Resumo
In this study, the toxic effects of melittin on Madin-Darby Bovine Kidney cells (MDBK) were analyzed with respect to mitochondrial functionality by reduction of MTT and flow cytometry, apoptosis potential, necrosis, oxygen reactive species (ROS) production, lipid peroxidation, and DNA fragmentation using flow cytometry and cell membrane destabilization by confocal microscopy. The toxicity presented dose-dependent characteristics and mitochondrial activity was inhibited by up to 78.24 ±3.59% (P<0.01, n = 6) in MDBK cells exposed to melittin (10µg/mL). Flow cytometry analysis revealed that melittin at 2µg/mL had the highest necrosis rate (P<0.05) for the cells. The lipoperoxidation of the membranes was also higher at 2µg/mL of melittin (P<0.05), which was further confirmed by the microphotographs obtained by confocal microscopy. The highest ROS production occurred when the cells were exposed to 2.5µg/mL melittin (P<0.05), and this concentration also increased DNA fragmentation (P<0.05). There was a significative and positive correlation between the lipoperoxidation of membranes with ROS (R=0.4158), mitochondrial functionality (R=0.4149), and apoptosis (R=0.4978). Thus, the oxidative stress generated by melittin culminates in the elevation of intracellular ROS that initiates a cascade of toxic events in MDBK cells.(AU)
Neste estudo, os efeitos tóxicos da melitina em células Madin-Darby Bovine Kidney (MDBK) foram analisados quanto à funcionalidade mitocondrial, por redução de MTT e citometria de fluxo, potencial de apoptose, necrose, produção de espécies reativas de oxigênio (ROS), peroxidação lipídica e fragmentação de DNA, utilizando-se citometria de fluxo e desestabilização da membrana celular, por microscopia confocal. A toxicidade apresentou características dose-dependentes e a atividade mitocondrial foi inibida até 78,24±3,59% (P<0,01, n = 6) em células MDBK expostas à melitina (10µg/mL). Análises por citometria de fluxo revelaram que a melitina a 2µg/mL apresentou o maior índice necrótico celular (P<0,05). A maior lipoperoxidação de membranas também foi na concentração de 2µg/mL de melitina (P<0,05), o que foi posteriormente confirmado por microscopia confocal. A maior produção de ROS aconteceu quando as células foram expostas a 2,5µg/mL de melitina (P<0,05), e essa concentração também aumentou a fragmentação de DNA (P<0,05). Houve uma significativa correlação positiva entre a lipoperoxidação de membranas e a produção de ROS (R=0,4158), funcionalidade mitocondrial (R=0,4149) e apoptose (R=0,4978). Portanto, o estresse oxidativo gerado pela melitina culminou na elevação de ROS intracelular, que inicia uma cascata de eventos tóxicos nas células MDBK.(AU)
Assuntos
Espécies Reativas de Oxigênio/efeitos adversos , Apoptose , Citotoxinas/análise , Meliteno/análise , Venenos de Abelha/análise , Microscopia Confocal , Citometria de FluxoResumo
The aim of this study was to obtain data on the morphology and morphometry of pre-ovigerous and post-ovigerous adults of the species Tanaisia (Paratanaisia) bragai, using confocal laser scanning microscopy to obtain tomographic images of the suckers and tegument. For morphometric analysis, 45 specimens (30 pre-ovigerous adults and 15 post-ovigerous adults) were measured with the aid of an ocular micrometer coupled to the objective of a photonic microscope. Pre-ovigerous and post-ovigerous adult individuals, stained with Mair carmalumen and mounted in permanent preparations, were analyzed by means of confocal laser scanning microscopy. Positive correlation was detected between the body length and ovary length of postovigerous adults (rs: 0.774; p< 0.05), the body length and the length of uterus (rs: 0.839; p< 0,01) and between the ovary width and egg length (rs: 0.777; p p<0.01). Morphological study of the pre-ovigerous adults demonstrated that the ovary and testes develop simultaneously before the development of the uterus and vitelline glands. The acetabulum was detected in pre-ovigerous adults stained with hematoxilin and observed using light microscopy. In these specimens, the acetabulum measured 36.7 ± 6.9 µm (25-50 µm) in width and 39.91 ± 6.8 µm (25-55 µm) in length. The acetabulum was not detected in post-ovigerous adults observed with light microscopy. However, this structure was detected using confocal miscrocopy. In the post-ovigerous specimens, the acetabulum presented a reduced size compared to the pre-ovigerous adults. This may imply that this structure has more functional significance in the larval and pre-ovigerous stages.(AU)
Assuntos
Animais , Trematódeos/anatomia & histologia , Trematódeos/crescimento & desenvolvimento , Microscopia Confocal/métodosResumo
The aim of this study was to obtain data on the morphology and morphometry of pre-ovigerous and post-ovigerous adults of the species Tanaisia (Paratanaisia) bragai, using confocal laser scanning microscopy to obtain tomographic images of the suckers and tegument. For morphometric analysis, 45 specimens (30 pre-ovigerous adults and 15 post-ovigerous adults) were measured with the aid of an ocular micrometer coupled to the objective of a photonic microscope. Pre-ovigerous and post-ovigerous adult individuals, stained with Mair carmalumen and mounted in permanent preparations, were analyzed by means of confocal laser scanning microscopy. Positive correlation was detected between the body length and ovary length of postovigerous adults (rs: 0.774; p< 0.05), the body length and the length of uterus (rs: 0.839; p< 0,01) and between the ovary width and egg length (rs: 0.777; p p<0.01). Morphological study of the pre-ovigerous adults demonstrated that the ovary and testes develop simultaneously before the development of the uterus and vitelline glands. The acetabulum was detected in pre-ovigerous adults stained with hematoxilin and observed using light microscopy. In these specimens, the acetabulum measured 36.7 ± 6.9 µm (25-50 µm) in width and 39.91 ± 6.8 µm (25-55 µm) in length. The acetabulum was not detected in post-ovigerous adults observed with light microscopy. However, this structure was detected using confocal miscrocopy. In the post-ovigerous specimens, the acetabulum presented a reduced size compared to the pre-ovigerous adults. This may imply that this structure has more functional significance in the larval and pre-ovigerous stages.
Assuntos
Animais , Trematódeos/anatomia & histologia , Trematódeos/crescimento & desenvolvimento , Microscopia Confocal/métodosResumo
Nodal glands are found in one third of the Polygalaceae genera and have valuable taxonomic, ecological and evolutionary significance. In Brazil, they occur in five of the eleven genera already registered. However, there is still a controversy regarding the origin of these structures. The objective of this study was to characterize the morphology and the origin of nodal glands inCaamembeca spectabilis, in order to increase the structural and functional knowledge of these glands in the genera. Samples of nodal regions were collected, fixed and processed according to the methods of light microscopy and electron scanning. Ants were observed and identified along the stem axis. The glucose in exudate allows us to classify these glands as extrafloral nectaries. They are located in pairs on the nodal region. However, its origin is in the leaf trace. In the longitudinal section, the nectaries were present in the apex of cells with anticlinal walls impregnated with suberin, which represents the first record for the family. In this region there is also the formation of a hole by lysis. The secretory tissue is surrounded by phloem. Xylem vessels were observed only on the basis of the nectary, where there are also idioblasts with crystals in druse type. We have studied the ontogeny of the glands nodal in Caamembeca spectabilis and unveiled that these glands are linked to the leaves as stipular nectaries. In addition, the new findings presented here may add support for the understanding of morphology and anatomy of nodal glands in Caamembeca.
Glândulas nodais são encontradas em um terço dos gêneros de Polygalaceae e possuem grande importância taxonômica, ecológica e evolutiva. No Brasil, ocorrem em cinco, dos onze gêneros já registrados. Contudo, ainda há controvérsias quanto à origem dessas estruturas. O objetivo deste trabalho foi caracterizar a morfologia e a origem das glândulas nodais em Caamembeca spectabilis, visando ampliar o conhecimento estrutural e funcional dessas glândulas no gênero. Amostras das regiões nodais foram coletadas, fixadas e processadas segundo os métodos em microscopia de luz e eletrônica de varredura. Formigas foram observadas e identificadas ao longo do eixo caulinar. A glicose no exsudato permite tratar as glândulas como nectários extraflorais. Esses estão localizados aos pares na região nodal. Porém, sua origem ocorre no traço foliar. Os nectários, em secção longitudinal, apresentam as células do ápice com paredes anticlinais impregnadas com suberina, sendo este o primeiro registro para a família. Nessa região também ocorre a formação de um orifício por lise. O tecido secretor do nectário é circundado pelo floema. Vasos de xilema foram observados apenas na base do nectário, onde também ocorrem idioblastos com cristais do tipo drusas. Dessa forma, este estudo confirma a trajetória ontogenética das glândulas nodais em C. spectabilis, as quais, de fato, estão ligadas às folhas, como nectários estipulares. Além de apresentar dados inéditos que auxiliam a compreensão morfológica e anatômica das mesmas em Caamembeca.
Assuntos
Estruturas Vegetais/anatomia & histologia , Néctar de Plantas , Polygalaceae , Formigas , Microscopia ConfocalResumo
Nodal glands are found in one third of the Polygalaceae genera and have valuable taxonomic, ecological and evolutionary significance. In Brazil, they occur in five of the eleven genera already registered. However, there is still a controversy regarding the origin of these structures. The objective of this study was to characterize the morphology and the origin of nodal glands inCaamembeca spectabilis, in order to increase the structural and functional knowledge of these glands in the genera. Samples of nodal regions were collected, fixed and processed according to the methods of light microscopy and electron scanning. Ants were observed and identified along the stem axis. The glucose in exudate allows us to classify these glands as extrafloral nectaries. They are located in pairs on the nodal region. However, its origin is in the leaf trace. In the longitudinal section, the nectaries were present in the apex of cells with anticlinal walls impregnated with suberin, which represents the first record for the family. In this region there is also the formation of a hole by lysis. The secretory tissue is surrounded by phloem. Xylem vessels were observed only on the basis of the nectary, where there are also idioblasts with crystals in druse type. We have studied the ontogeny of the glands nodal in Caamembeca spectabilis and unveiled that these glands are linked to the leaves as stipular nectaries. In addition, the new findings presented here may add support for the understanding of morphology and anatomy of nodal glands in Caamembeca.(AU)
Glândulas nodais são encontradas em um terço dos gêneros de Polygalaceae e possuem grande importância taxonômica, ecológica e evolutiva. No Brasil, ocorrem em cinco, dos onze gêneros já registrados. Contudo, ainda há controvérsias quanto à origem dessas estruturas. O objetivo deste trabalho foi caracterizar a morfologia e a origem das glândulas nodais em Caamembeca spectabilis, visando ampliar o conhecimento estrutural e funcional dessas glândulas no gênero. Amostras das regiões nodais foram coletadas, fixadas e processadas segundo os métodos em microscopia de luz e eletrônica de varredura. Formigas foram observadas e identificadas ao longo do eixo caulinar. A glicose no exsudato permite tratar as glândulas como nectários extraflorais. Esses estão localizados aos pares na região nodal. Porém, sua origem ocorre no traço foliar. Os nectários, em secção longitudinal, apresentam as células do ápice com paredes anticlinais impregnadas com suberina, sendo este o primeiro registro para a família. Nessa região também ocorre a formação de um orifício por lise. O tecido secretor do nectário é circundado pelo floema. Vasos de xilema foram observados apenas na base do nectário, onde também ocorrem idioblastos com cristais do tipo drusas. Dessa forma, este estudo confirma a trajetória ontogenética das glândulas nodais em C. spectabilis, as quais, de fato, estão ligadas às folhas, como nectários estipulares. Além de apresentar dados inéditos que auxiliam a compreensão morfológica e anatômica das mesmas em Caamembeca.(AU)
Assuntos
Polygalaceae , Néctar de Plantas , Estruturas Vegetais/anatomia & histologia , Formigas , Microscopia ConfocalResumo
This study aimed to map the stem biomass of an even-aged eucalyptus plantation in southeastern Brazil based on canopy height profile (CHPs) statistics using wall-to-wall discrete return airborne laser scanning (ALS), and compare the results with alternative maps generated by ordinary kriging interpolation from field-derived measurements. The assessment of stem biomass with ALS data was carried out using regression analysis methods. Initially, CHPs were determined to express the distribution of laser point heights in the ALS cloud for each sample plot. The probability density function (pdf) used was the Weibull distribution, with two parameters that in a secondary task, were used as explanatory variables to model stem biomass. ALS metrics such as height percentiles, dispersion of heights, and proportion of points were also investigated. A simple linear regression model of stem biomass as a function of the Weibull scale parameter showed high correlation (adj.R2 = 0.89). The alternative model considering the 30th percentile and the Weibull shape parameter slightly improved the quality of the estimation (adj.R2 = 0.93). Stem biomass maps based on the Weibull scale parameter doubled the accuracy of the ordinary kriging approach (relative root mean square error = 6 % and 13 %, respectively).
Assuntos
Biomassa , Eucalyptus , Eucalyptus/crescimento & desenvolvimento , Identificação Biométrica/métodos , Microscopia Confocal , Microscopia Confocal/estatística & dados numéricosResumo
This study aimed to map the stem biomass of an even-aged eucalyptus plantation in southeastern Brazil based on canopy height profile (CHPs) statistics using wall-to-wall discrete return airborne laser scanning (ALS), and compare the results with alternative maps generated by ordinary kriging interpolation from field-derived measurements. The assessment of stem biomass with ALS data was carried out using regression analysis methods. Initially, CHPs were determined to express the distribution of laser point heights in the ALS cloud for each sample plot. The probability density function (pdf) used was the Weibull distribution, with two parameters that in a secondary task, were used as explanatory variables to model stem biomass. ALS metrics such as height percentiles, dispersion of heights, and proportion of points were also investigated. A simple linear regression model of stem biomass as a function of the Weibull scale parameter showed high correlation (adj.R2 = 0.89). The alternative model considering the 30th percentile and the Weibull shape parameter slightly improved the quality of the estimation (adj.R2 = 0.93). Stem biomass maps based on the Weibull scale parameter doubled the accuracy of the ordinary kriging approach (relative root mean square error = 6 % and 13 %, respectively).(AU)
Assuntos
Biomassa , Eucalyptus , Eucalyptus/crescimento & desenvolvimento , Microscopia Confocal/estatística & dados numéricos , Microscopia Confocal , Identificação Biométrica/métodosResumo
A morfologia das papilas linguais da cabra doméstica (Capra hircus) foi estudada por Microscopia Eletrônica de Varredura (MEV) e Microscopia de luz. Papilasfiiliformes, cônicas e lentiformes possuem função mecânica, enquanto que as papilas fungiformes e as valadas possuem função gustativa. As papilasfiiliformes estavam distribuídas por toda a extensão da língua, nas superfícies dorsal e lateral. Possuem um formato cônico com a extremidade pontiaguda direcionada caudalmente. Possuem vários processos secundários, de tamanhos diferentes, com a mesma orientação. A histologia revelou abundante tecido queratinizado. As papilas fungiformes foram observadas nas superfícies dorsal e lateral do ápice e do corpo da língua. Possuem formato semelhante a cogumelos, com epitélio corni[1]icado espesso e botões gustativos distribuídos na superfície. As papilas lentiformes se distribuíam na linha mediana da parte mais dorsal do toro, com projeções elevadas além da superfície da língua. Algumas apresentavam o formato de pirâmide e outras um formato mais achatado. As papilas valadas foram encontradas na superfície lateral da parte mais caudal do toro. Possuem formato arredondado, envolvido por um sulco pouco profundo. Possuem botões gustativos na parede lateral que se abrem no sulco. As papilas cônicas foram observadas no toro da língua e possuem formato alongado, com base larga e ponta romba. Embora as principais características morfológicas e estruturais do epitélio lingual sejam específicas da espécie, o tipo de alimentação e os hábitos alimentares podem influenciar na sua estrutura. Assim, o presente trabalho fornece uma descrição histológica e ultraestrutural da língua de cabras domésticas submetidas à alimentação baseada em gramíneas e ração.(AU)
The morphology of the lingual papillae of domestic goat (Capra hircus) was studied by scanning electron microscopy (SEM) and light microscopy (LM). Filiform, conical and lentiform papillae exert mechanical function whereas fungiform and valate papillae have gustative function. Filliform papillae were distributed all over the lateral and dorsal surfaces of the tongue. They present a conical shape with the thin ends pointed caudally; and possess several secondary processes, of different sizes, with the same orientation. Histological examination of these papillae revealed abundant queratinized tissue. Fungiform papillae were observed on the dorsal and lateral surface of the apical region and body of the tongue. Present a fungi-like shape, with a thick cornified epithelium and taste buds distributed on the surface. Lentiform papillae were distributed over the median line of the dorsal torus, presenting projections above the surface of the tongue. Some displayed pyramidal shapes; other were more flattened. Valate papillae were observed on the lateral surface of the posterior part of the torus and present a rounding shape delimited by a deep groove. These papillae possess taste buds on their lateral wall that open up on the groove. Conical papillae were observed on the lingual torus and present an extended shape, with a wide base and a blunt tip. Although the main morphological and structural characteristics of the lingual epithelium are species-specific, some features may be influenced by the type of food and alimentary habits. Therefore, the present study provides a histological and ultrastructural description of the tongue of the domestic goat fed with mixed food (grass and concentrate).(AU)