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site-identification by ligand competitive saturation (SILCS)

Application of sample

INSTRUMENTATION

MP14:

Anti-inflammatory activity

Acute toxicity studies

Drugs and chemicals

Oral administration of doses

Animal handling and care

Animals

Preparation of distilled extracts

Harvest of germinated seed

Germination of N.sativaseeds

Collection of N.sativaseeds

Statisticalanalysis

ROS assay

Cytotoxicity assay by Sulphorhodamine B (SRB) me

Cytotoxicity assay by MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5 diphenyltetrazolium Bromide)method

Cytotoxicityscreenin

Cell culture

Drugs and chemicals

Preparation of distilled extracts

Harvest of germinated seeds

Germination of N. sativaseeds

Collection of N. sativaseeds

TLC study of alkaloids

TLC study of phenols

TLC study of alkaloids

Activation of TLC plate

Preparation of TLC plate

Study of Phytochemicals by Thin Layer Chromatography (Wagner, and Bladt, 1996)

Test for cardiac glycosides

Test for Terpenoids

Tests for Flavonoids

Tests for Phenolic compounds

Tests for Saponins

Tests for Tannins

Tests for Alkaloids

Test for Sterols

Qualitative study of phytochemicals of N. sativadurin

Inoculum preparation

Clinical bacterial strains used for the study

Preparation of distilled extracts

Harvest of germinated seeds

Germination of N.sativaseeds

Collection of N.sativaseeds

Testing of bioformulations

Preparation of bioformulations and determination of

Validation of bioformulations under laboratory conditions (in vitro

Sequence analysis

DNA sequencing of the 18S rDNA fragment

Purification of PCR product

Analysis of internal transcribed spacer region

RAPDand SSRscoring and data analysis

PCR amplification

Running of gel and visualization of DNA

Determination of the yield

Agarose gel electrophoresis

Qualitative and quantitative estimation of DNA

Determination of the yield

Procedure for DNA isolation

Reagents required for fungal DNA isolationand p

DNA isolation of Trichodermaisolate

Genetic variability analysis through RAPD and SSR

Photography, evaluation and documentation

Procedurefor SDS-PAGE

Materialsrequired for SDS-PAGE

Protein profiling of bioagent through SDS-PAGE

Biochemical analysis (Protein estimation)

Protein estimation through Kjeldahl method

Oxidative-Fermentative test

Active site prediction and docking study

Superposition of predicted structure and template (Dof

Validations

Three dimensional structure prediction, refinements and evaluation of Dof proteins

Cis-regulatory element analysis

Mapping of SbDof genes on sorghum chromosomes and its intron/exon gene structure prediction

In silico prediction of Dof gene family members in S. bicolor (L

Motif identification

Multiple sequence alignment and phylogenetic analysis

In silico analysis of sequenced Dof domain and Dof genes

Reaction resuspension

Post-reaction clean up

Sequencing PCR

Sequencing reaction

Digestion of Plasmid DNA

Minipreparation of plasmid DNA from transformed co

Screening of recombinant E. coli clone

Transformation of ligation mixture in electro-competent E. coli host cells (DH5ααααstrain)

Transformation of ligated product in chemically competent E. coli host cells (DH5αααα strain)

Ligation reactions

Dephosphorylation of vector

Restriction digestion

Ligation of eluted PCR product in pBSK vector

Cloning of gel eluted PCR produc

Gel elution of PCR Produ

Scoring of amplification data points and construction of a den

Analysis of PCR amplicons using agarose gel electrophoresis

Cycling condition

PCR reaction set up

Basic requirements for PC

PCR amplification of Dof domain and Dof genes from different

Primer designing for PCR amplification of Dof domain and D

Qualitative analysis of DNA by agarose gel electrophoresi

Spectrophotometric quantification of genomic DNA

DNA purification

DNA extraction procedure

Isolation of genomic DNA by CTAB method

Germination of seeds

Solutions used for cytokine assay

Assessment of Cytokine levels- IFN--12/IL-10 in lymphocytes of cured/endemic patients

Nitrite production in macrophages of hamsters

Assessment of Lymphocyte proliferative responses (LTT) in cured/exposed patients and hamsters

Immunological assays

Treatment of L. donovani infected hamsters and isolation of mononuclear cells (lymph node cells)

Patients and isolation of peripheral blood mononuclear cells (PB

Preparation of soluble L. donovani promastigote antigen

Animals

Parasites

Polyclonal antibody generation

Overexpression and Purification of recombinant protein (rLdTP

Amplification, Cloning and Sequencing

Cloning, Overexpression and purificationof TP

Production of polyclonal antibodies against rLdADHT and We

Purification of recombinant ADHT (rLdADHT)

Recombinant protein Expression

Clone confirmation

Transformation procedure

Transformation in E. coli Rosetta (DE3) cells

Subcloning of ADHT in pET-28a expression vector

Sequencing of ADHT gene

Restriction Digestion of Plasmid DNA

Colony PCR

Preparation of master plate and isolation of plasmid DNA from transformed E. coli (Mini Prep)

Confirmation of positive clone(s)

Transformation

Preparation of chemically competent Escherichia coli using calcium chloride method

Quantification of MMP or ΔΨmloss

Assessmentof mitochondrial membrane potential(MMP or ΔΨm)

Glutathione peroxidase (GPx) assay

Superoxide dismutase (SOD) assay

Catalase assay

Assay for reduced glutathione(GSH)

Assay for Lipid peroxidase(LPO)

Estimation of Total protein

Oxidative stress parameters

Quantification of ROS

Detection of intracellular ROS

Acridine orange/ethidium bromide (AO –EB) staining

Cytomorphological analysis

MTT [(4, 5–dimethylthiazol–2–yl)–2, 5-diphenyl tetrazolium Bromide] assay

Cell culture

Catalytic activity

Growth kinetic studies

Kirby-Bauer Disk diffusion method

Antimicrobial activity

Hydrogen peroxide quenching activity

Nitric oxide radical quenching activity

Superoxide radical scavenging activity

Reducing power assay

DPPH free radical quenching activity

In vitro free radical scavenging assays

TGDSC analysis

DLS particle size and zeta potential analysis

FTIR analysis

TEM-EDAX analysis

UV–visible spectral analysis

Characterization of AgNPs

Synthesis of AgNPs

GC-MS analysis

Qualitative analysis of compounds

Collection and preparation of extracts

The Posterior Probability of Mixing Dis-tribution

Algorithm

EM Procedure

Poisson Mixture

Data Sources

Poisson Model

We have analysed the Cancer data of patients in 155 wards of Chennai Cor-poration by the above described method. As preliminary analyses, we havecreated the Choropleth maps for Observed counts, Population of wards, ex-pected counts for patients and SMR's.The Choropleth map for the observed counts Figure 5.2 does not show anypattern. But the Choropleth map for the expected counts Figure: 5.4 indi-cate that the inner regions of the Chennai Corporations have lower expectedcounts and the regions along the border have larger counts of patients. As ameasure of spatial heterogeneity we have computed PSH= 0:7108:Hence ofthe total spatial random variation, nearly 71% is due to spatial heterogene-ity and the remaining 28:92% is due to Poisson variation. Thus the spatialvariation is present in the data.The Choropleth map for Empirical Bayes smoothed rates Figure 5.5 re-veals that only 13 sub regions have high risk values. The wards with numbers53, 64, 67, 70, 78, 93, 100, 103, 110, 117, 122, 147 and 151 have high riskvalues. Though this information could be used by the health managers toconcentrate their work on these regions, one can look for additional covariatesin these regions for further study

Empirical Bayesian Smoothing

Incidence Rate and SMR

. Statistical analysis

Inhibition of haemocytes spreading behavior

. Inhibition of haemocytes aggregation behavior

Haemolymph protein profiling

Total haemocyte count (THC)

Haemolymph collection