Optimization Of Promoter Sequences In Order To Mimic Physiological Pde6b Expression

Purpose: Many retinal degenerations result from defective retina-specific gene expressions. Thus, it is important to understand how the expression of a photoreceptor-specific gene is regulated in vivo in order to achieve successful gene therapy. The present study aims to design an AAV2/8 vector that can regulate the transcript level in a physiological manner to replace missing PDE6b in Rd1 and Rd10 mice. In previous studies (Ogieta, et al., 2000), the short 5' flanking sequence of the human PDE6b gene (350 bp) was shown to be photoreceptor-specific in transgenic mice. However, the efficiency and specificity of the 5' flanking region of the human PDE6b was not investigated in the context of gene therapy during retinal degeneration. In this study, two different sequences of the 5' flanking region of the human PDE6b gene were studied as promoter elements and their expression will be tested in wild type and diseased retinas (Rd 10 mice).Methods: Two 5' flanking fragments of the human PDE6b gene: (-93 to +53 (150 bp) and -297 to +53 (350 bp)) were cloned in different plasmids in order to check their expression in vitro and in vivo by constructing an AAV2/8 vector. These elements drove the activity of either luciferase (pGL3 plasmids) or EGFP. jetPEI transfection in Y 79 cells was used to evaluate gene expression through luciferase activity. Constructs encoding EGFP under the control of the two promoters were performed in AAV2.1-93 (or 297)-EGFP plasmids to produce AAV2/8 vectors.Results: When pGL3-93 (150 bp) or pGL3-297 (350 bp) were transfected in the Y-79 cells, the smaller fragment (150 bp) showed higher gene expression compared to the 350 bp element and to the SV40 control, as previously reported. The 350 bp drove similar levels of expression when compared to the SV40 promoter. In view of these results, the fragments (150 bp or 350 bp) were integrated into the AAV2.1-EGFP plasmid to produce AAV2/8 vector, and we are currently evaluating the efficiency and specificity of the produced constructs in vivo in normal and diseased retinas.Conclusions: Comparisons of these vectors with vectors bearing ubiquitous promoters should reveal which construct is the most suitable to drive efficient and specific gene expression in diseased retinas in order to restore a normal function on the long term.

Bagavadam ou Doctrine divine ; ouvrage indien canonique, sur l'être suprême, les dieux, les géants, les hommes...

[Purāṇa. Bhāgavata-Purāṇa (français). 1788]

State Public Defender's Office Performance Report, FY 2013

This report pursuant to Department of Management procedures provides an overview of the State Public Defender’s fiscal year 2013 operations related to court-appointed counsel, claims processing, and other indigent defense matters. This information, in accordance with the Accountable Government Act, is meant to help improve decision making and increase accountability to stakeholders and the citizens of Iowa. Iowa’s indigent defense system remains strong. The more than 200 employees of the State Public Defender System have been efficiently providing high quality representation on more cases than ever before. In Fiscal Year 2013, public defender offices closed more than 91,000 cases, at an average cost per case of $282.02, and there was a final finding of ineffective assistance of counsel in less than .002% of these cases. Similarly, more than 1,000 contract attorneys continue to provide high quality representation throughout all 99 of Iowa’s counties. In Fiscal Year 2013, 60,929 claims were submitted to the indigent defense fund, at an average cost per claim of $507.86. This was an decrease of nearly 350 claims from Fiscal Year 2012, and the total amount spent from the indigent defense fund increased slightly.

Genetic Association of Recovery from Eating Disorders: The Role of GABA Receptor SNPs.

Follow-up studies of eating disorders (EDs) suggest outcomes ranging from recovery to chronic illness or death, but predictors of outcome have not been consistently identified. We tested 5151 single-nucleotide polymorphisms (SNPs) in approximately 350 candidate genes for association with recovery from ED in 1878 women. Initial analyses focused on a strictly defined discovery cohort of women who were over age 25 years, carried a lifetime diagnosis of an ED, and for whom data were available regarding the presence (n=361 ongoing symptoms in the past year, ie, 'ill') or absence (n=115 no symptoms in the past year, ie, 'recovered') of ED symptoms. An intronic SNP (rs17536211) in GABRG1 showed the strongest statistical evidence of association (p=4.63 × 10(-6), false discovery rate (FDR)=0.021, odds ratio (OR)=0.46). We replicated these findings in a more liberally defined cohort of women age 25 years or younger (n=464 ill, n=107 recovered; p=0.0336, OR=0.68; combined sample p=4.57 × 10(-6), FDR=0.0049, OR=0.55). Enrichment analyses revealed that GABA (γ-aminobutyric acid) SNPs were over-represented among SNPs associated at p<0.05 in both the discovery (Z=3.64, p=0.0003) and combined cohorts (Z=2.07, p=0.0388). In follow-up phenomic association analyses with a third independent cohort (n=154 ED cases, n=677 controls), rs17536211 was associated with trait anxiety (p=0.049), suggesting a possible mechanism through which this variant may influence ED outcome. These findings could provide new insights into the development of more effective interventions for the most treatment-resistant patients.

Field Evaluation of Class A Subbase Using Fly Ash, MLR-87-3, 1988

This project consisted of slipforming a 4-inch thick econocrete subbase on a 6-mile section of US 63. The project location extends south from one mile south of Denver, Iowa to Black Hawk County Road C-66 and consisted of the reconstruction and new construction of a divided four-lane facility. The econocrete was placed 27.3 feet wide in a single pass. Fly ash was used in this field study to replace 30, 45 and 60 percent of the portland cement in three portland cement econocrete base paving mixes. The three mixes contained 300, 350 and 400 pounds of cementitious material per cubic yard. Two Class "C" ashes from Iowa approved sources were used. The ash was substituted on the basis of one pound of ash for each pound of cement removed. The work was done October 6-29, 1987 and May 25-June 9, 1988. The twelve subbase mixes were placed in sections 2500 to 3000 feet in length on both the north and southbound roadways. Compressive strengths of all mixes were determined at 3 and 28 days of age. Flexural strengths of all mixes were determined at 7 and 14 days. In all cases strengths were adequate. The freeze/thaw durability of the econocrete mixes used was reduced by increased fly ash levels but remained above acceptable limits. The test results demonstrate the feasibility of producing econocrete with satisfactory properties even using fly ash at substitution rates up to 45 percent.

Aplicação de fungicidas via água de irrigação por aspersão para o controle da ferrugem-do-feijoeiro-comum

Este trabalho objetivou avaliar os fungicidas bitertanol, mancozeb, benomyl e iprodione nas doses de 175, 1.600, 350 e 750 g i.a./ha, respectivamente, aplicados via irrigação, no controle da ferrugem (Uromyces appendiculatus) do feijoeiro (Phaseolus vulgaris L.). Em parcelas de 12 x 12 m, foram realizadas quatro aplicações de cada fungicida, por meio de aplicador portátil de produtos químicos, a intervalos de seis a dez dias, lâmina de água de 3,4 mm, e seis minutos de aplicação. A produção de grãos (kg/ha) e a porcentagem de infecção foram de 1.550,0 e 6,6; 1.238,0 e 20,5; 1.358,0 e 26,9; 1.014,0 e 42,0; e 1.088,0 e 52,2 com bitertanol, mancozeb, benomyl, iprodione e testemunha, respectivamente. Os resultados mostraram a viabilidade da fungigação e a eficácia do bitertanol no controle da ferrugem-do- feijoeiro.

Valoración de pacientes tratados con hemodiálisis según la Ley de Promoción de la Autonomía Personal y Atención a las personas en situación de dependencia

La valoración del grado de dependencia de los pacientes en diálisis ha sido objeto de interés como indicador clínico y organizativo. El objetivo del estudio es conocer el grado de dependencia que presentan las personas sometidas a tratamiento con diálisis en Catalunya, según los criterios de la Ley sobre Promoción de la Autonomía Personal y Atención a las personas en situación de dependencia. Se ha realizado un estudio descriptivo transversal en 42 centros de Cataluña sobre el grado de dependencia de los pacientes en hemodiálisis. La recogida de datos se hizo mediante una encuesta basada en el baremo de la Ley más datos sociodemográfico y características del tratamiento. De los pacientes que componían la población renal de Cataluña fueron considerados por los profesionales sanitarios con algún grado de dependencia 806, de ellos 425 eran hombres y 381 mujeres; un 61% tienen edades superiores a 70 años. El 53% viven en pareja y el 80,1% tenían uno o más hijos. El 77,4% de los pacientes habían sido trabajadores no cualifi cados, y el 65,4% dijeron no tener estudios. Se consideraron no dependientes 137 pacientes, con dependencia moderada 350, con dependencia

Alterations of the 5'untranslated region of SLC16A12 lead to age-related cataract.

PURPOSE. Knowledge of genetic factors predisposing to age-related cataract is very limited. The aim of this study was to identify DNA sequences that either lead to or predispose for this disease. METHODS. The candidate gene SLC16A12, which encodes a solute carrier of the monocarboxylate transporter family, was sequenced in 484 patients with cataract (134 with juvenile cataract, 350 with age-related cataract) and 190 control subjects. Expression studies included luciferase reporter assay and RT-PCR experiments. RESULTS. One patient with age-related cataract showed a novel heterozygous mutation (c.-17A>G) in the 5'untranslated region (5'UTR). This mutation is in cis with the minor G-allele of the single nucleotide polymorphism (SNP) rs3740030 (c.-42T/G), also within the 5'UTR. Using a luciferase reporter assay system, a construct with the patient's haplotype caused a significant upregulation of luciferase activity. In comparison, the SNP G-allele alone promoted less activity, but that amount was still significantly higher than the amount of the common T-allele. Analysis of SLC16A12 transcripts in surrogate tissue demonstrated striking allele-specific differences causing 5'UTR heterogeneity with respect to sequence and quantity. These differences in gene expression were mirrored in an allele-specific predisposition to age-related cataract, as determined in a Swiss population (odds ratio approximately 2.2; confidence intervals, 1.23-4.3). CONCLUSIONS. The monocarboxylate transporter SLC16A12 may contribute to age-related cataract. Sequences within the 5'UTR modulate translational efficiency with pathogenic consequences.

Physiological responses of two tropical weeds to shade: I. Growth and biomass allocation

Ipomoea asarifolia (Desr.) Roem. & Schultz (Convolvulaceae) and Stachytarpheta cayennensis (Rich) Vahl. (Verbenaceae), two weeds found in pastures and crop areas in Brazilian Amazonia, were grown in controlled environment cabinets under high (800-1000 µmol m-² s-¹) and low (200-350 µmol m-² s-¹) light regimes during a 40-day period. For both species leaf dry mass and leaf area per total plant dry mass, and leaf area per leaf dry mass were higher for low-light plants, whereas root mass per total plant dry mass was higher for high-light plants. High-light S. cayennensis allocated significantly more biomass to reproductive tissue than low-light plants, suggesting a probably lower ability of this species to maintain itself under shaded conditions. Relative growth rate (RGR) in I. asarifolia was initially higher for high-light grown plants and after 20 days started decreasing, becoming similar to low-light plants at the last two harvests (at 30 and 40 days). In S. cayennensis, RGR was also higher for high-light plants; however, this trend was not significant at the first and last harvest dates (10 and 40 days). These results are discussed in relation to their ecological and weed management implications.

Physiological responses of two tropical weeds to shade: II. Leaf gas exchange and nitrogen content

Ipomoea asarifolia (Desr.) Roem. & Schultz (Convolvulaceae) and Stachytarpheta cayennensis (Rich) Vahl. (Verbenaceae), two weeds found in pastures and crop areas in the Brazilian Amazonia, Brazil, were grown in controlled environment cabinets under high (800-1000 µmol m-² s-¹) and low (200-350 µmol m-² s-¹) light regimes during a 40-day period. The objective was to determine the effect of shade on photosynthetic features and leaf nitrogen content of I. asarifolia and S. cayennensis. High-irradiance grown I. asarifolia leaves had significantly higher dark respiration and light saturated rates of photosynthesis than low-irradiance leaves. No significant differences for these traits, between treatments, were observed in S. cayennensis. Low-irradiance leaves of both species displayed higher CO2 assimilation rates under low irradiance. High-irradiance grown leaves of both species had less nitrogen per unit of weight. Low-irradiance S. cayennensis had more nitrogen per unit of leaf area than high-irradiance plants; however, I. asarifolia showed no consistent pattern for this variable through time. For S. cayennensis, leaf nitrogen content and CO2 assimilation were inversely correlated to the amount of biomass allocated to developing reproductive structures. These results are discussed in relation to their ecological and weed management implications.

The effects of arbuscular mycorrhizal fungi inoculation on Euterpe oleracea mart. (açaí) seedlings

With the objective of verifying the response of Euterpe oleracea seedlings to seven arbuscular mycorrhizal fungi species, an experimental trial was carried out under greenhouse conditions. Seeds of E. oleracea were sown in carbonized rice husk. Germinating seeds were initially transferred to plastic cups, containing fumigated Reddish Yellow Quartz Sand and inoculated with arbuscular mycorrhizal fungi. Two months later, seedlings were transferred to 2 kg black plastic bags, containing the same soil without fumigation. Plant growth and mineral nutrients were evaluated nine months after mycorrhizal inoculation. Differential effects were observed among the species tested, with Scutellispora gilmorei being the most effective ones in promoting growth and nutrient content of E. oleracea seedlings. The increment resulted from inoculation with S. gilmorei were 92% in total plant height, 116% in stem diameter, 361% in dry matter production, 191% in N, 664% in P, 46% in K, 562% in Ca, 363% in Mg and 350% in Zn contents, comparing to uninoculated controls. Infected root length was positively correlated to nutrient content and plant growth. It was concluded that growth and nutrient uptake of E. oleracea seedlings could be significantly improved by inoculation of effective arbuscular mycorrhizal fungi.

Eficiência de diferentes volumes e concentrações de calda para quebra de dormência na macieira cultivar 'Gala'

Este trabalho teve como objetivo avaliar a eficiência dos diferentes volumes de calda --340, 450, 1.000, 1.350 L/ha -- em duas concentrações de óleo mineral (OM) + cianamida hidrogenada (CH) -- 30+3 e 50+5 L/ha, respectivamente -- em macieira da cultivar Gala. Os resultados mostraram que nos anos com, pelo menos, 800 unidades de frio, todos os volumes e concentrações foram eficientes, exceto o de 340 L/ha, na concentração de 30+3 L/ha de OM+CH. Nos anos com menos de 800 unidades de frio, os melhores resultados foram com a concentração de 50+5 L/ha de OM+CH, em todos os volumes testados.

Avaliação de espaçamentos e populações de plantas de feijão visando à colheita mecanizada direta

O trabalho foi conduzido na safra 1992/93, no Centro Agropecuário Palma (CAP)/Universidade Federal de Pelotas (UFPel), objetivando avaliar o efeito da variação do espaçamento entre linhas e da população de plantas sobre características agronômicas relacionadas com a colheita mecanizada de feijoeiro (Phaseolus vulgaris L.). Foram testados três espaçamentos entre linhas (25, 50 e 75 cm) e quatro populações de plantas/ha (100, 200, 350 e 500 mil), utilizando como reagente a cultivar Pampa, de hábito de crescimento indeterminado, tipo II. O delineamento experimental foi em blocos casualizados, com quatro repetições. Somente ocorreu interação entre os fatores para o índice de acamamento, onde o aumento da população tendeu a reduzi-lo. A combinação do maior espaçamento com a menor população provocou o maior acamamento. A redução no espaçamento entre linhas, apesar de reduzir a altura de plantas, a altura de inserção de vagens e o rendimento de grãos, provocou aumento na altura da ponta da vagem mais baixa até o solo, e redução na porcentagem de plantas com vagens encostando no solo. O aumento na população de plantas, apesar de não afetar a maioria das características agronômicas das plantas, provocou redução na porcentagem de plantas com vagens encostando no solo, e não provocou modificações no rendimento de grãos.