SlideShare a Scribd company logo
1 of 50
Download to read offline
“Biogenic synthesis of silver nanoparticles and its Pharmacotherapeutic
Applications using aqueous extract of Lepidium Sativum (Garden Cress
Seeds)”
Name of Research Scholar- P.Sree Vennela Rao
Supervisor- Prof.B.Veeresh
Scholarship- National Fellowship
Registration Num- 9009-12-891-003
Presentation- PROGRESS SEMINAR
Dept of Pharmacy,
University College of Technology,
Osmania University,Hyderabad,500007.
Overview of Presentation
1.Introduction and Literature review
2.Objectives
I) Synthesis and characterization of silver nanoparticles.
II) Anti bacterial, Anti oxidant and Anti inflammatory activities.
III) Anti cancer activity (in vitro)
IV) Anti inflammatory/Wound healing activity (in vivo).
3.Methodology
4.Expected outcome
5.Summary and Discussion.
6.Limitations and Future prospects
7.Publications
6.Certifications
INTRODUCTION&
LITERATURE REVIEW
Nanoparticles…
• A nanoparticle is a small particle that ranges between 1 to 100
nanometers in size. Undetectable by the human eye, nanoparticles can
exhibit significantly different physical and chemical properties to their
larger material counterparts.
• Nanoparticles have large surface area, which gives them more
pronounced effects.
Particle Type Diameter Size Range
Coarse particles 2500 to 10,000 nm
Fine particles/Dust 100 to 2500 nm
Nano particles 1 to 100 nm
Atoms and small molecules 0.1 nm
Nano particle and its Applications..
Types of nanoparticles…
• Sea spray
• Mineral composites
• Volcanic ash
• virus
NATURAL/
NATURE
• Cooking smoke
• Diesel exhaust
• Welding fumes
• Industrial effluents
• Sand blasting
INCIDENTAL/
MANMADE
• Metals (Ag,Au,Zn,Cu)
• Quantum dots
• Nanotubes/bulkyballs
(60 atoms)
• Suncreen pigments
• Nanocapsules/nanorods
ENGINEERED/
MANFACTURED
Applications of AgNP…
Nanoparticles Synthesis
Silver nanoparticles Specialty
• A great number of scientists have focused on silver nano-particles, which
are applied the most widely due to their well-known antibacterial and
antifungal effects, as well as their plasmonic (biosensors) and opto-
electrical properties (molecular diagnostics) (Pokhrel and Dubey 2013)
• Anti microbial assessment.
• Toxicity evaluation.
• Photo thermal ablation.
• Therapeutic efficacy.
• Anti cancerous activity.
• Wound healing capacity.
Why Green synthesis..?
• The growing production and exploitation of metal nanoparticles raise
concerns about their release to the atmosphere, as such release may have a
negative impact on the environment, and thus also on human health
(Cvjetko et al. 2018).
Biologically-prepared NPs -
• high yield, solubility, and stability
• simple, rapid, non-toxic, dependable, Eco friendly,Economical
• produce well-defined size and morphology useful for translational research.
• In the end, a green approach for the synthesis of NPs shows promising
outcome for future research.
LEPIDIUM SATIVUM (GARDEN CRESS)
Taxonomic classification
Kingdom: Plantae
Division: Tracheophyta
Class: Magnoliopsida,
Order: Brassicales,
Family: Brassicaceae
Genus: Lepidium, Species: Lepidium sativum [8].
OTHER NAMES-Garden Cress (English),Chandrashoor (Sanskrit),Adityalu or Aadhalu (Telugu).
DISTRIBUTION-It is distributed in Africa (Egypt, Ethiopia and Kenya), Asia (Kuwait, Oman, Saudi Arabia, United
Arab Emirates, Yemen, Afghanistan, Iran, Iraq, Palestine, Jordan, Lebanon, Syria, Turkey, Pakistan, China, Japan,
India).
Introduction to Lepidium Sativum Seeds
• Herbal medicine is the oldest form of healthcare known to mankind. Herbs had been used by
all cultures throughout history. Plants are a valuable source of a wide range of secondary
metabolites, which are used as pharmaceuticals, agrochemicals, flavours, fragrances, colours,
biopesticides and food additives .
• The phytochemical analysis of Lepidium sativum showed that it contained cardiac glycoside,
alkaloids, phenolic, flavonoids, cardiotonic glycosides, coumarins, glucosinolates,
carbohydrates, proteins and amino-acids, mucilage, resins, saponins, sterols, tannins, volatile
oils, triterpene, sinapic acid and uric acid.
Phytochemical screening of LS
Constituents Petroleum Chloroform Alchohol Aqueous
Alkaloids - + + +
carbohydrates - - - +
Phenolic
compounds
- + + -
Flavonoid - + + +
Protiens and amino
acids
- - + +
Saponins - - + +
Mucilage - - + -
Resins + + + -
Lipids/fats + - - -
Physiochemical characteristics of Lepidium Seeds -100g
physio chemical g/100g
Moisture 5.30± 0.08
Ash 4.25 ± 0.05
Protein 22.83 ± 0.72
Total fat 25.61 ± 1.04
Insoluble dietary fiber 5.68 ± 0.46
Soluble dietary fiber 1.59 ± 0.23
Total Dietary fiber 7.27 ± 0.60
Carbohydrate 34.67 ± 0.88
Energy
K.Cal
474.23
Water soluble Vitamins on fresh weight basis mg/100g
Sample
name
Ribof
lavin
(B2)
Niacin
(B3)
Pantoth
enic
acid
(B5)
Pyridox
ine (B6)
Folic
acid
(B9)
Vit-C
lepidium
seeds
0.28
±
0.01
0.97 ±
0.03
0.286 ±
0.01
-- 0.116
± 0.01
63 ±
5.26
GC-MS report presence of alkaloids of
Lepidium Sativum Seeds
• eicosatetraenoate (10.24%),
• guanosine (9.29%),
• dodecanamide, n-(2-hydroxyethyl) (7.48%),
• hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester (7.13%),
• 1-(1-adamantyl)-3-(1-piperidinyl)-1-propanone (6%),
• hexadecanoic acid (5.33%),
• 3-butylindolizidine (4.80%),
• 9,12-octadecadienoic acid (Z,Z)-,
• 2-hydroxy-1-hydroxymethyl (4.79%),
• 3-methyl alpha.-d-glucopyranoside (1.81%),
• stigmast-5-en-3-ol, (3. beta.) (3.58%),
• soyasapogenol B (1.15%),
• stigmasterol (1.07%),
• fucosterol (3.29%),
• gamma-tocopherol (5.04%)
• squalene (3.44%)
OBJECTIVE 1/CHAPTER 1
I) SYNTHESIS AND CHARACTERIZATION OF
SILVER NANO PARTICLES USING LEPIDIUM
SATIVUM.
Methodology
Preparation of 1% aqueous extract and 0.1mM AgNo3
• Seeds have been procured from local market identified and certified by
dept of botany Osmania university
• 100 gms of ls seeds coarsely grounded and from that 1 gram of
powder+100 ml distilled water is boiled at 80 c for 20 mins.
• This solution is filtered using Whatman filter NO 1 paper and aqueous
extract is collected the Final obtained solution was 50 ml.
• 1mM AgNo3 (0.168 mg) in 250 ml Milli Q water kept aside for
further evaluation.
Green synthesis of Silver Nanoparticles
40 ml of 1% LS extract
at pH 4 +10 ml of
0.1mM AgNO3 (1:4v/v)
Left at room
temperature 10
min-1hr
Nanoparticle
formed pale yellow
to brown colour
Nanoparticle extract mq water Agno3
Characterisation of Lepidium Sativum Silver
Nanoparticles (LsAgNp)
1. Uv-(Silver Nano Particle formation peak)
2. Ftir-(identification of functional groups)
3. Zeta potential-(charge)
4. Sem-(shape of nanoparticle)
Ultraviolet Spectrophotometry-404nM
Silver Nanoparticle formation was by
peak at 404 nM
FTIR Spectrum of silver nanoparticles
Band at 3728 cm-1 can be
associated with OH amide
vibrations.
• The bands at 2924 cm-1 and
794 cm-1 are CH amide groups
which confirms the presence
of alkenes
• The finger print regions at 765
and 588 are alkyl halides.
• The reduction,capping and
stabilization of nanoparticles
are involved in these bioactive
molecules
Scanning Electron Microscopes
The AgNp are polymorphic showing triangular, hexagonal, deformed
spherical.
Zeta potential is a measure of the effective electric charge on the nanoparticle surface. The
magnitude of the zeta potential provides information about particle stability.
PeakNo. Zeta
Potential
ElectrophoreticMobili
ty
1
-31.2 mV
-0.000242cm2/Vs
2 ---mV ---cm2/Vs
3 ---mV ---cm2/Vs
Zeta Potential(Mean) :-31.2mV
ElectrophoreticMobilitymean :-0.000242cm2/Vs
Results
• Synthesis of Nanoparticle-Sucessfully the silver nano particles are synthesized from Lepidium
sativum aqueous extract from Pale yellowish solution to dark brown solution.
• UV-Vis -Further to confirm AgNp through Uv vis spectrophotometer confirmed and the peak was
observed at 404 nm
• FTIR-The bands observed at 2924 cm-1 and 794 cm-1 are CH amide groups which confirms the
presence of alkenes,The finger print regions at 765 cm-1 and 588cm-1 are alkyl halides.The
reduction, capping and stabilization of nanoparticles are involved in these bioactive molecules.
• SCANNING ELECTRON MICROSCOPIC -The AgNp are polymorphic showing triangular,
hexagonal, deformed spherical.
• ZETA POTENTIAL -The small distribution of particulate size with z-average value of 40.5 nm
and low index of Polydistributor 0.31 is evident from of the Lepidium sativum seed extract
AgNPS.
Objective II
Anti Bacterial Activity of Lepidium
Sativum Silver Nanoparticles
Methodology
MEDIA PREPARATION:
1. 28 g of NA powder was Suspend in 1L of distilled water
in a 2 L conical flask.
2. The suspended NA mixture was digested and stirred to
completely dissolve.
3. It was autoclaved at 1210 c / 15 min.
4. When the flask was autoclaved it was permitted to cool
but not solidify
5. The medium solidified NA media was poured 25ml into
each petri plate, leaving the plates on the clean, uniform
surface at room temperature until the agar was solidified.
6. After the petri plates were solidified completely, these
were stored for further use.
Nutrient Agar Composition
Peptic digest of animal
tissue
5.000
Sodium chloride 5.000
Beef extract 1.500
Yeast extract 1.500
Agar 15.000
Final pH ( at 25°C) 7.4±0.2
Standard drug -OFLAXACIN
1 2 3
4 5 6
Gram Positive
Bacteria
1. Esterisia coli
2. Bacillus subtills
3. Klebsicella
pnenmoniae
4. Bacillus
stearothermoph
ilis
5. Staphylococus
aureus
6. Pseudomonas
aeruginosa
7 8 9
10 11 12
Gram Negative
Bacteria
7.Salmonella para
typhi
8.Salmonella typhi
9. Micrococcus
luteus
10.Proteus
vulgaris
11.Bacillus
megaterium
12. Bacillus cereus
Anti Bacterial Activity studies
Test Organisms Lepidium Silver
nanoparticles
AgNo3 Standard DW Extract
Zone of inhibition in mm
Salmonella typhi 23 18 09 ND ND
Salmonella Para typhi 24 16 20 ND ND
Staphylococcus aureus 19 10 06 ND ND
Proteus vulgaris 25 15 17 ND ND
Pseudomonas aeruginosa 21 16 30 ND ND
Micrococcus luteus 26 17 10 ND ND
klebsiella pneumoniae 21 09 22 ND ND
E.Coli 29 11 06 ND ND
Bacillus stearothermophilus 30 09 07 ND ND
Bacillus subtilis 31 18 07 ND ND
Bacillus megaterium 19 19 08 ND ND
Observations
1. The anti bacterial studies of LsAgNp indicate that the zone of
inhibition is more for Bacillus stereothermphilus (30), Bacillus
subtilis (31).
2. These above bacteria causes ANTHRAX bacteremia, endocarditis,
pneumonia, and septicemia.
3. Hence the LsAgNp may be used for treatment of anti microbial
resistant therapy providing maximum therapeutic effect with
minimal side effect.
4. The results confirm the potential uses of LS-AgNP as broad-
spectrum microbicides, disinfectants, and wound care products.
LsAgNP as Antimicrobial resistant treatment
Objective II-Anti Oxidant Activity Studies
• DPPH ASSAY
• NITRIC OXIDE
• ANTI OXIDANT REDUCING POWER ASSAY
DPPH ASSAY
METHOD
• DPPH-2,2-Diphenyl-1-Picrylhydrazyl (crystalline purple
compound,stable free radical,light and heat sensitive)
• Standard-Ascorbic acid
• Principle -DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free
radical method is an antioxidant assay based on electron-transfer that
produces a violet solution in ethanol . This free radical, stable at room
temperature, is reduced in the presence of an antioxidant molecule,
giving rise to colorless ethanol solution.
Procedure of DPPH Assay
1.Various concentrations of aqueous extract and standard (Ascorbic Acid) are been taken from (20 ug -100ug) and made
up to 3ml with methanol in test tubes.
2.Further 1 ml of 0.004% DPPH is been added and left at dark for 30mins and absorbance is calculated at 517 nm using
The below formula.
• The DPPH radical scavenging activity increased dependently on concentration
• Purple DPPH was converted to yellow DPPH
• The highest Inhibition percentage of lepidium sativum silver nanoparticles was 53.29±0.64 % in 100µg/ml
and the extract with 49.01±0.64%, while the standard ascorbic acid activity was 53.54± 0.20%.
Anti-inflammatory activity by bovine serum albumin denaturation
of lepidium sativum Silver nanoparticles.
0
20
40
60
80
100
120
1 2 3 4 5
indomethacin lsnp lsae
Concentr
ation
μg/ml
Indometh
acin
lepidium
sativum
seed
aqueous
mediated
silver
nanoparticl
es (LSNP)
lepidium sativum
seed aqueous
extracts (LSAE)
25 71.4 67 61
50 81.5 70.3 67.7
100 92.7 75.2 72.9
150 94.8 80.1 76.8
200 95.9 84.5 81.4
Results -Anti oxidant (DPPH,Nitric oxide,FRAP)
and Anti inflammatory (BSA Denaturation)
•Anti oxidant activity studies –
conducted by DPPH,NO,FRAP shows a dose dependent anti
oxidant activity with standard taken as ASCORBIC ACID
• Anti Inflammatory activity studies-
Conducted by Bovine Serum Albumin Denaturation method shows
dose dependent anti inflammatory activity with standard taken as
INDOMETHACIN.
Objective III (in-vitro)
ANTICANCER ACTIVITY STUDIES –MTT (3-(4,5-
Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)
ASSAY
Method –MTT stock solution
• MTT stock solution: Dissolve in the 10 mL phosphate buffer solution, 500 mg MTT powder. Stir the solution in the dark
place for around 1 hr with a magnet stirrer. Filter the sterilized solution with a 0.22 mm filter (Millipore, Ireland), then hold
at -20 ° C (van Meerloo et al., 2011) with 10-ml aliquots (50 mg / mL). On the day of the experiment dilution must produce
the working solution (5 mg / mL).
• ANTICANCER ACTIVITY (MTT ASSAY)
 Cells were seeded at a density of 1/104 cells/well in a 96-well flat-bottom microtiter plate and allowed to stay in a CO2
incubator for 24 hours at 37 ° C.
 Following 24 hrs of incubation, the media was replaced by a fresh medium.
 Cells were then treated in a CO2 incubator with different concentrations of the target compound for 24 hours at 37 ° C
 The culture media was replaced with a fresh medium after 24 hrs of incubation.
 10 μL of MTT working solution (5 mg / mL in phosphate buffer solution) was subsequently applied to each well and the
plate was incubated in a CO2 incubator for 4 hrs at 37 ° C.
 The formed formazan crystals were then solubilized by adding 50 μL DMSO per pool at 37 ° C for 30 min at a CO2
incubator.
 Eventually, with the ELISA plate reader at 570 nm, the pressure of the dissolved formazan crystals (purple colour) was
quantified.
Schematic Representation of MTT ASSAY
Cell viability
S.No Test Compound
Cell
Line Concentration
treated to cells
1 Untreated HeLa No treatment
2 Cisplatin HeLa 15ug/ml
3 Blank -
Only Media
without cells
4
lepidium
sativum seed
mediated Silver
nanoparticles.
HeLa
6.25,12.5,25,50
,100µg/ml 0
10
20
30
40
50
60
70
80
90
100
UNTREATED STD 6.25 12.5 25 50 100
%
of
Cell
Viability
Drug Concentration (ug/ml)
Comparative % of Cell viability of Silver NPs on HeLa cells
Graph : % cell viability values of SILVER NPS on HeLa cells
100 ug/ml Control Untreated
6.25 µg/ml 12.5ug/ml 25 ug/ml 50ug/ml
OVERALL CONCLUSIONS ..
• An successful eco-friendly and inexpensive method has been achieved for the
green synthesis of silver nanoparticles (AgNPs) by Lepidium sativum seed extract.
• Biosynthesized AgNPs is spherical in form, by antibacterial activity studies the
nanoparticle shows more inhibition on gram positive bacteria
• DPPH antioxidant studies shows more reducing power with increase in
concentration of AgNp extract.
• lepidium sativum seed mediated Silver nanoparticles as anticancer in nature on
HELA cells due to its lower IC50 values with 13.98 suggest that the lepidium
sativum seed mediated Silver nanoparticles is anticancer in nature and exhibiting
cytotoxic potency against Human cervical cancer cells (HeLa)
• This green biosynthesis approach is a non-toxic alternative to conventional
chemical and physical approaches and will be suitable for large-scale biological
processing and prospective therapies.
OBJECTIVE IV-(IN-VIVO)
• ANTI INFLAMMATORY AND WOUND HEALING IN ANIMAL MODEL
BENCH WORK COMPLETED BUT AWAITING FOR COMPILATION AND
CALCULATIONS.
UGC CARE LIST IOSR-Journal of Pharmacy
and Biological Sciences
Other publications (2014-2021)
1. Sree Vennela Rao1 and Veeresh1, “Biogenic synthesis and characterization of silver nanoparticles using aqueous extract
of Lepidium sativum assessing its antibacterial and antioxidant properties” IOSR Journal of Pharmacy and Biological
sciences (IOSR-JPBS),e-ISSN:2278-3008,Volume 16,Issue 1 Ser.II (jan.-feb.2021) pp (46-56).
2. P Amruth Rao1,Sree Vennela Rao2,Kavanshri N P,Devi CH V R, “Effect of probiotic curd (Indian dahi) Supplementation
in delaying the progression of prediabetes to diabetics Mellius-a pilot study, “International Journal of Diabetes and
Metabolic disorders-2021” ,ISSN 2475-5451,Vol 6.Issue 1,pg 136-139.
3. Amruth rao P,Shivaprakash M ,Madhavi G ,Sree Vennela Rao , MD.Khaleel Pasha , V Srinivas . R. Hemalatha. “Effect of
probiotic curd on cardiovascular disease (CVD) risk factors and body mass composition in People living with
HIV(PLWH)” - A pilot study Clinical Division,NIN, Hyderabad, India; BMC Infectious Diseases 2020, 20(Suppl
1):ISSHID.
4. Sree Vennela1*, Neharika Reddy, Rani Samyuktha, Poornachandar.G, T.Tamilanban, Venkataramana Devi.P “Therapeutic
Efficacy and Safety Profile of Tolvaptan Administered In Hyponatremia Patients” American Journal of Pharmatech
Research 2013 Oct issue 5.
5. Poorna chander G, Sree Vennela.P, Dr.Ramana Devi, Srinivas Naik.B “Invitro screening for different medicinal plants”
Journal of Phamacy and Biological Sciences” ISSN -2278-3008,volume 9,issue 2 ver II (mar-apr.2014).pg 68-79.
6. Amruth Rao P.*1 , Sree Vennela Rao P.3 , Khaleel Pasha MD.2 , Srinivas V.2 and Hemalatha R. “A study on
the effect of probiotic curd supplementation on HIV (PLWH) undergoing anti retroviral therapy (ART)” World Journal of
Pharmaceutical Research, ISSN 2277– 7105, Volume 10, Issue 5, 1492-1497,Apr 2021,pg 1492-1497.
THANK YOU…

More Related Content

Similar to Veenela.pptx

Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...
Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...
Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...KSDEEPAK2
 
Biological method for the preparation of nanoparticles(Sheersho)
Biological method for the preparation of nanoparticles(Sheersho)Biological method for the preparation of nanoparticles(Sheersho)
Biological method for the preparation of nanoparticles(Sheersho)Sheersha Pramanik 🇮🇳
 
Is Nano Medicine And Nano Technology The Most Trending Thing Now?
Is Nano Medicine And Nano Technology The Most Trending Thing Now?Is Nano Medicine And Nano Technology The Most Trending Thing Now?
Is Nano Medicine And Nano Technology The Most Trending Thing Now?science journals
 
Green synthesis of gold nanoparticles using various extract of plants and spices
Green synthesis of gold nanoparticles using various extract of plants and spicesGreen synthesis of gold nanoparticles using various extract of plants and spices
Green synthesis of gold nanoparticles using various extract of plants and spicesijsidonlineinfo
 
Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and Es...
Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and  Es...Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and  Es...
Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and Es...IJSIT Editor
 
Bombax ceiba thorn extract mediated synthesis of silver nanoparticles, eva...
Bombax ceiba thorn  extract mediated  synthesis of silver  nanoparticles, eva...Bombax ceiba thorn  extract mediated  synthesis of silver  nanoparticles, eva...
Bombax ceiba thorn extract mediated synthesis of silver nanoparticles, eva...roshan telrandhe
 
Green synthesis of silver nanoparticles and its application for mosquito
Green synthesis of silver nanoparticles and its application for mosquitoGreen synthesis of silver nanoparticles and its application for mosquito
Green synthesis of silver nanoparticles and its application for mosquitoAnh Vu
 
8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES
8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES
8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETESRavindragouda Patil
 
Isolation and Characterization of Osmolytes from Halophiles
Isolation and Characterization of  Osmolytes  from Halophiles Isolation and Characterization of  Osmolytes  from Halophiles
Isolation and Characterization of Osmolytes from Halophiles sachinhalladamani
 
Isolation, partial purification and characterization
Isolation, partial purification and characterizationIsolation, partial purification and characterization
Isolation, partial purification and characterizationeSAT Publishing House
 
Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...
Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...
Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...IRJET Journal
 
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...journal ijrtem
 
IRJET- Structure Prediction, Functional Characterization of Lectins and their...
IRJET- Structure Prediction, Functional Characterization of Lectins and their...IRJET- Structure Prediction, Functional Characterization of Lectins and their...
IRJET- Structure Prediction, Functional Characterization of Lectins and their...IRJET Journal
 
Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...
Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...
Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...Conference-Proceedings-CrimsonPublishers
 
IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)
IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)
IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)IRJET Journal
 

Similar to Veenela.pptx (20)

gupea_2077_38173_5
gupea_2077_38173_5gupea_2077_38173_5
gupea_2077_38173_5
 
Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...
Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...
Biogenic synthesis of Silver Nanoparticles fromOldenlandiya Cormybosa plant e...
 
Biological method for the preparation of nanoparticles(Sheersho)
Biological method for the preparation of nanoparticles(Sheersho)Biological method for the preparation of nanoparticles(Sheersho)
Biological method for the preparation of nanoparticles(Sheersho)
 
Is Nano Medicine And Nano Technology The Most Trending Thing Now?
Is Nano Medicine And Nano Technology The Most Trending Thing Now?Is Nano Medicine And Nano Technology The Most Trending Thing Now?
Is Nano Medicine And Nano Technology The Most Trending Thing Now?
 
Green synthesis of gold nanoparticles using various extract of plants and spices
Green synthesis of gold nanoparticles using various extract of plants and spicesGreen synthesis of gold nanoparticles using various extract of plants and spices
Green synthesis of gold nanoparticles using various extract of plants and spices
 
Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and Es...
Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and  Es...Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and  Es...
Determination of Anti-microbial Property of Gycyrrhiza lepidota roots and Es...
 
Bombax ceiba thorn extract mediated synthesis of silver nanoparticles, eva...
Bombax ceiba thorn  extract mediated  synthesis of silver  nanoparticles, eva...Bombax ceiba thorn  extract mediated  synthesis of silver  nanoparticles, eva...
Bombax ceiba thorn extract mediated synthesis of silver nanoparticles, eva...
 
Neem gold nanoparticles
Neem gold nanoparticlesNeem gold nanoparticles
Neem gold nanoparticles
 
Green synthesis of silver nanoparticles and its application for mosquito
Green synthesis of silver nanoparticles and its application for mosquitoGreen synthesis of silver nanoparticles and its application for mosquito
Green synthesis of silver nanoparticles and its application for mosquito
 
8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES
8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES
8- IJRANSS-ANTIMICROBIAL POTENTIAL OF MARINE ACTINOMYCETES
 
Isolation and Characterization of Osmolytes from Halophiles
Isolation and Characterization of  Osmolytes  from Halophiles Isolation and Characterization of  Osmolytes  from Halophiles
Isolation and Characterization of Osmolytes from Halophiles
 
Recent paper of dr.magapu solomon
Recent paper of dr.magapu solomonRecent paper of dr.magapu solomon
Recent paper of dr.magapu solomon
 
Isolation, partial purification and characterization
Isolation, partial purification and characterizationIsolation, partial purification and characterization
Isolation, partial purification and characterization
 
published
publishedpublished
published
 
Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...
Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...
Synthesis of the silver nanoparticles from Aloe barbadensis extract and its a...
 
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...
DNA Barcoding of Stone Fish Uranoscopus Oligolepis: Intra Species Delineation...
 
IRJET- Structure Prediction, Functional Characterization of Lectins and their...
IRJET- Structure Prediction, Functional Characterization of Lectins and their...IRJET- Structure Prediction, Functional Characterization of Lectins and their...
IRJET- Structure Prediction, Functional Characterization of Lectins and their...
 
Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...
Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...
Aqueous Stable Silver Nanoparticles Derived from Nimble Leaves of Psidium Gua...
 
ASSESSMENT OF UV-B INDUCED CHANGES IN GROWTH, PHOTOSYNTHETIC PIGMENTS AND BIO...
ASSESSMENT OF UV-B INDUCED CHANGES IN GROWTH, PHOTOSYNTHETIC PIGMENTS AND BIO...ASSESSMENT OF UV-B INDUCED CHANGES IN GROWTH, PHOTOSYNTHETIC PIGMENTS AND BIO...
ASSESSMENT OF UV-B INDUCED CHANGES IN GROWTH, PHOTOSYNTHETIC PIGMENTS AND BIO...
 
IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)
IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)
IRJET- Regeneration studies in chickpea genotypes (Cicer arietinum L.)
 

Veenela.pptx

  • 1. “Biogenic synthesis of silver nanoparticles and its Pharmacotherapeutic Applications using aqueous extract of Lepidium Sativum (Garden Cress Seeds)” Name of Research Scholar- P.Sree Vennela Rao Supervisor- Prof.B.Veeresh Scholarship- National Fellowship Registration Num- 9009-12-891-003 Presentation- PROGRESS SEMINAR Dept of Pharmacy, University College of Technology, Osmania University,Hyderabad,500007.
  • 2. Overview of Presentation 1.Introduction and Literature review 2.Objectives I) Synthesis and characterization of silver nanoparticles. II) Anti bacterial, Anti oxidant and Anti inflammatory activities. III) Anti cancer activity (in vitro) IV) Anti inflammatory/Wound healing activity (in vivo). 3.Methodology 4.Expected outcome 5.Summary and Discussion. 6.Limitations and Future prospects 7.Publications 6.Certifications
  • 4. Nanoparticles… • A nanoparticle is a small particle that ranges between 1 to 100 nanometers in size. Undetectable by the human eye, nanoparticles can exhibit significantly different physical and chemical properties to their larger material counterparts. • Nanoparticles have large surface area, which gives them more pronounced effects. Particle Type Diameter Size Range Coarse particles 2500 to 10,000 nm Fine particles/Dust 100 to 2500 nm Nano particles 1 to 100 nm Atoms and small molecules 0.1 nm
  • 5. Nano particle and its Applications..
  • 6. Types of nanoparticles… • Sea spray • Mineral composites • Volcanic ash • virus NATURAL/ NATURE • Cooking smoke • Diesel exhaust • Welding fumes • Industrial effluents • Sand blasting INCIDENTAL/ MANMADE • Metals (Ag,Au,Zn,Cu) • Quantum dots • Nanotubes/bulkyballs (60 atoms) • Suncreen pigments • Nanocapsules/nanorods ENGINEERED/ MANFACTURED
  • 7.
  • 10. Silver nanoparticles Specialty • A great number of scientists have focused on silver nano-particles, which are applied the most widely due to their well-known antibacterial and antifungal effects, as well as their plasmonic (biosensors) and opto- electrical properties (molecular diagnostics) (Pokhrel and Dubey 2013) • Anti microbial assessment. • Toxicity evaluation. • Photo thermal ablation. • Therapeutic efficacy. • Anti cancerous activity. • Wound healing capacity.
  • 11. Why Green synthesis..? • The growing production and exploitation of metal nanoparticles raise concerns about their release to the atmosphere, as such release may have a negative impact on the environment, and thus also on human health (Cvjetko et al. 2018). Biologically-prepared NPs - • high yield, solubility, and stability • simple, rapid, non-toxic, dependable, Eco friendly,Economical • produce well-defined size and morphology useful for translational research. • In the end, a green approach for the synthesis of NPs shows promising outcome for future research.
  • 12. LEPIDIUM SATIVUM (GARDEN CRESS) Taxonomic classification Kingdom: Plantae Division: Tracheophyta Class: Magnoliopsida, Order: Brassicales, Family: Brassicaceae Genus: Lepidium, Species: Lepidium sativum [8]. OTHER NAMES-Garden Cress (English),Chandrashoor (Sanskrit),Adityalu or Aadhalu (Telugu). DISTRIBUTION-It is distributed in Africa (Egypt, Ethiopia and Kenya), Asia (Kuwait, Oman, Saudi Arabia, United Arab Emirates, Yemen, Afghanistan, Iran, Iraq, Palestine, Jordan, Lebanon, Syria, Turkey, Pakistan, China, Japan, India).
  • 13. Introduction to Lepidium Sativum Seeds • Herbal medicine is the oldest form of healthcare known to mankind. Herbs had been used by all cultures throughout history. Plants are a valuable source of a wide range of secondary metabolites, which are used as pharmaceuticals, agrochemicals, flavours, fragrances, colours, biopesticides and food additives . • The phytochemical analysis of Lepidium sativum showed that it contained cardiac glycoside, alkaloids, phenolic, flavonoids, cardiotonic glycosides, coumarins, glucosinolates, carbohydrates, proteins and amino-acids, mucilage, resins, saponins, sterols, tannins, volatile oils, triterpene, sinapic acid and uric acid.
  • 14. Phytochemical screening of LS Constituents Petroleum Chloroform Alchohol Aqueous Alkaloids - + + + carbohydrates - - - + Phenolic compounds - + + - Flavonoid - + + + Protiens and amino acids - - + + Saponins - - + + Mucilage - - + - Resins + + + - Lipids/fats + - - -
  • 15. Physiochemical characteristics of Lepidium Seeds -100g physio chemical g/100g Moisture 5.30± 0.08 Ash 4.25 ± 0.05 Protein 22.83 ± 0.72 Total fat 25.61 ± 1.04 Insoluble dietary fiber 5.68 ± 0.46 Soluble dietary fiber 1.59 ± 0.23 Total Dietary fiber 7.27 ± 0.60 Carbohydrate 34.67 ± 0.88 Energy K.Cal 474.23
  • 16. Water soluble Vitamins on fresh weight basis mg/100g Sample name Ribof lavin (B2) Niacin (B3) Pantoth enic acid (B5) Pyridox ine (B6) Folic acid (B9) Vit-C lepidium seeds 0.28 ± 0.01 0.97 ± 0.03 0.286 ± 0.01 -- 0.116 ± 0.01 63 ± 5.26
  • 17. GC-MS report presence of alkaloids of Lepidium Sativum Seeds • eicosatetraenoate (10.24%), • guanosine (9.29%), • dodecanamide, n-(2-hydroxyethyl) (7.48%), • hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester (7.13%), • 1-(1-adamantyl)-3-(1-piperidinyl)-1-propanone (6%), • hexadecanoic acid (5.33%), • 3-butylindolizidine (4.80%), • 9,12-octadecadienoic acid (Z,Z)-, • 2-hydroxy-1-hydroxymethyl (4.79%), • 3-methyl alpha.-d-glucopyranoside (1.81%), • stigmast-5-en-3-ol, (3. beta.) (3.58%), • soyasapogenol B (1.15%), • stigmasterol (1.07%), • fucosterol (3.29%), • gamma-tocopherol (5.04%) • squalene (3.44%)
  • 18. OBJECTIVE 1/CHAPTER 1 I) SYNTHESIS AND CHARACTERIZATION OF SILVER NANO PARTICLES USING LEPIDIUM SATIVUM.
  • 19. Methodology Preparation of 1% aqueous extract and 0.1mM AgNo3 • Seeds have been procured from local market identified and certified by dept of botany Osmania university • 100 gms of ls seeds coarsely grounded and from that 1 gram of powder+100 ml distilled water is boiled at 80 c for 20 mins. • This solution is filtered using Whatman filter NO 1 paper and aqueous extract is collected the Final obtained solution was 50 ml. • 1mM AgNo3 (0.168 mg) in 250 ml Milli Q water kept aside for further evaluation.
  • 20. Green synthesis of Silver Nanoparticles 40 ml of 1% LS extract at pH 4 +10 ml of 0.1mM AgNO3 (1:4v/v) Left at room temperature 10 min-1hr Nanoparticle formed pale yellow to brown colour Nanoparticle extract mq water Agno3
  • 21. Characterisation of Lepidium Sativum Silver Nanoparticles (LsAgNp) 1. Uv-(Silver Nano Particle formation peak) 2. Ftir-(identification of functional groups) 3. Zeta potential-(charge) 4. Sem-(shape of nanoparticle)
  • 23. FTIR Spectrum of silver nanoparticles Band at 3728 cm-1 can be associated with OH amide vibrations. • The bands at 2924 cm-1 and 794 cm-1 are CH amide groups which confirms the presence of alkenes • The finger print regions at 765 and 588 are alkyl halides. • The reduction,capping and stabilization of nanoparticles are involved in these bioactive molecules
  • 24. Scanning Electron Microscopes The AgNp are polymorphic showing triangular, hexagonal, deformed spherical.
  • 25. Zeta potential is a measure of the effective electric charge on the nanoparticle surface. The magnitude of the zeta potential provides information about particle stability. PeakNo. Zeta Potential ElectrophoreticMobili ty 1 -31.2 mV -0.000242cm2/Vs 2 ---mV ---cm2/Vs 3 ---mV ---cm2/Vs Zeta Potential(Mean) :-31.2mV ElectrophoreticMobilitymean :-0.000242cm2/Vs
  • 26. Results • Synthesis of Nanoparticle-Sucessfully the silver nano particles are synthesized from Lepidium sativum aqueous extract from Pale yellowish solution to dark brown solution. • UV-Vis -Further to confirm AgNp through Uv vis spectrophotometer confirmed and the peak was observed at 404 nm • FTIR-The bands observed at 2924 cm-1 and 794 cm-1 are CH amide groups which confirms the presence of alkenes,The finger print regions at 765 cm-1 and 588cm-1 are alkyl halides.The reduction, capping and stabilization of nanoparticles are involved in these bioactive molecules. • SCANNING ELECTRON MICROSCOPIC -The AgNp are polymorphic showing triangular, hexagonal, deformed spherical. • ZETA POTENTIAL -The small distribution of particulate size with z-average value of 40.5 nm and low index of Polydistributor 0.31 is evident from of the Lepidium sativum seed extract AgNPS.
  • 27. Objective II Anti Bacterial Activity of Lepidium Sativum Silver Nanoparticles
  • 28. Methodology MEDIA PREPARATION: 1. 28 g of NA powder was Suspend in 1L of distilled water in a 2 L conical flask. 2. The suspended NA mixture was digested and stirred to completely dissolve. 3. It was autoclaved at 1210 c / 15 min. 4. When the flask was autoclaved it was permitted to cool but not solidify 5. The medium solidified NA media was poured 25ml into each petri plate, leaving the plates on the clean, uniform surface at room temperature until the agar was solidified. 6. After the petri plates were solidified completely, these were stored for further use. Nutrient Agar Composition Peptic digest of animal tissue 5.000 Sodium chloride 5.000 Beef extract 1.500 Yeast extract 1.500 Agar 15.000 Final pH ( at 25°C) 7.4±0.2 Standard drug -OFLAXACIN
  • 29. 1 2 3 4 5 6 Gram Positive Bacteria 1. Esterisia coli 2. Bacillus subtills 3. Klebsicella pnenmoniae 4. Bacillus stearothermoph ilis 5. Staphylococus aureus 6. Pseudomonas aeruginosa
  • 30. 7 8 9 10 11 12 Gram Negative Bacteria 7.Salmonella para typhi 8.Salmonella typhi 9. Micrococcus luteus 10.Proteus vulgaris 11.Bacillus megaterium 12. Bacillus cereus
  • 31. Anti Bacterial Activity studies Test Organisms Lepidium Silver nanoparticles AgNo3 Standard DW Extract Zone of inhibition in mm Salmonella typhi 23 18 09 ND ND Salmonella Para typhi 24 16 20 ND ND Staphylococcus aureus 19 10 06 ND ND Proteus vulgaris 25 15 17 ND ND Pseudomonas aeruginosa 21 16 30 ND ND Micrococcus luteus 26 17 10 ND ND klebsiella pneumoniae 21 09 22 ND ND E.Coli 29 11 06 ND ND Bacillus stearothermophilus 30 09 07 ND ND Bacillus subtilis 31 18 07 ND ND Bacillus megaterium 19 19 08 ND ND
  • 32. Observations 1. The anti bacterial studies of LsAgNp indicate that the zone of inhibition is more for Bacillus stereothermphilus (30), Bacillus subtilis (31). 2. These above bacteria causes ANTHRAX bacteremia, endocarditis, pneumonia, and septicemia. 3. Hence the LsAgNp may be used for treatment of anti microbial resistant therapy providing maximum therapeutic effect with minimal side effect. 4. The results confirm the potential uses of LS-AgNP as broad- spectrum microbicides, disinfectants, and wound care products.
  • 33. LsAgNP as Antimicrobial resistant treatment
  • 34. Objective II-Anti Oxidant Activity Studies • DPPH ASSAY • NITRIC OXIDE • ANTI OXIDANT REDUCING POWER ASSAY
  • 35. DPPH ASSAY METHOD • DPPH-2,2-Diphenyl-1-Picrylhydrazyl (crystalline purple compound,stable free radical,light and heat sensitive) • Standard-Ascorbic acid • Principle -DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) free radical method is an antioxidant assay based on electron-transfer that produces a violet solution in ethanol . This free radical, stable at room temperature, is reduced in the presence of an antioxidant molecule, giving rise to colorless ethanol solution.
  • 36. Procedure of DPPH Assay 1.Various concentrations of aqueous extract and standard (Ascorbic Acid) are been taken from (20 ug -100ug) and made up to 3ml with methanol in test tubes. 2.Further 1 ml of 0.004% DPPH is been added and left at dark for 30mins and absorbance is calculated at 517 nm using The below formula. • The DPPH radical scavenging activity increased dependently on concentration • Purple DPPH was converted to yellow DPPH • The highest Inhibition percentage of lepidium sativum silver nanoparticles was 53.29±0.64 % in 100µg/ml and the extract with 49.01±0.64%, while the standard ascorbic acid activity was 53.54± 0.20%.
  • 37. Anti-inflammatory activity by bovine serum albumin denaturation of lepidium sativum Silver nanoparticles. 0 20 40 60 80 100 120 1 2 3 4 5 indomethacin lsnp lsae Concentr ation μg/ml Indometh acin lepidium sativum seed aqueous mediated silver nanoparticl es (LSNP) lepidium sativum seed aqueous extracts (LSAE) 25 71.4 67 61 50 81.5 70.3 67.7 100 92.7 75.2 72.9 150 94.8 80.1 76.8 200 95.9 84.5 81.4
  • 38. Results -Anti oxidant (DPPH,Nitric oxide,FRAP) and Anti inflammatory (BSA Denaturation) •Anti oxidant activity studies – conducted by DPPH,NO,FRAP shows a dose dependent anti oxidant activity with standard taken as ASCORBIC ACID • Anti Inflammatory activity studies- Conducted by Bovine Serum Albumin Denaturation method shows dose dependent anti inflammatory activity with standard taken as INDOMETHACIN.
  • 39. Objective III (in-vitro) ANTICANCER ACTIVITY STUDIES –MTT (3-(4,5- Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) ASSAY
  • 40. Method –MTT stock solution • MTT stock solution: Dissolve in the 10 mL phosphate buffer solution, 500 mg MTT powder. Stir the solution in the dark place for around 1 hr with a magnet stirrer. Filter the sterilized solution with a 0.22 mm filter (Millipore, Ireland), then hold at -20 ° C (van Meerloo et al., 2011) with 10-ml aliquots (50 mg / mL). On the day of the experiment dilution must produce the working solution (5 mg / mL). • ANTICANCER ACTIVITY (MTT ASSAY)  Cells were seeded at a density of 1/104 cells/well in a 96-well flat-bottom microtiter plate and allowed to stay in a CO2 incubator for 24 hours at 37 ° C.  Following 24 hrs of incubation, the media was replaced by a fresh medium.  Cells were then treated in a CO2 incubator with different concentrations of the target compound for 24 hours at 37 ° C  The culture media was replaced with a fresh medium after 24 hrs of incubation.  10 μL of MTT working solution (5 mg / mL in phosphate buffer solution) was subsequently applied to each well and the plate was incubated in a CO2 incubator for 4 hrs at 37 ° C.  The formed formazan crystals were then solubilized by adding 50 μL DMSO per pool at 37 ° C for 30 min at a CO2 incubator.  Eventually, with the ELISA plate reader at 570 nm, the pressure of the dissolved formazan crystals (purple colour) was quantified.
  • 42. Cell viability S.No Test Compound Cell Line Concentration treated to cells 1 Untreated HeLa No treatment 2 Cisplatin HeLa 15ug/ml 3 Blank - Only Media without cells 4 lepidium sativum seed mediated Silver nanoparticles. HeLa 6.25,12.5,25,50 ,100µg/ml 0 10 20 30 40 50 60 70 80 90 100 UNTREATED STD 6.25 12.5 25 50 100 % of Cell Viability Drug Concentration (ug/ml) Comparative % of Cell viability of Silver NPs on HeLa cells Graph : % cell viability values of SILVER NPS on HeLa cells
  • 43. 100 ug/ml Control Untreated 6.25 µg/ml 12.5ug/ml 25 ug/ml 50ug/ml
  • 44. OVERALL CONCLUSIONS .. • An successful eco-friendly and inexpensive method has been achieved for the green synthesis of silver nanoparticles (AgNPs) by Lepidium sativum seed extract. • Biosynthesized AgNPs is spherical in form, by antibacterial activity studies the nanoparticle shows more inhibition on gram positive bacteria • DPPH antioxidant studies shows more reducing power with increase in concentration of AgNp extract. • lepidium sativum seed mediated Silver nanoparticles as anticancer in nature on HELA cells due to its lower IC50 values with 13.98 suggest that the lepidium sativum seed mediated Silver nanoparticles is anticancer in nature and exhibiting cytotoxic potency against Human cervical cancer cells (HeLa) • This green biosynthesis approach is a non-toxic alternative to conventional chemical and physical approaches and will be suitable for large-scale biological processing and prospective therapies.
  • 45. OBJECTIVE IV-(IN-VIVO) • ANTI INFLAMMATORY AND WOUND HEALING IN ANIMAL MODEL BENCH WORK COMPLETED BUT AWAITING FOR COMPILATION AND CALCULATIONS.
  • 46.
  • 47. UGC CARE LIST IOSR-Journal of Pharmacy and Biological Sciences
  • 48. Other publications (2014-2021) 1. Sree Vennela Rao1 and Veeresh1, “Biogenic synthesis and characterization of silver nanoparticles using aqueous extract of Lepidium sativum assessing its antibacterial and antioxidant properties” IOSR Journal of Pharmacy and Biological sciences (IOSR-JPBS),e-ISSN:2278-3008,Volume 16,Issue 1 Ser.II (jan.-feb.2021) pp (46-56). 2. P Amruth Rao1,Sree Vennela Rao2,Kavanshri N P,Devi CH V R, “Effect of probiotic curd (Indian dahi) Supplementation in delaying the progression of prediabetes to diabetics Mellius-a pilot study, “International Journal of Diabetes and Metabolic disorders-2021” ,ISSN 2475-5451,Vol 6.Issue 1,pg 136-139. 3. Amruth rao P,Shivaprakash M ,Madhavi G ,Sree Vennela Rao , MD.Khaleel Pasha , V Srinivas . R. Hemalatha. “Effect of probiotic curd on cardiovascular disease (CVD) risk factors and body mass composition in People living with HIV(PLWH)” - A pilot study Clinical Division,NIN, Hyderabad, India; BMC Infectious Diseases 2020, 20(Suppl 1):ISSHID. 4. Sree Vennela1*, Neharika Reddy, Rani Samyuktha, Poornachandar.G, T.Tamilanban, Venkataramana Devi.P “Therapeutic Efficacy and Safety Profile of Tolvaptan Administered In Hyponatremia Patients” American Journal of Pharmatech Research 2013 Oct issue 5. 5. Poorna chander G, Sree Vennela.P, Dr.Ramana Devi, Srinivas Naik.B “Invitro screening for different medicinal plants” Journal of Phamacy and Biological Sciences” ISSN -2278-3008,volume 9,issue 2 ver II (mar-apr.2014).pg 68-79. 6. Amruth Rao P.*1 , Sree Vennela Rao P.3 , Khaleel Pasha MD.2 , Srinivas V.2 and Hemalatha R. “A study on the effect of probiotic curd supplementation on HIV (PLWH) undergoing anti retroviral therapy (ART)” World Journal of Pharmaceutical Research, ISSN 2277– 7105, Volume 10, Issue 5, 1492-1497,Apr 2021,pg 1492-1497.
  • 49.