Comparative Proteomics And Metallomics Studies In Arabidopsis Thaliana Leaf Tissues: Evaluation Of The Selenium Addition In Transgenic And Nontransgenic Plants Using Two-dimensional Difference Gel Electrophoresis And Laser Ablation Imaging

The main goal of this work is to evaluate some differential protein species in transgenic (T) and nontransgenic (NT) Arabidopsis thaliana plants after their cultivation in the presence or absence of sodium selenite. The transgenic line was obtained through insertion of CaMV 35S controlling nptII gene. Comparative proteomics through 2D-DIGE is carried out in four different groups (NT × T; NT × Se-NT (where Se is selenium); Se-NT × Se-T, and T × Se-T). Although no differential proteins are achieved in the T × Se-T group, for the others, 68 differential proteins (by applying a regulation factor ≥1.5) are achieved, and 27 of them accurately characterized by ESI-MS/MS. These proteins are classified into metabolism, energy, signal transduction, disease/defense categories, and some of them are involved in the glycolysis pathway-Photosystems I and II and ROS combat. Additionally, laser ablation imaging is used for evaluating the Se and sulfur distribution in leaves of different groups, corroborating some results obtained and related to proteins involved in the glycolysis pathway. From these results, it is possible to conclude that the genetic modification also confers to the plant resistance to oxidative stress. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.1407/08/15904912Gilbert, N., A hard look at GM crops (2013) Nature, 497, pp. 21-23Ye, X., Al-Babili, S., Klöti, A., Zhang, J., Engineering the provitamin A (β-carotene) biosynthetic pathway into carotenoid-free rice endosperm (2000) Science, 287, pp. 303-305Paine, J.A., Shipton, C.A., Chaggar, S., Howells, R., Improving the nutritional value of Golden Rice through increased pro-vitamin A content (2005) Nat. Biothechnol., 23, pp. 482-487Fujisawa, M., Takita, E., Harada, H., Sakurai, N., Pathway engineering of Brassica napus seeds using multiple key enzyme genes involved in ketocarotenoid formation (2009) J. Exp. Bot., 60, pp. 1319-1332Qi, B., Fraser, T., Mugford, S., Dobson, G., Production of very long chain polyunsaturated omega-3 and omega-6 fatty acids in plants (2004) Nat. Biotechnol., 22, pp. 739-745Karunanandaa, B., Qi, Q., Hao, M., Baszis, S.R., Metabolically engineered oilseed crops with enhanced seed tocopherol (2005) Meta Eng, 7, pp. 384-400Mataveli, L.V.R., Fioramonte, M., Gozzo, F.C., Arruda, M.A.Z., Improving metallomics information related to transgenic and non-transgenic soybean seeds using 2D-HPLC-ICP-MS and ESI-MS/MS (2012) Metallomics, 4, pp. 373-378Barbosa, H.S., Arruda, S.C.C., Azevedo, R.A., Arruda, M.A.Z., New insights on proteomics of transgenic soybean seeds: evaluation of differential expressions of enzymes and proteins (2012) Anal. Bioanal. Chem., 402, pp. 299-314Arruda, S.C.C., Barbosa, H.S., Azevedo, R.A., Arruda, M.A.Z., Comparative studies focusing on transgenic through cp4EPSPS gene and non-transgenic soybean plants: an analysis of protein species and enzymes (2013) J. Proteomics, 93, pp. 107-116Gonçalves, D.C., (2012), Universidade Estadual de Campinas, Unicamp. Campinas, BrazilUnseld, M., Marienfeld, J., Brandt, P., Brennicke, A., The mitochondrial genome of Arabidopsis thaliana contains 57 genes in 366,924 nucleotides (1997) Nat. Genet., 15, pp. 57-61Meinke, D., Cherry, J., Dean, C., Rounsley, S., Koornneef, M., Arabidopsis thaliana: a model plant for genome analysis (1998) Science, 28, pp. 662-682Initiative, T.A.G., Analysis of the genome sequence of the flowering plant Arabidopsis thaliana (2000) Nature, 408, pp. 796-815Filkowski, J., Besplug, J., Burke, P., Kovalchuk, I., Kovalchuck, O., Genotoxicity of 2,4-D and dicamba revealed by transgenic Arabidopsis thaliana plants harboring recombination and point mutation markers (2003) Mutat. Res., 542, pp. 23-32Lin, Y., Ludlow, E., Kalla, R., Pallaghy, C., Organ-specific, developmentally-regulated and abiotic stress-induced activities of four Arabidopsis thaliana promoters in transgenic while clover (Trifolium repens L.) (2003) Plant Sci, 165, pp. 1437-1444Zupan, J.R., Zambryski, P., Transfer of T-DNA from Agrobacterium to the plant cell (1995) Plant Physiol., 107, pp. 1041-1047Valvekens, D., Van Montagu, M., Van Lijsebettens, M., Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection (1988) Botany, 85, pp. 5536-5540Wilkinson, J.E., Twell, D., Lindsey, K., Activities of CaMV 35S and nos promoters in pollen: implications for field release of transgenic plants (1997) J. Exp. Bot., 48, pp. 265-275Silva, M.A.O., Arruda, M.A.Z., Laser ablation (imaging) for mapping and determining Se and S in sunflower leaves (2013) Metallomics, 5, pp. 62-67Bechtold, N., Ellis, J., Pelletier, G., In plant Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants (1993) C. R. Acad. Sci., 316, pp. 1194-1199Clough, S.J., Bent, A.F., Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana (1998) Plant J., 16, pp. 735-743Borges, J.C., Peroto, M.C., Ramos, C.H.I., Molecular chaperone genes in the SugarCane Expressed Sequence Database (SUCEST) (2001) Gen. Mol. Biol., 24, pp. 85-92Borges, J.C., Cagliari, T.C., Ramos, C.H.I., Expression and variability of molecular chaperones in the sugarcane expressome (2007) J. Plant Physiol., 164, pp. 505-513da Silva, V.C.H., Cagliari, T.C., Lima, T.B., Gozzo, F.C., Ramos, C.H.I., Conformational and functional studies of a cytosolic 90 kDa heat shock protein Hsp90 from sugarcane (2013) Plant Physiol. Biochem., 68, pp. 16-22Koncz, C., Shell, J., The promoter of TL-DNA gene 5 controls the tissue-specific expression of chimeric genes carried by a novel type of Agrobacterium binary vector (1986) Mol. Gen. Genet., 204, pp. 383-396Silva, M.A.O., Garcia, J.S., Souza, G.H.M.F., Eberlin, M.N., Evaluation of sample preparation protocols for proteomic analysis of sunflower leaves (2010) Talanta, 80, pp. 1545-1551Lin, S.-T., Chou, H.-C., Chang, S.-J., Chen, Y.-W., Proteomic analysis of proteins responsible for the development of doxorubicin resistance in human uterine cancer (2012) J. Proteomics, 75, pp. 5822-5847Taiz, L., Zeiger, E., (2006) Plant Physiology, pp. 148-287. , 4th ed., Artmed Ltd., MA, USAXiong, L., Schumaker, K.S., Zhu, J.K., Cell signaling during cold, drought and salt stress (2002) The Plant Cell, 14, pp. S165-S183Ide, Y., Nagasaki, N., Tomioka, R., Suito, M., Molecular properties of a novel, hydrophilic cation-binding protein associated with the plasma membrane (2007) J. Exp. Botany, 58, pp. 1173-1183Kato, M., Nagasaki-Takeuchi, N., Ide, Y., Tomioka, R., Maeshima, M., PCaPs, possible regulators of PtdInsP signals on plasma membrane (2010) Plant Sign. Beh., 5, pp. 848-850Li, J., Wang, X., Qin, T., Zhang, Y., MDP25, a novel calcium regulatory protein, mediates hypocotyl cell elongation by destabilizing cortical microtubules in Arabidopsis (2011) The Plant Cell, 23, pp. 4411-4427Wu, B., Chen, Y., Becker, J.S., Study of essential element accumulation in the leaves of a Cu-tolerant plant Elsholtzia splendens after Cu treatment by imaging laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) (2009) Anal. Chim. Acta, 633, pp. 165-172Sors, T.G., Ellis, D.R., Salt, D.E., Selenium uptake, translocation, assimilation and metabolic fate in plants (2005) Photosynth. Res., 86, pp. 373-38

Eosinophilia In Peripheral Blood Of Women With Recurrent Vaginal Candidiasis [eosinofilia No Sangue Periférico De Mulheres Com Candidiase Vaginal Recorrente]

PURPOSE: To quantify the number of defense cells and immunoglobulin E (IgE) levels in peripheral blood sampled from women with recurrent vulvovaginal candidiasis. METHODS: A cross-sectional study was conducted on 60 women, 40 with vulvovaginal candidiasis and 20 controls. The defense cells were identified using an impedance system combined with flow cytometry and total and specific IgE was measured by chemiluminescence. The Mann-Whitney test was used for nominal variables and the Spearman test was used to determine the correlation of IgE concentration and eosinophils in peripheral blood. RESULTS: The number of eosinophils in peripheral blood from patients with recurrent vulvovaginal candidiasis, 302.60 (±253.07), was significantly higher compared to control, 175.75 (±109.24) (p=0.037). Serum levels of total and specific IgE were similar in the groups of women with and without recurrent vulvovaginal candidiasis (p=0.361). However, there was a moderate positive correlation between eosinophils and total serum IgE in the candidiasis group (r=0.25). CONCLUSION: Women with recurrent vulvovaginal candidiasis are more likely to have eosinophils in peripheral blood.3510453457Kauffman, C.A., Pappas, P.G., Sobel, J.D., Dismukes, W.E., (2011) Essentials of clinical mycology, , 2 nd ed. New York: SpringerFoxman, B., Muraglia, R., Dietz, J.P., Sobel, J.D., Wagner, J., Prevalence of recurrent vulvovaginal candidiasis in 5 European countries and the United States: Results from an internet panel survey (2013) J Low Genit Tract Dis., 17 (3), pp. 340-345Ilkit, M., Guzel, A.B., The epidemiology, pathogenesis, and diagnosis of vulvovaginal candidosis: A mycological perspective (2011) Crit Rev Microbiol., 37 (3), pp. 250-261Watson, C., Calabretto, H., Comprehensive review of conventional and non-conventional methods of management of recurrent vulvovaginal candidiasis (2007) Aust N Z J Obstet Gynaecol., 47 (4), pp. 262-272Garcia, R.B., Bonifaz, A., Chassin, A.O., Kuba, E.B., Ariza, J., Cabello, R.R., Correlación clínico-micológica de la candidiasis vulvovaginal (2007) Ginecol Obstet Mex., 75 (2), pp. 68-72Hanna, S., Etzioni, A., New host defense mechanisms against Candida species clarify the basis of clinical phenotypes (2011) J Allergy Clin Immunol., 127 (6), pp. 1433-1437Naglik, J.R., Moyes, D.L., Wächtler, B., Hube, B., Candida albicans interactions with epithelial cells and mucosal immunity (2011) Microbes Infect., 13 (12-13), pp. 963-976Fidel Jr., P.L., History and update on host defense against vaginal candidiasis (2007) Am J Reprod Immunol., 57 (1), pp. 2-12Blanco, J.L., Garcia, M.E., Immune response to fungal infections (2008) Vet Immunol Immunopathol., 125 (1-2), pp. 47-70Paulovičová, L., Paulovičová, E., Karelin, A.A., Tsvetkov, Y.E., Nifantiev, N.E., Bystrický, S., Humoral and cell-mediated immunity following vaccination with synthetic Candida cell wall mannan derived heptamannoside-protein conjugate: Immunomodulatory properties of heptamannoside-BSA conjugate (2012) Int Immunopharmacol., 14 (2), pp. 179-187Kim, J., Han, B.J., Kim, H., Lee, J.Y., Joo, I., Omer, S., Kim, Y.S., Han, Y., Th1 immunity induction by ginsenoside Re involves in protection of mice against disseminated candidiasis due to Candida albicans (2012) Int Immunopharmacol., 14 (4), pp. 481-486Yano, J., Noverr, M.C., Fidel Jr., P.L., Cytokines in the host response to Candida vaginitis: Identifying a role for non-classical immune mediators, , S100 alarminsYano, J., Noverr, M.C., Fidel Jr., P.L., Cytokines in the host response to Candida vaginitis: Identifying a role for non-classical immune mediators, S100 alarmins (2012) Cytokine., 58 (1), pp. 118-128Ramírez-Santos, A., Pereiro Jr., M., Toribio, J., Recurrent vulvovaginitis: Diagnostic assessment and therapeutic management (2008) Actas Dermosifiliogr., 99 (3), pp. 190-198. , SpanishWitkin, S.S., Immunologic factors influencing susceptibility to recurrent candidal vaginitis (1991) Clin Obstet Gynecol., 34 (3), pp. 662-668Neves, N.A., Carvalho, L.P., De Oliveira, M.A., Giraldo, P.C., Bacellar, O., Cruz, A.A., Association between atopy and recurrent vaginal candidiasis (2005) Clin Exp Immunol., 142 (1), pp. 167-171Guo, R., Zheng, N., Lu, H., Yin, H., Yao, J., Chen, Y., Increased diversity of fungal flora in the vagina of patients with recurrent vaginal candidiasis and allergic rhinitis (2012) Microb Ecol., 64 (4), pp. 918-927Tsunemi, Y., Kadono, T., Saeki, H., Kikuchi, K., Tamaki, K., Sato, S., Secondary cutaneous candidiasis with eosinophilia (2010) J Dermatol., 37 (2), pp. 175-178Imanaka, A., Inui, S., Itoh, H., Masahito, T., Masahito, K., Shigetoshi, S., Generalized candidiasis with eosinophilic infiltration and atypical clinical appearance (2003) Rinsho Derma (Tokyo)., 45 (9), pp. 1005-1006Mori, A., Ikeda, Y., Taniguchi, M., Aoyama, C., Maeda, Y., Hasegawa, M., IL-5 production by peripheral blood Th cells of adult asthma patients in response to Candida albicans allergen (2001) Int Arch Allergy Immunol., 125 (SUPPL. 1), pp. 48-50Fan, S.R., Liao, Q.P., Liu, X.P., Liu, Z.H., Zhang, D., Vaginal allergic response in women with vulvovaginal candidiasis (2008) Int J Gynaecol Obstet., 101 (1), pp. 27-30Ong, P.Y., Ferdman, R.M., Church, J.A., Late-onset of IgE sensitization to microbial allergens in young children with atopic dermatitis (2010) Br J Dermatol., 162 (1), pp. 159-161Carvalho, R.J.V., Cunha, C.M., Silva, D.A.O., Sopelete, M.C., Urzedo, J.E., Moreira, T.A., IgA, IgE e subclasses de IgG anti-Candida albicans no soro e lavado vaginal de pacientes com candidíase vulvovaginal (2003) Rev Assoc Med Bras., 49 (4), pp. 434-43