Glucosinolate-accumulating S-cells in Arabidopsis leaves and flower stalks undergo programmed cell death at early stages of differentiation

Plant secondary metabolites glucosinolates (GSL) have important functions in plant resistance to herbivores and pathogens. We identified all major GSL that are accumulated in S-cells in Arabidopsis by MALDI-TOF MS, and estimated by LC-MS that the total GSL concentration in these cells is above 130 mM. The precise locations of the S-cells outside phloem bundles in rosette and cauline leaves and in flower stalks were visualised using sulphur mapping by cryo-SEM/EDX. S-cells contain up to 40% of total sulphur in flower stalk tissues. S-cells in emerging flower stalks and developing leaf tissues show typical signs of Programmed Cell Death (PCD) or apoptosis, such as chromatin condensation in the nucleus and blebbing of the membranes. TUNEL staining for DNA double strand breaks confirmed PCD in S-cells in postmeristematic tissues in the flower stalk as well as in the leaf. Our results show that S-cells in postmeristematic tissues proceed to an extreme degree of metabolic specialisation besides PCD. Accumulation and maintenance of a high concentration of GSL in these cells are accompanied by degradation of a number of cell organelles. The substantial changes in the cell composition during S-cell differentiation indicate the importance of this particular GSL-based phloem defence system. The specific anatomy of the S-cells and ability to accumulate specialised secondary metabolites is similar to that of the non-articulated laticifer cells in latex plants and thus indicates a common evolutionary origin.

Genistein blocks homocysteine-induced alterations in the proteome of human endothelial cells

Dietary isoflavones from soy are suggested to protect endothelial cells from damaging effects of endothelial stressors and thereby to prevent atherosclerosis. In search of the molecular targets of isoflavone action, we analyzed the effects of the major soy isoflavone, genistein, on changes in protein expression levels induced by the endothelial stressor homocysteine (Hcy) in EA.hy 926 endothelial cells. Proteins from cells exposed for 24 h to 25 mu M Hcy alone or in combination with 2.5 mu M genistein were separated by two-dimensional gel electrophoresis and those with altered spot intensities were identified by peptide mass fingerprinting, Genistein reversed Hcy-induced changes of proteins involved in metabolism, detoxification, and gene regulation: and some of those effects can be linked functionally to the antiatherosclerotic properties of the soy isoflavone. Alterations of steady-state levels of cytoskeletal proteins by genistein suggested an effect oil apoptosis. As a matter of fact genistein caused inhibition of Hcy-mediated apoptotic cell death as indicated by inhibition of DNA fragmentation and chromatin condensation. In conclusion, proteome analysis allows the rapid identification of cellular target proteins of genistein action in endothelial cells exposed to the endothelial stressor Hcy and therefore enables the identification of molecular pathways of its antiatherosclerotic action

Bioprospecção de polissacarídeos sulfatados de macroalgas marinhas do litoral do Rio Grande do Norte: caracterização de uma heterofucana extraída da alga marron sargassum filipendula que induz apoptose em células hela

The coast of Rio Grande do Norte has more than 100 species of seaweed, mostly unexplored regarding their pharmacological potential. The sulfated polysaccharides (PS) are by far the more seaweed compounds studied, these present a range of biological properties, such as anticoagulant activity, anti-inflammatory, antitumor and antioxidant properties. In this study, we extract sulfated polysaccharide rich-extracts of eleven algae from the coast of Rio Grande do Norte (Dictyota cervicornis; Dictiopterys delicatula; Dictyota menstruallis; Dictyota mertensis; Sargassum filipendula; Spatoglossum schröederi; Gracilaria caudata; Caulerpa cupresoides; Caulerpa prolifera; Caulerpa sertularioides e Codim isthmocladum), and these were evaluated for the potential anticoagulant, antioxidant and antiproliferative. All polysaccharide extracts showed activity for anticoagulant, antioxidant and/or antiproliferative activity, especially D. delicatula and S. filipendula, which showed the most prominent pharmacological potential, thereby being chosen to have their sulfated polysaccharides extracted. By fractionating method were obtained six fractions rich in sulfated polysaccharides to the algae D. delicatula (DD-0,5V, DD-0, 7V, DD-1,0v, DD-1,3v, DD-1,5v and DD-2,0) and five fractions to the alga S. filipendula (SF-0,5V, SF-0,7V, SF-1,0v, SF-1,5v and SF-2,0v). For the anticoagulant assay only the fractions of D. delicatula showed activity, with emphasis on DD-1, 5v that presented the most prominent activity, with APTT ratio similar to clexane® at 0.1 mg/mL. When evaluated the antioxidant potential, all fractions showed potential in all tests (total antioxidant capacity, hydroxyl and superoxide radicals scavenging, ferrous chelation and reducing power), however, the ability to chelate iron ions appears as the main mechanism antioxidant of sulfated polysaccharides from seaweed. In antiproliferative assay, all heterofucanas showed dose-dependent activity for the inhibition of cell proliferation of HeLa, however, with the exception of SF-0,7V, SF- 1,0v and SF-1,5v, all fractions showed antiproliferative activity against MC3T3, a normal cell line. The heterofucana SF-1,5V had its antiproliferative mechanism of action evaluated. This heterofucan induces apoptosis in HeLa cells by a pathway caspase independent, promoting the release of apoptosis Inducing Factor (AIF) in the cytosol, which in turn induces chromatin condensation and DNA fragmentation into 50Kb fragments. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.

Ultrastructural analysis of spermiogenesis in the neotropical cichlid Cichla kelberi Kullander & Ferreira, 2006 (Perciformes: Cichlidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The ultrastructure of Sorubim lima (Teleostei, Siluriformes, Pimelodidae) spermatogenesis: premeiotic and meiotic periods

The ultrastructure of Sorubim lima spermatogenesis during the premeiotic and meiotic periods was studied. Our observations showed that the germ cells in the cysts are connected by cytoplasmic bridges and the mitotic and meiotic divisions are slightly asynchronous, the first and the last spermatogonial generations differ in the cellular and nuclear volume, nucleolus, chromatin condensation, distribution, size, density, and shape of the mitochondria, presence of 'lamellae anulata', amount and dimension of the 'nuages', and movement of the centrioles. In addition to the nuclear prophase structures, the spermatocyte I shows changes in all other cellular organelles and elongated vesicles appear in the cytoplasm. The accentuated cytoplasmic density and thickened walled vesicles are morphological characteristics that differentiate spermatocytes II from the other germ cells in the cysts of Sorubim lima testis. (C) 1999 Harcourt Publishers Ltd.

Ultrastructure of the ovary of Dermatobia hominis (Diptera: cuterebridae). I. Development during the 3rd larval instar

The ultrastructure and distribution of gonial and somatic cells in the ovary of Dermatobia hominis was studied during the 3rd larval instar. In larvae weighing between 400 and 500 mg, the ovary is partially divided into basal and apical regions by oblong somatic cells that penetrate from the periphery; these cells show ovoid nucleus and cytoplasm full of microtubules. In both regions, gonial cells with regular outlines, large nucleus and low electron-density cytoplasm are scattered among the interstitial somatic cells. These later cells have small nucleus and electrodense cytoplasm. Clear somatic cells with small nucleus and cytoplasm of very low electron-density are restrict to the apical region of the gonad. Degenerating interstitial somatic cells are seen in the basal portion close to the ovary peduncle. During all this larval period the morphological features of the ovary remain almost the same. At the end of the period there is a gradual deposition of glycogen in the cytoplasm of the somatic cells, increase in the number and density of their mitochondria plus nuclear modification as membrane wrinkling and chromatin condensation in masses.

Spermatic cell characteristics in Gymnotiformes (Teleostei: Ostariophysi) and their phylogenetic meaning

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Phosphorylation of Histone H3S10 in Animal Chromosomes: Is There a Uniform Pattern?

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ultrastructural analysis of the nucleolar aspects at spermiogenesis in triatomines (Heteroptera, Triatominae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Espermiogênese em Eupemphix nattereri (Anura, Leiuperidae): aspectos ultra-estruturais

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparative description and discussion of spermiogenesis and spermatozoal ultrastructure in some species of Heptapteridae and Pseudopimelodidae (Teleostei: Siluriformes)

Os dados obtidos no presente estudo sobre a ultraestrutura da espermiogênese e dos espermatozóides de Pseudopimelodidae e Heptapteridae mostram que eles compartilham algumas características, mas são bastante diferentes uns dos outros. As principais diferenças são a ocorrência de espermiogênese do tipo I em Pseudopimelodidae e do tipo III em Heptapteridae, a presença de fossa nuclear em Pseudopimelodidae e sua ausência em Heptapteridae, a presença de uma peça intermediária longa em Pseudopimelodidae e uma peça intermediária curta em Heptapteridae, a presença de um canal citoplasmático em Pseudopimelodidae e sua ausência em Heptapteridae, a presença de muitas vesículas grandes na peça intermediária de Pseudopimelodidae, e a presença de vesículas muito alongadas e dispostas em posição periférica distal em Heptapteridae e mitocôndrias distribuídas em toda a peça intermediária de Pseudopimelodidae e muito próximas ao núcleo em Heptapteridae. Heptapteridae e Pimelodidae compartilham várias características como a espermiogênese do tipo III, o mesmo padrão de condensação da cromatina e a ausência de fossa nuclear e projeções laterais ou fins. O espermatozóide de Pseudopimelodidae é mais similar aos dos Siluridae, porém a ausência de dados adicionais sobre a espermiogênese e o espermatozóide de outros siluriformes ainda limitam uma discussão mais ampla na ordem.

Morphological findings in the tracheal epithelium of dogs exposed to the inhalation of poorly conditioned gases under use of an endotracheal tube or laryngeal mask airway

OBJETIVO: Avaliar as alterações morfológicas no epitélio traqueal de cães expostos à inalação de gases pouco condicionados, sob ventilação com tubo traqueal (TT) ou máscara laríngea (ML). MÉTODOS: Doze cães adultos foram divididos aleatoriamente em dois grupos: grupo TT (n-6) e grupo ML (n-6), submetidos à anestesia venosa e ventilação mecânica, em sistema sem reabsorção de CO2. Foram registrados parâmetros hemodinâmicos e ventilatórios, temperatura timpânica, temperatura, umidade relativa e absoluta do ar ambiente e dos gases inalados durante 3 horas. Ao término do experimento, os animais foram submetidos a eutanásia e realizadas biópsias ao longo do segmento traqueal para estudo morfológico. Três cães saudáveis foram utilizados para controle morfológico. RESULTADOS: A temperatura dos gases inalados manteve-se entre 24ºC e 26ºC, a umidade relativa entre 10% e 12%, e umidade absoluta entre 2 -3 mg H2O.L-1 sem diferença significativa entre os grupos. em ambos os grupos a análise histológica evidenciou processo inflamatório epitelial e congestão no córion, e a microscopia eletrônica de varredura mostrou agrupamento e desorganização ciliar. A microscopia eletrônica de transmissão detectou maiores alterações no grupo TT do que no ML, como alargamento das junções intercelulares, desorientação ciliar, vacuolização citoplasmática, alterações nucleares como pcinose e condensação da cromatina. CONCLUSÃO: A máscara laríngea determinou alterações menos pronunciadas no epitélio traqueal de cães expostos à inalação de gases pouco condicionados.

ULTRASTRUCTURE OF THE HINNY (EQUUS-ASINUS X EQUUS-CABALLUS) SEMINIFEROUS EPITHELIUM

The gonads and the germinative cells of 3 male hinnies were studied with light and transmission electron microscopy with the aim to observe the development of germ cells and verify the morphological modifications due to the hybridization. The hinny seminiferous epithelium presented Sertoli cells and spermatogonia with normal features and anomalous spermatocytes I. The other cells from the spermatogenic sequence were not seen. Most of the alterations began to occur in the cytes I, which presented nuclear vacuolization and deposits of amorphous material between the carioteca and the nuclear lamina, forming vesicles, or exaggerated chromatin condensation, resulting in pyknosis. In the cytoplasm vacuolization was also observed, besides organelle destruction.The arrest of meiosis due to lock of chromosome homologies leads to germinative cell degeneration and, therefore, the spermatogenesis arrest. This fact causes a profound alteration in the seminiferous epithelium morphology in comparison with the parental species.

Cytological steps during spermiogenesis in the house sparrow (Passes domesticus, Linnaeus)

Spermiogenesis of the domestic sparrow was investigated with the light and electron microscopes and a step by step classification is proposed. Three cell populations corresponding to early, mid and late spermatids were easily divided according to their positions in the seminiferous epithelium. In addition to this initial separation, six steps were recognized, based on nuclear morphology and the degree of chromatin condensation, in association to their acrosomal and flagellar development. Early spermiogenesis is the period previous to chromatin condensation. The first step can be recognized by the extending flagellum and the second by the pro-acrosome development in contact with the nucleus. During the third or intermediate step, chromatin condenses and the cell becomes polarized with the pro-acrosomic vesicle and the tail occupying opposite sides of the nucleus. Late spermiogenesis, including steps IV-VI, is marked by complete chromatin condensation. The final cellular modifications lead to the formation of a spiraled spermatozoon. This shape is due to the twisting of the acrosome and nucleus, as well as the helical arrangement of mitochondria around the axoneme along most of the flagellum, making an exceptionally long middle piece. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Morphological Characterization of the Testicular Cells and Seminiferous Epithelium Cycle in Six Species of Neotropical Bats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)