro conditions, as they are in their natural environment. The objective of this chapter is to highlight those principles and procedures that have particular relevance to the use of cell culture in tissue engineering.

Summary

Idealistic theories of property are projected to offer general and explicit statements of the justification for deciding legal and moral questions about the status property claims. Debates over property rights in biotechnology were occasioned by specific legislative proposals such as the US Animal Patent Act of 1986, and by filing of patent applications for DNA sequences and processes in the early 1990s. While these debates make sporadic petition to philosophical theories of property, moral claims were entwined with questions about filing requirements, tests for worth, and the rules for licensing and defending patents. The comparative scarcity of discussion on intellectual property at that time dictated the general approach of the chapter: Review basic philosophical approaches to property rights, and speculate on how one might use these approaches in constructing an argument pertinent to agricultural biotechnology. This chapter deals with different approaches to intellectual property in the field of Plant Sciences especially the Biotechnology. It incorporate many different types of intellectual property and a variety of incommensurable values play a role in the justification of intellectual property.

Summary

Bioenergy is the subject of increasing attention around the globe and represents an important issue in the present context. The rise of commodities prices, the negative impact on food security, and climate change represent different challenges to be overcome before the full potential of bioenergy can be realized. Bioenergy issues related to agriculture needs special attention for its development. Bioenergy development is always linked with rural economy. Farmers, having the choice to convert their food crops to fuels crops, naturally expect a high return from their farmland, thereby generating a scenario where food production gets depleted. The optimal production of bioenergy can be generated by using the wasteland present in different area. This way, food security for the future generation can be maintained along with optimal growth of bioenergy.