NOVEL STRATEGIES TO DEVELOP BETTER BRUCELLOSIS VACCINES USING BRUCELLA ABORTUS RB51 AND B. NEOTOMAE

The genus Brucella consists of Gram-negative, facultative intracellular coccobacilli that can cause chronic infections in several mammals. Brucella spp. can exhibit a smooth or rough phenotype; smooth Brucella spp. contain a surface-exposed O-polysaccharide in their cell wall structure while the rough Brucella spp. are devoid of the O-polysaccharide. Acquired immunity against Brucella infection is primarily cell-mediated and involves both CD4+ T cells and CD8+ T cell responses. However, antibodies to the O-polysaccharide also play a role in enhancing the protection against infections by virulent Brucella species in some hosts. B. abortus strain RB51 is a stable rough attenuated mutant which is used as a licensed live vaccine for bovine brucellosis in the United States and several other countries. Previous studies have shown that the wboA gene, which encodes a glycosyltransferase required for the synthesis of O-polysaccharide in Brucella, is disrupted in B. abortus RB51 by an IS711 element. Although low-levels of intra-cytoplasmic O-polysaccharide were produced when RB51 was complemented with a functional wboA gene (strain RB51WboA), it did not result in a smooth phenotype. This suggests that mutations in several genes of the O-polysaccharide biosynthesis pathway contribute to the rough phenotype of RB51. However, nucleotide sequence analysis has revealed that there are no other gene-disrupting mutations that could affect the smooth LPS synthesis in strain RB51

APPLIED ANATOMY OF PRANVAHSROTAS W.S.R TO CARDIOPULMONARY SYSTEM

Srotas are the channel in the body through which various substances pass from one place to another. There are an infinite number of Srotas present in the body. All the physiological and pathological processes conducted in the body are dependent on Srotas. Pranvahasrotas are the most important Srotas in the body which is the pathway for Pranswaropivayu or we can say oxygen. Pranvahasrotas transport oxygen to all over the body and our life is dependent on oxygen. Like Pranvahsrotas, in contemporary science cardiopulmonary system transport oxygen. Pranavaha Srotas has its two Moolsthan from where it is regulated. Cardiopulmonary system is consists of two interdependent system. One is Respiratory System which is involved in transfer of oxygen to blood and make it oxygenated and cardiovascular system is responsible to carry it all over the body. Since heart and its entities and lungs are the principal organ of the cardiopulmonary system. Function, sign and symptoms of Pranvahasrotas and cardiopulmonary system has many similarities. Objective of the study is comprehensive study of anatomical feature of Pranvahsrotasmool in Ayurved to cardiopulmonary system in contemporary science

Dissecting the role of phospholipase A2 in the late stages of regulated exocytosis

This thesis is divided into five chapters. The overarching working hypothesis is published as a ‘perspective’ article in International Journal of Biochemistry and Cell biology, 2017 https://doi.org/10.1016/j.biocel.2017.01.011 and is also discussed in the last section of Chapter - 1. Hence, there are similarities in the published article and sections of Chapter - 1. The working hypothesis states that endogenous PLA2 near docking/ fusion sites provide basal levels of LPC that acts as a ‘molecular brake’ against spontaneous fusion, and FFA that aids in trans-SNARE complex formation to ensure efficient docking of secretory vesicles to the plasma membrane (PM) prior to triggering. The data in Chapters - 2 and - 3 then test the working hypothesis and are also published as original articles in cells, 2019 https://doi.org/10.3390/cells8040303 and Biochemical and Biophysical Research Communications, 2019 https://doi.org/10.1016/j.bbrc.2019.05.106, respectively. The data presented in Chapter - 2, indicate that vesicle-associated endogenous PLA2 isozymes – sPLA2 in the CV lumen and iPLA2 on the CV surface – maintain docking/priming steps in regulated exocytosis. The specific CV localization of sPLA2 and iPLA2 is also shown, using highly sensitive two-dimensional gel electrophoresis (2DE) immuno-blotting with selective antibodies on separated luminal and membrane fractions. Biochemical assays confirmed that these isozymes are catalytically active; sPLA2 showed higher efficiency in catalyzing PE than PC, while iPLA2 showed no such preference. Notably, a significant increase in the de novo metabolite TAG upon iPLA2 inhibition confirmed the role of this isozyme in maintaining membrane lipid homeostasis. Removal of CV surface proteins by treatment with trypsin, coupled with the use of a selective PLA2 substrate (PED6) also confirmed the presence of an active PLA2 isozyme in the outer CV membrane. Overall, the data presented in Chapter - 2, provide evidence in support of the working hypothesis. To test the hypothesis further, in Chapter - 3, exogenous arachidonic acid (ARA) and lysophosphatidylcholine (LPC), canonical PLA2 metabolites, are shown to impair docking/priming and actual membrane merger steps, respectively. The observed decrease in docking/priming induced by exogenous ARA is contradictory to part of the working hypothesis that was put forth based on existing literature. That exogenous LPC and ARA were to a significant extent converted into endogenous PC and FFA, respectively, indicated that endogenous enzymes maintain lipid homeostasis in the native vesicle membrane. With the intension of activating CV luminal sPLA2, Phospholipase A2 activating peptide (PLAP) - a melittin homolog, was used in Chapter – 4. The data presented indicates that PLAP inhibited CSC, and CV fusion by impairing docking/priming and also altered endogenous PE and FFA levels in the CV membrane. This suggests that PLAP caused no change in the CV luminal sPLA2 activity; an observation contrary to the earlier in vitro studies. To test further, PLAP was delivered to mouse chromaffin cells and was seen to reduce membrane capacitance and readily releasable pool (RRP) size, when triggered with depolarizing pulses. This substantiate that the PLAP blocked a common underlying mechanism associated with the late stages of Ca2+ triggered exocytosis. The Chapter - 4 is another manuscript in preparation and will be submitted to an appropriate journal. Finally, in Chapter - 5, the Discussion and Future Directions, all key finding in this thesis are reviewed and integrated with a critical evaluation of the existing literature, and prospective future experiments are outlined

Role of Essential Oils and Bioactive Components for Manufacturing Cosmetic Items

The cosmetics industry has been forced to search for new active chemicals as a result of increasing customer demand as well as stricter international regulations. Botanical extract offers a source of new active chemicals that is nearly endless and may be used to make cosmetics. Due to their olfactory character in the creation of fragrances and perfumes and the many beneficial properties of their individual components (EOCs), essential oils (EOs) are becoming increasingly popular natural ingredients in the cosmetics and toiletries industry. These properties include anti-inflammatory, antimicrobial, and antioxidant properties (e.g., moisturizers, lotions and cleanser in skin care cosmetics; conditioners, masks or antidandruff products in hair care products; lipsticks, or fragrances in perfumery). It is difficult to generalise the possible applications of essential oils in cosmetics and toiletries because each essential oil has its own unique chemical profile, which is connected with its own particular collection of benefits. Instead, formulators need to make an effort to discover acceptable mixes of essential oils or essential oil compounds in order to get the results they want from the completed goods. This paper presents a detailed examination of the existing research on the usage of essential oils and essential oil compounds (EOs and EOCs) in the cosmetics sector. In addition, we will focus on a few critical topics regarding the safety of essential oils and essential oil compounds used in the cosmetics sector. When formulators of cosmetics are working to perfect products that are based on botanical extracts, they are very likely to find the information offered in this review to be valuable

A Hydro-Geochemical Evaluation of Groundwater Suitability for Irrigation and Domestic Purposes: A Case Study from Narmada Mahi Inter-Stream Region of Gujarat, India

Groundwater Hydrology, Irrigatio

An Integrated Geochemical and Geospatial Approach for Assessing the Potential Ground Water Recharge Zones in Mahi -Narmada Inter Stream Doab Area, Gujarat, India

Mahi-Narmada inter-stream region (Doab) in Gujarat is a unique landscape bounded by rivers Mahi and Narmada in the north and south respectively followed by the physiographic truncation by the eastern highlands comprising mainly the Deccan flood basalts and abruptly ending along the western margin due to the Cambay gulf. These distinct hydro-geological boundaries have controlled the ground water regime in the region. The region being part of the Golden Quadrilateral has witnessed highest industrial growth in India. Use of groundwater for irrigation, industrial development and urban expansion have stressed the aquifers and even the surface water bodies are also over utilized; thus adversely affecting the quality and quantum of the water resource of the region. A study was carried out to understand the gravity of the problem stated and also to propose a strategy of monitoring and management of the water resource of the region. The pre- and post-monsoon groundwater sampling of 2002 was carried out and corresponding physical and chemical data generated has established that the groundwater in the study area has unusually high concentration of major, minor and trace elements. A multi-parametric dataset comprising satellite data and other conventional maps including Survey of India (SoI) topographic maps (1:50,000 scale), drainage density, landforms, slope, soil and land use maps, etc. were integrated into Geographic Information System (GIS) environment to generate various thematic maps such as; recharge and discharge zone maps, groundwater quality maps finally culminating into water resource management scheme for the region. The maps so generated have been very close to the field conditions, the study has thus helped in the development of ground water monitoring and managing strategy. The study indicates that periodic monitoring of the groundwater resource with the help of GIS will help in proper management of groundwater resource by identifying the zones/areas of recharge that shall be required to be protected from being converted into built up areas. Moreover, such area also needs to be protected for being used for extensive cultivation as this can leads to contamination of groundwater resource by use of insecticide, pesticides and other chemicals. The study puts emphasis on conjunctive use of surface and groundwater by applying proper management practices so that there is effective use of water. Keywords: Hydro geochemistry, GIS, groundwater, Total Dissolved Solids (TDS

The Roles of Phospholipase A₂ in Phagocytes

Phagocytic cells, such as macrophages, neutrophils, and dendritic cells, ingest particles larger than about 0.5 μM and thereby clear microbial pathogens and malignant cells from the body. These phagocytic cargoes are proteolytically degraded within the lumen of phagosomes, and peptides derived from them are presented on Major Histocompatibility Complexes (MHC) for the activation of T cells. Mammalian PLA2 isozymes belong to a large family of enzymes that cleave phospholipids at the second position of the glycerol backbone, releasing a free fatty acid and a lysolipid moiety. In human macrophages, at least 15 different PLA2 forms are expressed, and expression of many of these is dependent on pathogenic stimulation. Intriguing questions are why so many PLA2 forms are expressed in macrophages, and what are the functional consequences of their altered gene expression after encountering pathogenic stimuli. In this review, we discuss the evidence of the differential roles of different forms of PLA2 in phagocytic immune cells. These roles include: lipid signaling for immune cell activation, initial phagocytic particle uptake, microbial action for the killing and degradation of ingested microbes, and the repair of membranes induced by oxygen radicals. We also discuss the roles of PLA2 in the subsequent digestion of ingested phagocytic cargoes for antigen presentation to T cells

Focal exocytosis of Syntaxin 3 and TRPML1 at pseudopodia of nascent phagosomes

Macrophages clear invading pathogens by phagocytosis. Phagocytosis is a complex mechanism involving the local expansion of the membrane, cytoskeletal remodeling, and the delivery of phagosomal proteins to the nascent phagosomes. However, the organelle trafficking events underlying this are largely unclear. Here, we show in human blood monocyte-derived macrophages that TRPML1, a calcium channel involved in the phagocytic process, is delivered to phagosomes in Syntaxin 3-positive vesicles. Syntaxin 3 is a SNARE protein previously shown to mediate the secretion of IL-6 by macrophages. Total Internal Reflection Microscopy (TIRF) revealed that Syntaxin 3 positive compartments carry TRPML1 to pseudopodia for focal exocytosis at the nascent phagosomes during E. coli uptake. Using siRNA knockdown, we show that both Syntaxin 3 and TRPML1 are required for E. coli uptake. Moreover, using TRPML1 agonists we show that increased TRPML1 activity leads to increased E. coli uptake, whereas calcium chelation decreased intracellular E. coli load. Understanding the membrane trafficking pathways is critical for understanding how macrophages clear invading pathogens