Antileukemic effects of Didemnum psammatodes (Tunicata: Ascidiacea) constituents

Chemical investigation of the methanolic extract of the ascidian Didemnum psammatodes has led to the identification of fourteen known compounds: three methyl esters (methyl myristate, methyl palmitate and methyl stearate), four steroids (cholesterol, campesterol, stigmasterol and beta-sitosterol), two fatty acids (palmitic acid and stearic acid), three glyceryl ethers {(1,2-propanediol, 3-(heptadecyloxy), batyl alcohol and 1,2-propanediol, 3-[(methyloctadecyl)oxy]) and two nucleosides (thymidine and 2`-deoxyguanosine). Their structures were proposed by NMR and comparison with literature data and GC analysis in comparison with authentic sample. The cytotoxic activity of these compounds was evaluated against human leukemia cell line panel using the MTT assay. The mixture of the three methyl esters was the most active group of compounds, showing antiproliferative and cytotoxic effects. Further studies on their mode of action suggest that these activities are connected with inhibition of DNA synthesis and induction of both necrosis and apoptosis. (C) 2007 Elsevier Inc. All rights reserved.

Cytotoxic activity of a dichloromethane extract and fractions obtained from Eudistoma vannamei (Tunicata: Ascidiacea)

This study consists of the bioassay-guided fractionation of the dichloromethane extract from Eudistoma vannamei and the pharmacological characterization of the active fractions. The dried hydromethanolic extract dissolved in aqueous methanol was partitioned with dichloromethane and chromatographed on a silica gel flash column. The anti-proliferative effect was monitored by the MTT assay. Four of the latest fractions, numbered 14 to 17, which held many chemical similarities amongst each other, were found to be the most active. The selected fractions were tested for viability, proliferation and death induction on cultures of HL-60 promycloblastic leukemia cells. The results suggested that the observed cytotoxicity is related to apoptosis induction. (C) 2007 Elsevier Inc. All rights reserved.

The British Tunicata; an unfinished monograph, by the late Joshua Alder and the late Albany Hancock. Edited by John Hopkinson, with a history of the work by the Rev. A. M. Norman.

1

The British Tunicata; an unfinished monograph, by the late Joshua Alder and the late Albany Hancock. Edited by John Hopkinson, with a history of the work by the Rev. A. M. Norman.

2

The British Tunicata; an unfinished monograph, by the late Joshua Alder and the late Albany Hancock. Edited by John Hopkinson, with a history of the work by the Rev. A. M. Norman.

3

Estudos genéticos sôbre o milho tunicata

The study of pod corn seems still of much importance from different points of view. The phylogenetical importance of the tunicate factor as a wild type relic gene has been recently discussed in much detail by MANGELSDORF and REEVES (1939), and by BRIEGER (1943, 1944a e b). Selection experiments have shown that the pleiotropic effect of the Tu factor can be modified very extensively (BRIEGER 1944a) and some of the forms thus obtained permitt comparison of male and female inflorescences in corn and related grasses. A detailed discussion of the botanical aspect shall be given shortly. The genetic apect, finally, is the subject of the present publication. Pod corn has been obtained twice: São Paulo Pod Corn and Bolivia Pod Corn. The former came from one half ear left in our laboratory by a student and belongs to the type of corn cultivated in the State of São Paulo, while the other belongs to the Andean group, and has been received both through Dr. CARDENAS, President of the University at Cochabamba, Bolivia, and through Dr. H. C. CUTLER, Harvard University, who collected material in the Andes. The results of the studies may be summarized as follows: 1) In both cases, pod corn is characterized by the presence of a dominant Tu factor, localized in the fourth chromosome and linked with sul. The crossover value differs somewhat from the mean value of 29% given by EMERSON, BEADLE and FRAZER (1935) and was 25% in 1217 plants for São Paulo Pod Corn and 36,5% in 345 plants for Bolivia Pod Corn. However not much importance should be attributed to the quantitative differences. 2) Segregation was completely normal in Bolivia Pod Corn while São Paulo Pod Corn proved to be heterozygous for a new com uma eliminação forte, funcionam apenas 8% em vez de 50%. Existem cerca de 30% de "jcrossing-over entre o gen doce (Su/su) e o fator gametofítico; è cerca de 5% entre o gen Tu e o fator gametofítico. A ordem dos gens no cromosômio IV é: Ga4 - Tu - Sul. 3) Using BRIEGER'S formulas (1930, 1937a, 1937b) the following determinations were made. a) the elimination of ga4 pollen tubes may be strong or weak. In the former case only about 8% and in the latter 37% of ga4 pollen tubes function, instead of the 50% expected in normal heterozygotes. b) There is about 30,4% crossing-over between sul and ga4 and 5,3% between Tu and ga3, the order of the factors beeing Su 1 - Tu - Ga4. 4) The new gametophyte factor differs from the two others factors in the same chromosome, causing competition between pollen tubes. The factor Gal, ocupies another locus, considerably to the left of Sul (EMERSON, BEADLE AND FRAZSER, 1935). The gen spl ocupies another locus and causes a difference of the size of the pollen grains, besides an elimination of pollen tubes, while no such differences were observed in the case of the new factor Ga4. 5) It may be mentioned, without entering into a detailed discussion, that it seems remarquable that three of the few gametophyte factors, so far studied in detail are localized in chromosome four. Actuality there are a few more known (BRIEGER, TIDBURY AND TSENG 1938), but only one other has been localized so far, Ga2, in chromosome five between btl and prl. (BRIEGER, 1935). 6) The fourth chromosome of corn seems to contain other pecularities still. MANGELSDORF AND REEVES (1939) concluded that it carries two translocations from Tripsacum chromosomes, and BRIEGER (1944b) suggested that the tu allel may have been introduced from a tripsacoid ancestor in substitution of the wild type gene Tu at the beginning of domestication. Serious disturbances in the segregation of fourth chromosome factors have been observed (BRIEGER, unpublished) in the hybrids of Brazilian corn and Mexican teosinte, caused by gametophytic and possibly zygotic elimination. Future studies must show wether there is any relation between the frequency of factors, causing gametophyte elimination and the presence of regions of chromosomes, tranfered either from Tripsacum or a related species, by translocation or crossing-over.

Recruitment and colonization of colonial ascidians (Tunicata: Ascidiacea) on intertidal rocks in Northeastern Brazil

Recruitment and colonization of colonial ascidians, starting on natural scraped substratum, was observed throughout one year, in quadrats marked on sea- and continental-rock faces in the sublittoral fringe of an exposed beach in the Northeast of Brazil. The species of the Didemnidae, Didemnum duplicatum Monniot, 1983, D. psammathodes Sluiter, 1895, and Polysyncraton amethysteum Van Name, 1902, and an unidentified species of Polycitoridae, Eudistoma sp. 1, were recruited. Presenting continuous recruitment of all species, the density of living and dead recruits and colonies was similar on both faces of the rocks. The highest rainfall during winter, affected significantly the recruits density of D. duplicatum and D. psammathodes. Different permanence periods were observed for colonies of each species. The longest permanence periods were assigned to the colonies of P. amethysteum and D. psammathodes, extending up to four and five months, respectively.

Ascídias (Tunicata, Ascidiacea) introduzidas no Arquipélago de Alcatrazes, São Paulo

O Arquipélago de Alcatrazes (24º06'S, 45º42'W) localiza-se a 36 km da costa e constitui uma Estação Ecológica desde 1987. Estando próximo à região portuária de São Sebastião, São Paulo, está sujeito à introdução de espécies exóticas. Com o objetivo de detectar se está ocorrendo ou não introdução de espécies, foram coletados 40 exemplares de ascídias da principal ilha do arquipélago, a Ilha de Alcatrazes, sendo 15 amostras da Baía do Oratório e 25 do Saco do Funil. Foram encontradas 24 espécies em sete famílias. Dentre estas, apenas cinco espécies podem ser consideradas como nativas para o Atlântico; quatro são atlânticas, mas criptogênicas na região devido à distribuição disjunta; cinco são classificadas como criptogênicas de ampla distribuição mundial; e uma pode ser considerada como um caso certo de introdução, Ciona intestinalis (Linnaeus, 1767). Há ainda nove espécies que não puderam ser identificadas, podendo algumas se tratar de espécies novas. A presença de uma espécie exótica e a grande quantidade de espécies criptogênicas com forte evidência de introdução são indícios de que as ilhas estão sendo ameaçadas por uma fauna não nativa. A conservação do Arquipélago deve levar em consideração a presença do porto como fonte de estresse e o controle das espécies introduzidas como parte do plano de manejo da Unidade de Conservação.

In vivo antithrombotic properties of a heparin from the oocyte test cells of the sea squirt Styela plicata(Chordata-Tunicata)

In the ascidian Styela plicata, the oocytes are surrounded by two types of accessory cells named follicle cells and test cells. A heparin-like substance with an anticoagulant activity equivalent to 10% of mammalian heparin and about 5% as potent as the mammalian counterpart for the inhibition of thrombin by antithrombin was isolated from the oocyte test cells. In the present study, we compared the antithrombotic and hemorrhagic effects of sea squirt oocyte test cell heparin with those of porcine heparin in rat models of venous thrombosis and blood loss. Intravenous administration of the oocyte test cell heparin to Wistar rats (both sexes, weighing ~300 g, N = 4 in each group) at a dose of 5.0 mg/kg body weight, which produced a 1.8-fold increase in plasma activated partial thromboplastin time, inhibited thrombosis by 45 ± 13.5% (mean ± SD) without any bleeding effect. The same dose of porcine heparin inhibited thrombosis by 100 ± 1.4%, but produced a blood loss three times greater than that of the saline-treated control. However, 10-fold reduction of the dose of porcine heparin to 0.5 mg/kg body weight, which produced a 5-fold increase in plasma-activated partial thromboplastin time, inhibited thrombosis by 70 ± 13% without any bleeding effect. The antithrombotic properties of a new heparin isolated from test cells of the sea squirt S. plicata, reported here for the first time, indicate that, although sea squirt oocyte test cell heparin was a poor anticoagulant compared to porcine heparin, it had a significant antithrombotic effect without causing bleeding.

Larvacean (Chordata, Tunicata) abundance and inferred secondary production off southeastern Brazil

We studied the temporal and vertical variability in larvacean abundance and secondary production on a fixed station off southeast Brazil, from January 2007 to December 2008. Larvacean biomass was derived from length weight regressions, and growth rates were estimated from an empirical model. We identified eleven larvacean species. Oikopleura longicauda occurred throughout the studied period and was the most abundant species, followed by Oikopleura fusiformis. Fritillaria haplostoma, O. fusiformis and O. longicauda were found mainly above the thermocline, whereas Oikopleura dioica and Fritillaria pellucida preferred bottom layers. Higher abundance and biomass were observed in warmer months, when the water column was stratified as a result of the bottom intrusions of the cold and nutrient-rich South Atlantic Central Water. Secondary production mirrored the biomass seasonal pattern. Larvacean biomass equaled to less than 10% of copepod biomass during the same period, but larvacean production comprised on average 77% that of copepods, whereas the production of discarded houses and fecal pellets comprised up to 2800% of larvaceans secondary production. This confirms the potential significance of larvaceans in the carbon flux in tropical and subtropical coastal regions. (C) 2011 Elsevier Ltd. All rights reserved.

Spatial competition induces the mobilization of morula cells in the colonial ascidian Didemnum perlucidum (Tunicata: Didemnidae)

The effects of spatial competition among colonial marine organisms are often evident in the contact zones between colonies. These effects are especially pronounced when the interaction results in overgrowth or necrosis of one of the competitors. Ascidians, one of the dominant taxonomic groups in subtidal sessile communities, have specialized morula cells that provide a defense against microbial infections. Injuries resulting from interspecific competitive interactions might also act as a stimulus for this defensive mechanism. Therefore, we expected to see the recruitment of morula cells in tissues near competitor contact zones. To test the hypothesis that spatial competition elicits this immune response, we placed colonies of the ascidian Didemnum perlucidum from southeastern Brazil in four different types of competitive situations: (1) overgrowth of the competitor, (2) stand-off interactions, (3) overgrowth by the competitor, and (4) free of competitors. Our results indicate that competitive interactions increase the population of morula cells in contact zones, as more cells were observed in interactions that resulted in the overgrowth of individuals of D. perlucidum, and fewer cells were observed in colonies that were free of competitors. We identified the defensive function of the morula cells by showing the presence of the enzyme phenoloxidase within its vacuoles. Phenoloxidase is a widespread enzyme among animals and plants, and is frequently used in defense by synthesizing toxic quinones from polyphenol substrates. This is the first study to document the presence of morula cells in didemnid ascidians and the mobilization of these cells by spatial competition by heterospecifics, and one of the first studies to identify phenoloxidase activity in morula cells.

Lack of COI variation for Clavelina oblonga (Tunicata, Ascidiacea) in Brazil: Evidence for its human-mediated transportation?

Recent studies indicate that ascidians are efficiently dispersed by human transport. We have chosen the mitochondrial gene cytochrome c oxidase subunit I (COI) to address whether Clavelina oblonga is an introduced species in the Brazilian coast. Colonies of C. oblonga were sampled in different localities along Atlantic coasts of USA, Panama, and Brazil. The sequencing of 92 colonies resulted in three haplotypes for the species, two unique to Florida and the other shared by exemplars collected in Brazil and Panama; the latter haplotype is identical to the published sequence of Azores. Our evidence, including the absence of C. oblonga in the country's northern tropical waters, its association with artificial habitats and lack of COI variation suggest that the species has been introduced in the southeastern and southern Brazilian coasts. Previous records (85 years old) suggest that it could be a relatively long-term introduction.

Der bespelzte Mais (Zea Mays tunicata Larranhago.)

von L. Wittmack