Declining in efficacy of a three-day combination regimen of mefloquine-artesunate in a multi-drug resistance area along the Thai-Myanmar border

<p>Abstract</p> <p>Background</p> <p>Declining in clinical efficacy of artesunate-mefloquine combination has been documented in areas along the eastern border (Thai-Cambodian) of Thailand. In the present study, the clinical efficacy of the three-day combination regimen of artesunate-mefloquine as first-line treatment for acute uncomplicated falciparum malaria in Thailand was monitored in an area along the western border (Thai-Myanmar) of the country.</p> <p>Methods</p> <p>A total of 150 Burmese patients (85 males and 65 females) aged between 16 and 50 years who were attending the Mae Tao clinic, Mae-Sot, Tak Province, and presenting with symptomatic acute uncomplicated <it>Plasmodium falciparum </it>malaria were included into the study. Patients were treated initially (day 0) with 4 mg/kg body weight artesunate and 15 mg/kg body weight mefloquine. The dose regimen on day 2 was 4 mg/kg body weight artesunate and 10 mg/kg body weight mefloquine. On day 3, artesunate at the dose of 4 mg/kg body weight was given with 0.6 mg/kg body weight primaquine. Whole blood mefloquine and plasma artesunate and dihydroartemisinin (active plasma metabolite of artesunate) concentrations following treatment were determined by high performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LCMS), respectively.</p> <p>Results</p> <p>Thirty-four cases had recrudescence during days 7 and 42. Five and 5 cases, respectively had reinfection with <it>P. falciparum </it>and reappearance of <it>Plasmodium vivax </it>in their peripheral blood during follow-up. The Kaplan-Meier estimate of the 42-and 28-day efficacy rates of this combination regimen were 72.58% (95% CI: 63.20-79.07%) and 83.06 (95% CI 76.14-94.40%), respectively. Parasite clearance time (PCT) and fever clearance time (FCT) were significantly prolonged in patients with treatment failure compared with those with sensitive response [median (95% CI) values for PCT 32.0 (20.0-48.0) <it>vs </it>24.0 (14.0-32.0) hr and FCT 30.0 (22.0-42.0) <it>vs </it>26.0 (18.0-36.0) hr; <it>p </it>< 0.005]. Whole blood mefloquine concentrations on days 1, 7 and 14 in patients with sensitive and recrudescence response were comparable. Although plasma concentration of dihydroartemisinin at 1 hour of treatment was significantly lower in patients with recrudescence compared with sensitive response [mean (95% CI) 456 (215-875) <it>vs </it>525 (452-599) ng/ml; <it>p </it>< 0.001], the proportion of patients with recrudescence who had relatively low (compared with the lower limit of 95% CI defined in the sensitive group) was significantly smaller than that of the sensitive group.</p> <p>Conclusions</p> <p>Although pharmacokinetic (ethnic-related) factors including resistance of <it>P. falciparum </it>to mefloquine contribute to some treatment failure following treatment with a three-day combination regimen of artesunate-mefloquine, results suggest that artesunate resistance may be emerging at the Thai-Myanmar border.</p

Mifepristone Treatment Promotes Testicular Leydig Cell Tumor Progression in Transgenic Mice

Simple SummaryRecently, the antiprogestin activity of selective progesterone receptor (PR) modulator mifepristone (MF) has proven unsuccessful as a potential anti-cancer agent in various clinical trials. Herein, we analyzed the effects of MF treatment on Leydig cell tumor (LCT) progression in a transgenic mouse model (inhibin-alpha promoter-driven SV40 T-antigen), as well as on the proliferation of two Leydig tumor cell lines. MF significantly stimulated the proliferation of LCT in vitro. Similarly, a 1-mo MF or P4 treatment stimulated LCT tumor growth in vivo. Only the abundant membrane Pgrmc1 expression was found in LCTs, but no other classical Pgr or nonclassical membrane PRs. Functional analysis showed that PGRMC1 is required for MF and P4 to stimulate the proliferation and invasiveness of LCTs. Our findings provide novel information that the use of MF as an anti-cancer agent should be considered with caution due to its potential PGRMC1 tumor-promoting pathway activation in cancers.The selective progesterone receptor modulator mifepristone (MF) may act as a potent antiproliferative agent in different steroid-dependent cancers due to its strong antagonistic effect on the nuclear progesterone receptor (PGR). Hereby, we analyzed the effects of MF treatment on Leydig cell tumor (LCT) progression in a transgenic mouse model (inhibin-alpha promoter-driven SV40 T-antigen), as well as on LCT (BLTK-1 and mLTC-1) cell proliferation. MF significantly stimulated the proliferation of LCT in vitro. Similarly, a 1-mo MF or P4 treatment stimulated LCT tumor growth in vivo. Traceable/absent classical Pgr or nonclassical membrane PRs alpha, beta, gamma and Pgrmc2, but abundant membrane Pgrmc1 expression, was found in LCTs. MF did not activate glucocorticoid or androgen receptors in LCTs. Functional analysis showed that PGRMC1 is required for MF and P4 to stimulate the proliferation and invasiveness of LCTs. Accordingly, MF and P4 induced PGRMC1 translocation into the nucleus and thereby stimulated the release of TGF beta 1 in LCT cells. MF and P4 treatments upregulated Tgfbr1, Tgfbr2, and Alk1 expression and stimulated TGF beta 1 release in LCT cells. Our findings provide novel mechanistic insights into the action of MF as a membrane PR agonist that promotes LCT growth through PGRMC1 and the alternative TGF beta 1 signaling pathway

Coxiella burnetii Phagocytosis Is Regulated by GTPases of the Rho Family and the RhoA Effectors mDia1 and ROCK

The GTPases belonging to the Rho family control the actin cytoskeleton rearrangements needed for particle internalization during phagocytosis. ROCK and mDia1 are downstream effectors of RhoA, a GTPase involved in that process. Coxiella burnetii, the etiologic agent of Q fever, is internalized by the host´s cells in an actin-dependent manner. Nevertheless, the molecular mechanism involved in this process has been poorly characterized. This work analyzes the role of different GTPases of the Rho family and some downstream effectors in the internalization of C. burnetii by phagocytic and non-phagocytic cells. The internalization of C. burnetii into HeLa and RAW cells was significantly inhibited when the cells were treated with Clostridium difficile Toxin B which irreversibly inactivates members of the Rho family. In addition, the internalization was reduced in HeLa cells that overexpressed the dominant negative mutants of RhoA, Rac1 or Cdc42 or that were knocked down for the Rho GTPases. The pharmacological inhibition or the knocking down of ROCK diminished bacterium internalization. Moreover, C. burnetii was less efficiently internalized in HeLa cells overexpressing mDia1-N1, a dominant negative mutant of mDia1, while the overexpression of the constitutively active mutant mDia1-ΔN3 increased bacteria uptake. Interestingly, when HeLa and RAW cells were infected, RhoA, Rac1 and mDia1 were recruited to membrane cell fractions. Our results suggest that the GTPases of the Rho family play an important role in C. burnetii phagocytosis in both HeLa and RAW cells. Additionally, we present evidence that ROCK and mDia1, which are downstream effectors of RhoA, are involved in that processFil: Salinas Ojeda, Romina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Ortiz Flores, Rodolfo Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Distel, Jesús Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Aguilera, Milton Osmar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Colombo, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Beron, Walter. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Cienicas Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentin

Chance and necessity in the genome evolution of endosymbiotic bacteria of insects

[EN] An open question in evolutionary biology is how does the selection¿drift balance determine the fates of biological interactions. We searched for signatures of selection and drift in genomes of five endosymbiotic bacterial groups known to evolve under strong genetic drift. Although most genes in endosymbiotic bacteria showed evidence of relaxed purifying selection, many genes in these bacteria exhibited stronger selective constraints than their orthologs in free-living bacterial relatives. Remarkably, most of these highly constrained genes had no role in the host¿symbiont interactions but were involved in either buffering the deleterious consequences of drift or other host-unrelated functions, suggesting that they have either acquired new roles or their role became more central in endosymbiotic bacteria. Experimental evolution of Escherichia coli under strong genetic drift revealed remarkable similarities in the mutational spectrum, genome reduction patterns and gene losses to endosymbiotic bacteria of insects. Interestingly, the transcriptome of the experimentally evolved lines showed a generalized deregulation of the genome that affected genes encoding proteins involved in mutational buffering, regulation and amino acid biosynthesis, patterns identical to those found in endosymbiotic bacteria. Our results indicate that drift has shaped endosymbiotic associations through a change in the functional landscape of bacterial genes and that the host had only a small role in such a shiftThis work was supported by Science Foundation Ireland (12/IP/1637) and grants from the Spanish Ministerio de Economia y Competitividad (MINECO-FEDER; BFU2012-36346 and BFU2015-66073-P) to MAF. DAP and CT were supported by Juan de la Cierva fellowships from MINECO (references: JCI-2011-11089 and JCA-2012-14056, respectively). DAP is supported by funds from the University of Nevada, Reno, NV, USA.Sabater-Muñoz, B.; Toft, C.; Alvarez-Ponce, D.; Fares Riaño, MA. (2017). Chance and necessity in the genome evolution of endosymbiotic bacteria of insects. The ISME Journal. 11(6):1291-1304. https://doi.org/10.1038/ismej.2017.18S12911304116Aguilar-Rodriguez J, Sabater-Munoz B, Montagud-Martinez R, Berlanga V, Alvarez-Ponce D, Wagner A et al. (2016). The molecular chaperone DnaK is a source of mutational robustness. Genome Biol Evol 8: 2979–2991.Alvarez-Ponce D, Sabater-Munoz B, Toft C, Ruiz-Gonzalez MX, Fares MA . (2016). Essentiality is a strong determinant of protein rates of evolution during mutation accumulation experiments in Escherichia coli. Genome Biol Evol 8: 2914–2927.Anders S, Huber W . (2010). Differential expression analysis for sequence count data. Genome Biol 11: R106.Archibald J . (2014) One Plus One Equals One: Symbiosis and the Evolution of Complex Life. Oxford University Press: Oxford, UK.Aussel L, Loiseau L, Hajj Chehade M, Pocachard B, Fontecave M, Pierrel F et al. (2014). ubiJ, a new gene required for aerobic growth and proliferation in macrophage, is involved in coenzyme Q biosynthesis in Escherichia coli and Salmonella enterica serovar Typhimurium. J Bacteriol 196: 70–79.Baumann P, Baumann L, Clark MA . (1996). Levels of Buchnera aphidicola chaperonin groEL during growth of the aphid Schizaphis graminum. Curr Microbiol 32: 7.Benjamini Y, Yekutieli Y . (2005). False discovery rate controlling confidence intervals for selected parameters. J Am Stat Assoc 100: 10.Bennett GM, Moran NA . (2015). Heritable symbiosis: the advantages and perils of an evolutionary rabbit hole. Proc Natl Acad Sci USA 112: 10169–10176.Bermingham J, Rabatel A, Calevro F, Vinuelas J, Febvay G, Charles H et al. (2009). Impact of host developmental age on the transcriptome of the symbiotic bacterium Buchnera aphidicola in the pea aphid (Acyrthosiphon pisum. Appl Environ Microbiol 75: 7294–7297.Bogumil D, Dagan T . (2010). Chaperonin-dependent accelerated substitution rates in prokaryotes. Genome Biol Evol 2: 602–608.Carbon S, Ireland A, Mungall CJ, Shu S, Marshall B, Lewis S et al. (2009). AmiGO: online access to ontology and annotation data. Bioinformatics 25: 288–289.Chen Z, Wang Y, Li Y, Li Y, Fu N, Ye J et al. (2012). Esre: a novel essential non-coding RNA in Escherichia coli. FEBS Lett 586: 1195–1200.Clark JW, Hossain S, Burnside CA, Kambhampati S . (2001). Coevolution between a cockroach and its bacterial endosymbiont: a biogeographical perspective. Proc Biol Sci 268: 393–398.Dale C, Wang B, Moran N, Ochman H . (2003). Loss of DNA recombinational repair enzymes in the initial stages of genome degeneration. Mol Biol Evol 20: 1188–1194.Deatherage DE, Barrick JE . (2014). Identification of mutations in laboratory-evolved microbes from next-generation sequencing data using breseq. Methods Mol Biol 1151: 165–188.Douglas AE . (2003). The nutritional physiology of aphids. Adv Insect Physiol 31: 68.Fares MA, Barrio E, Sabater-Munoz B, Moya A . (2002a). The evolution of the heat-shock protein GroEL from Buchnera, the primary endosymbiont of aphids, is governed by positive selection. Mol Biol Evol 19: 1162–1170.Fares MA, Ruiz-Gonzalez MX, Moya A, Elena SF, Barrio E . (2002b). Endosymbiotic bacteria: groEL buffers against deleterious mutations. Nature 417: 398.Gancedo C, Flores CL, Gancedo JM . (2016). The expanding landscape of moonlighting proteins in yeasts. Microbiol Mol Biol Rev 80: 765–777.Gerardo NM, Altincicek B, Anselme C, Atamian H, Barribeau SM, de Vos M et al. (2010). Immunity and other defenses in pea aphids, Acyrthosiphon pisum. Genome Biol 11: R21.Gomez-Valero L, Latorre A, Silva FJ . (2004). The evolutionary fate of nonfunctional DNA in the bacterial endosymbiont Buchnera aphidicola. Mol Biol Evol 21: 2172–2181.Gomez-Valero L, Silva FJ, Christophe Simon J, Latorre A . (2007). Genome reduction of the aphid endosymbiont Buchnera aphidicola in a recent evolutionary time scale. Gene 389: 87–95.Gonzalez-Domenech CM, Belda E, Patino-Navarrete R, Moya A, Pereto J, Latorre A . (2012). Metabolic stasis in an ancient symbiosis: genome-scale metabolic networks from two Blattabacterium cuenoti strains, primary endosymbionts of cockroaches. BMC Microbiol 12 (Suppl 1): S5.Hansen AK, Moran NA . (2011). Aphid genome expression reveals host-symbiont cooperation in the production of amino acids. Proc Natl Acad Sci USA 108: 2849–2854.Hansen AK, Moran NA . (2014). The impact of microbial symbionts on host plant utilization by herbivorous insects. Mol Ecol 23: 1473–1496.Henderson B, Fares MA, Lund PA . (2013). Chaperonin 60: a paradoxical, evolutionarily conserved protein family with multiple moonlighting functions. Biol Rev Camb Philos Soc 88: 955–987.Humphreys NJ, Douglas AE . (1997). Partitioning of symbiotic bacteria between generations of an insect: a quantitative study of a Buchnera sp. in the pea aphid (Acyrthosiphon pisum reared at different temperatures. Appl Environ Microbiol 63: 3294–3296.International Aphid Genomics Consortium. (2010). Genome sequence of the pea aphid Acyrthosiphon pisum. PLoS Biol 8: e1000313.Kadibalban AS, Bogumil D, Landan G, Dagan T . (2016). DnaK-dependent accelerated evolutionary rate in prokaryotes. Genome Biol Evol 8: 1590–1599.Katoh K, Standley DM . (2013). MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 30: 772–780.Kelkar YD, Ochman H . (2013). Genome reduction promotes increase in protein functional complexity in bacteria. Genetics 193: 303–307.Koga R, Meng XY, Tsuchida T, Fukatsu T . (2012). Cellular mechanism for selective vertical transmission of an obligate insect symbiont at the bacteriocyte-embryo interface. Proc Natl Acad Sci USA 109: E1230–E1237.Kuo CH, Moran NA, Ochman H . (2009). The consequences of genetic drift for bacterial genome complexity. Genome Res 19: 1450–1454.Kuo CH, Ochman H . (2009). Deletional bias across the three domains of life. Genome Biol Evol 1: 145–152.Law R, Lewis DH . (1983). Biotic environments and the maintenance of sex-some evidence from mutualistic symbioses. Biol J Linnean Soc 20: 28.Liu XD, Xie L, Wei Y, Zhou X, Jia B, Liu J et al. (2014). Abiotic stress resistance, a novel moonlighting function of ribosomal protein RPL44 in the halophilic fungus Aspergillus glaucus. Appl Environ Microbiol 80: 4294–4300.Lohse M, Bolger AM, Nagel A, Fernie AR, Lunn JE, Stitt M et al. (2012). RobiNA: a user-friendly, integrated software solution for RNA-Seq-based transcriptomics. Nucleic Acids Res 40: W622–W627.Macdonald SJ, Lin GG, Russell CW, Thomas GH, Douglas AE . (2012). The central role of the host cell in symbiotic nitrogen metabolism. Proc Biol Sci 279: 2965–2973.McClure R, Balasubramanian D, Sun Y, Bobrovskyy M, Sumby P, Genco CA et al. (2013). Computational analysis of bacterial RNA-Seq data. Nucleic Acids Res 41: e140.McCutcheon JP, Moran NA . (2012). Extreme genome reduction in symbiotic bacteria. Nat Rev Microbiol 10: 13–26.McFall-Ngai M, Hadfield MG, Bosch TC, Carey HV, Domazet-Loso T, Douglas AE et al. (2013). Animals in a bacterial world, a new imperative for the life sciences. Proc Natl Acad Sci USA 110: 3229–3236.Mira A, Ochman H, Moran NA . (2001). Deletional bias and the evolution of bacterial genomes. Trends Genet 17: 589–596.Moran NA . (1996). Accelerated evolution and Muller's rachet in endosymbiotic bacteria. Proc Natl Acad Sci USA 93: 2873–2878.Moran NA, Dunbar HE, Wilcox JL . (2005). Regulation of transcription in a reduced bacterial genome: nutrient-provisioning genes of the obligate symbiont Buchnera aphidicola. J Bacteriol 187: 4229–4237.Moran NA, McCutcheon JP, Nakabachi A . (2008). Genomics and evolution of heritable bacterial symbionts. Annu Rev Genet 42: 165–190.Moran NA, McLaughlin HJ, Sorek R . (2009). The dynamics and time scale of ongoing genomic erosion in symbiotic bacteria. Science 323: 379–382.Nakabachi A, Ishida K, Hongoh Y, Ohkuma M, Miyagishima SY . (2014). Aphid gene of bacterial origin encodes a protein transported to an obligate endosymbiont. Curr Biol 24: R640–R641.Nilsson AI, Koskiniemi S, Eriksson S, Kugelberg E, Hinton JC, Andersson DI . (2005). Bacterial genome size reduction by experimental evolution. Proc Natl Acad Sci USA 102: 12112–12116.Patino-Navarrete R, Moya A, Latorre A, Pereto J . (2013). Comparative genomics of Blattabacterium cuenoti: the frozen legacy of an ancient endosymbiont genome. Genome Biol Evol 5: 351–361.Pettersson ME, Berg OG . (2007). Muller's ratchet in symbiont populations. Genetica 130: 199–211.Price DR, Feng H, Baker JD, Bavan S, Luetje CW, Wilson AC . (2014). Aphid amino acid transporter regulates glutamine supply to intracellular bacterial symbionts. Proc Natl Acad Sci USA 111: 320–325.Reyes-Prieto M, Vargas-Chavez C, Latorre A, Moya A . (2015). SymbioGenomesDB: a database for the integration and access to knowledge on host-symbiont relationships. Database 2015: bav109 (1–8).Robinson MD, McCarthy DJ, Smyth GK . (2010). edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26: 139–140.Sabater-Muñoz B, Prats-Escriche M, Montagud-Martinez R, Lopez-Cerdan A, Toft C, Aguilar-Rodriguez J et al. (2015). Fitness trade-offs determine the role of the molecular chaperonin groel in buffering mutations. Mol Biol Evol 32: 2681–2693.Schlicker A, Domingues FS, Rahnenfuhrer J, Lengauer T . (2006). A new measure for functional similarity of gene products based on Gene Ontology. BMC Bioinformatics 7: 302.Shigenobu S, Watanabe H, Hattori M, Sakaki Y, Ishikawa H . (2000). Genome sequence of the endocellular bacterial symbiont of aphids Buchnera sp. APS. Nature 407: 81–86.Supek F, Bosnjak M, Skunca N, Smuc T . (2011). REVIGO summarizes and visualizes long lists of gene ontology terms. PLoS ONE 6: e21800.Tamas I, Klasson L, Canback B, Naslund AK, Eriksson AS, Wernegreen JJ et al. (2002). 50 million years of genomic stasis in endosymbiotic bacteria. Science 296: 2376–2379.Toft C, Fares MA . (2008). The evolution of the flagellar assembly pathway in endosymbiotic bacterial genomes. Mol Biol Evol 25: 2069–2076.van Ham RC, Kamerbeek J, Palacios C, Rausell C, Abascal F, Bastolla U et al. (2003). Reductive genome evolution in Buchnera aphidicola. Proc Natl Acad Sci USA 100: 581–586.Wernegreen JJ . (2002). Genome evolution in bacterial endosymbionts of insects. Nat Rev Genet 3: 850–861.Wernegreen JJ . (2011). Reduced selective constraint in endosymbionts: elevation in radical amino acid replacements occurs genome-wide. PLoS One 6: e28905.Williams TA, Fares MA . (2010). The effect of chaperonin buffering on protein evolution. Genome Biol Evol 2: 609–619.Yang Z . (2007). PAML 4: phylogenetic analysis by maximum likelihood. Mol Biol Evol 24: 1586–1591

Apoptosis of human melanoma cells induced by inhibition of B-RAFV600E involves preferential splicing of bimS

Bim is known to be critical in killing of melanoma cells by inhibition of the RAF/MEK/ERK pathway. However, the potential role of the most potent apoptosis-inducing isoform of Bim, BimS, remains largely unappreciated. Here, we show that inhibition of the mutant B-RAFV600E triggers preferential splicing to produce BimS, which is particularly important in induction of apoptosis in B-RAFV600E melanoma cells. Although the specific B-RAFV600E inhibitor PLX4720 upregulates all three major isoforms of Bim, BimEL, BimL, and BimS, at the protein and mRNA levels in B-RAFV600E melanoma cells, the increase in the ratios of BimS mRNA to BimEL and BimL mRNA indicates that it favours BimS splicing. Consistently, enforced expression of B-RAFV600E in wild-type B-RAF melanoma cells and melanocytes inhibits BimS expression. The splicing factor SRp55 appears necessary for the increase in BimS splicing, as SRp55 is upregulated, and its inhibition by small interfering RNA blocks induction of BimS and apoptosis induced by PLX4720. The PLX4720-induced, SRp55-mediated increase in BimS splicing is also mirrored in freshly isolated B-RAFV600E melanoma cells. These results identify a key mechanism for induction of apoptosis by PLX4720, and are instructive for sensitizing melanoma cells to B-RAFV600E inhibitors

Comparative study of the functional properties of three legume seed isolates: adzuki, pea and soy bean

The aim of this work was to compare functional properties including solubility, emulsifying and foaming properties of native and thermally treated adzuki, soy and pea protein isolates prepared under the same conditions. These functional properties were tested at four pH values: pH 3.0, pH 5.0, pH 7.0 and pH 8.0. The lowest solubility at all pH values were obtained for isolate of adzuki whereas isolates of soybean had the highest values at almost all pHs. Thermal treatment reduced solubility of soy and pea isolates at all pH values, whereas solubility of adzuki isolate was unchanged, except at pH 8. Native isolate of adzuki had the best emulsifying properties at pH 7.0 whereas at the other pH values some of native pea and soybean protein isolates were superior. After thermal treatment, depending on tested pH and selected variety all of three species could be a good emulsifier. Native soy protein isolates formed the most stable foams at all pHs. Thermal treatment significantly improved foaming properties of adzuki isolate, whereas reduced foaming capacity of soy and pea isolates, but could improve foam stability of these isolates at specific pH. Appropriate selection of legume seed as well as variety could have great importance in achievement of desirable functional properties of final products. All three tested species could find specific application in wide range of food products

Hsa-miR-196a2 Rs11614913 Polymorphism Contributes to Cancer Susceptibility: Evidence from 15 Case-Control Studies

BACKGROUND: MicroRNAs (miRNAs) are a family of endogenous, small and noncoding RNAs that negatively regulate gene expression by suppressing translation or degrading mRNAs. Recently, many studies investigated the association between hsa-miR-196a2 rs11614913 polymorphism and cancer risk, which showed inconclusive results. METHODOLOGY/PRINCIPAL FINDINGS: We conducted a meta-analysis of 15 studies that included 9,341 cancer cases and 10,569 case-free controls. We assessed the strength of the association, using odds ratios (ORs) with 95% confidence intervals (CIs). Overall, individuals with the TC/CC genotypes were associated with higher cancer risk than those with the TT genotype (OR=1.18, 95% CI=1.03-1.34, P<0.001 for heterogeneity test). In the stratified analyses, we observed that the CC genotype might modulate breast cancer risk (OR=1.11, 95%CI=1.01-1.23, Pheterogeneity=0.210) and lung cancer risk (OR=1.25, 95%CI=1.06-1.46, Pheterogeneity=0.958), comparing with the TC/TT genotype. Moreover, a significantly increased risk was found among Asian populations in a dominant model (TC/CC versus TT, OR=1.24, 95% CI=1.07-1.43, Pheterogeneity=0.006). CONCLUSIONS: These findings supported that hsa-miR-196a2 rs11614913 polymorphism may contribute to the susceptibility of cancers

Dendritic cell density and activation status in human breast cancer – CD1a, CMRF-44, CMRF-56 and CD-83 expression

Low CD1a-positive putative dendritic cell numbers in human breast cancer has recently been described and may explain the apparent ‘poor immunogenicity’ previously reported in breast cancer. Little attention has been given to dendritic cell activation within the tumour microenvironment, which is another reason why the in-situ immune response may be severely deficient. We have therefore examined CD1a expression as a marker for dendritic cells, together with CMRF-44 and -56 as markers of dendritic cell activation status, in 40 human breast cancers. The results demonstrate few or no CD1a-positive putative dendritic cells and minimal or no expression of the dendritic cell activation markers. Both dendritic cell number and dendritic cell activation appear substantially deficient in human breast cancers, regardless of tumour histological grade

A functional alternative splicing mutation in human tryptophan hydroxylase-2

The brain serotonergic system has an essential role in the physiological functions of the central nervous system and dysregulation of serotonin (5-HT) homeostasis has been implicated in many neuropsychiatric disorders. The tryptophan hydroxylase-2 (TPH2) gene is the rate-limiting enzyme in brain 5-HT synthesis, and thus is an ideal candidate gene for understanding the role of dysregulation of brain serotonergic homeostasis. Here, we characterized a common, but functional single-nucleotide polymorphism (SNP rs1386493) in the TPH2 gene, which decreases efficiency of normal RNA splicing, resulting in a truncated TPH2 protein (TPH2-TR) by alternative splicing. TPH2-TR, which lacks TPH2 enzyme activity, dominant-negatively affects full-length TPH2 function, causing reduced 5-HT production. The predicted mRNA for TPH2-TR is present in postmortem brain of rs1386493 carriers. The rs13864923 variant does not appear to be overrepresented in either global or multiplex depression cohorts. However, in combination with other gene variants linked to 5-HT homeostasis, this variant may exhibit important epistatic influences