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Research & Reviews: A Journal of Life Sciences

Contents

1. Identification of Bacterial Strain from Waste Decay Material Using 16s rRNA Ribotyping Suresh Kumar, Bhawna Garg, Ankur R. Bhardwaj, Khaling Mikawlrawng

1

2. A Study on Physiological Effect of Yoga Asana on Sleep Sarika Yadav, Om Prakash Dadhich

5

3. The Interaction of Extracellular Matrix and Growth Factors Haile Nega Mulata, Gidey Gebremeskel

9

4. Morinda citrifolia Leaf Induced Leukemia Cells Apoptosis and Produced Synergistic Effects with Zerumbone Negin Ahmadi, Heshu Sulaiman Rahman, Rozita Rosli, Tengku Azmi Tengku Ibrahim, Swee Keong Yeap, Suhaila Mohamed

24

5. Significance of TE Domain and Genetic Variations in FASN Gene in Dairy Animals Manoj Kumar, Poonam Ratwan, Uday Kannegundla, Vikas Vohra

33


Research & Reviews: A Journal of Life Sciences

ISSN: 2249-8656(online), ISSN: 2348-9545(print) Volume 6, Issue 3 www.stmjournals.com

Identification of Bacterial Strain from Waste Decay Material Using 16s rRNA Ribotyping Suresh Kumar1,*, Bhawna Garg2, Ankur R. Bhardwaj1, Khaling Mikawlrawng1 1

2

Department of Botany, Ramjas College, University of Delhi, Delhi, India Department of Biotechnology, Chaudhary Charan Singh University, Meerut, Uttar Pradesh, India

Abstract

Ribotyping is a classical technique in molecular studies where conserved and unique DNA sequences, more specifically 16s RNA, are investigated to differentiate among strains of microorganisms. This method was utilized in the current study to identify the strains of bacteria present in waste decaying matter. Waste material was collected from Indian Institute of Technology (IIT), Delhi campus, Hauz Khas, New Delhi, and cultured for bacterial growth. Purified bacterial cultures were used to isolated genomic DNA which was subsequently used for PCR based amplification of 16s RNA gene. Sequencing and computational analysis of the generated sequences revealed the presence of two bacterial strains Bacillus pumulis strain IKMB13-518F and B. licheniformis strain MAC-110. Keywords: Ribotyping, decay waste, Bacillus pumulis, B. licheniformis

INTRODUCTION

Bacteria are simple, single-celled, microscopic organisms. They have prokaryotic cellular organization, which means an organized nucleus is absent, and genomic DNA is roughly localized in a region known as nucleoid. Unlike viruses, bacteria have autonomous transcription and translational machinery, which enables them for independent replication. Astonishingly, ecological niche of bacterial domain is highly diverse, ranging from human gut to hot sulfur springs and from deep oceans to arctic snow [1–4]. Bacterial species serve many important functions. These include maintenance of soil composition through cycling of nutrients, industrial scale fermentation and production of antibiotics. They have found numerous applications in biological research based on their small genome size and rapid growth/reproduction. The species of genus Bacillus are Gram-positive and appear rodshaped. These bacterial species can be either obligate aerobes or facultative anaerobes, and can be free-living or parasitic. It has been observed that bacterial species belonging to genus Bacillus can potentially produce significant amount of many enzymes. For instance, Bacillus amyloliquefaciens is used as a natural source of a ribonuclease protein barnase, a protease-subtilisin, and the widely

used restriction endonuclease BamH1 [5–7]. The cry genes have been isolated form Bacillus thuringiensis and incorporated in few economically important crop plants such as maize and cotton [8]. The origin of bacterial lineage is still a widely debatable field. It is understood that, unicellular life forms mainly consisting of archaeal and bacterial lines were predominant approximately 3.5 to 4 billion years ago [9]. But it is difficult to trace bacterial phylogeny owing to poor fossil records. In this context, it is commonsensical to use the information of genes and genomes to decipher evolutionary history. Today, we understand that rRNA genes had remained conserved through billions of years of evolutionary course [10]. Profiling of sequence homology of rRNA genes readily reveals the relatedness of an organism with its other counterparts and hence helps to sketch the position of the species in the evolutionary pedigree. Furthermore, a detailed enquiry of the rRNA gene sequence can also be used to identify a previously unknown bacterial species [11].

RRJoLS (2016) 1-4 © STM Journals 2016. All Rights Reserved

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Research & Reviews: A Journal of Life Sciences

ISSN: 2249-8656(online), ISSN: 2348-9545(print) Volume 6, Issue 3 www.stmjournals.com

A Study on Physiological Effect of Yoga Asana on Sleep Sarika Yadav*, Om Prakash Dadhich P.G. Department of Sharir Kriya, National Institute of Ayurveda, Jaipur, Rajasthan, India Abstract

Ahara, Nidra and Brahmcharya are the three main pillars of life. These play a vital role in maintenance of good health and longevity. Among these Ahara is responsible for physical, Brahmcharya for mental and Nidra for psychosomatic well-being. Mechanisation of world, advancement of science and technology has provided us a great range of comfort; this comfort has led to disturbances in normal Ritucharya and Dincharya. Increased use of various electronic gazettes such as laptop, mobiles, television has led to change in normal sleep cycle resulting sleep deprivation. Growing evidences suggests that these advancements take their toll on human functioning and health via their bad effects on sleep quality, quantity and timing. Chronic sleep deprivation may result in fatigue, muscle ache, day time sleepiness, clumsiness, weight loss or weight gain, memory lapses, depression, headaches, malaise etc. In Ayurveda various modalities have been described to maintain proper sleep such as Abhyanga, Dincharya Palana, and Yoga Asana etc. Yoga Asana not only help to keep the body young, strong and supple but also incorporate mental activities, discipline that help to develop attention and concentration and thus stimulate the total mental and physical calmness and further improve the quality and quantity of sleep. As disturbed sleep is quite common nowadays and performing Yoga Asana is supposed to play an important role in calmness of heart, mind and soul so hereby an emphasis is made to understand the role of Yoga Asana on physiology of sleep. Keywords: Nidra, physiology, Yoga Asana, Dincharya

INTRODUCTION

Ayurveda is the science of life having prime goal of prevention of disease in healthy individuals and cure of disease in diseased. Ahara, Nidra and Brahmcharya are the three main pillars of life and these play a great role in maintenance of health and long life span. Being supported by these Upstambh [1], the life is endowed with strength, complexion and growth. The mechanisation of world and advancement of science and technology have provided with enormous range of comforts. This has led to unwholesome behaviour or ignorance of basic health principles and thus resulting in failure of sustaining normal functions of these Upstambha. Theory of Trayopstambh is one of the important principles in preventive medicine as because if in wholesome way these Upastambhas are observed, there is no possibility of generation of diseases. Among Trayopstambha, Nidra affects both the factors—physical and mental equally [2]. It is one of the life-maintaining factors and if not used properly several fatal diseases may produce. But if it is practiced according to the prescribed regimen it

provides ‘Sukh’ which is considered as Arogya. The comfort driven mode of living has led to changes in lifestyle which further generated restlessness, tension, stress and a new horde of disease and disorders. The practice of Yoga not only keeps the young body strong and supple but also incorporate mental activities, discipline that help to develop attention and concentration, and the creative ability that are latent in the human mind itself. As per Aacharya Patanjali, Yoga is the name given to Chittavritinirodh that is to control the wayward flow of tendencies of the Chitta or mind. Thus performing Yoga Asana in routine may significantly enhance quality of sleep.

AIMS AND OBJECTIVES

1. To study the normal physiology of sleep. 2. To study the effects of Yoga Asana on sleep physiology.

RRJoLS (2016) 5-8 © STM Journals 2016. All Rights Reserved

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Research & Reviews: A Journal of Life Sciences

ISSN: 2249-8656(online), ISSN: 2348-9545(print) Volume 6, Issue 3 www.stmjournals.com

The Interaction of Extracellular Matrix and Growth Factors Haile Nega Mulata*, Gidey Gebremeskel Department of Medical Biochemistry, College of Health Sciences, Mekelle University, Mekelle, Ethiopia Abstract

The extracellular matrix (ECM) regulates cell behavior by influencing cell proliferation, survival, shape, migration and differentiation. ECM assembly is regulated by the 3D environment and the cellular tension that is transmitted through integrins. The hematopoietic microenvironment is a complex structure in which stem cells, progenitor cells, stromal cells, growth factors, and ECM molecules each interact to direct the coordinated regulation of blood cell development. Hematopoietic growth factors are any of several glycoproteins that regulate/promote the survival, self-renewal, proliferation, and differentiation of hematopoietic progenitor cells. Hematopoietic growth factors act by binding to specific members of a highly related family of single pass transmembrane proteins. They have a vital role in promoting hematopoietic cell growth, differentiation, and prevent apoptosis of progenitor cells. Each growth factor has a specific cell surface receptor, which activates both unique and shared signal transduction pathways. This interaction promotes signal transmission across the cell membrane and activates intercellular signaling cascades that are integrated at the gene expression level. Cell–cell and cell–ECM interactions through adhesion receptors play a major role in the hematopoietic process. Hematopoietic signaling is activated by growth factors and cellular interactions with the ECM. ECM functions as a part of the cell-controlled machinery to store and activate growth factors during development. Cell-mediated release of ECM-bound growth factors can occur via proteolytic cleavage of LTBP1. Keywords: extracellular matrix (ECM), growth factors, hematopoietic signaling

INTRODUCTION

Cells of higher multicellular organisms are surrounded by a highly organized matrix of proteins and glycoconjugates. Endothelial cells and their surrounding microenvironment are composed of neighboring cells and the extracellular matrix (ECM), which consists of a network of molecules, composed primarily of type IV collagen, laminins, heparan sulfate and proteoglycans. The understanding of ECM functions has been enhanced in recent years by the discoveries of novel ECM constituents, mapping sites crucial for the interactions of ECM constituents with each other and with cells, characterization of proteases and protease inhibitors that regulate ECM assembly, and turnover and identification of novel receptors and signaling mechanisms that mediate cellular responses in ECM constituents. Matrix components include ligands that activate intracellular signaling pathways within cells, thereby regulating cell

proliferation, survival, and differentiation. These functions can be mediated not only through interaction with cell receptors but also through direct interaction with growth factors. The ECM thereby helps to regulate the special and temporal properties of signals conveyed by these molecules [1].

THE EXTRACELLULAR MATRIX

The extracellular matrix (ECM) was originally defined morphologically as extracellular material visible as fibrils or sheets in the electron microscope. ECM can be defined as “the substance between cells” and the “material in the intercellular space”. It is also a substance produced by cells and secreted into the environment in which the cells are embedded; contains collagen, proteoglycans, glycosaminoglycans, and fluid; can influence the behavior of the cells. The ECM were thought to function only as a scaffold to maintain tissue and organ structure, and

RRJoLS (2016) 9-23 © STM Journals 2016. All Rights Reserved

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Research & Reviews: A Journal of Life Sciences

ISSN: 2249-8656(online), ISSN: 2348-9545(print) Volume 6, Issue 3 www.stmjournals.com

Morinda citrifolia Leaf Induced Leukemia Cells Apoptosis and Produced Synergistic Effects with Zerumbone Negin Ahmadi1,2,3, Heshu Sulaiman Rahman1,4, Rozita Rosli1,2, Tengku Azmi Tengku Ibrahim1, Swee Keong Yeap1, Suhaila Mohamed1,* 1

Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia 3 Petroleum Industry Health Organization, Masjedsoleiman, Imam Khomeini Hospital, Iran 4 College of Veterinary Medicine, University of Sulaimani, Sulaimani Nwe, Street 27, Sulaimani City, Kurdistan Region, Northern Iraq 2

Abstract

Leukemia is the most common cancer among children and the 7th most common cancer worldwide. The vegetable Morinda citrifolia leaves was investigated and compared with Zerumbone (a known anti-leukemia compound) separately and in combination, for their apoptotic effects on murine myelomonocytic leukaemia cell line (WEHI-3B). Zerumbone, a sesquiterpene from Zingiber zerumbet was anti-cancer via suppressing free radical generation, inflammation, cancer cell proliferation, metastasis, anti-apoptosis, NF-κβ, Iκβα kinase activation and invasion. The Morinda leaf extract (standardized to the coumarin scopoletin and the flavonoid epicatechin) dose-dependently inhibited WEHI-3B cells proliferation with an IC50 of 17.1±0.1 μg/ml after 72 h incubations. Comparatively, the leaf extract alone and in combination with Zerumbone significantly suppressed WEHI-3B cells with an IC50 of 11.0 and 9.4 μg/ml, respectively. The extract arrested leukemia cells at the G0/G1 phase, and induced apoptosis via the activation of caspase-3 and caspase-8 (death receptor extrinsic pathway). Zerumbone arrested WEHI-3B cell cycle at the G2/M phase and induced apoptosis via the activation of caspase-3 and caspase-9 (intrinsic pathway). When combined, the Zerumbone and Morinda leaf extract indicated synergistic anti-leukemia effects via both the intrinsic and extrinsic pathways, and showed no toxicity to normal cells. Dietary vegetable complementary therapy helps support the primary cancer treatments. Keywords: Morinda citrifolia, Zerumbone, synergy, leukemia, apoptosis

INTRODUCTION

The Morinda citrifolia L. leaves [Noni (Hawaii) or mengkudu (Malay)], are consumed as vegetable by the natives after blanching and demonstrated therapeutic effects against various cancers [1, 2]. Morinda leaf possess immune enhancing, antioxidative, liver-protective and wound healing properties without any observable acute, subacute and subchronic oral toxicity at 1 g extract/kg rodent bodyweight [3]. Zerumbone (C15H22O, MW: 218.34) is a sesquiterpene from Zingiber zerumbet Smith from Southeast Asia and known for its anti-leukemia effects. Zerumbone suppressed free radical generation, inflammation, cancer cell proliferation, metastasis, anti-apoptosis, NF-κβ and Iκβα

kinase activation leading to downregulation of cancer invasion [4]. The M. citrifolia leaf extract possess anti-inflammatory, immunomodulatory and anti-oxidant properties [5]. The leaves contained phytosterols, catechins, various polyphenols and flavonoids [5]. The epicatechin and scopoletin rich M. citrifolia leaves extract demonstrated immunostimulatory effects in whole animals by enhancing total WBC, immune cells CD4, T cells CD8, B cells CD19 and natural killer cells CD335 levels in the blood and spleen. The M. citrifolia leaves inhibited nonsmall cell lung cancer cell proliferation, inflammation and angiogenesis via mRNA signalling pathways by modifying MDM2/p53, RAF/MEK/ERK, PI3K/AKT/

RRJoLS (2016) 24-32 © STM Journals 2016. All Rights Reserved

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Research & Reviews: A Journal of Life Sciences

ISSN: 2249-8656(online), ISSN: 2348-9545(print) Volume 6, Issue 3 www.stmjournals.com

Significance of TE Domain and Genetic Variations in FASN Gene in Dairy Animals 1

Manoj Kumar1,*, Poonam Ratwan1, Uday Kannegundla1, Vikas Vohra2

Animal Genetics and Breeding Division, ICAR-National Dairy Research Institute, Karnal, Haryana, India 2 AGR Division, ICAR-National Bureau of Animal Genetic Resources, Karnal, Haryana, India

Abstract

The interest in the bovine fatty acid synthase (FASN) gene has increased during the last few years. FASN is a multifunctional enzyme that catalyzes de novo biosynthesis of long-chain saturated fatty acid, and alteration of this enzyme may affect economic traits including fatty acid composition. FASN is involved in fat metabolism and is an important candidate gene affecting fat composition of milk because of its central role in de-novo lipogenesis in mammals. This cytosolic enzyme catalyses palmitic acid synthesis from acetyl coenzyme-A and malonyl coenzyme-A in the presence of NADPH. Thioesterase (TE) domain of FASN gene regulates the termination of fatty acid synthesis and thus determines the quality of fat synthesized in milk through early termination of fatty acid chains. TE domain spreads from exon 39–41 within the FASN complex. The gene frequency of polymorphism in FASN gene has been found to be very high in dairy cattle. Some associated studies such as milk yield, fat content and protein yield have been carried out in dairy cattle. These associations will provide insight in to the underlying mechanism of FASN gene and polymorphisms that can be used for selection purposes in dairy animals. Keywords: Bovine, fat content, TE domain, polymorphism

INTRODUCTION

During the last decade, the progress in molecular genetics has led to the discovery of candidate genes, which exert large effects on traits of economic importance such as milk production and its composition (fat) traits. Candidate gene strategy has been proposed by direct search for quantitative trait loci (QTL). Genetic or marker based selection followed with appropriate breeding methods for higher milk fat can facilitate genetic selection in different bovine breed improvement programmes. In dairy animals, it is important to explore genetic variations at DNA level and subsequently, utilize them in breeding programmes. Recent studies carried by Stoop et al. in exotic cattle have revealed large genetic variation in bovine milk-fat as well as milk fat composition and many SNPs in these candidate genes, which play a role in fat synthesis, have been associated with milk-fat percentage or milk-fat yield [1]. Many such

candidate genes have been identified for fat synthesis such as fatty acid synthase (FASN) gene [2–4], oxidized low density lipoprotein receptor1 (OLR1) gene [5, 6], diacylglycerolacyl-transferase1 (DGAT1) gene [7], signal transducer and activator of transcription 1 (STAT1) gene [8, 9]. According to Asadollahpour et al. genetic research in farm animals focuses mainly on the identification of genes influencing economically important traits that could be useful in breeding [10]. In crossbred cattle, fast genetic progress can be made if animals are selected based on the milk production efficiency traits [11, 12]. Many studies reported that FASN gene is the candidate gene that influences milk fat content and fat yield in dairy animals. Fatty acid synthase (FASN) is a complex homodimeric enzyme that regulates de novo biosynthesis of long-chain fatty acids. This is a key function in the lipid as well as the general metabolism of all living animals. Mammalian FASN is not only an essential metabolic

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Research & Reviews: A

Journal of Life Sciences (RRJoLS) September–December 2016

STM Journals invites the papers from the National Conferences, International Conferences, Seminars conducted by Colleges, Universities, Research Organizations etc. for Conference Proceedings and Special Issue. xSpecial Issues come in Online and Print versions. xSTM Journals offers schemes to publish such issues on payment and gratis (online) basis as well.

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Research & Reviews: A Journal of Life Sciences vol 6 issue 3  

Research & Reviews: A Journal of Life Sciences (RRJoLS) is a print and online journal focused towards the rapid publication of fundamental r...

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