Effects of phytochemicals on ionization radiation-mediated carcinogenesis and cancer therapy

Ionizing radiation (IR)-induced cellular damage is implicated in carcinogenesis as well as therapy of cancer. Advances in radiation therapy have led to the decrease in dosage and localizing the effects to the tumor; however, the development of radioresistance in cancer cells and radiation toxicity to normal tissues are still the major concerns. The development of radioresistance involves several mechanisms, including the activation of mitogenic and survival signaling, induction of DNA repair, and changes in redox signaling and epigenetic regulation. The current strategy of combining radiation with standard cytotoxic chemotherapeutic agents can potentially lead to unwanted side effects due to both agents. Thus agents are needed that could improve the efficacy of radiation killing of cancer cells and prevent the damage to normal cells and tissues caused by the direct and bystander effects of radiation, without have its own systemic toxicity. Chemopreventive phytochemicals, usually non-toxic agents with both cancer preventive and therapeutic activities, could rightly fit in this approach. In this regard, naturally occurring compounds, including curcumin, parthenolide, genistein, gossypol, ellagic acid, withaferin, plumbagin and resveratrol, have shown considerable potential. These agents suppress the radiation-induced activation of receptor tyrosine kinases and nuclear factor-κB signaling, can modify cell survival and DNA repair efficacy, and may potentiate ceramide signaling. These radiosensitizing and counter radioresistance mechanisms of phytochemicals in cancer cells are also associated with changes in epigenetic gene regulation. Because radioresistance involves multiple mechanisms, more studies are needed to discover novel phytochemicals having multiple mechanisms of radiosensitization and to overcome radioresistance of cancer cells. Pre-clinical studies are needed to address the appropriate dosage, timing, and duration of the application of phytochemicals with radiation to justify clinical trials. Nonetheless, some phytochemicals in combination with IR may play a significant role in enhancing the therapeutic index of cancer treatment.     

Response of groundnut (Arachis hypogaea L) toRhizobium inoculation in the field: Problems and prospects

Summary A successfulRhizobium inoculant strain needs to be more competitive than native soil rhizobia in forming nodules and to be effective in fixing nitrogen. Persistence of the strain from one season to the next will be an added advantage, since it eliminates the need to inoculate every season. Earlier studies indicated that groundnut,Arachis hypogaea, seldom responds toRhizobium inoculation in soils already containing rhizobia which nodulate groundnut. However, the results of seven years of research at ICRISAT have opened up the prospect of inoculating groundnut in such soils. These experiments indicate that inoculating with sufficient numbers of an effectiveRhizobium strain, NC 92, applied as a liquid slurry below the seed, increased yields of certain groundnut cultivars. Similar results were reported from other research centres in India. Inoculation with NC 92 for two consecutive seasons increased the proportion of nodules formed by this strain, from 25–32% in the first season to 41–54% in the second season, indicating that this strain can persist in the field for the following season. This paper also discusses results of experiments relating to host cultivar specificity and some possible problems in applying this information to farmers' fields. Strain NC 92 also produces a siderophore, an iron chelating compound, which may help in the iron nutrition of the plant.

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Resumen Respuesta del maní (Arachis hypogaeaL.)a la inoculación con Rhizobiumen el campo. Problemas y perspectivas.La inoculación conRhizobium para tener exito debe de realizarse con una cepa que sea competitivamente mejor que las cepas nativas del suelo en cuestión, en relación a la habilidad para formar nódulos y a la eficacia en la fijación de nitrógeno. La persistencia de la cepa de una temporada para otra es una caracteristica deseable ya que elimina la necesidad de reinocular cada temporada. Estudios previos indicaban que el maní (Arachis hypogaeae) raras veces responde a la inoculación conRhizobium en suelos que ya contienenRhizobium especificos. Sin embargo los resultados de siete años de investigaciones en ICRISAT han abierto nuevas perspectivas para la inoculación del maní en dichos suelos. Esta experiencias indican que la inoculación, en cantidades suficientes de una cepa deRhizobium eficaz (NC 92) aplicada en forma de liquido viscoso bajo la semilla incrementa el rendimiento de ciertos cultivars de mani. Resultados semejantes se han observado en otros centros de investigación en India.La inoculación de la cepa NC 92 durante dos años consecutivos incrementó la proporción de nódulos formados desde 25–32% en la primera temporada hasta 41–54% en la segunda indicando pues la persistencia de dicha cepa en el suelo de una a otra temporada. Este trabajo también evalua los resultados de investigaciones relacionando la especificidad del inóculo con la posible problemática que conlleva la aplicación de esta informacion por el agricultor.

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Résumé Réponse de l'arachide (Arachis hypogeaL.)à l'inculation de Rhizobiumdans les champs. Problèmes et perspectives.Pour être couronnée de succès, une souche deRhizobium inoculée doit supplanter les rhizobiums natifs du sol en ce qui concerne la formation de nodules et doit fixer l'azote efficacement. La persistance de la souche d'une saison à l'autre, rendant inutile la répétition saisonnière de l'inoculation, est un avantage supplémentaire. Des études antérieures ont montré que l'arachide (Arachis hypogea) ne répond que rarement à l'inoculation dans des sols contenant déjà des rhizobiums capables de noduler cette plante. Cependant, les résultats de sept années de recherches éffectuées à l'ICRISAT ont ouvert des possibilités quant à l'inoculation dans ce type de sols. Ces expériences montrent qu'une bouillie liquide d'une souche efficace deRhizobium (NC 92), répandue en quantité suffisante sous les graines, augmente le rendement de certains cultivars d'arachide. Des résultats similaires ont été rapportés par d'autres centres de recherches en Inde. L'inoculation de NC 92 pendant deux saisons consécutives a augmenté la proportion des nodules formés par cette souche de 25 à 32% pour la première saison, et de 41 à 54% pour la seconde, ce qui indique que cette souche peut persister dans le sol d'une saison à l'autre. Dans cet article sont également discutés les résultats d'expériences concernant la spécificité de l'hôte et quelques problèmes concernant l'utilisation de ces informations par les agriculteurs. La souche NC 92 produit aussi un sidérophore, agent complexant du fer pouvant jouer un rôle dans le métabolisme de ce métal chez la plante.

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Submitted as ICRISAT J. A. No. 475.     

Detection of G-Quadruplex DNA Using Primer Extension as a Tool

DNA sequence and structure play a key role in imparting fragility to different regions of the genome. Recent studies have shown that non-B DNA structures play a key role in causing genomic instability, apart from their physiological roles at telomeres and promoters. Structures such as G-quadruplexes, cruciforms, and triplexes have been implicated in making DNA susceptible to breakage, resulting in genomic rearrangements. Hence, techniques that aid in the easy identification of such non-B DNA motifs will prove to be very useful in determining factors responsible for genomic instability. In this study, we provide evidence for the use of primer extension as a sensitive and specific tool to detect such altered DNA structures. We have used the G-quadruplex motif, recently characterized at the BCL2 major breakpoint region as a proof of principle to demonstrate the advantages of the technique. Our results show that pause sites corresponding to the non-B DNA are specific, since they are absent when the G-quadruplex motif is mutated and their positions change in tandem with that of the primers. The efficiency of primer extension pause sites varied according to the concentration of monovalant cations tested, which support G-quadruplex formation. Overall, our results demonstrate that primer extension is a strong in vitro tool to detect non-B DNA structures such as G-quadruplex on a plasmid DNA, which can be further adapted to identify non-B DNA structures, even at the genomic level.     

Enzyme-linked immunosorbent assay for riboflavin carrier protein and modified protein: application for epitope analysis.

Rabbit antibodies to native riboflavin carrier protein (RCP), are to a large extent directed towards the conformational epitopes and antibodies to disulphide bond reduced carboxymethylated riboflavin carrier protein (RCM-RCP) to the sequential epitopes. Taking advantage of this premise and in order to map the epitopes of RCP recognized by the antibodies, enzyme-linked immunosorbent assays were validated for RCP and RCM-RCP using the Avidin-Biotin system. The usefulness of these assays were illustrated when antigenicity of peptides derived from RCM-RCP following trypsinization were examined. Two major (T1,T2) and one minor peptide (T3) fractions were obtained when the tryptic peptides were fractionated on DEAE-cellulose. RCP has a blocked N-terminal. Tryptic peptides (T1 and T2) on microsequencing revealed the absence of an N-terminal amino acid, indicating that these fragments emanate from the N-terminal region of RCP. In support of this observation is the finding that antipeptide antibody to cRCP (10-24) of cRCP interacted with T1 as well as T2 indicating the presence of the sequential epitope (10-24) of cRCP in these fragments. In RCP-ELISA, only T2 displaced RCP and peptides T1 and T2 displaced RCM-RCP in RCM-RCP ELISA. Differences in the ability of these fragments (T1 and T2) to displace RCP and RCM-RCP reflect the subtle changes in the spatial structures of these epitopes in RCP and RCM-RCP.     

Marine mycoflora of south India with special emphasis on lignicolous marine fungi

A study dealing with the marine fungi associated with decaying wood samples in the brackish water mangrove ecosystem and shoreline ecosystem was carried out in south India. A total of 19 marine fungi were isolated from the brackish water mangrove ecosystem. They included 13 Ascomycetes, one Basidiomycete and five Mitosporic fungi. In terms of percent frequency of occurrence, the most frequent species obtained from the brackishwater were the Lignincola longirostris (16.60%) and Savoryella lignicola (12.09%). Nine species were found frequently. Five species were occasionally encoun-tered. Aigialus mangrovei, Aniptodera mangrovei and Halosarpheia marina were the rare species recorded. The average number of isolates per wood sample was 1.53. A total of 27 marine fungi including 15 ascomycetes, one basidiomycete and ten mitosporic fungi were recorded from the shoreline ecosystem. In terms of percent frequency of occurrence, the most frequent species obtained from Kanyakumari were the Arenariomyces trifurcates (13.66%), Corollospora maritima (12.44%), and Cirrenalia pygmea (10.98%). Seven species were found frequently. Fourteen species were occasionally encountered. Three species were found to be rare in occurrence. The average number of isolates per wood sample was 1.21.     

All-optical constant-force laser tweezers

Optical tweezers are a powerful tool for the study of single biomolecules. Many applications require that a molecule be held under constant tension while its extension is measured. We present two schemes based on scanning-line optical tweezers to accomplish this, providing all-optical alternatives to force-clamp traps that rely on electronic feedback to maintain constant-force conditions for the molecule. In these schemes, a laser beam is rapidly scanned along a line in the focal plane of the microscope objective, effectively creating an extended one-dimensional optical potential over distances of up to 8 microm. A position-independent lateral force acting on a trapped particle is created by either modulating the laser beam intensity during the scan or by using an asymmetric beam profile in the back focal plane of the microscope objective. With these techniques, forces of up to 2.69 pN have been applied over distances of up to 3.4 microm with residual spring constants of <26.6 fN/microm. We used these techniques in conjunction with a fast position measurement scheme to study the relaxation of lambda-DNA molecules against a constant external force with submillisecond time resolution. We compare the results to predictions from the wormlike chain model.     

Limiting an Insect Infestation of Nitrogen-Fixing Root Nodules of the Pigeon Pea (Cajanus cajan) by Engineering the Expression of an Entomocidal Gene in Its Root Nodules

A region of DNA which determined the production of the insecticidal toxin of Bacillus thuringiensis subsp. israelensis was cloned into a derivative of a broad-host-range group IncQ plasmid vector of gram-negative bacteria. The plasmid which we constructed was transferred by conjugative mobilization into a Bradyrhizobium species that nodulates pigeon peas. In this species the construction was maintained stably in the absence of selection and expressed the gene that was installed. Experiments in a greenhouse with the strain which we constructed indicated that this organism provides protection against root nodule damage by the larvae of the insect Rivellia angulata (Diptera).