Grx5 glutaredoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae

Glutaredoxins are members of a superfamily of thiol disulfide oxidoreductases involved in maintaining the redox state of target proteins. In Saccharomyces cerevisiae, two glutaredoxins (Grx1 and Grx2) containing a cysteine pair at the active site had been characterized as protecting yeast cells against oxidative damage. In this work, another subfamily of yeast glutaredoxins (Grx3, Grx4, and Grx5) that differs from the first in containing a single cysteine residue at the putative active site is described. This trait is also characteristic for a number of glutaredoxins from bacteria to humans, with which the Grx3/4/5 group has extensive homology over two regions. Mutants lacking Grx5 are partially deficient in growth in rich and minimal media and also highly sensitive to oxidative damage caused by menadione and hydrogen peroxide. A significant increase in total protein carbonyl content is constitutively observed in grx5cells, and a number of specific proteins, including transketolase, appear to be highly oxidized in this mutant. The synthetic lethality of the grx5 and grx2 mutations on one hand and ofgrx5 with the grx3 grx4 combination on the other points to a complex functional relationship among yeast glutaredoxins, with Grx5 playing a specially important role in protection against oxidative stress both during ordinary growth conditions and after externally induced damage. Grx5-deficient mutants are also sensitive to osmotic stress, which indicates a relationship between the two types of stress in yeast cells.

Desenvolvimento floral e produção de pessegueiros 'granada' sob distintas condições climáticas

A cultivar de pessegueiro 'Granada' vem apresentando baixa frutificação e irregularidade de produção nas principais regiões produtoras de pêssego no Estado do Rio Grande do Sul. Este trabalho teve como objetivo comparar o desenvolvimento floral e a produção de pessegueiros 'Granada' em duas regiões com distintas condições climáticas. Os pomares estudados, nas safras de 2004 e 2005, localizam-se nos municípios de Charqueadas e Canguçu, nas regiões Depressão Central e Sul do RS, respectivamente. Conclui-se que o pessegueiro 'Granada' mostra-se muito instável em termos de produção. A baixa produção e a viabilidade do pólen, aliada ao atraso no desenvolvimento dos óvulos, influenciadas sobretudo pela ocorrência de altas temperaturas na pré-floração e floração, foram as principais causas do baixo desempenho reprodutivo e produtivo do pessegueiro 'Granada' em Charqueadas, em 2004, e em Canguçu, em 2005.

Influence of floral structure and flower bud quality on productivity and fruit shape in different apple cultivars

This study examines the relationship between floral structure and bud quality with the productivity and fruit shape of Gala, Fuji and Daiane apple cultivars under the mild winter conditions in Southern Brazil. Six different types of floral structures were characterized in field growing plants, according to their nature and bud size: spurs, short and long twigs with weak and vigorous buds. Variables related to the phenology and the productivity for these different structures were evaluated. Gala and Fuji cvs. showed earlier phenological development in the twigs, and cv. Daiane in the spurs. For the three cvs. the highest percentage of buds in each phenological phase was observed in the long twigs. The long twigs also showed the highest sprout and fruit set index, floral number per cluster, and leaf area in the three cvs., while the bud abortion was higher in the spurs than in the twigs. No difference was observed among the structures in cvs. Gala and Fuji regarding to the fruit shape. In the cv. Daiane, however, a tendency to higher length diameter ratio of the fruits produced by the long twigs was observed.

A multi-contrast MRI study of microstructural brain damage in patients with mild cognitive impairment.

OBJECTIVES: The aim of this study was to investigate pathological mechanisms underlying brain tissue alterations in mild cognitive impairment (MCI) using multi-contrast 3 T magnetic resonance imaging (MRI). METHODS: Forty-two MCI patients and 77 healthy controls (HC) underwent T1/T2* relaxometry as well as Magnetization Transfer (MT) MRI. Between-groups comparisons in MRI metrics were performed using permutation-based tests. Using MRI data, a generalized linear model (GLM) was computed to predict clinical performance and a support-vector machine (SVM) classification was used to classify MCI and HC subjects. RESULTS: Multi-parametric MRI data showed microstructural brain alterations in MCI patients vs HC that might be interpreted as: (i) a broad loss of myelin/cellular proteins and tissue microstructure in the hippocampus (p ≤ 0.01) and global white matter (p < 0.05); and (ii) iron accumulation in the pallidus nucleus (p ≤ 0.05). MRI metrics accurately predicted memory and executive performances in patients (p ≤ 0.005). SVM classification reached an accuracy of 75% to separate MCI and HC, and performed best using both volumes and T1/T2*/MT metrics. CONCLUSION: Multi-contrast MRI appears to be a promising approach to infer pathophysiological mechanisms leading to brain tissue alterations in MCI. Likewise, parametric MRI data provide powerful correlates of cognitive deficits and improve automatic disease classification based on morphometric features.

Histologic evaluation of thermal damage produced on soft tissues by CO2, Er,Cr:YSGG and diode lasers

Objective: The aim of this in vitro experimental study was to perform histological evaluation of the thermal effect produced on soft tissue irradiated with CO2, Er,Cr:YSGG or diode lasers. Study design: Porcine oral mucosa samples were irradiated with Er,Cr:YSGG laser at 1 W with and without water / air spray, at 2 W with and without water / air spray, and at 4 W with water / air spray, with CO2 laser at 1 W, 2 W, 10 W, 20 W continuous mode and 20 W pulsed mode and diode laser at 2W, 5W, and 10W pulsed mode. The thermal effect was evaluated measuring the width of damaged tissue adjacent to the incision, stained positively for hyalinized tissue with Hematoxylin-Eosin and Masson Trichrome stains. Besides, histological changes in the irradiated tissue were described using subjective grading scales. Results: The evaluated lasers developed a wide range of thermal damage with significant differences between groups. The samples with lowest thermal effect were those irradiated with Er,Cr:YSGG laser using water / air spray, followed by CO2 and diode lasers. Conclusions: Emission parameters of each laser system may influence the thermal damage inflicted on the soft tissue, however, the wave length of each laser determines the absorption rate characteristics of every tissue and the thermal effect

BIOLOGIA FLORAL E VIABILIDADE DE PÓLEN EM CULTIVARES DE CAQUIZEIRO (Diospyros kaki L.) E Diospyros virginiania L.

RESUMO O caquizeiro é uma planta muito complexa na questão de sua biologia floral, já que pode apresentar três tipos de flor (feminina, masculina ou hermafrodita) ou apenas flores femininas, como no caso das cultivares comerciais. O objetivo deste trabalho foi estudar o período de florescimento, os tipos de flores e estimar a viabilidade dos grãos de pólen em nove cultivares de D. kaki e em D. virginiana. As cultivares Pomelo e Rama Forte foram precoces para o início do florescimento. A cultivar Pomelo e a espécie D. virginiana apresentaram produção de flores masculinas, com pólen viável. As nove cultivares de D. kakiestudadas apresentaram produção de flores femininas (também masculinas em Pomelo) como esperado em cultivares comerciais. Porém a cultivar Mikado apresentou, ao final do florescimento, flores com produção de pólen viável, o que ainda não havia sido relatado. A viabilidade dos grãos de pólen, em geral, foi acima de 90% para as cultivares Pomelo e Mikado, e para D. virginiana.

CARACTERIZAÇÃO MOLECULAR DE ISOLADOS DE Colletotrichum spp. ASSOCIADOS A PODRIDÃO FLORAL DOS CITROS

RESUMO A podridão floral dos citros (PFC) é uma importante doença dessa cultura, responsável por elevadas perdas de produção. Normalmente, essa doença mostra-se limitante quando ocorrem prolongados períodos chuvosos durante o florescimento das plantas ou quando existe intenso molhamento foliar. Duas espécies de Colletotrichum estão associadas à doença: C. acutatum e C. gloeosporioides. Entretanto, recentemente, tem-se verificado que, mesmo sob condições não tão propícias, a doença tem ocorrido com relativa frequência, suspeitando-se do envolvimento de outras espécies de Colletotrichumou de novas condições de adaptação das espécies descritas. Este trabalho teve como objetivo determinar se há ou não outra espécie de Colletotrichum associada a PFC e avaliar a viabilidade do emprego de marcadores moleculares ISSR na caracterização taxonômica de isolados de Colletotrichum spp. associados a sintomas de PFC em flores, assim como de tecidos foliares e frutos cítricos assintomáticos. Para tanto, foi empregada uma combinação de iniciadores específicos, levando em conta a região ITS e marcadores moleculares ISSR. Os marcadores ISSR mostraram-se eficientes na caracterização taxonômica dos isolados de Colletotrichum analisados. A população avaliada foi constituída apenas por C. acutatum e C. gloeosporioides, descartando o envolvimento de uma espécie adicional. Foi constatada alta diversidade genética entre os isolados analisados, o que também se mostra convergente quanto às diferenças fenotípicas observadas sob condições de campo. Entretanto, não foi encontrada relação quanto à origem e as espécies de Colletotrichum spp. associadas. De modo inédito, ainda que assintomaticamente, foi detectada a presença de um isolado de C. acutatum associado a frutos cítricos.

SUSCETIBILIDADE DE CLONES DE LIMA ÁCIDA ‘TAHITI’ À PODRIDÃO FLORAL DOS CITROS

RESUMO A podridão floral dos citros (PFC), causada por Colletotrichum acutatum Simmons e C. gloeosporioides, é a doença fúngica mais importante em limeira-ácida ‘Tahiti’, pois, leva à queda prematura de flores e frutos, acarretando a redução da produção. Avaliou-se a suscetibilidade à PFC dos clones de lima-ácida ‘Tahiti’ “IAC 5”, “IAC 5-1”, “CNPMF/EECB”, “CNPMF 2000” e “CNPMF 2001”, em Bebedouro-SP. Todos os clones são suscetíveis à doença. A maior incidência de PFC em plantas do clone “IAC 5-1” indica que este clone apresenta maior suscetibilidade. Mesmo com sintomas, plantas do clone “CNPMF/EECB” apresentam maior fixação de frutos.

Impact of the early use of immunomodulators or TNF antagonists on bowel damage and surgery in Crohn's disease.

BACKGROUND: The impact of early treatment with immunomodulators (IM) and/or TNF antagonists on bowel damage in Crohn's disease (CD) patients is unknown. AIM: To assess whether 'early treatment' with IM and/or TNF antagonists, defined as treatment within a 2-year period from the date of CD diagnosis, was associated with development of lesser number of disease complications when compared to 'late treatment', which was defined as treatment initiation after >2 years from the time of CD diagnosis. METHODS: Data from the Swiss IBD Cohort Study were analysed. The following outcomes were assessed using Cox proportional hazard modelling: bowel strictures, perianal fistulas, internal fistulas, intestinal surgery, perianal surgery and any of the aforementioned complications. RESULTS: The 'early treatment' group of 292 CD patients was compared to the 'late treatment' group of 248 CD patients. We found that 'early treatment' with IM or TNF antagonists alone was associated with reduced risk of bowel strictures [hazard ratio (HR) 0.496, P = 0.004 for IM; HR 0.276, P = 0.018 for TNF antagonists]. Furthermore, 'early treatment' with IM was associated with reduced risk of undergoing intestinal surgery (HR 0.322, P = 0.005), and perianal surgery (HR 0.361, P = 0.042), as well as developing any complication (HR 0.567, P = 0.006). CONCLUSIONS: Treatment with immunomodulators or TNF antagonists within the first 2 years of CD diagnosis was associated with reduced risk of developing bowel strictures, when compared to initiating these drugs >2 years after diagnosis. Furthermore, early immunomodulators treatment was associated with reduced risk of intestinal surgery, perianal surgery and any complication.

Visualizing the spatiotemporal dynamics of DNA damage in budding yeast.

Fluorescence microscopy has enabled the analysis of both the spatial distribution of DNA damage and its dynamics during the DNA damage response (DDR). Three microscopic techniques can be used to study the spatiotemporal dynamics of DNA damage. In the first part we describe how we determine the position of DNA double-strand breaks (DSBs) relative to the nuclear envelope. The second part describes how to quantify the co-localization of DNA DSBs with nuclear pore clusters, or other nuclear subcompartments. The final protocols describe methods for the quantification of locus mobility over time.

Decoupled evolution of floral traits and climatic preferences in a clade of Neotropical Gesneriaceae.

BACKGROUND: Major factors influencing the phenotypic diversity of a lineage can be recognized by characterizing the extent and mode of trait evolution between related species. Here, we compared the evolutionary dynamics of traits associated with floral morphology and climatic preferences in a clade composed of the genera Codonanthopsis, Codonanthe and Nematanthus (Gesneriaceae). To test the mode and specific components that lead to phenotypic diversity in this group, we performed a Bayesian phylogenetic analysis of combined nuclear and plastid DNA sequences and modeled the evolution of quantitative traits related to flower shape and size and to climatic preferences. We propose an alternative approach to display graphically the complex dynamics of trait evolution along a phylogenetic tree using a wide range of evolutionary scenarios. RESULTS: Our results demonstrated heterogeneous trait evolution. Floral shapes displaced into separate regimes selected by the different pollinator types (hummingbirds versus insects), while floral size underwent a clade-specific evolution. Rates of evolution were higher for the clade that is hummingbird pollinated and experienced flower resupination, compared with species pollinated by bees, suggesting a relevant role of plant-pollinator interactions in lowland rainforest. The evolution of temperature preferences is best explained by a model with distinct selective regimes between the Brazilian Atlantic Forest and the other biomes, whereas differentiation along the precipitation axis was characterized by higher rates, compared with temperature, and no regime or clade-specific patterns. CONCLUSIONS: Our study shows different selective regimes and clade-specific patterns in the evolution of morphological and climatic components during the diversification of Neotropical species. Our new graphical visualization tool allows the representation of trait trajectories under parameter-rich models, thus contributing to a better understanding of complex evolutionary dynamics.

Verbal emotional memory in a case with left amygdala damage.

The amygdala nuclei appear to be critically implicated in emotional memory. However, in most studies, encoding and consolidation processes cannot be analyzed separately. We thus studied the verbal emotional memory in a young woman with a ganglioglioma of the left amygdala and analyzed its impact (1) on each step of the memory process (encoding, retrieval, and recognition) (2) on short- and long-term consolidation (1-hour and 1-week delay) and (3) on processing of valence (positive and negative items compared to neutral words). Results showed emotional encoding impairments and, after encoding was controlled for, emotional long-term consolidation. Finally, although the negative words were not acknowledged as emotionally arousing by the patient, these words were specifically poorly encoded, recalled, and consolidated. Our data suggest that separate cerebral networks support the processing of emotional versus neutral stimuli.

Pattern of injury with a graded excitotoxic insult and ensuing chronic medial septal damage in the rat brain

Brain damage caused by an acute injury depends on the initial severity of the injury and the time elapsed after the injury. To determine whether these two variables activate common mechanisms, we compared the response of the rat medial septum to insult with a graded series of concentrations of a-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) with the time-course effects of a low dose of AMPA. For this purpose we conducted a dose-response study at concentrations of AMPA between 0.27 and 10.8 nmol to measure atrophy of the septal area, losses of cholinergic and GABAergic neurons, astroglial and microglial reactions, and calcification. Cholinergic neurons, whose loss paralleled the degree of septal atrophy produced by AMPA, are more sensitive than GABAergic neurons to the injury produced by AMPA. At doses of AMPA above 2.7 nmol, calcification and the degree of microglial reaction increased only in the GABAergic region of the septal area, whereas atrophy and neuronal loss reached a plateau. We chose the 2.7-nmol dose of AMPA to determine how these parameters were modified between 4 days and 6 months after injection. We found that atrophy and neuronal loss increased progressively through the 6-month study period, whereas astrogliosis ceased to be observed after 1 month, and calcium precipitates were never detected. We conclude that septal damage does not increase with the intensity of an excitotoxic insult. Rather, it progresses continuously after the insult. Because these two situations involve different mechanisms, short-term paradigms are inappropriate for interpreting the pathogenic mechanisms responsible for long-term neurodegenerative processes.

Retinal damage induced by commercial light emitting diodes (LEDs).

Spectra of "white LEDs" are characterized by an intense emission in the blue region of the visible spectrum, absent in daylight spectra. This blue component and the high intensity of emission are the main sources of concern about the health risks of LEDs with respect to their toxicity to the eye and the retina. The aim of our study was to elucidate the role of blue light from LEDs in retinal damage. Commercially available white LEDs and four different blue LEDs (507, 473, 467, and 449nm) were used for exposure experiments on Wistar rats. Immunohistochemical stain, transmission electron microscopy, and Western blot were used to exam the retinas. We evaluated LED-induced retinal cell damage by studying oxidative stress, stress response pathways, and the identification of cell death pathways. LED light caused a state of suffering of the retina with oxidative damage and retinal injury. We observed a loss of photoreceptors and the activation of caspase-independent apoptosis, necroptosis, and necrosis. A wavelength dependence of the effects was observed. Phototoxicity of LEDs on the retina is characterized by a strong damage of photoreceptors and by the induction of necrosis.