Infantile Encephaloneuromyopathy and Defective Mitochondrial Translation Are Due to a Homozygous RMND1 Mutation

Defects of mitochondrial protein synthesis are clinically and genetically heterogeneous. We previously described a male infant who was born to consanguineous parents and who presented with severe congenital encephalopathy, peripheral neuropathy, myopathy, and lactic acidosis associated with deficiencies of multiple mitochondrial respiratory-chain enzymes and defective mitochondrial translation. In this work, we have characterized four additional affected family members, performed homozygosity mapping, and identified a homozygous splicing mutation in the splice donor site of exon 2 (c.504+1G>A) of RMND1 (required for meiotic nuclear division-1) in the affected individuals. Fibroblasts from affected individuals expressed two aberrant transcripts and had decreased wild-type mRNA and deficiencies of mitochondrial respiratory-chain enzymes. The RMND1 mutation caused haploinsufficiency that was rescued by overexpression of the wild-type transcript in mutant fibroblasts; this overexpression increased the levels and activities of mitochondrial respiratory-chain proteins. Knockdown of RMND1 via shRNA recapitulated the biochemical defect of the mutant fibroblasts, further supporting a loss-of-function pathomechanism in this disease. RMND1 belongs to the sif2 family, an evolutionary conserved group of proteins that share the DUF155 domain, have unknown function, and have never been associated with human disease. We documented that the protein localizes to mitochondria in mammalian and yeast cells. Further studies are necessary for understanding the function of this protein in mitochondrial protein translation.

Invariant Natural Killer T Cell Agonist Modulates Experimental Focal and Segmental Glomerulosclerosis

A growing body of evidence demonstrates a correlation between Th2 cytokines and the development of focal and segmental glomerulosclerosis ( FSGS). Therefore, we hypothesized that GSL-1, a monoglycosylceramide from Sphingomonas ssp. with pro-Th1 activity on invariant Natural Killer T ( iNKT) lymphocytes, could counterbalance the Th2 profile and modulate glomerulosclerosis. Using an adriamycin( ADM)-based model of FSGS, we found that BALB/c mice presented albuminuria and glomerular degeneration in association with a Th2-like pro-fibrogenic profile; these mice also expressed a combination of inflammatory cytokines, such as IL-4, IL-1 alpha, IL-1 beta, IL-17, TNF-alpha, and chemokines, such as RANTES and eotaxin. In addition, we observed a decrease in the mRNA levels of GD3 synthase, the enzyme responsible for GD3 metabolism, a glycolipid associated with podocyte physiology. GSL-1 treatment inhibited ADM-induced renal dysfunction and preserved kidney architecture, a phenomenon associated with the induction of a Th1-like response, increased levels of GD3 synthase transcripts and inhibition of pro-fibrotic transcripts and inflammatory cytokines. TGF-beta analysis revealed increased levels of circulating protein and tissue transcripts in both ADM- and GSL-1-treated mice, suggesting that TGF-beta could be associated with both FSGS pathology and iNKT-mediated immunosuppression; therefore, we analyzed the kidney expression of phosphorylated SMAD2/3 and SMAD7 proteins, molecules associated with the deleterious and protective effects of TGF-beta, respectively. We found high levels of phosphoSMAD2/3 in ADM mice in contrast to the GSL-1 treated group in which SMAD7 expression increased. These data suggest that GSL-1 treatment modulates the downstream signaling of TGF-beta through a renoprotective pathway. Finally, GSL-1 treatment at day 4, a period when proteinuria was already established, was still able to improve renal function, preserve renal structure and inhibit fibrogenic transcripts. In conclusion, our work demonstrates that the iNKT agonist GSL-1 modulates the pathogenesis of ADM-induced glomerulosclerosis and may provide an alternative approach to disease management.

Evaluation of the dental arch asymmetry in natural normal occlusion and Class II malocclusion individuals

OBJECTIVE: To verify the presence and degree of asymmetry of dental arches in Brazilian individuals with natural normal occlusion and Class II, Divisions 1 and 2 malocclusions. METHODS: The study evaluated the symmetry of the maxillary and mandibular dental arches of 180 pairs of dental casts, divided into: Group I = 60 pairs of natural normal occlusion individuals; Group II = 60 pairs of Class II, Division 1 malocclusion individuals; and Group III = 60 pairs of Class II, Division 2 malocclusion individuals. A device was used to measure dental midline deviation and the canine tip in the dental arches (in degrees). It was also verified the distance of the upper canines from the palatal suture, intercanine distance, and anteroposterior upper and lower first molar position. RESULTS: Dental arches of individuals from all groups presented asymmetry, regardless of the presence of malocclusion. Group I showed a lower asymmetry degree in relation to Groups II and III. The asymmetry in Groups II and III was similar. CONCLUSION: The dental arches of individuals with natural normal occlusion and with Class II, Division 1 and Division 2 malocclusions showed asymmetry. The asymmetry degree was higher in the mandibular dental arches than in the maxillary dental arches in all 3 evaluated groups.

Index of Complexity, Outcome and Need scored on plaster and digital models

The aim of this study was to compare standard plaster models with their digital counterparts for the applicability of the Index of Complexity, Outcome, and Need (ICON). Generated study models of 30 randomly selected patients: 30 pre- (T(0)) and 30 post- (T(1)) treatment. Two examiners, calibrated in the ICON, scored the digital and plaster models. The overall ICON scores were evaluated for reliability and reproducibility using kappa statistics and reliability coefficients. The values for reliability of the total and weighted ICON scores were generally high for the T(0) sample (range 0.83-0.95) but less high for the T(1) sample (range 0.55-0.85). Differences in total ICON score between plaster and digital models resulted in mostly statistically insignificant values (P values ranging from 0.07 to 0.19), except for observer 1 in the T(1) sample. No statistically different values were found for the total ICON score on either plaster or digital models. ICON scores performed on computer-based models appear to be as accurate and reliable as ICON scores on plaster models.

ROLES FOR BRAF KINASE ACTIVATING MUTATIONS IN MELANOMA: MICROENVIRONMENTAL IMMUNOSUPPRESSION

The BRAF oncogene demonstrates a characteristic mutation (V600E) in a significant fraction of cutaneous melanomas, leading to constitutive activation of the MAP kinase pathway. This genetic lesion endows tumor cells with proliferative and survival advantages, and metastatic melanoma patients treated with the BRAF(V600E)-specific inhibitor, Vemurafenib, have shown dramatic clinical responses. Here, I show that BRAF(V600E) induces transcription of the IL-1α and IL-1β genes in both melanocytes and melanoma cell lines and that this upregulation is specifically abrogated by targeted BRAF(V600E) inhibitors. Furthermore, treatment of melanoma tumor-associated fibroblasts (TAFs) with IL-1α/β significantly enhanced the ability of TAFs to suppress the proliferation and function of melanoma antigen-specific cytotoxic T cells. IL-1α/β treatment of TAFs upregulated multiple immunosuppressive factors, including COX-2 and the PD-1 ligands PD-L1 and PD-L2. Specific BRAF(V600E) inhibitors largely abrogated the ability of melanoma cells to confer T cell-suppressive properties on TAFs. These results support a model in which BRAF(V600E) promotes immune suppression in the melanoma tumor environment through an IL-1-mediated mechanism involving resident stromal fibroblasts. Based on these findings, combination therapies involving targeted BRAF inhibition and T cell-based immunotherapies are warranted.

The role of Ski/Sno oncoproteins as negative regulators of the TGF-beta/SMAD signaling pathway in B -cell non -Hodgkin's lymphoma (NHL-B)

Non-Hodgkin's lymphomas are common tumors of the human immune system, primarily of B cell lineage (NHL-B). Negative growth regulation in the B cell lineage is mediated primarily through the TGF-β/SMAD signaling pathway that regulates a variety of tumor suppressor genes. Ski was originally identified as a transforming oncoprotein, whereas SnoN is an isoform of the Sno protein that shares a large region of homology with Ski. In this study, we show that Ski/SnoN are endogenously over-expressed both in patients' lymphoma cells and NHL-B cell lines. Exogenous TGF-β1 treatment induces down-regulation of Ski and SnoN oncoprotein expression in an NHL-B cell line, implying that Ski and SnoN modulate the TGF-β signaling pathway and are involved in cell growth regulation. Furthermore, we have developed an NHL-B cell line (DB) that has a null mutation in TGF-β receptor type II. In this mutant cell line, Ski/SnoN proteins are not down-regulated in response to TGF-β1 treatment, suggesting that downregulation of Ski and SnoN proteins in NHL-B require an intact functional TGF-β signaling pathway Resting normal B cells do not express Ski until activated by antigens and exogenous cytokines, whereas a low level of SnoN is also present in peripheral blood Go B cells. In contrast, autonomously growing NHL-B cells over-express Ski and SnoN, implying that Ski and SnoN are important cell cycle regulators. To further investigate a possible link between reduction of the Ski protein level and growth inhibition, Ski antisense oligodeoxynucleotides were transfected into NHL-B cells. The Ski protein level was found to decrease to less than 40%, resulting in restoring the effect of TGF-β and leading to cell growth inhibition and G1 cell cycle arrest. Co-immunoprecipitation experiments demonstrated that Ski associates with Smad4 in the nucleus, strongly suggesting that over-expression of the nuclear protein Ski and/or SnoN negatively regulates the TGF-β pathway, possibly by modulating Smad-mediated tumor suppressor gene expression. Together, in NHL-B, the TGF-β/SMAD growth inhibitory pathway is usually intact, but over-expression of the Ski and/or SnoN, which binds to Smad4, abrogates the negative regulatory effects of TGF-β/SMAD in lymphoma cell growth and potentiates the growth potential of neoplastic B cells. ^

Modeled integrated responses of ppERK1 and ppERK2 of normal and elevated ERK1 and ERK2 levels (Figure 5b)

The present dataset contain source data for Figure 5b from Schilling et al., 2009. Cell fate decisions are regulated by the coordinated activation of signalling pathways such as the extracellular signal-regulated kinase (ERK) cascade, but contributions of individual kinase isoforms are mostly unknown. The authors combined quantitative data from erythropoietin-induced pathway activation in primary erythroid progenitor (colony-forming unit erythroid stage, CFU-E) cells with mathematical modelling, in order to predict and experimentally confirmed a distributive ERK phosphorylation mechanism in CFU-E cells. The authors found evidences that double-phosphorylated ERK1 attenuates proliferation beyond a certain activation level, whereas activated ERK2 enhances proliferation with saturation kinetics. They show integrated responses of double-phosphorylated ERK1 and ERK2 that were calculated for different Epo concentrations for the original model as well as for models with elevated ERK1 or ERK2 levels.

Proliferation of retrovirally transduced CFU-E cells after incubation with increasing Epo-concentration (Figure 5c)

Data contain source data for Figure 5c from Schilling et al., 2009. Cell fate decisions are regulated by the coordinated activation of signalling pathways such as the extracellular signal-regulated kinase (ERK) cascade, but contributions of individual kinase isoforms are mostly unknown. The authors combined quantitative data from erythropoietin-induced pathway activation in primary erythroid progenitor (colony-forming unit erythroid stage, CFU-E) cells with mathematical modelling, in order to predict and experimentally confirmed a distributive ERK phosphorylation mechanism in CFU-E cells. The authors found evidences that double-phosphorylated ERK1 attenuates proliferation beyond a certain activation level, whereas activated ERK2 enhances proliferation with saturation kinetics. Retrovirally transduced CFU-E cells were incubated with increasing Epo concentrations for 14 h and proliferation was measured by [3H]-thymidine incorporation.

In Situ effects of low pH and elevated HCO3 on juvenile massive Porites spp. in Moorea, French Polynesia

Juvenile colonies of massive Porites spp. were exposed to manipulated pH and bicarbonate ([HCO3-]) in situ to test the hypothesis that ocean acidification (OA) does not affect respiration and calcification. Incubations lasted 28 h and exposed corals to ambient temperature and light with ecologically relevant water motion. Three treatments were applied: (1) ambient conditions of pH 8.04 and 1751 µmol HCO3- kg(-1) (Treatment 1), (2) pCO2-induced ocean acidification of pH 7.73 and 2011 µmol HCO3- kg(-1) (Treatment 2), and (3) pCO2 and HCO3--enriched seawater of pH 7.69 and 2730 µmol HCO3- kg(-1) (Treatment 3). The third treatment providing elevated [HCO3-] was used to test for stimulatory effects of dissolved inorganic carbon on calcification under low pH and low saturation of aragonite (Omega arag), but it does not reflect conditions expected to occur under CO2-driven OA. Calcification of juvenile massive Porites spp. was affected by treatments, with an 81% elevation in Treatment 3 versus Treatment 1, but no difference between Treatments 1 and 2; respiration and the metabolic expenditure concurrent with calcification remained unaffected. These findings indicate that juvenile massive Porites spp. are resistant to short exposures to OA in situ, and separately, that they can increase calcification at low pH and low Omega arag if [HCO3-] is elevated. Juvenile Porites spp. may therefore be limited by dissolved inorganic carbon under ambient pCO2 conditions

Efecto de la fertilización orgánica y mineral en las propiedades del suelo, la emisión de los principales gases de efecto invernadero y en las diferentes fases fenológicas del cultivo de café (Coffea arabica L.)

El cultivo de café es de gran importancia a nivel mundial (ICO, 2011), y en el Ecuador ha sido uno de los cultivos más importantes en la generación de divisas (COFENAC, 2011). Sin embargo en los sistemas productivos de este país se puede apreciar el uso inapropiado de fertilizantes, lo que conlleva a una pérdida de nutrientes, por lo que es importante estudiar las dosis adecuadas para la fertilización tanto mineral como orgánica. El objetivo del trabajo fue evaluar el efecto de la fertilización mineral y orgánica en diferentes dosis en un monocultivo de café en la provincia de Loja, sobre las propiedades del suelo, la emisión de los principales gases que provocan el efecto invernadero y la fenología y productividad del cultivo. En la provincia de Loja (Ecuador) se seleccionó un área de 2.520 m2 en la que se establecieron 21 parcelas de café arábigo (Coffea arabica L.) var. caturra y se aplicó tres tratamientos con tres repeticiones de fertilización mineral y tres orgánicos con dosis: bajas minerales (MIN 1= 157 Kg NPK ha-1 año-1 para el primer año y 425 Kg NPK ha-1 año-1 para el segundo año), medias minerales (MIN 2= 325 Kg NPK ha-1 año-1 para el primer año y 650 Kg NPK ha-1 año-1 en el segundo año) y altas minerales (MIN 3= 487 y 875 Kg NPK ha-1 año-1 para el primer y segundo año respectivamente), bajas orgánicas (ORG 1= 147 Kg NPK ha-1 año-1 en el primer año y 388 Kg NPK ha-1 año-1 en el año dos), medias orgánicas (ORG 2= 265 Kg NPK ha-1 año-1 para el primer año y 541 Kg NPK ha-1 año-1 en el segundo año), altas orgánicas (ORG 3= 368 Kg NPK ha-1 año-1 para el primer año y 727 Kg NPK ha-1 año-1 en el segundo año) y fertilización cero (TES = sin fertilización). Se usó urea, roca fosfórica y muriato de potasio en la fertilización mineral y humus (Bioabor) en la orgánica, más un tratamiento testigo, cada tratamiento tuvo tres repeticiones. El tiempo de evaluación de los fertilizantes aplicados fue de dos años consecutivos, la fertilización se la realizó dos veces por año y en base a análisis del suelo y demandas nutricionales del cultivo. para determinar las características del suelo se realizó muestreos de suelos en cada parcela a una profundidad de 20 cm de estas muestras los parámetro iniciales determinados fueron: color (Munsell), textura (método del hidrómetro), pH (relación 1:2,5 suelo-agua), Materia orgánica (Walkey y Black), Nitrógeno (Micro Kjendahl), Fósforo (Bray y Kurtz), Potasio (Olsen), estos procesos se repitieron cada seis meses para poder evaluar los cambios de que se producen debido a la fertilización mineral y orgánica en el cultivo. Las emisiones de gases efecto invernadero desde el suelo al ambiente se determinaron por el método de cámara cerrada (Rondón, 2000) y la concentración por cromatografía de gases. Las mediciones fisiológicas (altura de planta, ancho de copa, grosor de tallo y producción) se las evaluó cada dos meses, a excepción de la producción que fue anual al término de cada cosecha. Además se realizó el análisis económico de la productividad del cultivo. El análisis estadístico de datos se lo realizó con el programa SPSS v. 17.0. Las medias fueron comprobadas mediante ANOVAS de un factor con test de Tukey (P < 0,05). El beneficio económico se estimó en términos de ingresos y gastos totales que se presentaron en el ensayo. Los resultados obtenidos al término del ensayo indican que los tratamientos MIN 2 y MIN 3 produjeron cambios más significativos en comparación con los otros tratamientos establecidos en la mejora de fertilidad del suelo, el pH ha sido menos afectado en la acidificación en comparación con los tratamientos orgánicos que se han acidificado mayormente; la materia orgánica (MO) tuvo incrementos considerablemente bueno en estos dos tratamientos, sin embargo fueron superados por los tratamientos de fertilización orgánica; el nitrógeno total (Nt )y el potasio (K) también presentaron mejores valores al termino del ensayo y el fósforo (P) mostro incrementos buenos aunque un poco menores que los de los tratamientos ORG 2 y ORG 3. En lo que respecta a las emisiones de gases efecto invernadero, los flujos acumulados de óxido nitroso (N2O) en los dos años han aumentado en todos los tratamientos en comparación con el tratamiento Testigo, pero de manera considerable y con mayores flujos en el tratamiento MIN 3 y MIN 2 que se podrían considerarse los de mayor contaminación por N2O al ambiente lo que se le atribuye a las dosis de fertilización mineral aplicadas en el periodo de investigación, los tratamiento MIN 1 y todos los tratamientos orgánicos muestran menores emisiones al ambiente. Las emisiones de metano (CH4) no muestran mayores diferencias de emisiones entre tratamientos, siendo los mayores emisores los tratamientos ORG 3 y ORG 2 posiblemente debido al abono orgánico y añadido al suelo; para las emisiones de dióxido de carbono (CO2) de manera similar al CH4 el tratamiento ORG 3 fue el que presento mayores emisiones, los flujos de CO2 al ambiente de los otros tratamientos fueron menores y no presentaron diferencias significativas entre ellos. La variables fisiológicas en todos los casos apoyaron al desarrollo de las plantas de café, esto al ser comparadas con el tratamiento Testigo, sin embargo las que alcanzaron las mayores altitudes, anchos de copas y diámetro de tallo fueron las plantas del tratamiento MIN 3, seguido del MIN 3, no mostrando significancia entre ellos, y para los tratamientos orgánicos el que presento muy buenos resultados en estas variables ha sido el ORG 3, el cual no presento diferencias significativas con el MIN 2, lo cual comprueba que la fertilización mineral es más efectiva en este caso frente a la orgánica. Para el primer año de producción el tratamiento mineral con fertilización MIN 3 es el que obtuvo mayor producción no presentando diferencia estadística con el tratamiento con el MIN 2, no obstante fueron significativamente mayores que los otros tratamientos. Vale indicar que también el tratamiento MIN 1 y el tratamiento ORG 3 han presentado una producción considerable de café no mostrando diferencias estadísticas entre ellos. Para el segundo año la producción el cultivo mostró mayores rendimientos que el primer año de evaluación en todos los tratamientos, esto debido a la fisiología propia del cultivo y por otra parte se atribuye a la adición de fertilizantes que se ha realizado durante todo el ensayo; de manera similar al anterior los tratamientos MIN 3 y MIN 2 obtuvieron mejores rendimientos, no enseñando diferencias estadísticas significativas entre ellos, no obstante el tratamiento mineral dosis MEDIA no presentó significancia estadística con el ORG 3. El benéfico económico ha resultado mayor en el tratamiento MIN 3 y MIN 2, aunque el tratamiento MIN 2, es el que obtiene la mejor relación costo-beneficio; los tratamientos ORG 2 y ORG 3 y Testigo has producido beneficios negativos para el productor. En cuanto a la parte ambiental se considera que los mejores tratamientos en cuanto ha cuidado ambiental serían los tratamientos MIN 1 y ORG 1, sin embargo a nivel de producción y rentabilidad para el productor baja. ABSTRACT Coffee growing has great importance worldwide (ICO, 2011), and in Ecuador, it has been one of the most important crops to generate income (COFENAC, 2011). However, in the productive systems of this country, the inappropriate use of fertilizers has been observed which produces loss of nutrients, thus it is important to study suitable doses for mineral and organic fertilizing. The purpose of the study was to evaluate the effect of mineral and organic fertilizing at different doses in a coffee monoculture in the province of Loja on soil characteristics, emission of the main gasses that produce the greenhouse effect and the phenology and productivity of crops. In the province of Loja (Ecuador) an area of 2.520 m2 was chosen, where 21 plots of Arabica coffee (Coffea arabica L.), the caturra variety were cultivated and three treatments with three repetitions each one for mineral and organic fertilization were used with doses that ranged from: mineral low (MIN 1= 157 Kg NPK ha-1 año-1 for the first year y 425 Kg NPK ha-1 año-1 for the second year), mineral medium (MIN 2= 325 Kg NPK ha-1 año-1 for the first year y 650 Kg NPK ha-1 año-1 I the second year) y mineral high (MIN 3= 487 y 875 Kg NPK ha-1 año-1 for the first and second year respectively), organic low (ORG 1= 147 Kg NPK ha-1 año-1 in the first year y 388 Kg NPK ha-1 año-1 in the second year), organics medium (ORG 2= 265 Kg NPK ha-1 año-1 for the first year y 541 Kg NPK ha-1 año-1 in the second year), organics high (ORG 3= 368 Kg NPK ha-1 año-1 for the first year and 727 Kg NPK ha-1 año-1 in the second year) y fertilization zero (TES = no fertilization).; urea, phosphoric rock and muriate of potash were used in the mineral fertilization and humus (Bioabor) in the organic, plus a blank treatment. Time to evaluate the applied fertilizers was for two consecutive years, fertilization was done twice per year based on soil analysis and nutritional requirements of the crops. In order to determine the characteristics of the soil, samples of soil in each plot with a depth of 20 cm were done; from these samples, the determined initial parameters were: color (Munsell), texture (hydrometer method), pH (soil-water 1:2,5 relation), organic matter (Walkey y Black), nitrogen (Micro Kjendahl), phosphorus (Bray y Kurtz), potassium (Olsen); these processes were repeated each six months in order to evaluate the changes that are produced due to mineral and organic fertilization in the crops. The emissions of greenhouse gasses from the soil to the atmosphere were determined by using enclosure method (Rondón, 2000) and the concentration, by using gas chromatography during the whole testing. The physiological measures (plant height, width of the top of the tree, thickness of the stem and production) were evaluated each two months, except for production which was annual at the end of each harvest. Moreover, the economic analysis of the productivity of the crops was done. The statistical analysis of the data was done using SPSS v. 17.0. The means were proved by ANOVAS with a factor of a Tukey test (P < 0,05). The economic benefit was estimated in terms of incomes and total expenses which were presented in the essay. The results obtained at the end of the essay show that the MIN 2 and MIN 3 treatments produced more meaningful changes in comparison with the other treatments used to improve soil fertility; pH was less affected in the acidification compared with the organic treatments which were greatly acidified; organic matter (MO) had increased considerably in these two treatments; however, they were surpassed by the organic treatments of fertilization; total nitrogen (Nt) and potassium (K) also presented better results at the end of the essay and phosphorus (P) showed good increasing figures although a little lower compared with ORG 2 and ORG 3 treatments. Regarding the emission of the greenhouse gasses, the fluxes accumulated from nitrous oxide (N2O) in two years increased in all the treatments in comparison with the blank treatment, but in a greater form and with higher fluxes in the MIN 3 and MIN 2 treatments which can be considered as the ones with greater contamination of N2O in the atmosphere, this can be due to the applied mineral doses to fertilize during the process; MIN 1 treatments and all the organic ones showed lower emission to the atmosphere. Methane emissions (CH4) did not show major differences in emissions in the treatments, being the greater emissions the ORG 3 and ORG 2 treatments; this is possibly due to the organic compost added to the soil; regarding carbon dioxide (CO2) emissions, in a similar way to CH4, the ORG 3 treatment was the one that presented greater emissions, the CO2 emissions to the atmosphere in the other treatments were lower and did not present meaningful differences among them. The physiological variables in all the cases helped coffee crops grow, this was observed when compared with the blank treatment; however, plants that reached the greatest height, width of top and diameter of stem were the plants of the MIN 3 treatment, followed by MIN 3, which did not show much significance among them, and for the organic treatments, the one that presented great results in these variables was ORG 3, which did not show meaningful differences compared with MIN 2, which proves that mineral fertilization is more effective in this case compared with the organic. In the first year of production, the mineral treatment with MIN 3 fertilization obtained greater production and thus did not show statistical difference with MIN 2 treatment, although the other treatments were greater. It is worth mentioning that MIN 1 treatment and ORG 3 treatment presented a meaningful production of coffee, not showing statistical differences among them. For the second year, the production of the crops showed greater profits than in the first year of evaluation in all the treatments, this was due to the physiological properties of the crops and on the other hand, it might be due to the addition of fertilizers during the whole essay; in a similar way, MIN 3 and MIN 2 performed better, not showing greater statistical differences among them, although the mineral treatment MEDIUM doses did not show statistical difference compared with ORG 3. The economic benefit was greater in the MIN 3 and MIN 2 treatments, although MIN 2 treatment is the one that shows the best cost-benefit ratios; ORG 2 and ORG 3 treatments and the blank produced negative benefits for the producer. Regarding the environment, the best treatments to care for the atmosphere are considered to be MIN 1 and ORG 1 treatments; however, regarding production volume and profitability they were low for the producer.

ANÁLISE COMPARATIVA DO NÚMERO DE CONTATOS OCLUSAIS AO FINAL DA FASE DE CONTENÇÃO, E APÓS UM PERÍODO DE 5 ANOS, EM PACIENTES COM MÁ-OCLUSÃO DE CLASSE II, TRATADOS COM EXTRAÇÕES DE 4 PRÉ- MOLARES.

Realizou-se um estudo comparativo do número de pontos de contato oclusais na posição de máxima intercuspidação habitual em uma amostra composta por 14 pacientes leucodermas, sendo 9 do sexo feminino e 5 do sexo masculino, com máoclusão de Classe II, divisão 1a de Angle, tratados ortodonticamente pela técnica de Edgewise, com extrações dos 4 primeiros pré-molares. Estes pontos foram registrados em dois tempos: T1 - ao final da fase de contenção superior e T2 - após um período médio de 5,2 anos. A contagem dos contatos oclusais foram realizadas nos arcos superior e inferior, separadamente, para as regiões anterior e posteriores. Depois da análise estatística, pôde-se concluir que não houve diferença estatisticamente significante entre o número médio de contatos oclusais nos diferentes períodos estudados.(AU)

ANÁLISE COMPARATIVA DO NÚMERO DE CONTATOS OCLUSAIS AO FINAL DA FASE DE CONTENÇÃO, E APÓS UM PERÍODO DE 5 ANOS, EM PACIENTES COM MÁ-OCLUSÃO DE CLASSE II, TRATADOS COM EXTRAÇÕES DE 4 PRÉ- MOLARES.

Realizou-se um estudo comparativo do número de pontos de contato oclusais na posição de máxima intercuspidação habitual em uma amostra composta por 14 pacientes leucodermas, sendo 9 do sexo feminino e 5 do sexo masculino, com máoclusão de Classe II, divisão 1a de Angle, tratados ortodonticamente pela técnica de Edgewise, com extrações dos 4 primeiros pré-molares. Estes pontos foram registrados em dois tempos: T1 - ao final da fase de contenção superior e T2 - após um período médio de 5,2 anos. A contagem dos contatos oclusais foram realizadas nos arcos superior e inferior, separadamente, para as regiões anterior e posteriores. Depois da análise estatística, pôde-se concluir que não houve diferença estatisticamente significante entre o número médio de contatos oclusais nos diferentes períodos estudados.(AU)

Avaliação de assimetria nas arcadas dentárias de indivíduos com oclusão normal e má oclusão de Classe II

A assimetria das arcadas dentárias constitui um assunto de grande importância estudado por profissionais de Ortodontia na elaboração de um diagnóstico correto, planejamento e execução de um tratamento ortodôntico bem sucedido. O objetivo deste estudo foi avaliar o grau de assimetria das arcadas dentárias em indivíduos com oclusão normal e má oclusão de Classe II, divisão 1 e 2, bem como o dimorfismo sexual existente. Foram avaliados 180 pares de modelos de estudo de indivíduos do sexo masculino e feminino, na faixa etária de 12 a 21 anos, divididos em 3 grupos de 60 pares de modelos, de acordo com a má oclusão. Os grupos foram classificados em: Grupo 1 - indivíduos com oclusão normal, Grupo 2 - indivíduos com má oclusão de Classe II divisão 1 (Cl II 1), e Grupo 3 - indivíduos com má oclusão de Classe II divisão 2 (Cl II 2). Os modelos foram medidos utilizando-se um aparelho inédito, idealizado e fabricado exclusivamente para a análise de assimetria das arcadas dentárias. Para a análise de assimetria foram realizadas 2 medições angulares desvio de linha média (DLM), posicionamento dos caninos (PC) e 3 lineares distância dos caninos à rafe palatina (DC), distância inter-caninos (DIC), posicionamento dos primeiros molares no sentido ântero-posterior (PM). Concluiu-se que a ocorrência de assimetria nas arcadas dentárias independe da má oclusão. O Grupo 1 apresentou um menor grau de assimetria nas arcadas dentárias em relação aos grupos 2 e 3, os quais apresentaram um grau de assimetria semelhante. O grau de assimetria nas arcadas dentárias inferiores, nos 3 grupos, foi maior do que nas arcadas dentárias superiores. A direção do desvio da linha média apresentou uma correlação inversamente proporcional do lado em que o molar se apresentava mesializado, nas arcadas superior e inferior dos três grupos, com exceção da arcada superior no Grupo 2 (Classe II divisão 1). Houve dimorfismo sexual estatisticamente significante para algumas medidas, porém é importante considerar os baixos valores e a disposição, destas diferenças, entre as medidas realizadas, a qual revela ter se tratado de dados obtidos ao acaso.

Avaliação de assimetria nas arcadas dentárias de indivíduos com oclusão normal e má oclusão de Classe II

A assimetria das arcadas dentárias constitui um assunto de grande importância estudado por profissionais de Ortodontia na elaboração de um diagnóstico correto, planejamento e execução de um tratamento ortodôntico bem sucedido. O objetivo deste estudo foi avaliar o grau de assimetria das arcadas dentárias em indivíduos com oclusão normal e má oclusão de Classe II, divisão 1 e 2, bem como o dimorfismo sexual existente. Foram avaliados 180 pares de modelos de estudo de indivíduos do sexo masculino e feminino, na faixa etária de 12 a 21 anos, divididos em 3 grupos de 60 pares de modelos, de acordo com a má oclusão. Os grupos foram classificados em: Grupo 1 - indivíduos com oclusão normal, Grupo 2 - indivíduos com má oclusão de Classe II divisão 1 (Cl II 1), e Grupo 3 - indivíduos com má oclusão de Classe II divisão 2 (Cl II 2). Os modelos foram medidos utilizando-se um aparelho inédito, idealizado e fabricado exclusivamente para a análise de assimetria das arcadas dentárias. Para a análise de assimetria foram realizadas 2 medições angulares desvio de linha média (DLM), posicionamento dos caninos (PC) e 3 lineares distância dos caninos à rafe palatina (DC), distância inter-caninos (DIC), posicionamento dos primeiros molares no sentido ântero-posterior (PM). Concluiu-se que a ocorrência de assimetria nas arcadas dentárias independe da má oclusão. O Grupo 1 apresentou um menor grau de assimetria nas arcadas dentárias em relação aos grupos 2 e 3, os quais apresentaram um grau de assimetria semelhante. O grau de assimetria nas arcadas dentárias inferiores, nos 3 grupos, foi maior do que nas arcadas dentárias superiores. A direção do desvio da linha média apresentou uma correlação inversamente proporcional do lado em que o molar se apresentava mesializado, nas arcadas superior e inferior dos três grupos, com exceção da arcada superior no Grupo 2 (Classe II divisão 1). Houve dimorfismo sexual estatisticamente significante para algumas medidas, porém é importante considerar os baixos valores e a disposição, destas diferenças, entre as medidas realizadas, a qual revela ter se tratado de dados obtidos ao acaso.

Human recombinant soluble guanylyl cyclase: Expression, purification, and regulation

The α1- and β1-subunits of human soluble guanylate cyclase (sGC) were coexpressed in the Sf9 cells/baculovirus system. In addition to the native enzyme, constructs with hexahistidine tag at the amino and carboxyl termini of each subunit were coexpressed. This permitted the rapid and efficient purification of active recombinant enzyme on a nickel-affinity column. The enzyme has one heme per heterodimer and was readily activated with the NO donor sodium nitroprusside or 3-(5′-hydroxymethyl-2′furyl)-1-benzyl-indazole (YC-1). Sodium nitroprusside and YC-1 treatment potentiated each other in combination and demonstrated a remarkable 2,200-fold stimulation of the human recombinant sGC. The effects were inhibited with 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1one (ODQ). The kinetics of the recombinant enzyme with respect to GTP was examined. The products of the reaction, cGMP and pyrophosphate, inhibited the enzyme. The extent of inhibition by cGMP depended on the activation state of the enzyme, whereas inhibition by pyrophosphate was not affected by the enzyme state. Both reaction products displayed independent binding and cooperativity with respect to enzyme inhibition. The expression of large quantities of active enzyme will facilitate structural characterization of the protein.