Cell wall maturation of Arabidopsis trichomes is dependent on exocyst subunit EXO70H4 and involves callose deposition.

Arabidopsis (Arabidopsis thaliana) leaf trichomes are single-cell structures with a well-studied development, but little is understood about their function. Developmental studies focused mainly on the early shaping stages, and little attention has been paid to the maturation stage. We focused on the EXO70H4 exocyst subunit, one of the most up-regulated genes in the mature trichome. We uncovered EXO70H4-dependent development of the secondary cell wall layer, highly autofluorescent and callose rich, deposited only in the upper part of the trichome. The boundary is formed between the apical and the basal parts of mature trichome by a callose ring that is also deposited in an EXO70H4-dependent manner. We call this structure the Ortmannian ring (OR). Both the secondary cell wall layer and the OR are absent in the exo70H4 mutants. Ecophysiological aspects of the trichome cell wall thickening include interference with antiherbivore defense and heavy metal accumulation. Ultraviolet B light induces EXO70H4 transcription in a CONSTITUTIVE PHOTOMORPHOGENIC1-dependent way, resulting in stimulation of trichome cell wall thickening and the OR biogenesis. EXO70H4-dependent trichome cell wall hardening is a unique phenomenon, which may be conserved among a variety of the land plants. Our analyses support a concept that Arabidopsis trichome is an excellent model to study molecular mechanisms of secondary cell wall deposition.

Ultrastructural study of spermiogenesis and the spermatozoon of the proteocephalidean cestode Barsonella lafoni de Chambrier et al., 2009, a parasite of the catfish Clarias gariepinus (Burchell, 1822) (Siluriformes, Clariidae)

Spermiogenesis in the proteocephalidean cestode Barsonella lafoni de Chambrier et al., 2009 shows typical characteristics of the type I spermiogenesis. These include the formation of distal cytoplasmic protrusions forming the differentiation zones, lined by cortical microtubules and containing two centrioles. An electron-dense material is present in the apical region of the differentiation zone during the early stages of spermiogenesis. Each centriole is associated to a striated rootlet, being separated by an intercentriolar body. Two free and unequal flagella originate from the centrioles and develop on the lateral sides of the differentiation zone. A median cytoplasmic process is formed between the flagella. Later these flagella rotate, become parallel to the median cytoplasmic process and finally fuse proximodistally with the latter. It is interesting to note that both flagellar growth and rotation are asynchronous. Later, the nucleus enlarges and penetrates into the spermatid body. Finally, the ring of arching membranes is strangled and the young spermatozoon is detached from the residual cytoplasm. The mature spermatozoon presents two axonemes of the 9 +"1" trepaxonematan pattern, crested body, parallel nucleus and cortical microtubules, and glycogen granules. Thus, it corresponds to the type II spermatozoon, described in almost all Proteocephalidea. The anterior extremity of the gamete is characterized by the presence of an apical cone surrounded by the lateral projections of the crested body. An arc formed by some thick and parallel cortical microtubules appears at the level of the centriole. They surround the centriole and later the first axoneme. This arc of electron-dense microtubules disorganizes when the second axoneme appears, and then two parallel rows of thin cortical microtubules are observed. The posterior extremity of the male gamete exhibits some cortical microtubules. This type of posterior extremity has never been described in proteocephalidean cestodes. The ultrastructural features of the spermatozoon/spermiogenesis of the Proteocephalidea species are analyzed and compared.

Características físico-químicas, polifenóis e flavonoides amarelos em frutos de espécies de pitaias comerciais e nativas do Cerrado

Apesar do grande potencial comercial da pitaia, ainda são escassos os estudos de caracterização físico-química de frutos da pitaia, principalmente considerando espécies nativas do Cerrado. Neste trabalho, objetivou-se analisar a caracterização físico-química, polifenóis e flavonoides amarelos totais de frutos de espécies de pitaia Hylocereus costaricensis, Hylocereus undatus, Selenicereus setaceus e Selenicereus megalanthus. Para as avaliações físico-químicas, foram realizadas as análises de sólidos solúveis, pH e acidez total titulável. Para a determinação dos compostos fenólicos, realizaram-se as análises de polifenóis extraíveis totais e flavonoides amarelos. Foram observadas diferenças significativas entre as espécies de pitaia e entre as partes basal, mediana e apical dos frutos, quanto às características físico-químicas e a concentração de compostos fenólicos. A espécie S. megalanthus apresentou maior quantidade de sólidos solúveis, apresentando, assim, a polpa mais doce. Tal característica foi mais pronunciada na parte mediana do fruto de todas as espécies. Houve diferença significativa entre o pH, com valores variando de 4,84 a 5,67, classificando-se como alimentos pouco ácidos. A acidez variou de 0,10 % a 0,15 % de ácido cítrico. H. costaricensis merece destaque pela presença de maior quantidade de polifenóis totais e de flavonoides amarelos, diferenciando-se significativamente das demais espécies.

Evaluation of winter temperatures on apple budbreak using grafted twigs

Temperature is the main climate factor related to induction, maintenance and dormancy release in apple (Malus domestica Borkh.). The inadequate chilling exposure in apples causes budbreak problems, resulting in decrease in yield potential. Thus, the knowledge of physiological principles and environmental factors determining the dormancy phenomenon, especially winter temperature effects, it is necessary for the efficient selection of cultivars in a productive region. In addition, it is indispensable to adapt the orchard management aiming to decrease the problems caused by lack chilling during winter. The objective of this study was to evaluate the influence of different thermal conditions during the dormancy period on budbreak of apple cultivars. One-year-old twigs of 'Castel Gala' and 'Royal Gala' cultivars, grafted on M7 rootstock, were submitted to temperatures of 5, 10 and 15ºC for different exposure periods (168; 336; 672; 1,008 and 1,344 hours). After treatments execution, the plants were kept in a greenhouse at 25ºC. Budbreak was quantified when accumulated 3,444; 6,888; 10,332; 13,776; 17,220 and 20,664 GDHºC after temperature treatments. The cultivars responded differently to temperature effect during the winter period. The temperature of 15ºC during winter shows a greater effectiveness on 'Castel Gala' apple budbreak while in the 'Royal Gala' apples the temperatures of 5 and 10ºC show better performance. 'Castel Gala' cultivar (low chilling requirement) may supply its physiological necessities, may be capable to budburst, even when subjected to higher temperatures in relation to 'Royal Gala' apples (high chilling requirement).

Ultrastructural study of spermiogenesis and the spermatozoon of the proteocephalidean cestode Barsonella lafoni de Chambrier et al., 2009, a parasite of the catfish Clarias gariepinus (Burchell, 1822) (Siluriformes, Clariidae)

Spermiogenesis in the proteocephalidean cestode Barsonella lafoni de Chambrier et al., 2009 shows typical characteristics of the type I spermiogenesis. These include the formation of distal cytoplasmic protrusions forming the differentiation zones, lined by cortical microtubules and containing two centrioles. An electron-dense material is present in the apical region of the differentiation zone during the early stages of spermiogenesis. Each centriole is associated to a striated rootlet, being separated by an intercentriolar body. Two free and unequal flagella originate from the centrioles and develop on the lateral sides of the differentiation zone. A median cytoplasmic process is formed between the flagella. Later these flagella rotate, become parallel to the median cytoplasmic process and finally fuse proximodistally with the latter. It is interesting to note that both flagellar growth and rotation are asynchronous. Later, the nucleus enlarges and penetrates into the spermatid body. Finally, the ring of arching membranes is strangled and the young spermatozoon is detached from the residual cytoplasm. The mature spermatozoon presents two axonemes of the 9 +"1" trepaxonematan pattern, crested body, parallel nucleus and cortical microtubules, and glycogen granules. Thus, it corresponds to the type II spermatozoon, described in almost all Proteocephalidea. The anterior extremity of the gamete is characterized by the presence of an apical cone surrounded by the lateral projections of the crested body. An arc formed by some thick and parallel cortical microtubules appears at the level of the centriole. They surround the centriole and later the first axoneme. This arc of electron-dense microtubules disorganizes when the second axoneme appears, and then two parallel rows of thin cortical microtubules are observed. The posterior extremity of the male gamete exhibits some cortical microtubules. This type of posterior extremity has never been described in proteocephalidean cestodes. The ultrastructural features of the spermatozoon/spermiogenesis of the Proteocephalidea species are analyzed and compared.

Propagação da gravioleira por miniestaquia

Avaliou-se o enraizamento de miniestacas de gravioleira (Annona muricata L.), utilizando-se de estacas apicais e subapicais herbáceas coletadas em plantas cultivadas no campo e em casa de vegetação. O experimento foi conduzido em delineamento inteiramente casualizado, em esquema fatorial 4 x 2, sendo 4 tipos de miniestacas (apical herbácea do campo e viveiro, subapical herbácea do campo e de viveiro) e 2 níveis de reguladores de crescimento (0 e 6.000 mg kg-1), totalizando 8 tratamentos e 3 repetições, com 12 estacas por parcela. As miniestacas foram padronizadas com sete centímetros de comprimento e tratadas ou não com ácido indolbutírico, colocadas em tubetes contendo substrato comercial e mantidas em câmara de nebulização. Aos 80 dias após o estaqueamento, foi observado que as miniestacas subapicais e apicais de casa de vegetação e subapicais do campo apresentaram percentual de enraizamento de 97,3%, 88% e 88% respectivamente. Não foi detectado efeito da aplicação de AIB no enraizamento das miniestacas.

Intensidades da poda seca e do desbaste de cacho na composição da uva Cabernet Sauvignon

A pesquisa foi realizada durante quatro anos, num vinhedo de Cabernet Sauvignon (Vitis vinifera L.), em Bento Gonçalves-RS. O objetivo foi determinar o efeito das intensidades da poda seca e do desbaste de cacho em variáveis relacionadas aos componentes de produção da videira e à composição do mosto de uva. Os tratamentos consistiram em dois níveis de poda seca - curta e longa - e quatro de desbaste de cacho (%) - 0; 25; 50 e 75 -; com cinco repetições. O delineamento experimental foi o em blocos casualizados, com os tratamentos em parcelas subdivididas. Os resultados mostram que a poda seca e o desbaste de cacho tiveram efeito altamente significativo na produtividade do vinhedo que, na média dos quatro anos, variou de 10.971 kg/ha − poda curta, 75% de desbaste − a 32.819 kg/ha − poda longa, 0% de desbaste. Houve, também, efeito significativo na produtividade por gema, peso de ramos podados por gema e por hectare, área foliar/peso fresco do fruto e produtividade/peso de ramos podados. Entretanto, o efeito nas variáveis relacionadas a açúcar e acidez do mosto da uva foi pouco expressivo. O componente 1 da análise de componentes principais discriminou o tratamento poda curta a 75% de desbaste de cacho dos tratamentos poda longa-0% de desbaste e poda longa-25% de desbaste.

Executive functions profile in extreme eating/weight conditions: from anorexia nervosa to obesity

Extreme weight conditions (EWC) groups along a continuum may share some biological risk factors and intermediate neurocognitive phenotypes. A core cognitive trait in EWC appears to be executive dysfunction, with a focus on decision making, response inhibition and cognitive flexibility. Differences between individuals in these areas are likely to contribute to the differences in vulnerability to EWC. The aim of the study was to investigate whether there is a common pattern of executive dysfunction in EWC while comparing anorexia nervosa patients (AN), obese subjects (OB) and healthy eating/weight controls (HC).

Fertilidade e reservas de carbono e nitrogênio em gemas de ramos das viníferas 'carbernet sauvignon' e 'nebbiolo'

O trabalho objetivou avaliar a fertilidade e as reservas de C e N em gemas de ramos, na entrada da dormência e dormência plena, das variedades 'Cabernet Sauvignon' e 'Nebbiolo'. Em um vinhedo de 'Cabernet Sauvignon' e outro de 'Nebbiolo', em São Joaquim (SC), foram coletadas gemas em ramos de ano, em duas épocas: na entrada da dormência (maio de 2012) e na dormência plena (agosto de 2012). Os tratamentos foram: (T1) gemas basais (1ª a 5ª); (T2) gemas medianas (6ª a 10ª), e (T3) gemas distais (11ª a 15ª). Em cada planta, foram coletados três ramos, um localizado no lado esquerdo, outro no lado direito e o terceiro no centro da planta. A presença ou ausência de primórdios de inflorescência foi realizada apenas nas gemas dos ramos coletados na dormência plena, usando um estereomicroscópio. Foram quantificados em gemas na entrada da dormência e na dormência plena, os teores de carboidratos solúveis totais, amido, proteínas totais, carbono orgânico total, nitrogênio total e relação C/N. A 'Cabernet Sauvignon' apresentou o maior número de gemas férteis na porção mediana dos ramos de ano. Mas, na cultivar 'Nebbiolo', o maior número de gemas férteis foi observado nas porções medianas e distais do ramo. As maiores concentrações de reservas de carbono nas gemas da 'Cabernet Sauvignon' podem ser responsáveis pela maior fertilidade de gemas. A carga de gemas férteis na 'Cabernet Sauvignon' pode ser aumentada com a poda de inverno média a longa e, na 'Nebbiolo', com poda longa, acima da 6ª gema, como o Guyot.

AVALIAÇÃO FENOLÓGICA DA PEREIRA ‘TRIUNFO’ CULTIVADA EM CLIMA SEMIÁRIDO NO NORDESTE DO BRASIL NA SAFRA DE 2012

A fenologia representa uma ferramenta primordial para a inserção de culturas em áreas que apresentam condições diferentes daquelas nas quais estão adaptadas. Opresente trabalho objetivou caracterizar fenológica e produtivamente pereira ‘Triunfo’, sob condiçãosemiárida tropical, no Vale do São Francisco, Nordeste brasileiro. O experimento foi conduzido de 10 de fevereiro a 2 julho de 2012, em pomar experimental, na Estação Experimental de Bebedouro da Embrapa Semiárido,em Petrolina-PE, Brasil. Foram realizadas observações diárias, do estádio de gema dormente até o amadurecimento dos frutos, e foram avaliados os seguintes parâmetros de produção: i) vingamento: a relação entrenúmero de flores e de frutos (%); ii) o número de frutos por planta; iii) produção de frutos por planta (kg); e iv) produtividade (t/ha). O ciclo fenológico da pereira ‘Triunfo’ durou 144 dias, o vingamento obtido foide 11,43%, o número de frutos por planta de 162, produção por planta de 26,34 kg e produtividade de 26,33 t ha-1. Baseado nos resultados obtidos, foi constatada a possibilidade de produção de frutos de pereira emcondição semiárida tropical.

AVALIAÇÃO FENOLÓGICA DAS DIFERENTES ESTRUTURAS DE FRUTIFICAÇÃO DAS MACIEIRAS 'GALA' E 'FUJI' NA REGIÃO DE CAÇADOR-SC

RESUMO A macieira apresenta duas fases que caracterizam seu ciclo anual: a de repouso hibernal e a de crescimento vegetativo e reprodutivo. A importância em se conhecer ainfluência das condições climáticas sobre os processos reprodutivos das plantas e em desenvolver tecnologias que minimizem os possíveis efeitos aumenta à medida que as projeções de aquecimento global se tornampreocupantes. O objetivo deste trabalho foi caracterizar o comportamento dos principais eventos fenológicos da macieira, da brotação à colheita, nas diferentes estruturas de frutificação, sob as condiçõesclimáticas do Sul do Brasil. O estudo experimental foi conduzido na Estação Experimental da Epagri (26°50’S, 50°58’W, altitude 950 m), durante os ciclos de 2011/12 e 2013/2014. As cultivares estudadas foram Gala e Fuji,com nove anos de idade, ambas sobre o porta-enxerto M9. O desenvolvimento fenológico foi observado nas diferentes estruturas de frutificação da macieira: gema de esporão, gema terminal de brindila e gemaaxilar de brindilas. As mesmas foram comparadas através dos dias e da exigência térmica necessários para a transição dos estádios, sendo contabilizados a partir do tratamento de quebra de dormência. As diferençasencontradas no início da brotação e do florescimento, entre as estruturas, dependeram consideravelmente das condições climáticas do ano em questão. Sob condições de altas temperaturas após o tratamento de quebra dedormência, houve maior sincronia fenológica entre as estruturas, sendo que sob baixas temperaturas se observou maior variabilidade dos estádios entre as mesmas. A partir do tratamento de quebra de dormência,gemas de esporões necessitam de menor acúmulo térmico para brotar, principalmente esporões de ‘Gala’ e gemas terminais de brindilas necessitam de maior acúmulo térmico que gemas de esporões para dar início aoflorescimento.

Changes of Root Length and Root-to-Crown Ratio after Apical Surgery: An Analysis by Using Cone-beam Computed Tomography.

INTRODUCTION: Apical surgery is an important treatment option for teeth with post-treatment periodontitis. Although apical surgery involves root-end resection, no morphometric data are yet available about root-end resection and its impact on the root-to-crown ratio (RCR). The present study assessed the length of apicectomy and calculated the loss of root length and changes of RCR after apical surgery. METHODS: In a prospective clinical study, cone-beam computed tomography scans were taken preoperatively and postoperatively. From these images, the crown and root lengths of 61 roots (54 teeth in 47 patients) were measured before and after apical surgery. Data were collected relative to the cementoenamel junction (CEJ) as well as to the crestal bone level (CBL). One observer took all measurements twice (to calculate the intraobserver variability), and the means were used for further analysis. The following parameters were assessed for all treated teeth as well as for specific tooth groups: length of root-end resection and percentage change of root length, preoperative and postoperative RCRs, and percentage change of RCR after apical surgery. RESULTS: The mean length of root-end resection was 3.58 ± 1.43 mm (relative to the CBL). This amounted to a loss of 33.2% of clinical and 26% of anatomic root length. There was an overall significant difference between the tooth groups (P < .05). There was also a statistically significant difference comparing mandibular and maxillary teeth (P < .05), but not for incisors/canines versus premolars/molars (P = .125). The mean preoperative and postoperative RCRs (relative to CEJ) were 1.83 and 1.35, respectively (P < .001). With regard to the CBL reference, the mean preoperative and postoperative RCRs were 1.08 and 0.71 (CBL), respectively (P < .001). The calculated changes of RCR after apical surgery were 24.8% relative to CEJ and 33.3% relative to CBL (P < .001). Across the different tooth groups, the mean RCR was not significantly different (P = .244 for CEJ and 0.114 for CBL). CONCLUSIONS: This CBCT-based study demonstrated that the RCR is significantly changed after root-end resection in apical surgery irrespective of the clinical (CBL) or anatomic (CEJ) reference levels. The lowest, and thus clinically most critical, postoperative RCR was observed in maxillary incisors. Future clinical studies need to show the impact of resection length and RCR changes on the outcome of apical surgery.

Thermal increment due to ErCr: YSGG and CO2 laser irradiation of different implant surfaces. A pilot study

Objective: An evaluation and comparison is made of the thermal increment at different implant surfaces during irradiation with CO2 and ErCr:YSGG lasers. Study design: Five threaded and impacted implants with four types of surfaces were inserted in an adult pig rib: two implants with a hydroxyapatite surface (HA)(impacted and threaded, respectively), a machined titanium surface implant (TI mach), a titanium plasma spray surface implant (TPS), and a sandblasted, acid-etched surface implant (SBAE). A 0.5-mm diameter bone defect was made in the implant apical zone, and a type-K thermocouple (Termopar)® was placed in contact with the implant. The implants were irradiated in the coronal zone of each implant with a CO2 (4 W continuous mode) and an ErCr:YSGG laser (1.5 W, pulsed mode) first without and then with refrigeration. The temperature variations at the implant apical surface were recorded. Results: An apical temperature increase was recorded in all cases during CO2 and ErCr:YSGG laser irradiation without refrigeration. However, when the ErCr:YSGG was used with a water spray, a decrease in temperature was observed in all implants. The acid-etched and sandblasted surfaces were those most affected by the thermal changes. Conclusions: The ErCr:YSGG laser with a water spray applied to the sealing cap or coronal zone of the implants does not generate thermal increments in the apical surface capable of adversely affecting osseointegration and the integrity of the peri-implant bone tissue

Cadmium uptake and induction of metallothionein synthesis in a renal epithelial cell line (LLC-PK1).

LLC-PK1 cells, an established cell line from pig kidney with proximal tubule properties, were cultivated in vitro at confluence on plastic dishes. They were then exposed (apical side) to inorganic cadmium (CdCl2, 5 microM) for periods ranging between 1 to 24 h. Analysis of the cell supernatant after homogenisation and ultracentrifugation indicated that Cd taken up in the first 3 h was bound to cytosolic high molecular weight proteins, but was redistributed to low molecular weight proteins at later stages. Induction of Cd-metallothionein (Cd-Mt) synthesis, as judged from Cd-Mt binding to a specific anti-Cd-Mt antibody and from the rate of 35S-cys incorporation into a specific protein fraction, was apparent 3-6 h after the addition of Cd to the incubation medium.

Polarized distribution of inducible nitric oxide synthase regulates activity in intestinal epithelial cells.

Inducible nitric oxide synthase (iNOS) functions as a homodimer. In cell extracts, iNOS molecules partition both in cytosolic and particulate fractions, indicating that iNOS exists as soluble and membrane associated forms. In this study, iNOS features were investigated in human intestinal epithelial cells stimulated with cytokines and in duodenum from mice exposed to flagellin. Our experiments indicate that iNOS is mainly associated with the particulate fraction of cell extracts. Confocal microscopy showed a preferential localization of iNOS at the apical pole of intestinal epithelial cells. In particulate fractions, iNOS dimers were more abundant than in the cytosolic fraction. Similar observations were seen in mouse duodenum samples. These results suggest that, in epithelial cells, iNOS activity is regulated by localization-dependent processes.