Synthesis, x-ray crystal structures and biological evaluation of some mono- and bi-cyclic 1,3-diazetidin-2-ones: non-natural β -lactam analogues

Mono- and bi-cyclic 1,3-diazetidin-2-ones (aza-β -lactams) are synthesised and evaluated as non-natural analogues of β -lactams. The aza-β -lactams are designed on the principle that their reaction with active site serine hydroxy will form a carbamoyl-enzyme intermediate that is sluggish to hydrolysis. The synthesis of racemic mono- and bi-cyclic aza-β -lactams is carried out starting from pyrimidinone 18 which is transformed to the densely functionalised substrate 20. The chemical reactivity of tricarbonyl 20 for selective functional group manipulation was first assessed and then it was transformed to amino alcohol 24. Cyclisation of 24 affords aza-carbapenams and its homologation followed by aldol cyclisation provides access to aza-carbacephams. The X-ray structures of aza-carbapenam 35 and aza-carbacepham 42 suggest that the structural requirements for biological activity in β -lactams are fulfilled. An unexpected ozonolysis product, phenol 52 resolves spontaneously during crystallisation and its crystal structure was also determined. The biological activity of the novel mono- and bi-cyclic aza-β -lactams was evaluated with potent gram-positive bacterial strain, Bacillus subtilis and compared with β -lactam antibiotics, ampicillin and penicillin G. Of the 19 aza-β -lactams tested, eight compounds show inhibition better than the standards while another eight are of comparable activity. This study shows that aza-β -lactams represent a novel and non-natural lead towards serine peptidase inhibitors

Enrichment of Genetic Linkage Maps and Mapping QTLs Specific to Seed Strength-Hardness/Softness-In Guava (Psidium guajava L.)

The present research focuses mainly on molecular mining and morphological evaluation of guava genome within a full-sib population and, thereby, mapping of quantitative trait loci related to fruit quality traits, viz., seed strength (hardness/softness) and average fruit weight. Linkage maps were enriched for both parental lines, 'Kamsari' and 'Purple Local' using a set of 60 RAPD markers following the pseudo-testcross strategy on a panel of 94 progeny. A total of 480 scorable markers were identified, of which 131 were specific to 'kamsari' and 28 to 'Purple Local', segregating as test cross markers, and, 321 showing intercross pattern common to both. 'Kamsari' spanned a total length of 1959.1cM with average marker interval distance of 3.93cM, while 'Purple Local' spanned a length of 1537.9cM with average marker interval distance of 3.29cM, by forming 11 linkage groups. Estimated genome length observed was 93.02% and 92.77% in 'Kamsari' and 'Purple Local', respectively. Composite Interval Mapping (CIM) was computed at significance of 0.05 and LOD threshold greater than 3.0, which led to detection of one major QTL for the trait of average fruit weight, and, four QTLs for the trait of seed strength (hardness/softness). Of these, two were major and two minor QTLs. Our study provides molecular mapping information on marker-assisted selection for improvement of guava in a breeding program

Size--sensitive melting characteristics of gallium clusters: Comparison of Experiment and Theory for Ga17+_{17}{}^{+} and Ga20+_{20}{}^{+}

Experiments and simulations have been performed to examine the finite-temperature behavior of Ga$_{17}{}^{+}$ and Ga$_{20}{}^{+}$ clusters. Specific heats and average collision cross sections have been measured as a function of temperature, and the results compared to simulations performed using first principles Density--Functional Molecular--Dynamics. The experimental results show that while Ga$_{17}{}^{+}$ apparently undergoes a solid--liquid transition without a significant peak in the specific--heat, Ga$_{20}{}^{+}$ melts with a relatively sharp peak. Our analysis of the computational results indicate a strong correlation between the ground--state geometry and the finite--temperature behavior of the cluster. If the ground--state geometry is symmetric and "ordered" the cluster is found to have a distinct peak in the specific--heat. However, if the ground--state geometry is amorphous or "disordered" the cluster melts without a peak in the specific--heat.Comment: 6 figure

Integrated Physiological, Biochemical, and Molecular Analysis Identifies Important Traits and Mechanisms Associated with Differential Response of Rice Genotypes to Elevated Temperature

In changing climate, heat stress caused by high temperature poses a serious threat to rice cultivation. A multiple organizational analysis at physiological, biochemical and molecular level is required to fully understand the impact of elevated temperature in rice. This study was aimed at deciphering the elevated temperature response in eleven popular and mega rice cultivars widely grown in India. Physiological and biochemical traits specifically membrane thermostability (MTS), antioxidants, and photosynthesis were studied at vegetative and reproductive phases which were used to establish a correlation with grain yield under stress. Several useful traits in different genotypes were identified which will be important resource to develop high temperature tolerant rice cultivars. Interestingly, Nagina22 emerged as best performer in terms of yield as well as expression of physiological and biochemical traits at elevated temperature. It showed lesser relative injury, lesser reduction in chlorophyll content, increased super oxide dismutase, catalase and peroxidase activity, lesser reduction in net photosynthetic rate (PN), high transpiration rate (E) and other photosynthetic/ fluorescence parameters contributing to least reduction in spikelet fertility and grain yield at elevated temperature. Further, expression of 14 genes including heat shock transcription factors and heat shock proteins was analyzed in Nagina22 (tolerant) and Vandana (susceptible) at flowering phase, strengthening the fact that N22 performs better at molecular level also during elevated temperature. This study shows that elevated temperature response is complex and involves multiple biological processes which are needed to be characterized to address the challenges of future climate extreme conditions

The glutathione biosynthetic pathway of Plasmodium is essential for mosquito transmission

1Infection of red blood cells (RBC) subjects the malaria parasite to oxidative stress. Therefore, efficient antioxidant and redox systems are required to prevent damage by reactive oxygen species. Plasmodium spp. have thioredoxin and glutathione (GSH) systems that are thought to play a major role as antioxidants during blood stage infection. In this report, we analyzed a critical component of the GSH biosynthesis pathway using reverse genetics. Plasmodium berghei parasites lacking expression of gamma-glutamylcysteine synthetase (γ-GCS), the rate limiting enzyme in de novo synthesis of GSH, were generated through targeted gene disruption thus demonstrating, quite unexpectedly, that γ-GCS is not essential for blood stage development. Despite a significant reduction in GSH levels, blood stage forms of pbggcs− parasites showed only a defect in growth as compared to wild type. In contrast, a dramatic effect on development of the parasites in the mosquito was observed. Infection of mosquitoes with pbggcs− parasites resulted in reduced numbers of stunted oocysts that did not produce sporozoites. These results have important implications for the design of drugs aiming at interfering with the GSH redox-system in blood stages and demonstrate that de novo synthesis of GSH is pivotal for development of Plasmodium in the mosquito

Deep sequencing of small RNAs reveals ribosomal origin of microRNAs in Oryza sativa and their regulatory role in high temperature

MicroRNAs are small noncoding regulatory RNAs which control gene expression by mRNA degradation or translational repression. They are significant molecular players regulating important biological processes such as developmental timing and stress response. We report here the discovery of miRNAs derived from ribosomal DNA using the small RNA datasets of 16 deep sequencing libraries of rice. Twelve putative miRNAs were identified based on highly stringent criteria of novel miRNA prediction. Surprisingly, 10 putative miRNAs (mi_7403, mi_8435, mi_12675, mi_4266, mi_4758, mi_4218, mi_8200, mi_4644, mi_14291, mi_16235) originated from rDNA of rice chromosome 9. Expression analysis of putative miRNAs and their target genes in heat tolerant and susceptible rice cultivars in control and high temperature treated seedlings revealed differential regulation of rDNA derived miRNAs. This is the first report of rDNA derived miRNAs in rice which indicates their role in gene regulation during high temperature stress in plants. Further studies in this area will open new research challenges and opportunities to broaden our knowledge on gene regulation mechanisms

Study of the properties of thermoset materials derived from epoxidized soybean oil and protein fillers

[EN] Novel bio-based thermoset formulations were prepared by using epoxidized soybean oil (ESBO), nadic methyl anhydride as a hardener and with different types of proteins as fillers. In the first part of the study, the effect of the protein-type (wheat gluten, soy protein, casein and ovalbumin) on cured ESBO materials was investigated. Thermal and mechanical properties were characterized by flexural tests, Shore D hardness, Charpy impact tests, Vicat softening temperature and heat deflection temperature. In addition, a study of the morphology of fractured surfaces by scanning electron microscopy was carried out. In general, the addition of protein-based fillers improved the mechanical and thermal properties. It was found that the highest increase of thermal and mechanical properties was achieved by ovalbumin. In the second part of the work, the effect of the total amount of ovalbumin filler was studied. Bio-based thermoset materials from ESBO and 15 wt % ovalbumin improved flexural modulus more than 150 % when compared to the unfilled material. Similar evolution was observed for other mechanical properties. Moreover, the brittleness of this composition was the minimum from the studied systems. A direct relationship between energy absorption capacity and morphologies of the failure surface was evidenced by SEM.This work is a part of the project IPT-310000-2010-037,"ECOTEXCOMP: Research and development of textile structures useful as reinforcement of composite materials with marked ecological character" funded by the "Ministerio de Ciencia e Innovacion", with an aid of 189540.20 euros, within the "Plan Nacional de Investigacion Cientifica, Desarrollo e Innovacion Tecnologica 2008-2011" and funded by the European Union through FEDER funds, Technology Fund 2007-2013, "Operational Programme on R+D+I for and on behalf of the companies". Also, Generalitat Valenciana ACOMP/2012/087 is acknowledged for financial support.Fombuena Borrás, V.; Sánchez Nacher, L.; Samper Madrigal, MD.; Juárez Varón, D.; Balart Gimeno, RA. (2013). Study of the properties of thermoset materials derived from epoxidized soybean oil and protein fillers. Journal of the American Oil Chemists' Society. 90(3):449-457. https://doi.org/10.1007/s11746-012-2171-2S449457903Alonso MV, Oliet M, Garcia J, Rodriguez F, Echeverria J (2006) Gelation and isoconversional kinetic analysis of lignin-phenol-formaldehyde resol resins cure. Chem Eng J 122:159–166Altuna FI, Esposito LH, Ruseckaite RA, Stefani PM (2011) Thermal and mechanical properties of anhydride-cured epoxy resins with different contents of bio-based epoxidized soybean oil. J Appl Polym Sci 120:789–798Boquillon N, Fringant C (2000) Polymer networks derived from curing of epoxidised linseed oil: influence of different catalysts and anhydride hardeners. Polymer 41:8603–8613Boquillon N, Elbez G, Schonfeld U (2004) Properties of wheat straw particleboards bonded with different types of resin. J Wood Sci 50:230–235Chakrapani S, Crivello JV (1998) Synthesis and photoinitiated cationic polymerization of epoxidized castor oil and its derivatives. J Macromol Sci-Pure Appl Chem A35:1–20Chen F, Zhang JW (2009) A new approach for morphology control of poly(butylene adipate-co-terephthalate) and soy protein blends. Polymer 50:3770–3777Cuq B, Contard N, Guilbert S (1998) Proteins as agricultural polymers for packaging production. Am Assoc Cereal Chem 75:1–9Czub P (2006) Application of modified natural oils as reactive diluents for epoxy resins. Macromol Symp 242:60–64DdS Martini, Braga BA, Samios D (2009) On the curing of linseed oil epoxidized methyl esters with different cyclic dicarboxylic anhydrides. Polymer 50:2919–2925Dogan E, Kuesefoglu S (2008) Synthesis and in situ foaming of biodegradable malonic acid ESO polymers. J Appl Polym Sci 110:1129–1135Espinosa-Perez J, Wiesenborn DP, Tostenson K, Ulven CA, Tatlari M (2007) Preparation and partial characterization of canola-based epoxy resins for bio-based plastic composites. ASABE Annual International Meeting, 076079, Minneapolis, MNJin H, Zhang L, Chen F (2003) Effects of lignin as a filler on properties of soy protein plastics. I Lignosulfonate. J Appl Polym Sci 88:3284–3290Liu ZS, Erhan SZ, Calvert PD (2007) Solid freeform fabrication of epoxidized soybean oil/epoxy composite with bis or polyalkyleneamine curing agents. Compos Part A Appl Sci Manuf 38:87–93Matejka L, Lovy J, Pokorny S, Bouchal K, Dusek K (1983) Curing epoxy-resins with anhydrides—model reactions and reaction-mechanism. J Polym Sci Part A Polym Chem 21:2873–2885Miyagawa H, Mohanty AK, Drzal LT, Misra M (2005) Nanocomposites from bio-based epoxy and single-wall carbon nanotubes: synthesis, and mechanical and thermo-physical properties evaluation. Nanotechnology 16:118–124Mohamed A, Finkenstadt VL, Gordon SH, Palmquist DE (2010) Thermal and mechanical properties of compression-molded pMDI-reinforced PCL/gluten composites. J Appl Polym Sci 118:2778–2790Montero de Espinosa L, Ronda JC, Galià M, Cádiz V (2008) A new enone-containing triglyceride derivative as precursor of thermosets from renewable resources. J Polym Sci Part A Polym Chem 46:6843–6850Park SJ, Jin FL, Lee JR (2004) Synthesis and thermal properties of epoxidized vegetable oil. Macromol Rapid Commun 25:724–727Pfister DP, Baker RJ, Henna HP, Lu Y, Larock CR (2008) Preparation and properties of tung oil-based composites using spent germ as a natural filler. J Appl Polym Sci 108:3618–3625Reiznautt QB, Garcia ITS, Samios D (2009) Oligoesters and polyesters produced by the curing of sunflower oil epoxidized biodiesel with cis-cyclohexane dicarboxylic anhydride: synthesis and characterization. Mater Sci Eng C Mater Biol Appl 29:2302–2311Rüsch Gen Klaas M, Warwel S (1999) Complete and partial epoxidation of plant oils by lipase-catalyzed perhydrolysis. Ind Crops Prod 9:125–132Sailaja RRN, Girija BG, Madras G, Balasubramanian N (2008) Effect of compatibilization on mechanical and thermal properties of polypropylene—soy flour composites. J Mater Sci 43:64–67Samper MD, Fombuena V, Boronat T, García-Sanoguera D, Balart R (2012) Thermal and mechanical characterization of epoxy resins (ELO and ESO) cured with anhydrides. J Am Oil Chem Soc 89(8):1521–1528Sharma S (2008) Fabrication and characterization of polymer blends and composites derived from biopolymers. Philosophy Materials Science and Engineering. Graduate School of Clemson University, ClemsonSharma BK, Liu Z, Adhvaryu A, Erhan SZ (2008) One-pot synthesis of chemically modified vegetable oils. J Agric Food Chem 56:3049–3056Sue HJ, Wang S, Jane J (1997) Morphology and mechanical behaviour of engineering soy plastics. J Polym 38:5035Wang S, Sue HJ, Jane J (1996) Effects of polyhydric alcohols on the mechanical properties of soy protein plastics. J Macromol Sci Pure Appl Chem A33:557–569Wazzan AA, Al-Turaif HA, Abdelkader AF (2006) Influence of submicron TiO2 particles on the mechanical properties and fracture characteristics of cured epoxy resin. Polym Plastics Technol Eng 45:1155–116

Hundreds of variants clustered in genomic loci and biological pathways affect human height

Most common human traits and diseases have a polygenic pattern of inheritance: DNA sequence variants at many genetic loci influence the phenotype. Genome-wide association (GWA) studies have identified more than 600 variants associated with human traits, but these typically explain small fractions of phenotypic variation, raising questions about the use of further studies. Here, using 183,727 individuals, we show that hundreds of genetic variants, in at least 180 loci, influence adult height, a highly heritable and classic polygenic trait. The large number of loci reveals patterns with important implications for genetic studies of common human diseases and traits. First, the 180 loci are not random, but instead are enriched for genes that are connected in biological pathways (P = 0.016) and that underlie skeletal growth defects (P < 0.001). Second, the likely causal gene is often located near the most strongly associated variant: in 13 of 21 loci containing a known skeletal growth gene, that gene was closest to the associated variant. Third, at least 19 loci have multiple independently associated variants, suggesting that allelic heterogeneity is a frequent feature of polygenic traits, that comprehensive explorations of already-discovered loci should discover additional variants and that an appreciable fraction of associated loci may have been identified. Fourth, associated variants are enriched for likely functional effects on genes, being over-represented among variants that alter amino-acid structure of proteins and expression levels of nearby genes. Our data explain approximately 10% of the phenotypic variation in height, and we estimate that unidentified common variants of similar effect sizes would increase this figure to approximately 16% of phenotypic variation (approximately 20% of heritable variation). Although additional approaches are needed to dissect the genetic architecture of polygenic human traits fully, our findings indicate that GWA studies can identify large numbers of loci that implicate biologically relevant genes and pathways.

Discovery and Fine-Mapping of Adiposity Loci Using High Density Imputation of Genome-Wide Association Studies in Individuals of African Ancestry: African Ancestry Anthropometry Genetics Consortium

Genome-wide association studies (GWAS) have identified \u3e 300 loci associated with measures of adiposity including body mass index (BMI) and waist-to-hip ratio (adjusted for BMI, WHRadjBMI), but few have been identified through screening of the African ancestry genomes. We performed large scale meta-analyses and replications in up to 52,895 individuals for BMI and up to 23,095 individuals for WHRadjBMI from the African Ancestry Anthropometry Genetics Consortium (AAAGC) using 1000 Genomes phase 1 imputed GWAS to improve coverage of both common and low frequency variants in the low linkage disequilibrium African ancestry genomes. In the sex-combined analyses, we identified one novel locus (TCF7L2/HABP2) for WHRadjBMI and eight previously established loci at P \u3c 5×10−8: seven for BMI, and one for WHRadjBMI in African ancestry individuals. An additional novel locus (SPRYD7/DLEU2) was identified for WHRadjBMI when combined with European GWAS. In the sex-stratified analyses, we identified three novel loci for BMI (INTS10/LPL and MLC1 in men, IRX4/IRX2 in women) and four for WHRadjBMI (SSX2IP, CASC8, PDE3B and ZDHHC1/HSD11B2 in women) in individuals of African ancestry or both African and European ancestry. For four of the novel variants, the minor allele frequency was low (\u3c5%). In the trans-ethnic fine mapping of 47 BMI loci and 27 WHRadjBMI loci that were locus-wide significant (P \u3c 0.05 adjusted for effective number of variants per locus) from the African ancestry sex-combined and sex-stratified analyses, 26 BMI loci and 17 WHRadjBMI loci contained ≤ 20 variants in the credible sets that jointly account for 99% posterior probability of driving the associations. The lead variants in 13 of these loci had a high probability of being causal. As compared to our previous HapMap imputed GWAS for BMI and WHRadjBMI including up to 71,412 and 27,350 African ancestry individuals, respectively, our results suggest that 1000 Genomes imputation showed modest improvement in identifying GWAS loci including low frequency variants. Trans-ethnic meta-analyses further improved fine mapping of putative causal variants in loci shared between the African and European ancestry populations