Analysis of BAC-end sequences (BESs) and development of BES-SSR markers for genetic mapping and hybrid purity assessment in pigeonpea (Cajanus spp.)

<p>Abstract</p> <p>Background</p> <p>Pigeonpea [<it>Cajanus cajan </it>(L.) Millsp.] is an important legume crop of rainfed agriculture. Despite of concerted research efforts directed to pigeonpea improvement, stagnated productivity of pigeonpea during last several decades may be accounted to prevalence of various biotic and abiotic constraints and the situation is exacerbated by availability of inadequate genomic resources to undertake any molecular breeding programme for accelerated crop improvement. With the objective of enhancing genomic resources for pigeonpea, this study reports for the first time, large scale development of SSR markers from BAC-end sequences and their subsequent use for genetic mapping and hybridity testing in pigeonpea.</p> <p>Results</p> <p>A set of 88,860 BAC (bacterial artificial chromosome)-end sequences (BESs) were generated after constructing two BAC libraries by using <it>Hin</it>dIII (34,560 clones) and <it>Bam</it>HI (34,560 clones) restriction enzymes. Clustering based on sequence identity of BESs yielded a set of >52K non-redundant sequences, comprising 35 Mbp or >4% of the pigeonpea genome. These sequences were analyzed to develop annotation lists and subdivide the BESs into genome fractions (e.g., genes, retroelements, transpons and non-annotated sequences). Parallel analysis of BESs for microsatellites or simple sequence repeats (SSRs) identified 18,149 SSRs, from which a set of 6,212 SSRs were selected for further analysis. A total of 3,072 novel SSR primer pairs were synthesized and tested for length polymorphism on a set of 22 parental genotypes of 13 mapping populations segregating for traits of interest. In total, we identified 842 polymorphic SSR markers that will have utility in pigeonpea improvement. Based on these markers, the <it>first </it>SSR-based genetic map comprising of 239 loci was developed for this previously uncharacterized genome. Utility of developed SSR markers was also demonstrated by identifying a set of 42 markers each for two hybrids (ICPH 2671 and ICPH 2438) for genetic purity assessment in commercial hybrid breeding programme.</p> <p>Conclusion</p> <p>In summary, while BAC libraries and BESs should be useful for genomics studies, BES-SSR markers, and the genetic map should be very useful for linking the genetic map with a future physical map as well as for molecular breeding in pigeonpea.</p

Rosiglitazone synergizes anticancer activity of cisplatin and reduces its nephrotoxicity in 7, 12-dimethyl benz{a}anthracene (DMBA) induced breast cancer rats

<p>Abstract</p> <p>Background</p> <p>Antineoplastic drug cisplatin remains the drug of choice for various solid tumours including breast cancer. But dose dependent nephrotoxicity is the major drawback in majority of platinum based chemotherapy regimens. Recent reports have shown that inflammatory pathways are the main offender for cisplatin induced nephrotoxicity. The present study was undertaken to assess the effect of rosiglitazone, a PPARγ agonist and an anti-inflammatory agent, on cisplatin induced nephrotoxicity, and its anticancer activity in DMBA induced breast cancer rats.</p> <p>Methods</p> <p>Mammary tumours were induced in female Sprague-Dawley rats by feeding orally with dimethylbenz [a]anthracene (DMBA) (60 mg/kg). Cisplatin induced nephropathy was assessed by measurements of blood urea nitrogen, albumin and creatinine levels. Posttranslational modifications of histone H3, mitogen-activated protein (MAP) kinase p38 expression and PPAR-γ expression were examined by western blotting.</p> <p>Results</p> <p>Our data shows involvement of TNF-α in preventing cisplatin induced nephrotoxicity by rosiglitazone. Rosiglitazone pre-treatment to cisplatin increases the expression of p38, PPAR-γ in mammary tumours and shows maximum tumour reduction. Furthermore, cisplatin induced changes in histone acetylation, phosphorylation and methylation of histone H3 in mammary tumours was ameliorated by pre-treatment of rosiglitazone. Suggesting, PPAR-γ directly or indirectly alters aberrant gene expression in mammary tumours by changing histone modifications.</p> <p>Conclusion</p> <p>To best of our knowledge this is the first report which shows that pre-treatment of rosiglitazone synergizes the anticancer activity of cisplatin and minimizes cisplatin induced nephrotoxicity in DMBA induced breast cancer.</p

Influenza A Virus Nucleoprotein Exploits Hsp40 to Inhibit PKR Activation

BACKGROUND: Double-stranded RNA dependent protein kinase (PKR) is a key regulator of the anti-viral innate immune response in mammalian cells. PKR activity is regulated by a 58 kilo Dalton cellular inhibitor (P58(IPK)), which is present in inactive state as a complex with Hsp40 under normal conditions. In case of influenza A virus (IAV) infection, P58(IPK) is known to dissociate from Hsp40 and inhibit PKR activation. However the influenza virus component responsible for PKR inhibition through P58(IPK) activation was hitherto unknown. PRINCIPAL FINDINGS: Human heat shock 40 protein (Hsp40) was identified as an interacting partner of Influenza A virus nucleoprotein (IAV NP) using a yeast two-hybrid screen. This interaction was confirmed by co-immunoprecipitation studies from mammalian cells transfected with IAV NP expressing plasmid. Further, the IAV NP-Hsp40 interaction was validated in mammalian cells infected with various seasonal and pandemic strains of influenza viruses. Cellular localization studies showed that NP and Hsp40 co-localize primarily in the nucleus. During IAV infection in mammalian cells, expression of NP coincided with the dissociation of P58(IPK) from Hsp40 and decrease PKR phosphorylation. We observed that, plasmid based expression of NP in mammalian cells leads to decrease in PKR phosphorylation. Furthermore, inhibition of NP expression during influenza virus replication led to PKR activation and concomitant increase in eIF2α phosphorylation. Inhibition of NP expression also led to reduced IRF3 phosphorylation, enhanced IFN β production and concomitant reduction of virus replication. Taken together our data suggest that NP is the viral factor responsible for P58(IPK) activation and subsequent inhibition of PKR-mediated host response during IAV infection. SIGNIFICANCE: Our findings demonstrate a novel role of IAV NP in inhibiting PKR-mediated anti-viral host response and help us understand P58(IPK) mediated inhibition of PKR activity during IAV infection

Bifurcation analysis of a plankton model with Discrete Delay(#Third Revision, paper code JRNL6919)

In this paper, a delayed plankton-nutrient interac- tion model consisting of phytoplankton, zooplankton and dis- solved nutrient is considered. It is assumed that some species of phytoplankton releases toxin (known as toxin producing phyto- plankton(TPP)) which is harmful for zooplankton growth and this toxin releasing process follows a discrete time variation. Using delay as bifurcation parameter, the stability of interior equilibrium point is investigated and it is shown that time delay can destabilise the otherwise stable non-zero equilibrium state by inducing Hopf-bifurcation when it crosses a certain threshold value. Explicit results are derived for stability and direction of the bifurcating periodic solution by using normal form theory and center manifold arguments. Finally, outcomes of the system are validated through numerical simulations

Contrasting Responses of Guar Genotypes Shed Light on Multiple Component Traits of Salinity Tolerance Mechanisms

Guar (Cyamopsis tetragonoloba (L.) Taub.) is a legume crop, and gum derived from its seeds has various industrial applications. Due to its tolerance to various abiotic stresses, guar can be grown under water-deficit or high-salinity conditions. In this investigation, four diverse guar genotypes that performed at a similar level in field conditions were evaluated in a salinity experiment in the greenhouse lysimeter system. Based on the salt tolerance index (STI) for shoot biomass, root biomass, shoot length, and root length, Matador and PI 268229 were classified as salt-tolerant, and PI 340261 and PI 537281 as salt-sensitive. Leaf Na concentrations were 4- to 5.5-fold higher, and leaf Cl concentrations were 1.6- to 1.9-fold higher in salt-sensitive lines than salt-tolerant lines under salinity. The strong associations between the leaf K concentrations under salinity compared to the control (K-salinity/K-control) ratio and STI for stem and root length advocate higher importance of K-salinity/K-control than total leaf K concentrations. The expression analyses of genes involved in Na+ and Cl− transport revealed the importance of different component traits of salinity tolerance mechanisms, such as the exclusion of Na+/Cl− from the root, sequestration of Cl− in root vacuoles, retrieval of Na+/Cl− from xylem during salinity stress, root-to-shoot Na+/Cl− translocation, and K+-Na+ homeostasis

Comparative Evaluation of Post Operative Analgesic Effects of Intraperitoneal Levobupivacaine Plus Dexmedetidomine and Levobupivacaine Plus Clonidine in Patients Undergoing Laparoscopic Cholecystectomy.

Background and Aims: Local anaesthetics have been used widely through the intraperitoneal route, either alone or with various adjuvants, to effectively decrease the&nbsp; post operative pain after laparoscopic cholecystectomy and increase the duration of analgesia.&nbsp; Our study compared the post operative&nbsp; analgesic efficacy&nbsp; of&nbsp; intraperitoneal&nbsp; Dexmedetomidine&nbsp; or&nbsp; Clonidine&nbsp; added&nbsp; to Levobupivacaine&nbsp; in patients undergoing laparoscopic cholecystectomy. Methods: &nbsp;90&nbsp;&nbsp; ,&nbsp; ASA&nbsp; I-II&nbsp; patients , who were randomly&nbsp; divided&nbsp; into three&nbsp;&nbsp; groups were enrolled in the study.&nbsp; Group I&nbsp;&nbsp; ( L )&nbsp; received&nbsp; 38 ml of 0.25%&nbsp; Levobupivacaine&nbsp; + 2 ml saline, Group II&nbsp;&nbsp; ( LD ) received 38 ml of 0.25%&nbsp; Levobupivaciane + 0.5 microgm/kg&nbsp; Dexmedetomidine diluted in 2 ml normal saline&nbsp; and&nbsp; Group&nbsp; III ( LC ) received 38ml of 0.25%&nbsp; Levobupivacaine&nbsp; + 1 microgm/kg&nbsp; Clonidine, diluted in 2 ml normal saline&nbsp; making a total solution of 40 ml.&nbsp; The study solution was given intraperitoneally before removal of trocar in Trendelenberg’s position into the hepatodiaphragmatic space, near and above the hepato -duodenal ligament and in the gallbladder fossa. Visual analogue scale score (VAS ) was used to assess the quality of analgesia. Time to the first request of analgesia, total dose of analgesic in the first 24 hrs, hemodynamic parameters at various post operative intervals&nbsp; and adverse effects were noted. Results: VAS score at various intervals&nbsp; post operative, overall VAS in 24 h was significantly lower (1.57 ± 0.27,&nbsp;&nbsp; 1.63 ± 0.24,&nbsp; 2.25 ± 0.18&nbsp; ), time to first request of analgesia (min) was longest (8.56 ± 2.15 hrs,&nbsp; 6.26 ± 4.28 hrs,&nbsp; 1.25 ± 0.89 hrs ) and&nbsp; mean of the total&nbsp; analgesic&nbsp; doses consumption&nbsp; was lowest (0.77 ± 0.63, 0.90 ± 0.66 ,&nbsp; 2.37 ± 0.56)&nbsp; in Group LD than Group LC than Group L. Conclusion: Postoperative analgesic efficacy of intraperitoneal instillation of&nbsp; Levobupivacaine&nbsp; with Dexmedetomidine was found to be&nbsp; superior to&nbsp; Clonidine&nbsp; group and much superior to instillation of&nbsp; levobupivacaine alone