A rapid and green GC-MS method for the sampling of volatile organic compounds in spices and flowers by concurrent headspace single-drop microextraction and solid-phase microextraction.

The equilibrium rather than the exhaustive nature of headspace single-drop microextraction (HS-SDME) and headspace solid-phase microextraction (HS-SPME) allowed the concurrent sampling of volatile organic compounds (VOCs) on the same sample in the same vial in a dual extraction configuration. This has avoided the necessity of conducting a separate set of experiments and was found to produce results in the time duration of a single sample preparation experiment. The results obtained by HS-SDME were validated against those found by the standard method of HS-SPME. Rectilinear calibration was made for certain VOCs tested as analytes over the range of 0.01-8 µg g-1, and the average values of R2, LOD and LOQ were found to be, respectively, 0.9992, 1.9 ng g-1 and 5.7 ng g-1 in HS-SDME, and 0.9991, 3.1 ng g-1 and 9.1 ng g-1 in HS-SPME. The spiked recoveries and RSD were, respectively, 100.5% and 3.3% in HS-SDME and 98.1% and 3.6% in HS-SPME. HS-SDME is convenient to perform and produce results in a much cheaper way than HS-SPME and free from the inconveniences of memory effects. With GC-MS, this method has also been implemented as a rapid, reliable and green procedure (by GAPI and AGREE tools) for the sampling of VOCs in real samples of spices, flowers, and a beetle nut chewing sample illicitly containing tobacco.

Non-viral 2A-like sequences for protein coexpression.

Simultaneous coexpression of multiple proteins is essential for biotechnology and synthetic biology. Currently, the most popular polyprotein coexpression system utilizes the foot-and-mouth disease virus (FMDV) 2A peptide that mediates translational ribosome-skipping events. However, due to unfavorable consumer acceptance of transgenic products containing animal-virus sequences, novel non-viral 2A-like peptides from purple sea urchin (Strongylcentrotus purpuratus) and California sea slug (Aplysia californica) were investigated for polyprotein coexpression in this study. We demonstrated that these non-viral 2A sequences functioned similarly to their viral counterpart in polyprotein processing, in both plant and mammalian cells, and were successfully used to express a functional recombinant antibody. The new non-viral 2A-like sequences offer an alternative tool for engineering multigenic traits or production of protein complexes as biomedicine via coexpression of protein subunits.

What are mycorrhizal traits?

Traits are inherent properties of organisms, but how are they defined for organismal networks such as mycorrhizal symbioses? Mycorrhizal symbioses are complex and diverse belowground symbioses between plants and fungi that have proved challenging to fit into a unified and coherent trait framework. We propose an inclusive mycorrhizal trait framework that classifies traits as morphological, physiological, and phenological features that have functional implications for the symbiosis. We further classify mycorrhizal traits by location - plant, fungus, or the symbiosis - which highlights new questions in trait-based mycorrhizal ecology designed to charge and challenge the scientific community. This new framework is an opportunity for researchers to interrogate their data to identify novel insights and gaps in our understanding of mycorrhizal symbioses.

Dispersive liquid-liquid microextraction and diffuse reflectance-Fourier transform infrared spectroscopy for iodate determination in food grade salt and food samples.

A novel method based on diffused reflectance Fourier-transform infrared spectroscopy (DRS-FTIR) was employed for iodate determination in food grade salt and food products. The method attained sensitivity that was comparable to or better than that in most of the contemporary spectrophotometric methods. This was realized through a combination of azo dye formation and dispersive liquid-liquid microextraction of dye when a 37-fold enrichment was obtained. FT-IR enabled integrating alternative target peak, and freedom in sample solvent composition relative to UV-visible spectrophotometry where the solvent polarity, pH, and presence of ions may affect the spectral properties of the measurable coloured species. Food samples containing iodide or covalently bonded iodine were oxidized with alkaline permanganate for mineralization and iodate formation. Optimization of both reaction conditions was carried out by means of response surface methodology. The method had a linear range 0.04-10 mg kg-1 iodate and limit of detection of 4.4 µg kg-1.

World symbiosis day webinar - when living together is a win-win.

The following information is intended for those who were unable to attend the first webinar of the International Symbiosis Society (ISS) on - 'When living together is a win-win' - a celebration of 'World Symbiosis Day' on 30 July 2020. The objective of the webinar was to disseminate information about the Society, to gather feedback, and to encourage the audience to join the Society. This introduction presents a summary of the webinar, highlighting the keynote presentations, the panel discussion, the journal Symbiosis, and the next ISS conference to be held in Lyon in 2022. In addition, we report on the discussions and feedback from participants that were collected through polls and other aspects of the webinar.

The Pharmacokinetics of Triheptanoin and Its Metabolites in Healthy Subjects and Patients With Long-Chain Fatty Acid Oxidation Disorders.

Long-chain fatty acid oxidation disorders (LC-FAODs) are a group of life-threatening autosomal recessive disorders caused by defects in nuclear genes encoding mitochondrial enzymes involved in the conversion of dietary long-chain fatty acids into energy. Triheptanoin is an odd-carbon, medium-chain triglyceride consisting of 3 fatty acids with 7 carbons each on a glycerol backbone developed to treat adult and pediatric patients with LC-FAODs. The pharmacokinetics of triheptanoin and circulating metabolites were explored in healthy subjects and patients with LC-FAODs using noncompartmental analyses. Systemic exposure to triheptanoin following an oral administration was negligible, as triheptanoin is extensively hydrolyzed to glycerol and heptanoate in the gastrointestinal tract. Multiple peaks for triheptanoin metabolites were observed in the plasma following oral administration of triheptanoin, generally coinciding with the time that meals were served. Heptanoate, the pharmacologically active metabolite of triheptanoin supplementing energy sources in patients with LC-FAODs, showed the greatest exposure among the metabolites of triheptanoin in human plasma following oral administration of triheptanoin. The exposure of heptanoate was approximately 10-fold greater than that of beta-hydroxypentoate, a downstream metabolite of heptanoate. Exposure to triheptanoin metabolites appeared to increase following multiple doses as compared with the single dose, and with the increase in triheptanoin dose levels.

Prophylactic administration of podophyllotoxin and rutin combination assists the revival of radiation-induced hematopoietic suppression in lethally irradiated mice.

Hematopoietic syndrome contributes to mortality after exposure to high doses of low LET radiation. In this context, we have earlier demonstrated the potential of G-003 M (a combination of podophyllotoxin and rutin) in alleviating radiation-induced bone marrow suppression. Similarly, we here demonstrate that G-003 M protected mice from death (>83% protection) and increased the populations of CD 34 (Cluster of differentiation 34) as well as CD 117 (Cluster of differentiation 117) positive cell population and their colony forming capacity. This was accompanied with increase in the serum titre of granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF). Interestingly, G-003 M lowered down the titre of fms-like tyrosine kinase (Flt-3) ligands. Our results furthermore demonstrates that G-003 M facilitated the nuclear translocation of ß-catenin and upregulated the expression of Wnt 10b. Conditioning of animal with G-003 M activated the expression of survivin, inhibited the activation of Caspase-3 in CD 34/117+ progenitor stem cells and protected the bone marrow vascularity and splenic colonies in lethally irradiated animals, which collectively promoted hemopoietic recovery in lethally irradiated mice.

Transcription Terminator-Mediated Enhancement in Transgene Expression in Maize: Preponderance of the AUGAAU Motif Overlapping With Poly(A) Signals.

The selection of transcription terminators (TTs) for pairing with high expressing constitutive promoters in chimeric constructs is crucial to deliver optimal transgene expression in plants. In this study, the use of the native combinations of four polyubiquitin gene promoters and corresponding TTs resulted in up to >3-fold increase in transgene expression in maize. Of the eight polyubiquitin promoter and TT regulatory elements utilized, seven were novel and identified from the polyubiquitin genes of Brachypodium distachyon, Setaria italica, and Zea mays. Furthermore, gene expression driven by the Cassava mosaic virus promoter was studied by pairing the promoter with distinct TTs derived from the high expressing genes of Arabidopsis. Of the three TTs studied, the polyubiquitin10 gene TT produced the highest transgene expression in maize. Polyadenylation patterns and mRNA abundance from eight distinct TTs were analyzed using 3'-RACE and next-generation sequencing. The results exhibited one to three unique polyadenylation sites in the TTs. The poly(A) site patterns for the StPinII TT were consistent when the same TT was deployed in chimeric constructs irrespective of the reporter gene and promoter used. Distal to the poly(A) sites, putative polyadenylation signals were identified in the near-upstream regions of the TTs based on previously reported mutagenesis and bioinformatics studies in rice and Arabidopsis. The putative polyadenylation signals were 9 to 11 nucleotides in length. Six of the eight TTs contained the putative polyadenylation signals that were overlaps of either canonical AAUAAA or AAUAAA-like polyadenylation signals and AUGAAU, a top-ranking-hexamer of rice and Arabidopsis gene near-upstream regions. Three of the polyubiquitin gene TTs contained the identical 9-nucleotide overlap, AUGAAUAAG, underscoring the functional significance of such overlaps in mRNA 3' end processing. In addition to identifying new combinations of regulatory elements for high constitutive trait gene expression in maize, this study demonstrated the importance of TTs for optimizing gene expression in plants. Learning from this study could be applied to other dicotyledonous and monocotyledonous plant species for transgene expression. Research on TTs is not limited to transgene expression but could be extended to the introduction of appropriate mutations into TTs via genome editing, paving the way for expression modulation of endogenous genes.

A combined prophylactic modality of podophyllotoxin and rutin alleviates radiation induced injuries to the lymphohematopoietic system of mice by modulating cytokines, cell cycle progression, and apoptosis.

The present study was conceptualized to delineate radioprotective efficacy of a formulation G-003M (a combination of podophyllotoxin and rutin) against radiation-induced damage to the lymphohematopoietic system of mice. C57BL/6J mice, treated with G-003M 1 h prior to 9 Gy lethal dose, were assessed for reactive oxygen species (ROS)/nitric oxide (NO) generation, antioxidant alterations, Annexin V/PI and TUNEL staining for apoptosis, modulation of apoptotic proteins, cell proliferation, histological alterations in thymus and cell cycle arrest in bone marrow cells. Induction of granulocyte colony-stimulating factor (G-CSF), granulocytes macrophage colony-stimulating factor (GM-CSF), interleukin-IL-6, IL-10, IL-1α, and IL-1ß in response to G-003M was also evaluated in different groups of mice. Haematopoietic reconstitution with G-003M was explored by examining endogenous spleen colony-forming units (CFU-S) in irradiated animals. G-003M significantly inhibited ROS/NO, malondialdehyde (MDA) and restored cellular antioxidant glutathione in the thymus of irradiated animals. G-003M pre-treatment significantly (p < 0.001) restrained apoptosis in thymocytes via upregulation of Bcl2 and down-regulation of Bax, p53 and caspase-3. Stimulation of cell proliferation and inhibition of apoptosis by G-003M, restored architecture of thymus in irradiated animals within 30 days as evaluated by histological analysis. G-003M arrested cells at the G2/M phase by inducing reversible cell cycle arrest. Peak expression of G-CSF (45-fold) and IL-6 (60-fold) as well as moderate induction of GM-CSF, IL-10, IL-1α by G-003M helped in haematopoietic recovery of irradiated mice. A higher number of endogenous CFU-S in G-003M pre-treated irradiated mice suggested haematopoietic recovery. Data obtained from the current study affirms that G-003M can be proved as a potential radioprotective agent against radiation damage.

Combination of podophyllotoxin and rutin modulate radiation-induced alterations of jejunal proteome in mice.

Purpose: Gastrointestinal (GI) injuries post ionizing radiation (IR) becomes a crucial factor in survival. Thus, the current study was aimed to explore the molecular mechanisms behind IR produced GI proteome alterations and their amelioration by a safe radioprotective formulation candidate, G-003M (podophyllotoxin+rutin).Materials and method: C57BL/6 mice were administered with G-003M 1 h before 9 Gy whole body γ irradiation. 2DE-MS analysis was conducted to identify differential expression of jejunum proteins with fold change >1.5 (p < .05) at various time-points. Results: G-003M pre-administration decreased total number of differential proteins. It mediated protection to cytoskeleton, modulated stress, apoptosis and inflammatory proteins. Direct effect on eukaryotic translation initiation factor 4H (Eif4h), thioredoxin domain-containing protein 17 (Txndc17) and interferon-induced protein 35 (Ifi35) was observed. Bioinformatics depicted transcription factor-MYC, was also positively modulated by G-003M. Further, it also enhanced level of citrulline (ELISA analysis), and restored crypts and villi lengths (histological analysis) against severe damage caused by lethal irradiation.Conclusion: Current findings reveal that G-003M may be an efficient candidate in protecting key proteins of metabolic and biochemical pathways assisting in the rapid recovery of GI proteome. This fairly improved the chances of animal survival exposed to lethal doses of whole body radiation.

Intein-ubiquitin chimeric domain for coordinated protein coexpression.

Efficient coproduction of multiple proteins or their subunits is important in many facets of life sciences and biotechnology. Here, we report a novel approach that exploits the synergy between an engineered mini-intein and an ubiquitin variant to achieve coordinated coexpression of multiple proteins in eukaryotic hosts, from a single open reading frame that encodes a polyprotein precursor consisting of proteins of interest (POIs) connected by an intervening intein-ubiquitin fusion domain. The intein variant mediates highly active autocatalytic cleavage at its N-terminus, whereas the endogenous deubiquitinases cleave at ubiquitin's C-terminus, leading to the release of the POIs. Using fluorescent reporter proteins for proof-of-concept, utility of the intein-ubiquitin domain was validated in higher plants and yeast systems. Essentially complete release of the POIs was achieved as demonstrated with western blots. Proteins expressed using the intein-ubiquitin system potentially preserve their intended sequences, which is important for preventing alteration of POI function.

Podophyllotoxin and Rutin Modulate M1 (iNOS+) Macrophages and Mitigate Lethal Radiation (LR) Induced Inflammatory Responses in Mice.

Accidental exposure to lethal doses of Gamma radiation leads to the systemic inflammatory syndrome which causes mortality. In view of this, management of hemopoietic syndrome by modulating pro-inflammatory response in clinically manageable time period seems to be the most appropriate strategy for encountering radiation induced damage and recovery. As both tissue and peripheral macrophages are critical for the management of radiation induced injuries, we have unraveled the immunomodulatory potential of radioprotective formulation (G-003M) on peripheral macrophages populations in this study. G-003M inhibited lethal radiation induced NO and Th1 effector cytokines in the exposed macrophages indicating its M1 dim polarizing capacity. In similar lines, conditioning of mice with G-003M before lethal irradiation (LR) inhibited LR induced titre of Th1 effector cytokines in both serums as well as in lung, small intestine, and spleen tissue confirming its immunomodulatory potential. G-003M potentially down modulated inflammatory response in LPS induced inflammatory model and enhanced M2 polarization of iNOS+ M1 effector macrophages providing a molecular hint on G-003M mechanism of action on macrophages. These observations revealed that G-003M potentially modulate pro-inflammatory programming of macrophages and mitigate radiation-induced inflammatory stress which is believed to contribute significantly to radioprotective attribute of G-003M. In this study, we demonstrate that Rutin and Podophyllotoxin drive M1dim/M2 polarization of LR primed macrophages apart from protecting DNA from radiation. These drugs have the capacity to programme innate immune cells like macrophages which may be involved in homeostasis during recovery.

Combination treatment of podophyllotoxin and rutin promotes mouse Lgr5+ ve intestinal stem cells survival against lethal radiation injury through Wnt signaling.

It has been well established that radiation-induced gastrointestinal injury is manifested through loss of intestinal crypt stem cells and disruption of the mucosal layers, resulting in diarrhoea, weight loss, electrolyte imbalance, infection and mortality. Podophyllotoxin and rutin in combination (G-003M) has been reported to regulate endogenous cellular antioxidant defense systems and inflammatory response. However, the mechanism by which G-003M ameliorates radiation-induced intestinal stem cell (ISC) injury remains unclear. Here, we hypothesize the radioprotective potential of G-003M would amplify the intestinal crypt stem cells through upregulation of Wnt/ß-catenin signaling and accelerate the reconstitution of the irradiated intestine. Our results showed significant functional and structural intestine regeneration in irradiated animals following G-003M treatment which resulted in improved animal survival. Immunohistochemical examination revealed an enhancement in Lgr5+ ve crypt stem cells. Increased ß-catenin nuclear translocation resulted in upregulation of ß-catenin target genes that supported ISC renewal and expansion in G-003M-treated mice, as compared to IR-treated mice. However, G-003M could not rescue the Wnt knockdown cohorts (XAV939 treated) which exhibited greater incidence of intestinal apoptosis, DNA damage and crypt depopulation upon radiation exposure. These findings suggest the involvement of Wnt pathway during G-003M mediated amelioration of IR-induced ISC injury. G-003M also minimised acute inflammation by restricting the infiltration of immune cells into the intestinal venules. Furthermore, G-003M treated animals showed improved anti-tumor response compared to FDA approved Amifostine. Taken together, our findings suggest that G-003M may be used as a potential countermeasure for radiation injuries as well as an adjuvant during anti-cancer therapy.

Overview of Biotechnology-Derived Herbicide Tolerance and Insect Resistance Traits in Plant Agriculture.

Biotechnology has been central for the acceleration of crop improvement over the last two decades. Since 1994, when the first commercial biotechnology-derived tomato crop was commercialized, the cultivated area for genetically modified crops has reached 185.1 million hactares worldwide. Both the number of crops and the number of traits developed using biotechnology have accounted for this increase. Among the most impactful biotechnology-derived traits are insect resistance and herbicide tolerance, which have greatly contributed to the worldwide increase in agricultural productivity and stabilization of food security. In this chapter, we provide an overview of the history of the biotechnology-derived input traits, the existing genetically engineered commercial crop products carrying insect resistance and herbicide tolerance traits, as well as a perspective on how new technologies could further impact the development of new traits in crops. With the projection of the world population to increase to 9.8 billion by the year 2050 and reduction in available farmland, one of the biggest challenges will be to provide sustainable nourishment to the projected population. Biotechnology will continue to be the key enabler for development of insect resistance and herbicide tolerance traits to overcome that imminent challenge.

Urbanization and biodiversity of arbuscular mycorrhizal fungi: The case study of Delhi, India / Urbanización y biodiversidad de hongos micorrízicos arbusculares -Caso de estudio de Delhi, India

Abstract Increasing urbanisation is widely associated with decline in biodiversity of all forms. The aim of the present study was to answer two questions: (i) Does rapid urbanization in Delhi (India) affect biodiversity of arbuscular mycorrhizal (AM) fungi? (ii) If so, how? We measured the AM fungal diversity at nine sites located in Delhi forests, which had different types of urban usage in terms of heavy vehicular traffic pollution, littering, defecation and recreational activities. The study revealed a significant decrease in AM fungal diversity (alpha diversity) and abundance measured as spore density, biovolume, mean infection percentage (MIP) in roots, soil hyphal length and easily extractable glomalin related soluble proteins (EE-GRSP) at polluted sites. Non-metric multidimensional scaling (NMDS) and nested PERMANOVA, revealed significant differences in AM fungal community structure which could be correlated with variations in soil moisture, temperature, pH, carbon, and nitrogen and phosphorus levels. BEST (biota and environmental matching) analysis of biological and environmental samples revealed that soil temperature and moisture accounted for 47.6 % of the total variations in the samples. The study demonstrated how different forms of human activities in urban ecosystems of Delhi are detrimental to the diversity and abundance of AM fungi.(AU)

Resumen El incremento en la urbanización está ampliamente asociado con una disminución de la biodiversidad de todas las formas. El objetivo del presente estudio fue responder dos preguntas: (i) ¿Afecta la urbanización rápida en Delhi (India) la biodiversidad de hongos micorrízicos arbusculares (MA)?, y (ii) si es así, ¿Cómo? Medimos la diversidad de hongos MA en nueve sitios ubicados en los bosques de Delhi, los cuales tenían diferentes tipos de uso urbano en términos de contaminación por alto tráfico vehicular, basura, defecación y actividades recreacionales. El estudio reveló una disminución significativa en la diversidad de hongos MA (diversidad alfa) y abundancia medida como densidad de esporas, biovolumen, porcentaje medio de infección (PMI) en raíces, longitud de las hifas del suelo y glomalina fácilmente extraible relacionada con proteínas solubles (EE-GRSP) en sitios contaminados. Análisis de escalamiento multidimensional no métrico (NMDS) y PERMANOVA anidados revelaron diferencias significativas en la estructura de hongos MA, que puede estar relacionada con la variación en humedad, temperatura, pH y niveles de carbono, nitrógeno y fósforo del suelo. El análisis BEST (biota y correspondencia ambiental) de muestras biológicas y ambientales mostró que la temperatura y humedad del suelo explican un 47.6 % del total de la variación en las muestras. Este estudio demostró cómo las diferentes actividades humanas en ecosistemas urbanos de Delhi son perjudiciales para la diversidad y abundancia de hongos MA.(AU)

Coexpression of octopine and succinamopine Agrobacterium virulence genes to generate high quality transgenic events in maize by reducing vector backbone integration.

Agrobacterium-mediated transformation is a complex process that is widely utilized for generating transgenic plants. However, one of the major concerns of this process is the frequent presence of undesirable T-DNA vector backbone sequences in the transgenic plants. To mitigate this deficiency, a ternary strain of A. tumefaciens was modified to increase the precision of T-DNA border nicking such that the backbone transfer is minimized. This particular strain supplemented the native succinamopine VirD1/VirD2 of EHA105 with VirD1/VirD2 derived from an octopine source (pTi15955), the same source as the binary T-DNA borders tested here, residing on a ternary helper plasmid containing an extra copy of the succinamopine VirB/C/G operons and VirD1. Transformation of maize immature embryos was carried out with two different test constructs, pDAB101556 and pDAB111437, bearing the reporter YFP gene and insecticidal toxin Cry1Fa gene, respectively, contained in the VirD-supplemented and regular control ternary strains. Molecular analyses of ~ 700 transgenic events revealed a significant 2.6-fold decrease in events containing vector backbone sequences, from 35.7% with the control to 13.9% with the VirD-supplemented strain for pDAB101556 and from 24.9% with the control to 9.3% with the VirD-supplemented strain for pDAB111437, without compromising transformation efficiency. In addition, while the number of single copy events recovered was similar, there was a 24-26% increase in backbone-free events with the VirD-supplemented strain compared to the control strain. Thus, supplementing existing VirD1/VirD2 genes in Agrobacterium, to recognize diverse T-DNA borders, proved to be a useful tool to increase the number of high quality events in maize.

Podophyllotoxin and rutin in combination prevents oxidative stress mediated cell death and advances revival of mice gastrointestine following lethal radiation injury.

Intestinal injury is inevitable during exposure to high radiation doses and is a common side effect observed during abdominal/pelvic radiotherapy. Yet, no radiation countermeasures are available for gastrointestine (GI) injury management. The aim of this study is to determine the effects of podophyllotoxin and rutin in combination (G-003M) on ionising radiation induced GI injury. We prophylactically administered G-003M to C57BL/6J mice exposed to 9 Gy total body radiation (TBI) and assessed for morphological changes, loss in absorption, fluid retention, biochemical alterations, immunohistochemical analysis to study cPARP, caspase-3, PCNA expression, and TUNEL staining. The irradiated intestine demonstrated extensive loss in crypts and villi, disrupted mucosal lining with reduced xylose uptake and enhanced fluid level post 7-day radiation. Mice receiving G-003M before radiation showed significant protection to intestinal epithelium, better allocation of secretory goblet cells, recovery in absorption, and reduced intestinal oedema. Additionally, G-003M administration also prevented radiation induced ROS generation, lipid peroxidation (MDA levels) and maintained the intestinal glutathione pool compared to the irradiated animals. G-003M supplementation also resulted in restoration of intestinal mitochondrial membrane potential, which was otherwise depolarised by radiation treatment. Immunohistochemical analysis demonstrated decrease in c-PARP and caspase-3 expression in jejuna cross sections and upregulation of PCNA in G-003M treated crypt cells as compared to 9 Gy irradiated mice. Our findings show that G-003M augment survival of mice against lethal radiation by promoting structural and functional regeneration in intestinal tissue. This combination therefore can be effectively explored for preventing radiation induced GI toxicity.

Targeting DNA Repair through Podophyllotoxin and Rutin Formulation in Hematopoietic Radioprotection: An in Silico, in Vitro, and in Vivo Study.

Drug discovery field has tremendously progressed during last few decades, however, an effective radiation countermeasure agent for the safe administration to the victims of radiation exposure is still unavailable. This multi-model study is aimed at elucidating the mechanistic aspects of a novel podophyllotoxin and rutin combination (henceforth referred as G-003M) in the hematopoietic radioprotection and its involvement in the DNA damage and repair signaling pathways. Using in silico study, we identified the binding sites and structural components of G-003M and validated in vitro. We further studied various in vivo endpoints related to the DNA repair and cell death pathways in mice pre-administered with G-003M, irradiated and subsequently euthanized to collect blood and bone marrow cells. In silico study showed the binding of podophyllotoxin to ß-tubulin and presence of a functional hydroxyl group in the rutin, suggested their involvement in G2/M arrest and the free radical scavenging respectively. This experimentation was further validated through in vitro studies. In vivo mice studies confirmed that G-003M pre-administration attenuated DNA damage and enhanced repair after whole body exposure. We further noticed a decrease in the levels of γH2AX, p53BP1, and ATM kinase and an increase in the levels of DNA pk, Ku 80, Ligase IV, Mre 11, Rad 50 and NBS 1 in the blood and bone marrow cells of the G-003M pre-administered and irradiated mice. We noticed an overall increase in the pro-survival factors in the G-003M pre-treated and irradiated groups establishing the radioprotective efficacy of this formulation. The lead obtained from this study will certainly help in developing this formulation as a safe and effective radioprotector which could be used for humans against any planned or emergency exposure of radiation.