1. Curriculum Vitae
Ramesh Kumar, Ph.D
Born January 3rd
1977, Gorakhpur, Uttar-Pradesh, India
Address for Correspondence
Ramesh Kumar, Ph.D
Department of Arthritis and Clinical Immunology,
Oklahoma Medical Research Foundation (OMRF),
825 13th
Street Oklahoma City, Ok 73104.
Mobile No. +1(405) 488-4835
E-mail: ramesh.kumar1@hotmail.com & ramesh-kumar@omrf.org
Current Position
Post-Doctoral Research Fellow (January 2012-Present)
Department of Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation
(OMRF), Oklahoma City, OK-73104, USA
Title of the project: “The role of lupus susceptible genes in the development of autoimmune
disease”
Principal Investigator: Dr. Marta E. Alarcon Riquelme
Academic Qualifications
Post-Doctoral Research Fellow at Umea (May 2009-May 2011)
Department of Molecular Biology, Umeå University, Sweden
Title of the project: “The study of interaction of activation induced cytidine deaminase
(AID) with transcription factor (TFs) proteins and regulation of B Cell development”
Principal Investigator: Professor Thomas Grundström
Ph.D.: Life Sciences (2009), Jawaharlal Nehru University (JNU) New Delhi, India
Thesis title: “Chemotherapy of Visceral leishmaniasis: Molecular and Biochemical
approach”
M.Sc. (1997-1999): Botany (Specialization in Molecular Genetics)
D. D. U. Gorakhpur University, Gorakhpur, India
B.Sc. (1994-1997): Zoology, Botany and Chemistry
D. D. U. Gorakhpur University, Gorakhpur, India
Scholastic Achievements and Awards
1. Post-doctoral Fellowship Award from the “Conseil Scientifique de la Ville de Paris”
September 2009 to September 2010, Declined.
2. Qualified pre-Ph.D. programme for Ph.D registration at JNU, New Delhi conducted by
JNU cell, CDRI, Lucknow, India 2004.
3. Teaching experience at graduate and post-graduate level at Botany Department, D. D. U.
Gorakhpur University, Gorakhpur, India 2002.
4. Qualified Ph.D. programme conducted by D. D. U. Gorakhpur University, Gorakhpur,
India 2001, Declined.
5. Qualified All India National Eligibility Test (NET) conducted by the Council of Scientific
& Industrial Research – University Grant Commission (CSIR-UGC), for the award of Junior
Research Fellowship 30th
December, 2001.

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Research Publications
Peer-reviewed Original Research Articles (Submitted)
1. Ying – Yu Wu*, Ramesh Kumar*, Ryuji Iida, Harini Bagavant, and Marta E. Alarcon-
Riquelme (2015). BANK1 controls the development of SLE modulating TLR7 signaling and
Type I IFN-induced Translation Initiation B cell pathway. Manuscript Submitted to Journal
of Exp Med (* Shared First Authorship).
2. Jannek Hauser, Christine Grundstrom, Ramesh Kumar and Thomas Grundstrom (2015).
Regulated localization of an AID complex with E2A, PAX5 and IRF4 at the Igh locus.
Manuscript Submitted to The Journal of Immunol, ID No. 15-01390-FL.
3. Simanta Pathak, Yuyuan Guo, Ramesh Kumar, Huihua Ding, Yong Du, Soyoun Min,
Kamala Vanarsa, Quanzhen Li, Sergey Kozyrev, Marta E. Alarcón-Riquelme, and Chandra
Mohan (2015). BANK1 transgenic mice exhibit lupus-associated B cell phenotypes
(Manuscript Under Preparation).
Peer-reviewed Original Research Articles (Published)
4. Biji T. Kurien, Valerie M. Harris, Syed Quadri, Patricia Coutinho-de Souza, Joshua Cavett,
Amanda Moyer, Bilal Ittiq, Angela Metcalf, Husayn F. Ramji, Dat Truong, Ramesh Kumar,
Kristi A. Koelsch, Michael Centola, Adam Payne, Debashish Danda, R. Hal Scofield (2015).
Significantly reduced lymphadenopathy, salivary gland infiltrates and proteinuria in MRL-
lpr/lpr mice treated with ultrasoluble curcumin/turmeric: increased survival with curcumin
treatment. Lupus Science & Medicine, 2:e000114.
5. Ying Yu Wu, Ina Georg, Alejandro Diaz-Barreiro, Nieves Verela, Bernard Lauwerys,
Ramesh Kumar, Harini Bagavant, Mireia Castillo-Martin, Fadi El Salem, Concepcion
Maranon-Lizana and Marta E. Alarcon- Riquelme (2015). Concordance of increased B1 cell
subset and lupus phenotypes in mice and humans is dependent on BLK expression levels. The
Journal of Immunol 194 (12); 5692-702.
6.Ying-Yu Wu, Ramesh Kumar, Mohammed Shamsul Haque, Casimiro Castillejo-Lopez
and Marta E. Alarcon-Riquelme (2013). Bank1 controls IL-6 secretion via a p38 and
MNK1/2-eIF4E translation initiation pathway. The Journal of Immunol 191; 6110-6116.
7. Ramesh Kumar, Gupta S., Srivastava R., Sahasrabuddhe A.A., Gupta C.M. (2010).
Expression of a PTS2-truncated hexokinase produces glucose toxicity in Leishmania
donovani. Mol Biochem Parasitol 170 (1); 41-4.
8. P. K. Murthy, S. Dixit, R. L. Gaur, Ramesh Kumar, M. K. Sahoo, N. Shakya, S. K.
Joseph, S. Palne & S. Gupta (2008). Influence of Brugia malayi life stages and BmAFII
fraction on experimental Leishmania donovani infection in hamsters. Acta Tropica 106; 81-
89.
9. Ashutosh, S. Gupta, Ramesh, S. Sunder, & N. Goyal (2005). Use of Leishmania donovani
Field Isolates Expressing the Luciferase Reporter Gene in In vitro Drug Screening.
Antimicrobial Agents and Chemotherapy 49 (9); 3776-3783.
10. S. Gupta, Ramesh, S. C. Sharma & V. M. L. Srivastava (2005). Efficacy of Picroliv in
combination with Miltefosine, an orally effective antileishmanial drug, against experimental
visceral leishmaniasis. Acta Tropica 94; 41-47.
11. N. Chandra, S. Pandey, Ramesh, S. N. Suryawanshi & S. Gupta (2006). Chemotherapy of
Leishmaniasis Part III: Synthesis and bioevaluation of novel aryl substituted terpenyl
pyrimidines as antileishmanial agents. European Journal of Medicinal Chemistry 41, 779-
785.

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12. A. Agarwal, Ramesh, Ashutosh, N. Goyal, P. M. S. Chauhan & S. Gupta (2005).
Dihydropyrido (2, 3-d) pyrimidines as a new class of antileishmanial agents. Bioorganic and
medicinal Chemistry 13; 6678-6684.
13. N. Chandra, Ramesh, Ashutosh, N. Goyal, S. N. Suryawanshi & S. Gupta (2005).
Antileishmanial Agents Part-IV: Synthesis and antileishmanial activity of novel terpenyl
pyrimidines. European Journal of Medicinal Chemistry 40; 552-556.
14. N. Tewari, Ramesh, R. C. Mishra, R. P. Tripathi, V. M. L. Srivastava & S. Gupta (2004).
Leishmanicidal activity of phenylene bridged C2 symmetric glycosyl ureides. Bioorganic and
Medicinal Chemistry Letters 14; 4055-4059.
Patents
1. Marta E. Alarcon Riquilme, Ying-Yu Wu and Ramesh Kumar. Treatments for Systemic
Lupus Erythematosus. U.S. Provisional Patent Application No. 61/885,859 Filed on
October 2013.
2. S. N. Suryavanshi, S. Gupta, Ramesh & N. Chandra. 5-[2-(2, 6, 6-Trimethyl-cyclohex-2-
enyl)-ethenyl]-isoxaazole, Patent No. 0212 NF 2005/IN, Application No.0529DEL2006 Filed
on; 12/07/06.
Presentations in Scientific Conferences and Workshop
1. Ying Yu Wu, Ina Georg, Alejandro Diaz-Barreiro, Nieves Verela, Bernard Lauwerys,
Ramesh Kumar, Harini Bagavant, Mireia Castillo-Martin, Fadi El Salem, Concepcion
Maranon-Lizana and Marta E. Alarcon- Riquelme (2015). Concordance of increased B1 cell
subset and lupus phenotypes in mouse and human dependent on BLK expression genotypes.
The Journal of Immunology (BA3P.114).
2. Ying-Yu Wu, Ramesh Kumar, Mikhail Dozmorov, Mohammed Shamsul Haque, Jonathan
Wren, Casimiro Castillejo-Lopez and Marta E. Alarcon-Riquelme (2013). Bank1 controls IL-
6 secretion and Type I interferon signature via a p38 and MNK1/2-eIF4E translation initiation
pathway. The Journal of Immunology, 190: 44.15.
3. Ramesh Kumar, S. Gupta & Amogh A. Sahasrabuddhe (2009). Glycosomal targeting of
hexokinase is essential in Leishmania donovani. 4th World Congress on leishmaniasis,
Central Drug Research Institute, Lucknow, India, February 3-7.
4. Ramesh, N. Chandra, S. Pandey, S. N. Suryavanshi & S. Gupta (2005). Bioevaluationof
novel aryl substituted terpenyl pyrimidines against Leishmania donovani. 17th
National
Congress of Parasitology, RMRC, Dibrugarh, Assam, October 24-26.
5. Ramesh, N. Tiwari, R.P.Tripathi, V.M.L. Srivastava & S. Gupta (2004).
Diureidylphenylene bridged glycosyl urinates- a new class of antileishmanial agent. 3rd
global
meet on parasitic diseases, Bangalore University, Bangalore, India, January 12-16.
Oral Presentation
Title “Bank1 controls IL-6 secretion and Type I interferon signature via a p38 and MNK1/2-
eIF4E translation initiation pathway” at Oklahoma Medical Research Foundation Retreat,
23-15th
January 2013.
Post-Doctoral Research Work at OMRF
In my current Post-Doctoral research I have been dealing with the two projects viz., Bank1
and BLK:
Systemic Lupus Erythematosus (SLE) is characterized by spontaneous proliferation,
expansion of auto-reactive B and T cells, and production of polyclonal autoantibodies against

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numerous nuclear antigens govern by the interplay of several genes and environmental
factors. In recent years, many lupus susceptible genes including Bank1 and BLK have been
identified, but their impact on disease progression has yet to be defined.
BANK1 is a B cell scaffold protein with ankyrin repeats1. Bank1 acts as an adaptor protein,
expressed in B cells. Because Bank1 contains an N-terminal putative Toll-interleukin 1
receptor (TIR) domain, we asked whether Bank1 affects signaling induced by the TLR9
agonist CpG? To discern the signaling cascades affected by Bank1, first we tested if Bank1
deficiency altered B cell proliferation after CpG stimulation by labeling purified splenic B
cells with CFSC (2µM) and data were acquired with LSRII. We did not observe any
difference in B cell proliferation. Then we checked phosphorylation of NF-kB and MAPKs
ERK1/2, JNK1/2 and p38 upon CpG stimulation by Western blot and found that BANK1
deficiency reduced p38 phosphorylation without affecting ERK1/2 or JNK1/2 and NF-kB. We
checked IL-6 cytokine in B cells culture supernatants, which was reduced in Bank1 deficient
B cells. Our Taqman real-time PCR data of Il6 and Tlr9 gene expression was not affected
between Bank1+/+ and Bank1-/- B cells. Next we used TLR4 (LPS) and TLR7/8 agonists
(R848) which did not show any difference suggesting Bank1 acts specifically through TLR9
modulates IL-6 secretion by reducing p38 phosphorylation.
Since, p38 directly controls translation initiation and/or mRNA stability through the kinases
MNK1/2 and MK2 respectively. First, we tested CGP57380 and Cercosporamide (MNK1/2
inhibitors) and PF3644022 (MK2 inhibitor) on B cells and stimulated with CpG which
suppressed IL-6 secretion in the culture supernatants and this suppression was not due to
cytotoxicity checked on FACSCalibur by Annexin V and PI staining. Next, we observed
intact Il6 mRNA stability by using actinomycin-D (1µg/ml) treatment after CpG stimulation.
Subsequently, we checked phosphorylation of MNK1/2 and eIF4E by Western blot, which
were significantly reduced in Bank1 deficient B cells suggesting an effect on translation
initiation without affecting Il6 mRNA stability via MK2 signaling. We checked activation of
Akt, mTOR and 4E-BP1 by Western blot following stimulation with CpG, anti-CD40 and a
combination of both, which were not affected by Bank1 deficiency. Together these data
established that Bank1 modulates p38 signaling and control of translation initiation of IL-6
via MNK1/2 and eIF4E in B cells on CpG induced responses in vitro (The Journal of
Immunol (2013), 191:6110-6116).
Having shown that Bank1 regulates p38 signaling as well as IL-6 cytokine production, and
others have reported that B6.Sle1zz.Yaa mouse through exacerbation of TLR7 expression
develops severe disease with splenomegaly, T and B cells activation, production of
autoantibodies and developing glomerulonephritis. We hypothesize that Bank1 might be
involved in the disease progression. To test our hypothesis we crossed Bank1-/- with
B6.Sle1zz.Yaa mice and intercrossed the resultant offsprings to get B6.Sle1zz.Yaa.Bank1+/+
and B6.Sle1zz.Yaa.Bank1-/- genotypes. Here we investigated the effects of the adaptor
Bank1 in TLR7 signaling. FACS data show that B6.Sle1zz.Yaa.Bank1-/- mice maintained
several B and myeloid cell phenotypes close to normal B6 WT levels. Our most striking
ELISAs results were in the reduction in total serum IgG antibodies, but not of IgM, and
reduced serum levels of autoantibodies, IL-6 and BAFF, features accompanied by reduced
mortality in B6.Sle1zz.Yaa.Bank1-/- mice in comparison with B6.Sle1zz.Yaa.Bank1+/+ mice.
Expression of CXCR4 in Tfh cells was down regulated in absence of Bank1. Purified B cells
from B6.Sle1zz.Yaa.Bank1-/- mice had strongly reduced IFNb, Irf7, Aicda and Stat1 gene
expression following TLR7 agonist (R837) stimulation. Furthermore, phosphorylation of the
transcription factor STAT1 was impaired in B6.Sle1zz.Yaa.Bank1-/- mice as was the
cytosolic and nuclear expression and nuclear translocation of IRF7, a key molecule in TLR7
signaling. As the optimal function of B cells depends on type I interferon, we investigated if
BANK1 had effects on the IFNAR signaling pathway. We observed that BANK1 controls
activation of the eIF4E translation initiation pathway induced by type I IFN, hence

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controlling interferon-inducible genes. Our results demonstrate that BANK1 controls TLR7
and IFN-inducible eIF4E signaling in B cells contributing to autoimmune disease
development (Manuscript Submitted to Journal of Exp Med (* Shared First Authorship)).
Since, I joined OMRF I have been assisting the colleague in BLK KO project where I have
shown no significant impairment of MZ B cells development in BLK+/- and BLK-/- mice.
Since, B1a cells are natural source of anti-IgM antibodies, which may differentiate into anti-
ds DNA IgG autoantibodies producing cells; we checked B1a B cells based on expression of
CD5+
(B1a cells) and CD5-
(B1b cells) marker. Our data show that splenic B1a cells, which
mostly localize inside the follicles, are significantly increased in BLK-/- mice. We checked
anti-ds DNA IgG autoantibodies in serum, which was increased in BLK-/- mice. The Journal
of Immunol (2015), 194 (12); 5692-702.
Post-Doctoral Research Work at Umea University
During my Post-Doctoral Research at Umea University, I have dealt with two major projects:
In the first project I studied the interaction of activation induced cytidine deaminase (AID)
with transcription factors (TFs) of E-proteins in the splenic B cells. Activation induced
cytidine deaminase (AID), a mutagenic antibody diversification enzyme, that is essential for
both somatic hypermutation (SHM) and class switch recombination (CSR) has to be tightly
controlled otherwise it will lead to disruptions in the B cell development. This disruption
could be controlled by interactions of TFs and the gene regulatory sequences, i.e. promoter
and enhancer sequences. To dissect this we used mouse splenic B cells purified by magnetic
anti-biotin beads, MidiMACS Separator, and LS columns (Miltenyi Biotec). These B cells
were used to show interactions of AID with TFs by proximity ligation assay (PLA) and the
signals were captured with fluorescent microscope. Afterward, the regulatory effects of DNA
binding of E-proteins have been compared between splenic B cells infected with retroviral
constructs (MSCV-IRES-GFP) expressing either Wt or CaMR
E12 (a spliced variant of E2A)
before and after anti-IgM stimulations at different time points. DNA binding experiment has
been performed by chromatin immunoprecipitation (ChIP) assay using FLAG-tag instead of
GFP in the above constructs (this experiment was done in association of lab mate). The
interactions of TFs proteins have been validated with recombinant proteins in questions by
using cynogen bromide (CNBr) activated Sepharose 4B beads. The above beads were
dissolved in 2x protein loading buffer, supernatant loaded on 10% SDS-PAGE and the
Western blot was analyzed (Manuscript Submitted to The Journal of Immunol, Manuscript
ID No. 15-01390-FL).
In the next project, I have been associated with mouse E2A-BAC (mE2A-BAC) cloning to
generate transgenic mouse. Here we have created three point mutations in the E12 (a spliced
variant of E2A gene) exon region. (I would be one of the authors of this manuscript).
Doctoral Research Work at CDRI, Lucknow
My entire Ph.D. work was focused on drug target validation, in an effort to develop suitable
in vitro screening techniques for antileishmanial screening, and combination therapy using
immunomodulators.
Under target validation L. donovani-hexokinase (Ld-HK) 1.4 kb gene was amplified by PCR,
cloned in pET-21d prokaryotic expression vector and overexpressed in E. coli, BL21 (DE3,
strain). The 47kDa molecular mass of L. donovani HK protein, found by coomassaie-blue
staining, was confirmed by Western blot analysis. Since, rLd-HK was insoluble, the inclusion
bodies protein prepared and rLd-HK protein was purified by SDS-PAGE gel excision method.
This rLd-HK protein was used for generation of anti-Ld-HK antibodies in rats and rabbit. The
monospecific, polyclonal antibodies were purified by affinity purification. The sub-cellular
study showed distribution of Ld-HK throughout in the cytoplasm in vesicle-like structures, the

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typical “microbodies”, which contain glycolytic enzymes and by using a glycosomal marker
“XPRT-GFP” vector we confirmed that the hexokinase was localized in the glycosomes. In
Trypanosoma brucei, various RNAi experiments have suggested that inhibition of glycolytic
import to glycosomes. Here we showed generation of peroxisome targeting signal mutant
(PTS-2ΔHK) Leishmania cell lines induce toxic effects in presence of sugars and established
compartmentation of glycolysis and its importance in Leishmania donovani glucose toxicity
which could be used as a drug target in trypanosomatids (MBP 2010, 170 (1); 41-4.
The next objective of the thesis was to establish transgenic cell lines for rapid in vitro
screening of antileishmanial compounds by using firefly luciferase gene expressing
Leishmania donovani promastigotes. The results obtained by luciferase assay were very
reliable, rapid and suitable for antileishmanial screening (Antimicrob Agents Chemother
2005, 49 (9); 3776-3783).
In the rest part of my thesis work I studied adjunctive effect of immunomodulators (phenylene
bridged C2 symmetric glycosyl ureides) with Sodium stibogluconate (SSGV
) (BMCL 2004,
14; 4055-4059). I also studied Picroliv (Picrorhiza kurooa) in combination of miltefosine in
lower doses to minimize its cytotoxicity against L. donovani in hamsters (Acta Tropica 2005,
94; 41- 47). We also examined the immunological influence of Brugia malayi and their
fraction extracts on L. donovani in hamster model and the reduction in parasite’s burden (Acta
Tropica 2008, 106, 81-89). Apart from these works, several synthetic and naturally occurring
antileishmanial compounds have also been studied and an Indian Patent has been filed
(Patent No. 0212 NF 2005/IN, Application No. 0529DEL2006). Rest of the works has been
published in reputed international journals (BMC 2005, 13; 6678-6684; E JMC 2005, 40;
552-556; E JMC 2006, 41, 779-785).
Animal Handling
Excellent in handling of regular laboratory animals – mice (Swiss, BALB/c, and C57BL/6),
hamster (Golden), rabbit and rat
Cell Culture
Maintenance of different strains of Leishmania species and mammalian cell lines viz. 293T,
J774A.1, U937, 3T3 and THP1, extraction of peritoneal macrophages and lymphocytes
Technical Exposures
Molecular Biology and genetic engineering
 BAC cloning
 Sequencing PCR
 PCR amplification and gene cloning
 Genomic DNA and total RNA isolation
 Analysis of gene expression by qRT-PCR
 Cloning and expression of recombinant proteins in prokaryotes (E. coli)
 Purification of recombinant proteins through metal chelating affinity chromatography,
ion exchange chromatography
 Generation of mutants by PCR
Immunological techniques
 Generation of polyclonal antibody against recombinant protein in rat, rabbit and mice
 Purification of monospecific polyclonal antibody by affinity chromatography
 ELISA and Western blot analysis
 Immunoprecipitation of antigen
 Immunofluorescence microscopy
 Thymidine (3H) incorporation and analysis of cellular immune response (LTT) by
liquid scintillation counter

7. Ramesh Kumar
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Cell biological techniques
 Thymocytes and peritoneal macrophage isolation, splenic B cells and Dendritic cells
purification from C57BL/6 mice by magnetic anti-biotin beads, MidiMACS Separator,
and LS columns (Miltenyi Biotec) and their cultivation
 Multicolor staining of B cells and their analyses by means of FACSCalibur and LSRII
 Production of MSCV-IRES-GFP based retrovirus in mouse fibroblast 293T cell lines
 Retroviral infection of splenic B cells
 Transfection of E.coli by electroporation method
 Transfection of Leishmania by electroporation method and selection of stable
transfectants against drugs geneticin, hygromycin and tunicamycin
 Sub-cellular distribution and co-localization of proteins in the cell through
immunofluorescence and confocal microscopy
Biochemical techniques
 Exposed to ChIP experiment
 Cytosolic and nuclear localization of proteins, e.g. IRF7
 Protein-Protein interaction by PLA (Proximity Ligation Assay) method by using
immunofluorescence microscope
 Purification of antibody by using ProteinA/G beads
 Labeling of antibody with biotin and digoxigenin
 Measurement of enzyme activities
 Digitonin fractionation of cell organelle
 Measurement of nitric oxide (NO), hydrogen peroxide (H2O2) concentration
Drug screening techniques
 In vitro screening of new antileishmanials against Leishmania promastigotes and
macrophage-amastigotes. IC50 and IC90 evaluation
 In vivo maintenance of Leishmania strains in Syrian golden hamster (Mesocricetus
auratus), assessment of infection and parasitic burden by spleen biopsy through
Giemsa staining method, and assessment of therapeutic/adjunct efficacy of compounds
Computer Knowledge
In addition to regular programs for primer design and other gene analyses like Gene runner &
Oligo have good hand on Word processor (Microsoft Word), spreadsheet (Microsoft Excel),
Powerpoint, in Windows & Mac environments. Also on statistical analysis and graphs by
GraphPad Prism® and Origin, and working knowledge of photography software- Adobe
Photoshop, Adobe Illustrator. Exposed to on line database search viz. EMBL, MEDLINE and
NCBI-BLAST.
Profile
Skilled researcher with strong background in basic immunology, molecular biology, cell
biology, biochemistry and microbiology. Broad range of experience in Molecular & Cellular
Biology, Genetic Engineering & Immunology viz. cloning, expression & purification of
recombinant protein, protein-protein interactions, antibody generation, qRT-PCR, FACS,
ELISA, blotting, cell culture techniques, Confocal & Immunofluorescence Microscopy etc.
Experience in troubleshooting in PCR, Cloning technique, strong computational and research
data analyzing skills. Expertise in handling of regular laboratory animals. I believe in positive
and result oriented working, comprehensive problem solving abilities, excellent verbal and
written communication skills, ability to deal with people, willingness to learn. Experienced in
managing research projects from conception to completion.
References: References will be provided when need arises.