1. The document discusses a study that examines the effects of two Ayurvedic formulations - Amalaki Rasayana (AR), an herbal derivative, and Rasa-Sindoor (RS), an organo-metallic derivative of mercury - on stress response in Drosophila melanogaster. 2. The study aims to analyze the effects of the formulations on longevity, development, fecundity, and stress tolerance in the flies. 3. The methodology describes preparation of the formulations, fly stocks, experimental assays to test thermotolerance, starvation resistance, and more, as well as analysis using immunostaining, confocal microscopy, and real-time PCR.

1. Modulation effects on stress response ofModulation effects on stress response of
Ayurvedic Formulations inAyurvedic Formulations in DrosophilaDrosophila
melanogastermelanogaster
Presented by:
Indramani Tripathi
[MSc-Biochemistry]
Session 2015-2017
Veer Bahadur Singh Purvanchal University , Jaunpur

2. INTRODUCTION
•Ayurveda represents the traditional medicine system of
India which is widely practiced uninterruptedly at least from
beginning of the Buddhist period in India.
•Ayurveda signifies “knowledge relating to life”, and regards
diseases and medicine as no more than facets of the
variegated theme of life.
•The two major groups of Ayurvedic drugs are Herbal
preparations and Herbo-bio-mineral-metallic preparations.
•AR is a Rasayana prepared from fruits of Amla or Indian
gooseberry (Phyllanthus emblica).

3. OBJECTIVE
The proposed work is design to study two different categories
of formulations, a Rasayana (Amalaki Rasayana or AR, an herbal
derivative) and a Bhasma (Rasa-Sindoor or RS, an organo-
metallic derivative of mercury), for effects on Drosophila
melanogaster using at least 200 larvae or flies for each assay.
•Longevity,
•Development,
•Fecundity,
•Stress-tolerance

4. METHODOLOGY
Preparation of formulations
• Amalaki Rasayana
• Rasa-Sindoor
Fly stocks: Oregon R+: A wild type strain of
Drosophila melanogaster
Drosophila food (2.5 L): Requirements for per unit of
simple Drosophila food.
Antibodies
Primary antibodies: Anti Hsp70 (7Fb): Anti-Hsp90 (SPA
830, Stress gen, USA)
Secondary antibodies: Alexa Flour-488 goat anti-rat IgG,
Alexa Flour -488 goat anti –mouse IgG

5. Experimental Methodology
•Stress Tolerance Assay
•Thermotolerance assay
•Starvation resistance assay
•Paraquat treatment
•E.coli killing assay
Immunostaining in Drosophila third instar larvae
Confocal microscopy and documentation
RNA isolation and Real Time PCR

6. EXPECTED OUTCOME AND SIGNIFICANCE OF
PROPOSED WORK
A 0.5% (weight/volume) supplement of AR or RS affect life-
history and other physiological traits in distinct ways. While
the size of salivary glands, hnRNP levels in larval tissues,
and thermotolerance of larvae/adult flies could improve
following feeding either of the two formulations, the median
life span and starvation resistance could improve with AR.
Feeding on AR or RS supplement food could improve
fecundity differently.

7. Feeding of larvae and adults with AR could increase the
fecundity while the same with RS has opposite effect.
The present study with two Ayurvedic formulations will reveals
formulation-specific effects on several parameters of the fly's
life, which seem to generally agree with their recommended
human usages in Ayurvedic practices. Thus, Drosophila, with its
very rich genetic tools and well-work-out developmental
pathways promises to be a very good model for examining the
cellular and molecular bases of the effects of different
Ayurvedia formulations