CSIR-National Botanical Research Institute

24 Pages·2013·2.12 MB·English

ISSN 0409-7467

CSIR News
N C s & i r
ewsletter of the ouNCil of CieNtifiC Ndustrial esearCh
Volume 63 No. 13 & 14 website: http://www.csir.res.in July 2013
In The News
I n T h i s I s s u e
Herbi-Chew: Tobacco and Chemical-free Mixture
developed by CSIR-CIMAP and CSIR-NBRI
145
In The News
• Herbi-Chew: Tobacco and
Tobacco-based products,
Chemical-free Mixture
Released especially gutkha, are
• OSDD to set up Chemically
popular with all age and
Diverse Compound Library
• MEMS Technology-based socioeconomic groups.
Gyroscope Unfortunately, they are a
potent cause of oral and
148
R&D Highlights throat cancer. In India,
• Paper with 1000+ Citations over 90 percent cases of
Published in
oral cancer are caused by
CSIR-NISCAIR Journal
• Beta-glucosidase Inhibitors tobacco use. CSIR-CIMAP
Hidden in the Microbial
and CSIR-NBRI have
World
now jointly developed a
154 tobacco- and nicotine-
MoUs
free herbal formulation Dr. P.K. Seth and Dr. C.S. Nautiyal (in centre) and team
155 with the aroma and taste of CSIR-CIMAP & CSIR-NBRI releasing Herbi-Chew formulation
Workshops
gutkha, but without its side
Dayanandan Mani,
156 effects.
National Technology Anirban Pal, Mahendra
The herbal mixture,
Day Celebrations Pandurang Darokar and
called Herbi-Chew, is a mouth
Alok Kalra from CSIR-
freshener with antioxidants,
159 National Science Day CIMAP and Ajay Kumar
anti-bacterial and digestive
Singh Rawat, Sanjeev Ojha
Celebrations
properties. Feedback from
and Chanana Venkat Rao
users has shown that the
160 from CSIR-NBRI.
National Safety Week
product has good tobacco
The new product was
Celebrations
de-addiction properties –
released on the occasion of
users stopped or considerably
161 the National Technology
Lectures
reduced the use of tobacco-
Day at CSIR-CIMAP by
based formulations.
164 the Chief Guest Dr. P.K.
Training Programmes
The formulation has
Seth, CEO, Biotech Park,
been developed by a team led
165 Lucknow. Leading gutkha
Honours and Awards
by Dr. C.S. Nautiyal, Director,
manufacturing companies
and comprising Dinesh
168 are vying for the Herbi-
Obituary
Kumar, Sudha Agarwal,
Chew technology.
CSIR News 145
JULY 2013
OSDD to set up Chemically Diverse
In The News
Compound Library
The Open Source Drug Discovery (OSDD), Under this programme, OSDD
a Council for Scientific and Industrial Chemistry Outreach Centres across various
Research (CSIR) venture, is planning to build CSIR laboratories train students in synthetic
a chemical library of chemically diverse chemistry and the compounds synthesized
compounds for successful drug discovery in the universities, institutes and colleges are
programmes. The novel initiative could help submitted to the OSDDChem database and
OSDD researchers discover novel drug sent to CSIR-CDRI. These molecules are
leads from the currently unexplored chemical then taken up for screening at CSIR-CDRI
space. for anti-TB and anti-malarial activity.
CSIR launched the OSDD programme Different projects from 34 academic
in 2008 with the objective of discovering institutions across the country have been
drugs for neglected diseases like TB, malaria, submitted to OSDDChem.
etc. Since then, OSDD has emerged as a Following the successful launch of the
successful model of open innovation hailed OSDDChem, which had envisioned the
by Science (24 February 2012, Vol. 335 no. development of a national online repository
6071 p. 909) and Forbes (9 April 2012) of small drug like molecules, OSDD is now
magazines. venturing into building chemical libraries with
OSDD works in all fields of drug diverse compounds for driving successful
discovery and development from in silico drug discovery programmes. This aligns with
predications to actual clinical trials and the OSDD policy of ‘no molecule will be
possesses robust biology and chemistry left behind’ for screening against neglected
components to support discovery and diseases and the assurance that the molecules
development modules. One of the novel submitted to OSDD will be taken up for
initiatives of OSDD during the 11th Plan was screening against neglected tropical
an open innovation model in chemistry called infections like TB and malaria.
the Chemistry Outreach Program Lack of chemically diverse compounds
(OSDDChem). has been recognized as a crucial factor for
146 CSIR News
JULY 2013
the poor success rates of anti-infective drug of compounds has been built up.
discovery and development in the past years. OSDD aims to make use of the vast
Diversity of chemical compounds used for and talented pool of chemists available
screening increases the chances of identifying across different CSIR laboratories to achieve
novel chemical scaffolds and hence this task of design and synthesis of novel
availability of compound libraries with compounds. Chemistry expertise available in
diverse set of molecules forms a critical different CSIR laboratories including CSIR-
component of successful drug discovery IICT, CSIR-NIIST, CSIR-NCL, CSIR-
programmes. CLRI, CSIR-CDRI, CSIR-IIIM and CSIR- While pharma
While pharma companies invest in NEIST are being tapped by OSDD to
companies invest in
therapeutic areas with larger revenues, achieve this purpose.
infectious diseases particularly tropical The compound library is to be created therapeutic areas
infections like TB, malaria and leishmaniasis using three important synthetic approaches with larger revenues,
are neglected and the modern compound – target-oriented synthesis, focused synthesis
infectious diseases
libraries often lack diverse molecule pool and diversity-oriented synthesis to generate
with potential antibacterial properties. OSDD novel compounds belonging to different particularly tropical
aims to bridge this gap in drug discovery classes.
infections like TB,
through a chemically diverse compound The compounds generated under this
malaria and
library, which could be used for screening programme will be received and stored in
various drug targets of discovery the dedicated space of OSDD within MOL leishmaniasis are
programmes of OSDD. bank facility of CSIR-IICT and CSIR-CDRI.
neglected and the
The initiative aims at the synthesis, To ensure purity and identity, quality control
modern compound
characterization, storage and management of is carried out and well characterized
chemically diverse compounds for screening compounds are to be registered with unique libraries often lack
against pathogens like Mycobacterium tuberculosis accession codes. OSDD also employs expert
diverse molecule
and research under OSDD umbrella. The personnel for management of samples,
compounds can also serve as probes for handling of data and appropriate distribution pool with potential
chemical biology research, which will, in turn, of plates/compounds for screening. antibacterial
help solve the biologically relevant questions OSDD has over 7500 participants across
properties. OSDD
in tuberculosis. 130 countries. It aims to acquire at least 2000
All the compounds deposited under this compounds every year from the CSIR aims to bridge this
initiative will be available for screening against laboratories with chemically diverse
gap in drug
M. tb, malaria, filariasis, and leishmaniasis. characteristics under this initiative.
discovery through a
OSDD is planning to make the library
available for screening against other Haridas Rode and chemically diverse
neglected diseases once a significant number Geetha Sugumaran
compound library,
which could be used
MEMS Technology-based Gyroscope for screening various
drug targets of
Developed at CSIR-CEERI
discovery
CSIR-Central Electronics Engineering Rate angle Signal programmes of
Research Institute (CEERI) has, for the Vports – tt2 ddm OSDD.
+ 1 m
first time in India, designed and developed
an MEMS gyroscope for aerospace Driving
electrode
applications. A strategic requirement for the Monitoring
Signal
country, the task was sponsored by the
Aeronautics Development Agency (ADA), V Vd2
d2
Bangalore.
y
The gyroscope design is based on x
Lagrangian analysis. Based on this analysis, Proposed structural view of nickel Gyroscope
CSIR News 147
JULY 2013
Complete structure of fabricated SEM microphotograph showing proof mass
nickel micro Gyroscope and cantilever springs of Gyroscope
lumped differential equations for both sense and excitation
modes of the device have been constructed and then
solved analytically for both transient and frequency
responses.
The design parameters considering nickel as a
structural material are: 47.76 kHz resonance frequency,
1.45 µgm proof mass value and 0.25 mV/degrees/s
2
sensitivity. The total chip size is 4x4 mm .
Considering nickel-specific designed parameters, mask
layouts were generated and masks were made
subsequently. The basic structure of nickel for the
gyroscope was then built by electro-forming technique
using UV-LIGA process.
Measured frequency-amplitude characteristics and
Detailed SEM microphotograph of corresponding phase of Gyro device showed resonance
actuation fingers of Gyroscope frequency of 45.6 kHz, which is close to design frequency
(47.76 kHz). The bandwidth estimated is 450 Hz.
R&D Highlights
Paper with 1000+ Citations Published in
CSIR-NISCAIR Journal
A recent study had revealed that there are paper published in a CSIR-National Institute
36 papers with at least one author from India of Science Communication and Information
that have received 1000 or more citations. Resources (NISCAIR) journal is by P. Kakkar,
As of April 2013, such papers have gone up B. Das and P.N. Viswanathan [A modified
to 38 with two more Indian papers receiving spectrophotometric assay of superoxide-
1000 or more citations. dismutase, Indian Journal of Biochemistry and
Incidentally, out of the 38 papers, there Biophysics, 21 (2) (1984) 130-132].
is only one paper that has been published in The paper describes a simple and rapid
an Indian journal. All the other papers that method for the assay of superoxide dismutase
have received 1000+ citations have been in biological samples. The method takes
published in foreign journals. The lone Indian advantage of the inhibition of NADH-
148 CSIR News
JULY 2013
Fig. 1. Growth of citations
dependent-nitroblue tetrazolium reduction 48 hours. The paper has
by the enzyme. The study found that As per the Science Citation Index-
facilitated estimation
inhibition of the formazan formation by Expanded database of the Web of Science,
superoxide dismutase was linear with increase the paper has received 1070 citations as on 4 of superoxide
in enzyme concentration and the formazan April 2013. The citation count is higher in
dismutase, an
extracted in butanol was stable even up to Google Scholar and Scopus owing to the
important
antioxidant enzyme
In conversation with the first author Dr. Poonam Kakkar, Scientist ‘G’ and Head, Herbal Research
Section, CSIR-Indian Institute of Toxicology Research (IITR), Lucknow whose level is found
to be altered in
GM: Please give us a brief history of the paper.
PK: The paper describes a simple and sensitive method for estimation of superoxide many disease
dismutase. This modified method was developed during the period when I was trying to
conditions. The
purify superoxide dismutase from lung tissue as a part of my PhD work. It was difficult to
estimate the enzyme in more than hundred samples in a day. Hence, we thought of developing method has been
an endpoint measurement assay to measure the formazan formed in large number of
used even in
samples with convenience.
laboratories where
GM: Did you expect your paper to become so hugely cited? sophisticated
PK: The paper was published in 1984 and soon after its publications we started getting
equipments are
large number of reprint requests. I had no idea that this method would become so popular.
However, with passage of time, I came across many people who were using this method. available.
GM: What are the major findings of the manuscript?
Dr. Poonam Kakkar
PK: The method described in the manuscript for estimation of an antioxidant enzyme
CSIR-IITR
superoxide dismutase, is a rapid, simple and sensitive method. Large number of samples
can be assayed simultaneously in test tubes. The formazan formed at the end of the
reaction once extracted in butanol is stable up to 48 hours, thus providing freedom to
measure it at a convenient time after stopping the reaction.
GM: What has been the impact of your paper?
PK: The paper has facilitated estimation of superoxide dismutase, an important antioxidant
enzyme whose level is found to be altered in many disease conditions. The method has
been used even in laboratories where sophisticated equipments are available.
CSIR News 149
JULY 2013
Table 1. Citations from Table 2. Citations from top 10
R&D Highlights
top 10 countries organizations
Countries/Territories Citations Organizations Citations
India 909 Annamalai Univ 303
USA 40 University of Kerala 79
Peoples R China 35 Indian Institute of Toxicology 56
Iran 32 Research, CSIR
Bose Institute 39
Pakistan 26
Central Drug Research Institute, 34
Saudi Arabia 26
CSIR
South Korea 20
Jadavpur University 34
Egypt 15
Metabolic Disorders Research Centre 32
France 9
All India Institute of Medical Sciences 30
Japan 6
Defence Research Development 30
Establishment
Gujarat University 26
Med College Hospital, Trivandrum 26
As per the Science
Citation Index-
Table 3. Citations from top 10 journals
Expanded database
Journals Papers JIF
of the Web of
Science, the paper Journal of Ethnopharmacology 41 3.014
Food and Chemical Toxicology 33 2.999
has received 1070
Indian Journal of Experimental Biology 26 1.295
citations as on
Molecular and Cellular Biochemistry 25 2.057
4 April 2013. The
European Journal of Pharmacology 22 2.516
citation count is Chemico Biological Interactions 21 2.865
higher in Google International Journal of Neuroscience 21 0.967
Journal of Pharmacy and Pharmacology 20 2.175
Scholar and Scopus
Toxicology 19 3.681
owing to the wider
Clinica Chimica Acta 16 2.535
coverage of these
two sources.
wider coverage of these two sources. countries but more than 90% of the citations
However, here we look at the citations as (909 citations) are from India followed by
available in the Science Citation Index- USA (40), China (35), Iran (32) and so on.
Expanded. Table 1 gives the top ten countries from
It can be seen from Figure 1 that the where the article has been most cited.
1984 paper received its first two citations in We also looked at the institutions that
1989, nearly five years after its publication. have cited the paper and it is seen that
In the first decade of the paper’s publication, researchers from the Annamalai University
it received less than ten citations per year have cited the paper 303 times followed by
and about 50% of the citations were received University of Kerala (79 papers). The Indian
in the first 24 years. Since 2009, the number Institute of Toxicology Research, the
of citations has surged with the paper institute that published the hugely cited
receiving more than hundred citations every paper, is the third (56 times) in the list of
year. institutions that have cited the paper the most
The paper has been cited from 39 (Table 2).
150 CSIR News
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The 1070 citations are of papers that The average IF of all the citing journals
have been published in 315 journals with is 2.148 and the journal with the highest IF
Journal of Ethonopharmacology citing the IJBB that has cited the article is Biomaterials
paper 41 times followed by Food and Chemical (2 citations, IF 7.404) followed by International
Toxicology (33) and another CSIR-NICAIR Journal of Cardiology (3 citations, IF 7.078).
journal, Indian Journal of Experimental Biology
citing the paper 26 times. Incidentally, the G. Mahesh and Yatish Panwar
source journal, Indian Journal of Biochemistry at CSIR-National Institute of Science
and Biophysics, has self-cited the paper only Communication and Information
11 times. Resources
Beta-glucosidase Inhibitors Hidden in the
Microbial World
Glucosidase inhibitors are responsible for inhibition tests and
disruption of the activity of enzyme interpreting the results.
glucosidase. These inhibitors have played a The methods employed,
vital role in revealing the functions of till date, indirectly assess
glucosidases in living systems, leading to the inhibition activity by
practical applications in agriculture and measuring the intensity
medicine. of a glucosidase positive
The antibiotics nojirimycin and reaction. PNPG
validamycin derived from the microbial (p-nitrophenyl-β-D-
species of Streptomyces and Bacillus are being glucopyranoside) has
extensively used as β-glucosidase inhibitors. been the most commonly
Owing to their therapeutic potential in the used substrate in either
treatment of certain metabolic disorders like TLC autographic or
diabetes, obesity, lysosomal storage disease microplate screening Fig. 1. β-glucosidase inhibition using the agar plate
and viral infections of HIV, the quest for assay. The reaction method developed in this study
new glucosidase inhibitors is crucial. Yet, the product, a nitrophenyl chromophore, is faint
search for β-glucosidase inhibitors in yellow and not in contrast with the
microorganisms is extremely limited, though surroundings for clear visual distinction of
natural sources like plant extracts, microalgae, the inhibitor.
cyanobacteria and mushrooms have been
explored.
Microorganisms, particularly marine
microorganisms, have an unparalleled
distinction of producing valuable
compounds because of their metabolic
diversity. Moreover, the microbial
production process is scalable using
biotechnological methods, so as to
circumvent the supply problem associated
with marine invertebrates and plankton.
Screening microbial culture extracts for
uncovering novel inhibitors is therefore of
immense interest. However, few beta-
glucosidase inhibitors have so far been
Fig. 2. Side-by-side comparison of agar plate method with TLC autography
reported from natural sources, maybe due method. Samples labelled as 1, 2, 3, 4, 5, 6, 7 and 8 are the methanol extracts
to the practical difficulties in performing the of marine microorganisms and, C is for control-0.75µg conduritol β-epoxide
CSIR News 151
JULY 2013
a Another TLC autographic method,
R&D Highlights
using esculin as substrate, was a better
alternative to the methods using PNPG, since
esculetin released from esculin, due to the
enzyme substrate reaction, formed a blackish
brown precipitate with FeCl that could be
3
clearly observed. However, TLC
autographic methods are not suitable for high
throughput screening as they are more
laborious, time consuming, and uniform
separation of compounds in all extracts
cannot be achieved with single solvent system.
b
Hence, spotting all the extracts on one TLC
plate to rapidly perform the assay would be
frustrating.
For screening a large number of natural
extracts, TLC autography was performed
without developing the plate so that activities
resulting from synergistic action of multiple
components of extracts are detected. In this
context, we considered the use of TLC plate
to be unnecessary because it increases the
time, cost and labour, but does not give
satisfactory results. The redundance of TLC
plate in this method is more pronounced
since the enzyme-agar solution does not
Fig. 3: Conduritol β-epoxide in different doses spread uniformly on the TLC plate, making
in: a) agar plate method – samples spot it difficult to observe the inhibition activity
inoculated on the agar surface b) TLC autogra-
as clear spots in contrast to the brown
phy method C1 - 2.5µg, C2 - 1.0µg, C3 -
0.50µg, C4 - 0.10µg and C5 – 0.05µg surrounding area. Although some zones were
visible, chances of losing a few promising
natural extracts were there as it was difficult
to ascertain certain samples as positive or
negative.
Actinobacteria Another important drawback of this
method was that the assay required around
4.5 to 5 hours, including an incubation time
of two hours each for the enzyme-inhibitor
and enzyme-substrate reaction. In a nutshell,
accurate assessment of glucosidase inhibition
ααααα-Proteobacteria activity in several extracts at a time is difficult
t and time consuming by conventional
1
βββββ-Proteobacteria methods.
In an effort to investigate microbial
Bacteroidetes
extracts from marine sponge and sediment
γ-Proteobacteria associated bacteria for β-glucosidase
inhibitors, we developed a novel agar plate
Fig. 4. Distribution of β-glucosidase
inhibiting marine isolates in different based method. The enzyme β-glucosidase
bacterial phyla showing Firmicutes making (0.01 U/ml) was incorporated in 0.7% agar
up the major fraction followed by solution along with the indicator 0.6%
Actinobacteria, Proteobacteria and
weight/volume FeCl and poured into petri
γ
Bacteroidetes. Within Proteobacteria, the - 3
dishes. Then the samples of microbial
Proteobacteria constitute the major fraction.
152 CSIR News
JULY 2013
extracts were spot inoculated on the agar which is pictorially depicted in Figure 4.
surface for the enzyme-inhibitor reaction and The inhibitors are abundant in bacterial
air dried for fifteen minutes. The plate was associates of marine sponge Aka coralliphaga,
flooded with 0.2% esculin solution and left and majorly belong to phylum Firmicutes
at room temperature for 15-30 minutes for followed by the phylum Actinobacteria.
the enzyme-substrate reaction. Species of Bacillus and Streptomyces are already
With the new agar plate method, pale known for their ability to produce diverse
yellowish zones against the blackish brown bioactive compounds; our finding is more
background could be visually observed with important in the context of genera like
more clarity in sample extracts where β- Staphylococcus, Stenotrophomonas, Pseudochrobactrum,
glucosidase inhibitor was present (Figure 1). Advenella, Dietzia and Chryseomicrobium, which
The new method was compared with the are hitherto unknown as producers of
closest existing method and established antimicrobial and anti-enzyme compounds.
clearly. The clarity and contrast of zones in This work fills a huge gap in the
agar plate based method, when compared literature concerning the use of
We believe our
side by side with TLC autographic method microorganisms as producers of beta-
is quite prominent in Figure 2 and Figure 3. glucosidase inhibitors. It was being speculated revelations form only
Conduritol β-epoxide, an irreversible that compounds producing glucosidase
the tip of an iceberg,
inhibitor of beta-glucosidase, was tested in inhibitors exist in the microbial world, but
a dose dependent order to confirm the there are discrete and few published reports. and this work will set
effectiveness of this method. Imidazole Many authors have emphasized that a new path to search
derivatives – 1-(3-aminopropyl)-imidazole phylogenetic diversity is the source of varied
for these compounds
and 2-aminobenzimidazole – were also biological activity.
tested as reversible inhibitors of β- Thus, within the realm of possibility we and find many more
glucosidase. This whole procedure required may assume these microorganisms have a
“cell factories”
about one hour for minimum 12 samples potential to produce new glucosidase
hidden in the deep
and the throughput could be increased with inhibitor compounds. Probably the missing
the size of the agar gel plate or by using a link between the phylogenetic diversity and sea. The search for
multiple of plates. The new protocol was functional diversity can be established, if all
better molecules is
simple, rapid and effective in detecting beta- these inhibitors are structurally identified and
never ending, so as
glucosidase inhibitors in a large number of their reaction mechanisms established.
microbial extracts in a short span of time. We believe our revelations form only the to replace or
A large number of methanol extracts tip of an iceberg, and this work will set a
augment the ones
of microorganisms were screened using the new path to search for these compounds
new method, and we found 98 extracts and find many more “cell factories” hidden which are already on
(32%) containing inhibitors out of 304 in the deep sea. The search for better the stands.
extracts tested. The significant number of molecules is never ending, so as to replace
glucosidase inhibiting microorganisms found or augment the ones which are already on
in this study prompted us to look into the the stands.
taxonomic diversity of microbial associates
of marine sponges and sediments producing More details can be found in:
β-glucosidase inhibitors. We found 41 1. Pandey et al. Diversity of marine bacteria
(22.7%) out of 181 bacteria, produced such producing beta-glucosidase inhibitors.
inhibitors. Microbial Cell Factories 2013 12:35.
All the active isolates were identified by doi:10.1186/1475-2859-12-35 (IF: 3.55).
partial 16S rDNA sequencing in the V1-V3 2. Pandey et al. A novel method for
variable region and confirmed by culture screening beta-glucosidase inhibitors.
characterization. The phylogenetic tree BMC Microbiology 2013 13:55. doi:10.
constructed shows active isolates are 1186/1471-2180-13-55 (IF: 3.04).
distributed in different bacterial phyla –
Firmicutes (23), Actinobacteria (9), Sony Pandey and collaborators at
Proteobacteria (7) and Bacteroidetes (1), CSIR-IMMT
CSIR News 153
JULY 2013
MoUs
MoUs
MoU between CSIR-CRRI and Volvo
Construction Equipment
MoU being signed between CSIR-CRRI and Volvo
CSIR-Central Road Research Institute common interest for engineering
(CRRI), New Delhi has entered into a MoU professionals.
with Volvo Construction Equipment • To conduct short term courses for
Division, Volvo India Private Limited, manpower skill development in areas
Bengaluru. Dr. S. Gangopadhyay, Director, of machine operation, thereby
CSIR-CRRI and Shri A.M. Muralidharan, addressing prevailing concern of
President, Volvo signed the MoU on 30 April shortage of skilled machine operators
2013 at CSIR-CRRI, New Delhi. for road construction equipment.
The broad objectives and terms and
• To come out with a white paper on
conditions of the MoU are as follows:
Construction and Maintenance
• To establish a working relationship Machinery for the Road Sector in the
between CSIR-CRRI and Volvo in areas form of catalogues, technical papers,
of road construction equipment and manuals, etc.
technology and skill development of
• To explore opportunities for
involved manpower so as to achieve
collaborative projects for development
overall productivity improvement for
of instrumentation for road
road construction industry.
construction equipment.
• To enhance awareness of latest road
• To jointly undertake projects in India
construction equipment amongst
and abroad.
trainees of CSIR-CRRI.
• To organize joint conferences and
• To conduct training programmes of
seminars in India and abroad.
154 CSIR News
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