In vivo bioimaging with tissue-specific transcription factor activated luciferase reporters.

The application of transcription factor activated luciferase reporter cassettes in vitro is widespread but potential for in vivo application has not yet been realized. Bioluminescence imaging enables non-invasive tracking of gene expression in transfected tissues of living rodents. However the mature immune response limits luciferase expression when delivered in adulthood. We present a novel approach of tissue-targeted delivery of transcription factor activated luciferase reporter lentiviruses to neonatal rodents as an alternative to the existing technology of generating germline transgenic light producing rodents. At this age, neonates acquire immune tolerance to the conditionally responsive luciferase reporter. This simple and transferrable procedure permits surrogate quantitation of transcription factor activity over the lifetime of the animal. We show principal efficacy by temporally quantifying NFκB activity in the brain, liver and lungs of somatotransgenic reporter mice subjected to lipopolysaccharide (LPS)-induced inflammation. This response is ablated in Tlr4(-/-) mice or when co-administered with the anti-inflammatory glucocorticoid analogue dexamethasone. Furthermore, we show the malleability of this technology by quantifying NFκB-mediated luciferase expression in outbred rats. Finally, we use somatotransgenic bioimaging to longitudinally quantify LPS- and ActivinA-induced upregulation of liver specific glucocorticoid receptor and Smad2/3 reporter constructs in somatotransgenic mice, respectively

Total blood lymphocyte counts in hemochromatosis probands with HFE C282Y homozygosity: relationship to severity of iron overload and HLA-A and -B alleles and haplotypes

BACKGROUND: It has been reported that some persons with hemochromatosis have low total blood lymphocyte counts, but the reason for this is unknown. METHODS: We measured total blood lymphocyte counts using an automated blood cell counter in 146 hemochromatosis probands (88 men, 58 women) with HFE C282Y homozygosity who were diagnosed in medical care. Univariate and multivariate analyses of total blood lymphocyte counts were evaluated using these variables: sex; age, transferrin saturation, and serum ferritin concentration at diagnosis; units of blood removed by phlebotomy to achieve iron depletion; and human leukocyte antigen (HLA)-A and -B alleles and haplotypes. RESULTS: The mean age at diagnosis was 49 ± 14 years (range 18 – 80 years) in men and 50 ± 13 years (range 22 – 88 years) in women. The correlations of total blood lymphocyte counts with sex, age, transferrin saturation, and serum ferritin concentration at diagnosis, and units of blood removed by phlebotomy to achieve iron depletion were not significant at the 0.05 level. Univariate analyses revealed significant associations between total blood lymphocyte counts and presence of the HLA-A*01, -B*08, and -B*14 alleles, and the A*01-B*08 haplotype. Presence of the A*01 allele, B*08 allele, or A*01-B*08 haplotype were associated with a lower total blood lymphocyte count, whereas presence of the B*14 allele was associated with a greater total blood lymphocyte count. There was an inverse association of total blood lymphocyte count with units of phlebotomy to achieve iron depletion, serum ferritin concentration, and with presence of the A*01-B*08 haplotype. CONCLUSION: We conclude that there is a significant inverse relationship of total blood lymphocyte counts and severity of iron overload in hemochromatosis probands with HFE C282Y homozygosity. The presence of the HLA-A*01 allele or the -B*08 allele was also associated with significantly lower total blood lymphocyte counts, whereas presence of the -B*14 allele was associated with significantly higher total blood lymphocyte counts. In univariate and multivariate analyses, total blood lymphocyte counts were significantly lower in probands with the HLA-A*01-B*08 haplotype than in probands without this haplotype

Identification of Intracellular and Plasma Membrane Calcium Channel Homologues in Pathogenic Parasites

Ca2+ channels regulate many crucial processes within cells and their abnormal activity can be damaging to cell survival, suggesting that they might represent attractive therapeutic targets in pathogenic organisms. Parasitic diseases such as malaria, leishmaniasis, trypanosomiasis and schistosomiasis are responsible for millions of deaths each year worldwide. The genomes of many pathogenic parasites have recently been sequenced, opening the way for rational design of targeted therapies. We analyzed genomes of pathogenic protozoan parasites as well as the genome of Schistosoma mansoni, and show the existence within them of genes encoding homologues of mammalian intracellular Ca2+ release channels: inositol 1,4,5-trisphosphate receptors (IP3Rs), ryanodine receptors (RyRs), two-pore Ca2+ channels (TPCs) and intracellular transient receptor potential (Trp) channels. The genomes of Trypanosoma, Leishmania and S. mansoni parasites encode IP3R/RyR and Trp channel homologues, and that of S. mansoni additionally encodes a TPC homologue. In contrast, apicomplexan parasites lack genes encoding IP3R/RyR homologues and possess only genes encoding TPC and Trp channel homologues (Toxoplasma gondii) or Trp channel homologues alone. The genomes of parasites also encode homologues of mammalian Ca2+ influx channels, including voltage-gated Ca2+ channels and plasma membrane Trp channels. The genome of S. mansoni also encodes Orai Ca2+ channel and STIM Ca2+ sensor homologues, suggesting that store-operated Ca2+ entry may occur in this parasite. Many anti-parasitic agents alter parasite Ca2+ homeostasis and some are known modulators of mammalian Ca2+ channels, suggesting that parasite Ca2+ channel homologues might be the targets of some current anti-parasitic drugs. Differences between human and parasite Ca2+ channels suggest that pathogen-specific targeting of these channels may be an attractive therapeutic prospect

Quality Of Life And Sleep In Obese Adolescents [qualidade De Vida E Do Sono De Adolescentes Obesos]

Obesity and sleep disorders in adolescence are strongly associated, and they impact both on the health and on quality of life (QL) in this age group. Objective: To comparatively assess QL and sleep in obese and eutrophic adolescents, who are older than ten years of age. Methods: Cross-sectional study including obese adolescents. This analysis was carried out between August 2009 and August 2010. The Control (eutrophic) Group comprised schoolchildren recruited from State schools in the city of Americana, in São Paulo State, Brazil. Instruments applied included a standardized questionnaire, the Sleep Behavior Questionnaire (SBQ), and the Pediatric QL Inventory (PedsQL™ 4.0). Results: The obese adolescents had poorer QL on physical (p<0.001), emotional (p=0.03), and social (p=0.002) functioning domains. A difference in mean psychosocial functioning was found between the groups (p=0.009) with obese subjects having a mean score of 69.5 (±16.0) and in eutrophic individuals of 76.2 (±16.7). A greater number of sleep problems was evident in the Obese Group (p=0.03). Conclusions: The obese adolescents had an impaired QL and a higher frequency of sleep problems compared to the eutrophic subjects.7127882(2008) Obesidade na infância e adolescência: Manual de orientação, p. 116. , Sociedade Brasileira de Pediatria. São Paulo: Sociedade Brasileira de Pediatria, Departamento de NutrologiaLuiz, A.M.G., Gorayeb, R., Liberatore Jr., R.D.R., Avaliação de depressão, problemas de comportamento e competência social em crianças obesas (2010) Est Psicol, 27, pp. 41-48Cataneo, C., Carvalho, A.M.P., Galindo, E.M.C., Obesidade e Aspectos Psicológicos: Maturidade emocional, auto-conceito, locus de controle e ansiedade (2005) Psicol Reflex Crit, 18, pp. 39-46Mesquita, G., Reimao, R., Nightly use of computer by adolescents: Its effect on quality of sleep (2007) Arq Neuropsiquiatr, 65, pp. 428-432Moreira, P., Santos, S., Padrão, P., Food patterns according to sociodemographics, physical activity, sleeping and obesity in Portuguese children (2010) Int J Environ Res Public Health, 7, pp. 1121-1138Rocha, C.R., Rossini, S., Reimão, R., Sleep disorders in high school and pre-university students (2010) Arq Neuropsiquiatr, 68, pp. 903-907Crispim, C.A., Zalcman, I., Dáttilo, M., Padilha, H.G., Tufik, S., Mello, M.T., Relação entre sono e obesidade: Uma revisão de literatura (2007) Arq Bras Endocrinol Metab, 51, pp. 1041-1049Chang, S.J., Chae, K.Y., Obstructive Sleep Apnea Syndrome in Children: Epidemiology, pathophisiology, diagnosis and sequelae (2010) Korean J Pediatr, 53, pp. 863-871Stein, M.A., Mendelsohn, J., Obermeyer, W.H., Amromin, J., Benca, R., Sleep and behavior problems in school-aged children (2001) Pediatrics, 107, p. 60Ravens-Sieberer, U., Redegeld, M., Bullinger, M., Quality of life after in-patient rehabilitation in children with obesity (2001) Int J Obes Relat Metab Disord, 25 (SUPPL. 1), pp. S63-S65(2007) Child growth standards: Methods and development. Head circumference-for-age, arm circumference-for-age, triceps skinfold-for-age and subscapular skinfold-for-age, p. 217. , World Health Organization. Geneva: World Health OrganizationCritério de Classificação Econômica Brasil (CCEB), , http://www.abep.org/novo/Content.aspx?ContentID=302, Associação Brasileira de Empresas de Pesquisa (ABEP). [citado 2 abr 2010]. Disponível emCortesi, F., Giannotti, F., Ottaviano, S., Sleep problems and daytime behavior in childhood idiopathic epilepsy (1999) Epilepsia, 40, pp. 1557-1565Batista, B.H.B., Nunes, M.L., Language validation of two scales to evaluate sleep quality in children (2006) J Epilepsy Clin Neurophysiol, 12, pp. 143-148Varni, J.W., Seid, M., Rode, C.A., The PedsQL: Measurement model for the pediatric quality of life inventory (1999) Med Care, 37, pp. 126-139Klatchoian, D.A., Len, C.A., Terreri, M.T., Quality of life of children and adolescents from São Paulo: Reliability and validity of the Brazilian version of the Pediatric Quality of Life InventoryTM version 4.0 Generic Core Scales (2008) J Pediatr, 84, pp. 308-315(2009) Avaliação nutricional da criança e do adolescente-Manual de Orientação/Sociedade Brasileira de Pediatria, p. 112. , Sociedade Brasileira de Pediatria. Departamento de Nutrologia. São Paulo: Sociedade Brasileira de PediatriaLytle, L.A., Pasch, K.E., Farbakhsh, K., The relationship between sleep and weight in a sample of adolescents (2011) Obesity, 19, pp. 324-331Carter III, R., Watenpaugh, D.E., Obesity and obstructive sleep apnea: Or is it OSA and obesity? (2008) Pathophysiology, 15, pp. 71-77Verhulst, S.L., Rooman, R., van Gaal, L., de Backer, W., Desager, K., Is sleep-disordered breathing an additional risk factor for the metabolic in obese children and adolescents? (2009) Int J Obes, 33, pp. 8-13Nguyen-Rodriguez, S.T., McClain, A.D., Spruijt-Metz, D., Anxiety mediates the relationship between sleep onset latency and emotional eating in minority children (2010) Eat Behav, 11, pp. 297-300Crabtree, V.M., Varni, J.W., Gozal, D., Health-related quality of life and depressive symptoms in children with suspected sleep-disordered breathing (2004) Sleep, 27, pp. 1131-1138Wang, Y., Monteiro, C., Popkin, B.M., Trends of obesity and underweight in older children and adolescents in the United States, Brazil, China and Russia (2002) Am J Clin Nutr, 75, pp. 971-977Schwimmer, J.B., Burwinkle, T.M., Varni, J.W., Health-related quality of life of severely obese children and adolescents (2003) JAMA, 289, pp. 1813-1819Johannsen, D.L., Johannsen, N.M., Specker, B.L., Influence of parents' eating behaviors and child feeding practices on children's weight status (2006) Obesity, 14, pp. 431-439Barros Filho, A.A., A visibilidade da criança ao longo da história (2010) Cienc Saúde Coletiva, 15, pp. 332-33

Antiprotozoan activity of Brazilian marine cnidarian extracts and of a modified steroid from the octocoral Carijoa riisei

In the present investigation, we have evaluated the antileishmanial and antitrypanosomal activity of methanolic crude extracts obtained from eight species of cnidarians and of a modified steroid isolated from the octocoral Carijoa riisei. The antileishmanial activity of cnidarians crude extracts showed 50% inhibitory concentration ( IC50) values in the concentration range between 2.8 and 93.3 mu g/mL. Trypomastigotes of Trypanosoma cruzi were less susceptible to the crude extracts, with IC50 values in the concentration range between 40.9 and 117.9 mu g/mL. The steroid (18-acetoxipregna-1,4,20-trien-3-one) displayed a strong antileishmanial activity, with an IC50 value of 5.5 mu g/mL against promastigotes and 16.88 mu g/mL against intracellular amastigotes. The steroid also displayed mammalian cytotoxicity (IC50 of 10.6 mu g/mL), but no hemolytic activity was observed at the highest concentration of 12.5 mu g/mL. The antileishmanial effect of the steroid in macrophages suggested other mechanism than macrophage activation, as no upregulation of nitric oxide was observed. The antitrypanosomal activity of the steroid resulted in an IC50 value of 50.5 mu g/mL. These results indicate the potential of cnidarian natural compounds as antileishmanial drug candidates.Instituto Adolfo LutzInstituto Adolfo LutzFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[06/05241-2]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[05/00974-9]Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[05/60175-2]CNPq[300194/1994-3]Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq